Betulinic acid nucleoside derivatives for the prevention or treatment of cancer

Abstract

The present application belongs to the field of medicine, and particularly relates to betulinic acid nucleoside derivatives for preventing or treating cancer. Some compounds provided by the present application exhibit excellent inhibitory activity on test cancer cells, and particularly have unexpected inhibitory activity on colon cancer or prostate cancer cells. The compounds provided by the present application are expected to be used for preventing or treating colon cancer, prostate cancer and other cancers or symptoms related thereto.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to betulinic acid nucleoside derivatives for preventing or treating cancer. BACKGROUND

[0002] It is known that colon cancer is not sensitive to most chemotherapeutic drugs, and the current drug treatment effect is very poor. Colon cancer is a common digestive tract malignant tumor occurring in the colon, which is most common in the junction of the rectum and sigmoid colon, and the incidence rate is highest in the age group of 40-50 years old, with a male to female ratio of 2-3:1. The incidence rate of colon cancer accounts for the third place of gastrointestinal tumors, and its general morphology is polypoid, ulcerative, etc. Colon cancer can develop along the intestinal wall in a circular manner, spread along the longitudinal axis of the intestinal canal upward and downward, or infiltrate the deep layer of the intestinal wall. In addition to metastasis through lymphatic vessels, blood flow and local invasion, it can also implant into the abdominal cavity or spread and metastasize along the suture line and incision surface. Patients with chronic colitis, colon polyps, obese men, etc. are susceptible populations.

[0003] Prostate cancer refers to an epithelial malignant tumor occurring in the prostate. In 2004, WHO "Pathology and Genetics of Tumors of the Urinary System and Male Reproductive Organs" includes adenocarcinoma (acinar adenocarcinoma), ductal adenocarcinoma, urothelial carcinoma, squamous cell carcinoma, and adenosquamous carcinoma in the pathological type of prostate cancer. Among them, prostate adenocarcinoma accounts for more than 95%, therefore, the prostate cancer we usually refer to is prostate adenocarcinoma. The incidence rate is low before the age of 55, and gradually increases after the age of 55, with the peak age being 70-80 years old. Patients with familial genetic prostate cancer have a slightly earlier age of onset, and patients aged ≤55 years old account for 43%. At present, early-stage prostate cancer patients can achieve good treatment effect through radical surgery or radical radiotherapy. For locally advanced (without metastasis) and metastatic prostate cancer, androgen removal therapy is generally selected to prolong the survival period of patients and improve the quality of life; some patients can choose surgical resection, or multiple means of comprehensive treatment based on radiotherapy. However, hormone therapy has many known side effects on the human body.

[0004] The inventors have disclosed betulinic acid nucleoside derivatives with anti-HIV activity in their published paper, Qiang Wang, et al., ACS Med. Chem. Lett. 2020, 11, 2290-2293. However, the aforementioned literature did not investigate the anticancer activity of these derivatives, particularly their inhibitory activity against cancer cells such as colon and prostate cancer. Furthermore, the inventors' prior Chinese patent applications 201810483852.9, 201810483941.3, and 201810700317.4 investigated the inhibitory activity of some compounds against A549 (lung cancer), SGC-7901 (gastric cancer), MCF-7 (breast cancer), and LUC-7721 (liver cancer), but did not investigate the inhibitory activity of these compounds against colon and prostate cancer cells.

[0005] Therefore, there is a need to develop new drugs with inhibitory activity against colon cancer cells, prostate cancer cells, and other cancer cells. Summary of the Invention

[0006] To address the aforementioned technical problems, the present invention provides the use of a compound of formula (I) in the preparation of a pharmaceutically acceptable salt, tautomer, stereoisomer, nitride, hydrate, or prodrug, wherein the pharmaceutically acceptable salt, tautomer, stereoisomer, nitride, hydrate, or prodrug is used for the prevention or treatment of colon cancer, prostate cancer, or lung cancer.

[0007]

[0008] Wherein, X is selected from O, unsubstituted, or optionally substituted by one, two, or more R. a The following groups are substituted: C 1-40 Alkyl, NH;

[0009] R1 is selected from H, is unsubstituted, or is optionally composed of one, two, or more R1s. b The following groups are substituted: C 1-40 Alkyl, C 3-20 Cycloalkyl, 3-20 membered heterocyclic groups, C 6-20 Aryl or 5-20 heteroaryl groups;

[0010] Het1 is selected from one, two or more R... m Substituted 5-6 membered heterocyclic groups, for example

[0011] Het2 is selected from one, two or more R... n Replacement

[0012] Het1 bonds with the cyclic carbon or nitrogen atoms in Het2 through its cyclic carbon or nitrogen atoms;

[0013] Het1 is bonded to the N atom of the triazole group through its cyclic carbon or nitrogen atom groups;

[0014] Y is selected from H, halogen, OH, SH, CN, OH, unsubstituted or optionally substituted by one, two or more R. c The following groups are substituted: C 1-40 Alkyl, C 3-20 cycloalkyl, C 1-40 Alkoxy, 3-20 membered heterocyclic groups, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclic oxygen, C 6-20 Aryloxy group, 5-20 heteroaryloxy group, NH2, -C(O)OR3, -OC(O)R4, -OP(O)(OR5)2, -OS(O)2R6;

[0015] Z is selected from O, S, halogen, OH, SH, CN, unsubstituted or optionally surrounded by one, two or more R. d The following groups are substituted: C 1-40 Alkyl, C 3-20 cycloalkyl, C 1-40 Alkoxy, 3-20 membered heterocyclic groups, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclic oxygen, C 6-20 aryloxy group, 5-20 heteroaryloxy group, NH2, -C(O)OR2, -OC(O)R3, -OP(O)(OR4)2, -OS(O)2R5; where, when When it is a double bond, Z is selected from O or S; when When it is a single bond, Z is selected from groups other than O and S as defined above;

[0016] Each R3, R4, R5, and R6 may be the same or different, and is independently selected from H, without substitution, or arbitrarily assigned to one, two, or more Rs. e The following groups are substituted: C 1-40 Alkyl, C 3-20 Cycloalkyl, 3-20 membered heterocyclic groups, C 6-20 Aryl, 5-20 heteroaryl;

[0017] R m R n They may be identical or different, independently selected from H, halogens, OH, SH, CN, without substitution or optionally by one, two or more Rs. f The following groups are substituted: C 1-40 Alkyl or C 1-40 Alkoxy;

[0018] R a R b R cR d R e R f They may be the same or different, and are independently selected from halogens, OH, CN, NO2, oxo (=O), thio (=S), and C. 1-40 Alkyl, C 2-40 alkenyl, C 2-40 alkynyl group, C 3-40 cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclic, C 1-40 Alkyloxy, C 2-40 alkenyloxy group, C 2-40 alkynyloxy group, C 3-40 Cycloalkyloxy, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy group, C 6-20 aryloxy group, 5-20 membered heteroaryloxy group, 3-20 membered heterocyclic oxy group, C 1-40 Alkyl thio, C 2-40 alkenyl thio, C 2-40 alkynyl thioyl, C 3-40 cycloalkylthio, C 3-40 cycloalkenylthio, C 3-40 Cycloalkynylthio, C 6-20 Arylthio, 5-20 membered heteroarylthio, 3-20 membered heterocyclic thio, NH2, -C(O)C 1-40 Alkyl group, -C(O)NH2, -C(O)NHC 1-40 Alkyl, -C(O)-NH-OH, -COOC 1-40 Alkyl, -COOH, -OC(O)C 1-40 Alkyl, -OC(O)H, -S(O)2C 1-40 Alkyl group, S(O)2H, -S(O)2OC 1-40 Alkyl, -OS(O)2C 1-40 Alkyl group, -P(O)(OH)2, -B(OH)2.

[0019] According to embodiments of the present invention, the compound represented by formula (I) may have the structure represented by formula (I-1) or formula (I-2):

[0020]

[0021] Wherein, X is selected from O, unsubstituted, or optionally substituted by one, two, or more R. a Substituted NH or CH2;

[0022] Y is selected from halogen, OH, CN, NH2, or C.1-6 alkyl;

[0023] R1 is selected from H, is unsubstituted, or is optionally composed of one, two, or more R1s. b The following groups are substituted: C 1-6 Alkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic groups, C 6-14 Aryl or 5-14 heteroaryl groups;

[0024] R2 and R3 may be the same or different, and are independently selected from H, halogens, OH, SH, and CN, and are unsubstituted or optionally substituted by one, two, or more Rs. f The following groups are substituted: C 1-6 Alkyl, C 1-6 Alkoxy;

[0025] R4 and R5 may be the same or different, and are independently selected from H, halogens, OH, SH, and CN, and are either unsubstituted or optionally substituted by one, two, or more Rs. f The following groups are substituted: C 1-6 Alkyl, C 1-6 Alkoxy;

[0026] Z is selected from O, halogen, OH, SH, CN, unsubstituted or optionally surrounded by one, two or more R. d Replacement C 1-6 Alkyl or C 1-6 Alkoxy, when When it is a double bond, Z is selected from O; when When it is a single bond, Z is selected from groups other than O as defined above;

[0027] R a Selected from C 1-6 alkyl;

[0028] R b R f They may be the same or different, and are independently selected from halogens, OH, CN, NO2, and C. 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclic oxygen, C 6-14 Aryloxy or 5-14 heteroaryloxy.

[0029] According to embodiments of the present invention, the compound represented by formula (I) may have the structure represented by formula (I-3) or formula (I-4):

[0030]

[0031] Among them, R1, R2, R3, R4, R5, X, and Y independently have the definitions described above.

[0032] According to an embodiment of the present invention, X is selected from O.

[0033] According to an embodiment of the present invention, Y is selected from OH, NH2, or C. 1-6 alkyl;

[0034] According to an embodiment of the present invention, R1 is selected from H or C. 1-6 alkyl.

[0035] According to embodiments of the present invention, R2 and R3 may be the same or different, and are independently selected from F, Cl, Br, OH or C. 1-6 alkyl.

[0036] According to embodiments of the present invention, R4 and R5 may be the same or different, and are independently selected from H, halogen, OH, SH, CN, or C. 1-6 alkyl.

[0037] According to an embodiment of the present invention, It is a double bond, Z is O; or, It is a single bond, and Z is selected from OH.

[0038] As examples, the compounds represented by formulas (I), (I-1), (I-2), (I-3), and (I-4) can be selected from the following compounds:

[0039]

[0040]

[0041] The present invention also provides a method for preparing the betulinic acid derivative, comprising reacting a compound of formula (1) with a compound of formula (2) to obtain the betulinic acid derivative shown in formula (I):

[0042]

[0043] Among them, R1, Het1, Het2, X, Z, It independently possesses the definition described above.

[0044] According to an embodiment of the present invention, the reaction can be carried out in a solvent; the solvent can be a mixture of water and an organic solvent; the organic solvent can be methanol, ethanol, isopropanol, n-butanol, tert-butanol, tetrahydrofuran, dioxane, or N,N-dimethylformamide.

[0045] According to an embodiment of the present invention, the volume ratio of water to organic solvent is 1:0.2-5, for example 1:0.5-3, and exemplarily 1:1;

[0046] According to an embodiment of the present invention, the reaction is carried out in the presence of a catalyst, which forms monovalent copper ions during the reaction, such as a combination of Cu and CuSO4, CuCl, or CuI.

[0047] According to an embodiment of the present invention, the molar ratio of compound 1 to catalyst in the reaction is 1:0.1-1, for example, 1:0.2-0.8;

[0048] According to an embodiment of the present invention, the molar ratio of compound 1 to compound 2 in the reaction is 1:0.5-2, for example, 1:0.8-1.2.

[0049] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the above-described compound, a pharmaceutically acceptable salt thereof, a tautomer, a stereoisomer, a nitride, a hydrate, or a prodrug.

[0050] According to an embodiment of the present invention, the drug may be a pharmaceutical composition.

[0051] According to embodiments of the present invention, the pharmaceutical composition further includes one or more pharmaceutically acceptable excipients, such as carriers or excipients.

[0052] According to embodiments of the present invention, the pharmaceutical composition may further contain or not contain one or more additional therapeutic agents.

[0053] According to embodiments of the present invention, the pharmaceutical composition further includes an excipient by which the pharmaceutical composition is diluted or packaged into a carrier, such as a capsule, pouch, paper, or other container. When the excipient is used as a diluent, it can be a solid, semi-solid, or liquid substance, serving as a solvent, carrier, or medium for the active ingredient. Therefore, the pharmaceutical composition can be in the following forms: tablets, pills, powders, lozenges, pouches, capsules, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or soluble in liquid solvents); for example, ointments containing up to 10% by weight of the active ingredient, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

[0054] Preferably, the suitable excipient is selected from lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose.

[0055] According to embodiments of the present invention, the present invention also provides the use of a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer, nitride, hydrate, or prodrug, in the preparation of a medicament for inhibiting cancer cells selected from: colon cancer cells, prostate cancer cells, and lung cancer cells such as non-small cell lung cancer cells.

[0056] According to embodiments of the present invention, the present invention also provides a method for preventing or treating cancer, comprising administering to a patient in need a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer, nitride, hydrate, or prodrug thereof, wherein the cancer is selected from colon cancer, prostate cancer, or lung cancer.

[0057] According to an embodiment of the present invention, the lung cancer may be selected from non-small cell lung cancer.

[0058] The present invention also provides a method for inhibiting cancer cells, comprising contacting a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer, nitride, hydrate, or prodrug, with cancer cells, wherein the cancer cells are selected from the following cancer cells: colon cancer cells, prostate cancer cells, and lung cancer cells such as non-small cell lung cancer cells.

[0059] As an example, the cancer cells are selected from, for example, the following cancer cells: human colon cancer cells SW620, human prostate cancer cells PC-3, and human non-small cell lung cancer A549.

[0060] According to embodiments of the present invention, the present invention also provides the use of compounds represented by formula (I-2) or (I-4), especially compounds 8c, 9c or 10c, for the preparation of medicaments for the prevention or treatment of tumors, such as cancer.

[0061] According to an embodiment of the present invention, the cancer may be selected from cervical cancer, lung adenocarcinoma, breast cancer, liver cancer, esophageal cancer, prostate cancer, colon cancer, or lung cancer such as non-small cell lung cancer.

[0062] According to embodiments of the present invention, the present invention also provides the use of compounds represented by formula (I-2) or (I-4), especially compounds 8c, 9c or 10c, for the preparation of medicaments for inhibiting tumor cells, such as cancer cells.

[0063] According to embodiments of the present invention, the cancer cells may be cervical cancer cells, lung adenocarcinoma cells, breast cancer cells, liver cancer cells, esophageal cancer cells, prostate cancer cells, colon cancer cells, or lung cancer cells such as non-small cell lung cancer cells.

[0064] As an example, the cancer cells are selected from one of the following cell lines: human cervical cancer cells HeLa, human liver cancer cells HepG2, human liver cancer cells SMMC-7721, human breast cancer cells MCF-7, human prostate cancer cells PC-3, human esophageal cancer cells KYSE-150, human colon cancer cells SW620, and human non-small cell lung cancer cells A549.

[0065] Beneficial effects

[0066] The inventors unexpectedly discovered that some compounds provided by this invention exhibit excellent inhibitory activity against tested cancer cells, particularly showing surprising inhibitory activity against colon cancer or prostate cancer cells. Furthermore, compounds 8c, 9c, or 10c exhibit even more superior inhibitory activity against certain tested cancer cells. Therefore, the compounds provided by this invention hold promise for improving the prevention or treatment of cancers such as colon cancer and prostate cancer, or related symptoms.

[0067] Terminology Definitions and Explanations

[0068] Unless otherwise stated, the definitions of groups and terms recorded in this application specification and claims, including their 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 shall fall within the scope of this application specification.

[0069] Unless otherwise stated, the numerical ranges described in this specification and claims are equivalent to describing at least each specific integer value therein. For example, the numerical range "1-40" is equivalent to describing each integer value in the numerical range "1-10", namely 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and each integer value in the numerical range "11-40", namely 11, 12, 13, 14, 15, ..., 35, 36, 37, 38, 39, 40. Furthermore, when certain numerical ranges are defined as "numbers", they should be understood to describe the two endpoints of the range, each integer within the range, and each decimal within the range. For example, "numbers from 0 to 10" should be understood to describe not only each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, but also at least the sum of each of these integers with 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, and 0.9.

[0070] It should be understood that in the description of 1, 2 or more, "more" should refer to an integer greater than 2, such as an integer greater than or equal to 3, such as 3, 4, 5, 6, 7, 8, 9 or 10.

[0071] The term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0072] Term "C" 1-40 "Alkyl" should be understood as referring to a straight-chain or branched saturated monovalent hydrocarbon group having 1 to 40 carbon atoms. For example, "C 1-6 "Alkyl" means a straight-chain or branched alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, or their isomers.

[0073] Term "C" 2-40 "Alkenyl" should be understood to preferably represent a straight or branched monovalent hydrocarbon group containing one or more double bonds and having 2 to 40 carbon atoms, preferably "C". 2-10 "Alkenyl". "C" 2-10 "Alkenyl" should be understood to preferably represent a straight or branched monovalent hydrocarbon group containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, for example, having 2, 3, 4, 5, or 6 carbon atoms (i.e., C...). 2-6 alkenyl), having 2 or 3 carbon atoms (i.e., C24, C34, C4 ... 2-3Alkenyl). It should be understood that when the alkenyl group contains more than one double bond, the double bonds may be separable or conjugated. The alkenyl group is, for example, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z)-pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)- Pentyl-1-enyl, (Z)-pentyl-1-enyl, hex-5-enyl, (E)-hex-4-enyl, (Z)-hex-4-enyl, (E)-hex-3-enyl, (Z)-hex-3-enyl, (E)-hex-2-enyl, (Z)-hex-2-enyl, (E)-hex-1-enyl, (Z)-hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl 2-Methylprop-1-enyl, (E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2-enyl, (Z)-1-methyl But-2-enyl, (E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl, (E)-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl, (E)-1-methylbut-1-enyl, (Z)-1-methylbut-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.

[0074] Term "C" 2-40 "Alkyne group" should be understood as representing a straight or branched monovalent hydrocarbon group containing one or more triple bonds and having 2 to 40 carbon atoms, preferably "C". 2-10 "Alkyne group". The term "C" 2-10 "Alkyne" should be understood to preferably represent a straight or branched monovalent hydrocarbon group containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, for example, having 2, 3, 4, 5, or 6 carbon atoms (i.e., "C"). 2-6 The alkynyl group ("C") has 2 or 3 carbon atoms ("C") 2-3The alkynyl group is, for example, ethynyl, prop-1-alkynyl, prop-2-alkynyl, but-1-alkynyl, but-2-alkynyl, but-3-alkynyl, pent-1-alkynyl, pent-2-alkynyl, pent-3-alkynyl, pent-4-alkynyl, hex-1-alkynyl, hex-2-alkynyl, hex-3-alkynyl, hex-4-alkynyl, hex-5-alkynyl, 1-methylprop-2-alkynyl, 2-methylbut-3-alkynyl, 1-methylbut-3-alkynyl, 1-methylbut-2-alkynyl, 3-methylbut-1-alkynyl, 1-ethylprop-2-alkynyl, 3-methylpent-4-alkynyl, 2-methylpent-4-alkynyl, 1-methylpent-4-alkynyl -Alynyl, 2-methylpentan-3-ynyl, 1-methylpentan-3-ynyl, 4-methylpentan-2-ynyl, 1-methylpentan-2-ynyl, 4-methylpentan-1-ynyl, 3-methylpentan-1-ynyl, 2-ethylbutan-3-ynyl, 1-ethylbutan-3-ynyl, 1-ethylbutan-2-ynyl, 1-propylpropan-2-ynyl, 1-isopropylpropan-2-ynyl, 2,2-dimethylbutan-3-ynyl, 1,1-dimethylbutan-3-ynyl, 1,1-dimethylbutan-2-ynyl, or 3,3-dimethylbutan-1-ynyl. In particular, the ynyl group is ethynyl, propan-1-ynyl, or propan-2-ynyl.

[0075] Term "C" 3-20 "Cycloalkyl" should be understood to refer to saturated monovalent monocyclic, bicyclic, or bridged cycloalkanes having 3 to 20 carbon atoms, preferably "C". 3-10 cycloalkyl. The term "C" 3-10 "Cycloalkyl" should be understood to refer to saturated monovalent monocyclic, bicyclic, or bridged cycloalkanes having 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The C... 3-10 Cycloalkyl groups can be monocyclic hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl, or bicyclic hydrocarbon groups such as decahydronaphthalene ring.

[0076] The term "3-20 membered heterocyclic group" refers to a saturated monovalent monocyclic, bicyclic, or bridged cyclic alkane comprising 1-5 heteroatoms independently selected from N, O, and S, forming a non-aromatic cyclic group with a total number of 3-20 atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, etc.), preferably a "3-10 membered heterocyclic group". The term "3-10 membered heterocyclic group" also refers to a saturated monovalent monocyclic, bicyclic, or bridged cyclic alkane comprising 1-5, preferably 1-3, heteroatoms selected from N, O, and S. The heterocyclic group can be attached to the remainder of the molecule via any one of the carbon atoms or a nitrogen atom (if present). Specifically, the heterocyclic group may include, but is not limited to: 4-membered rings, such as azirrobutyl and oxobutyl; 5-membered rings, such as tetrahydrofuranyl, dioxacyclopentenyl, pyrrolyl, imidazoyl, pyrazolyl, or pyrrololinyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazinyl, or trithiaalkyl; or 7-membered rings, such as diazacycloheptyl. Optionally, the heterocyclic group may be benzofused. The heterocyclic group may be bicyclic, for example, but not limited to, 5,5-membered rings, such as hexahydrocyclopenta[c]pyrrolo-2(1H)-yl rings, or 5,6-membered bicyclic rings, such as hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl rings. The nitrogen-containing ring can be partially unsaturated, meaning it can contain one or more double bonds, such as, but not limited to, 2,5-dihydro-1H-pyrrole, 4H-[1,3,4]thiadiazinyl, 4,5-dihydrooxazolyl, or 4H-[1,4]thiazinyl, or it can be benzofused, such as, but not limited to, dihydroisoquinolinyl. According to the invention, the heterocyclic group is non-aromatic. When the 3-20 membered heterocyclic group is linked to other groups to form the compounds of the invention, the carbon atom on the 3-20 membered heterocyclic group can be linked to other groups, or the heterocyclic atom on the 3-20 membered heterocyclic ring can be linked to other groups. For example, when the 3-20 membered heterocyclic group is selected from piperazine, the nitrogen atom on the piperazine group can be linked to other groups. Or when the 3-20 membered heterocyclic group is selected from piperidinyl, the nitrogen atom on the piperidinyl ring and its para-carbon atom can be linked to other groups.

[0077] Term "C" 6-20 "Aryl" should be understood to preferably represent a monocyclic, bicyclic, or tricyclic hydrocarbon ring having 6 to 20 carbon atoms and possessing monovalent aromaticity or partial aromaticity, preferably "C". 6-14 "Aromatic". The term "C" 6-14 "Aryl" should be understood to preferably represent a monovalent aromatic or partially aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring ("C") having 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon atoms. 6-14Aryl), particularly a ring with 6 carbon atoms (“C6 aryl”), such as phenyl; or biphenyl, or a ring with 9 carbon atoms (“C9 aryl”), such as indenyl or indenyl, or a ring with 10 carbon atoms (“C9 aryl”). 10 Aryl groups, such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl, or rings with 13 carbon atoms (“C”). 13 Aryl groups, such as fluorene groups, or rings with 14 carbon atoms (“C”). 14 Aryl), for example, anthracene. When the C 6-20 When the aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution site; for example, it can be ortho, para, or meta substituted.

[0078] The term "5-20-membered heteroaryl" should be understood to include monovalent monocyclic, bicyclic, or tricyclic aromatic ring systems having 5 to 20 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O, and S, such as "5-14-membered heteroaryl". The term "5-14-membered heteroaryl" should also be understood to include monovalent monocyclic, bicyclic, or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 5, 6, 9, or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3, heteroatoms independently selected from N, O, and S, and in each case, may be benzofused. Specifically, the heteroaryl group is selected from thienyl, furanyl, pyrroleyl, oxazolyl, thiazolyl, imidazoleyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl, and their benzo[derivatives], such as benzofuranyl, benzothienyl, benzoxazolyl, benzoisoxazolyl, benzoimidazolyl, benzotriazolyl, indazole, indolyl, isindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and their benzo[derivatives], such as quinolinyl, quinazolinyl, isoquinolinyl, etc.; or acrylonitrileyl, inazinyl, purinyl, and their benzo[derivatives]; or terpineyl, phthalazinyl, quinazolinyl, quinoxolinyl, naphthidyl, pteridinyl, carbazolyl, acrylonitrileyl, phenazinyl, phenothiazinyl, phenothiazinyl, phenothiazinyl, etc. When the 5-20 membered heteroaryl group is linked with other groups to form the compound of the present invention, the carbon atom on the 5-20 membered heteroaryl ring may be linked with other groups, or the heteroatom on the 5-20 membered heteroaryl ring may be linked with other groups. When the 5-20 membered heteroaryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution site; for example, the hydrogen atom linked to the carbon atom on the heteroaryl ring may be substituted, or the hydrogen atom linked to the heteroatom on the heteroaryl ring may be substituted.

[0079] Unless otherwise stated, heterocyclic, heteroaryl, or heteroaryl groups include all possible isomers, such as their positional isomers. Thus, for some illustrative, non-limiting examples, forms may include those in which one, two, or more of the following positions (if present) are substituted or bonded to other groups, including pyridin-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-3-yl, pyridin-4-yl, and pyridin-4-yl; thiophene or thiophene groups include thiophene-2-yl, thiophene-2-yl, thiophene-3-yl, and thiophene-3-yl; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

[0080] Unless otherwise stated, the definitions of terms in this document also apply to groups containing the term, such as C. 1-40 The definition of alkyl also applies to C 1-40 Alkyl groups, etc.

[0081] Those skilled in the art will understand that the compounds shown in Formula I can exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they can form acid addition salts; if these compounds have an acidic center, they can form base addition salts; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they can also form inner salts. Acid addition salts include, but are not limited to: hydrochlorides, hydrofluorides, hydrobroms, hydroiodates, sulfates, pyrosulfonates, phosphates, nitrates, methanesulfonates, ethanesulfonates, 2-hydroxyethanesulfonates, benzenesulfonates, toluenesulfonates, aminosulfonates, 2-naphthalenesulfonates, formates, acetoacetic acid, pyruvic acid, ceramides, cinnamates, benzoates, acetates, dihydroxyacetates, trifluoroacetates, trimethylacetates, propionates, butyrates, hexanoates, heptanoates, undecanoates, stearates, ascorbic acid salts, camphorates, camphor sulfonates. Citrate, fumarate, malate, maleate, hydroxymaleate, oxalate, salicylate, succinate, gluconate, quinate, dihydroxynaphthylate, glycolate, tartrate, lactate, 2-(4-hydroxybenzoyl)benzoate, cyclopentanepropionate, digluconate, 3-hydroxy-2-naphthylcarboxylate, nicotinate, pyruvate, pectin ester, 3-phenylpropionate, picrate, pentylate, itaconic acid, trifluoromethanesulfonate, dodecyl sulfate, p-toluenesulfonate, naphthalene disulfonate, malonate. Adipate, alginate, mandelate, gluconate, glycerol phosphate, sulfosalicylate, hemisulfate or thiocyanate, aspartate, etc.; base addition salts such as alkali metal salts, alkaline earth metal salts, and ammonium salts, specifically including but not limited to: sodium salts, lithium salts, potassium salts, ammonium salts, aluminum salts, magnesium salts, calcium salts, barium salts, iron salts, ferrous salts, manganese salts, manganese salts, zinc salts, ammonium salts (including salts formed with NH3 and organic amines (NH4 salts), methylammonium salts, trimethylammonium salts, diethylammonium salts, triethylammonium salts, propanemonium salts, tripropylammonium salts, isopropylammonium salts, tert-butylammonium salts, N,N '-Dibenzyl ethylenediamine salt, dicyclohexanediamine salt, 1,6-hexammoniadiamine salt, benzyl ammonium salt, ethanol ammonium salt, N,N-dimethyl ethanol ammonium salt, N,N-diethyl ethanol ammonium salt, triethanolammonium salt, tromethamine salt, lysine salt, arginine salt, histidine salt, glucosamine salt, N-methylglucosamine salt, dimethylglucosamine salt, ethylglucosamine salt, glucosamine, betaine salt, caffeine salt, chloroprocaine salt, procaine salt, lidocaine salt, pyridine salt, methylpyridine salt, piperidine salt, morpholine salt, piperazine salt, purine salt, theobromine salt, choline salt, etc.

[0082] Based on their molecular structure, the compounds of the present invention are chiral and therefore may exist in various enantiomeric forms. Thus, these compounds can exist in racemic or optically active forms. The compounds of the present invention or their intermediates can be isolated as enantiomers by chemical or physical methods known to those skilled in the art, or used in this form for synthesis. 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 enantiomeric resolution 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.

[0083] The term "tautomerism" includes those tautomer forms known to those skilled in the art, such as tautomers selected from enol-keto, amide-imino, lactam-lactamimide, enamine-iminoenamine-enamine, etc.

[0084] The corresponding stable isomers can be separated using known methods, such as extraction, filtration, or column chromatography.

[0085] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred.

[0086] The term “therapeutic effective amount” as used herein refers to the amount of an active compound or drug that researchers, veterinarians, physicians, or other clinicians are searching for in tissues, systems, animals, individuals, or humans to elicit a biological or medical response. It includes one or more of the following: (1) prevention of disease: e.g., prevention of disease, disorder, or condition in individuals who are susceptible to disease, disorder, or symptom but have not yet experienced or developed the pathology or symptoms of the disease. (2) suppression of disease: e.g., suppression of disease, disorder, or symptom in individuals who are experiencing or developing the pathology or symptoms of the disease, disorder, or symptom (i.e., prevention of further development of the pathology and / or symptoms). (3) relief of disease: e.g., relief of disease, disorder, or symptom in individuals who are experiencing or developing the pathology or symptoms of the disease, disorder, or symptom (i.e., reversal of the pathology and / or symptoms). Detailed Implementation

[0087] 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.

[0088] I. Preparation Example

[0089] The following compounds 8a-e, 9a-e, and 10a-e are known in the prior art and can be obtained, for example, by the methods described in the inventor's published paper Qiang Wang, et al., ACS Med. Chem. Lett. 2020, 11, 2290-2293, and in the inventor's prior Chinese patent applications 201810483852.9, 201810483941.3, 201810700317.4, 201911216447.1, and 202010647506.7:

[0090]

[0091] Alternatively, the above compounds can also be prepared by reacting the raw materials AFC, AFU, AZT, AZC, and AZU with compounds 8, 9, or 10, respectively, based on the previously published paper by Qiang Wang, et al., ACSMed. Chem. Lett. 2020, 11, 2290-2293.

[0092]

[0093] II. Activity Test Examples

[0094] 1. Experimental Materials

[0095] 1.1 Cell lines

[0096] Human cervical cancer cells HeLa, human liver cancer cells HepG2, human liver cancer cells SMMC-7721, human breast cancer cells MCF-7, human prostate cancer cells PC-3, human esophageal cancer cells KYSE-150, human colon cancer cells SW620, and human non-small cell lung cancer cells A549 were all passaged and preserved in our laboratory.

[0097] 1.2 Experimental reagents

[0098] The above compounds are 8a-e, 9a-e, and 10a-e.

[0099] Betulinic acid (BA), purchased from Aladdin, lot number L1614002.

[0100] Cisplatin, purchased from Yuanye Biotechnology Co., Ltd., batch number B21M11L110000.

[0101] 1.3 Experimental Reagents

[0102]

[0103] 1.4 Experimental Apparatus

[0104]

[0105]

[0106] 2. Experimental Methods

[0107] 2.1 Preparation of relevant reagents

[0108] (1) DMEM complete medium: Fetal bovine serum, DMEM medium and double antibiotics were placed in a sterile 50mL EP tube at a volume ratio of 1:9:0.1, mixed evenly and stored at 4℃.

[0109] (2) 1640 complete medium: Fetal bovine serum, 1640 medium and double antibiotics were placed in a sterile 50 mL EP tube at a volume ratio of 1:9:0.1, mixed well and stored at 4℃.

[0110] (3) Preparation of the test drug: Accurately weigh a certain mass of the test drug, put it into a 1.5 mL EP tube, dissolve it in a certain volume of DMSO solution to make the drug concentration 100 mmol / L, and store it in a refrigerator at 4℃.

[0111] 2.2 Cell resuscitation

[0112] Remove the frozen cells and thaw them rapidly in a 37°C water bath. Centrifuge, discard the supernatant, resuspend the cells in complete culture medium, transfer them to a cell culture flask, and continue culturing.

[0113] 2.3 Cell passage

[0114] When the cells proliferated to 75% in the culture flask, wash them three times with PBS, add 2 mL of trypsin, incubate for 2 min, then add 2 mL of complete culture medium, centrifuge, discard the supernatant, add complete culture medium and continue culturing.

[0115] 2.4 Cell cryopreservation

[0116] When the cells proliferate to 75% in the culture flask, digest the cells, centrifuge, discard the supernatant, add cell cryopreservation solution and place them in cell cryovials, label with cryopreservation information, then place the cryovials in a cryopreservation box and store in a -80°C freezer, and then transfer to a liquid nitrogen tank.

[0117] 2.5 Cell Count

[0118] Digest the cells, then centrifuge, discard the supernatant, and add complete culture medium to dilute the cell suspension. Add 10 μL of the cell suspension to a cell counting plate, ensuring no air bubbles are present, and count the cells using an inverted microscope. Determine the original cell count based on the counted cells and the dilution factor.

[0119] Cell counting formula:

[0120] 2.6 CCK-8 assay for cell growth inhibition rate

[0121] (1) When the cells proliferate to 75% in the culture flask, collect the cells, centrifuge, discard the supernatant, resuspend the cells in complete culture medium, and adjust the cell density to 8 × 10⁻⁶. 4 Cells were seeded at a density of 100 μL / mL in 96-well plates, with 100 μL of cell suspension in each well. 100 μL of sterile PBS solution was added to the peripheral wells to prevent marginalization.

[0122] (2) After culturing in a cell culture incubator for 12 hours, the culture medium was discarded, and 100 μL of drug solution was added to each well to make the final concentrations of the drug solution in the experimental groups 0.75, 1.5, 3, 6, 12.5, 25, 50, 75 and 100 μmol / L, respectively. At the same time, a BA control group, a positive control group of cisplatin, a negative control group and a PBS zeroing group were set up, and three replicates were set up for each concentration.

[0123] (3) After the 96-well plate was incubated in the incubator for 48 hours, the 96-well plate was taken out and placed on a sterile operating table. 20 μL of CCK-8 solution was added to each well, and then the plate was placed in the incubator for 4 hours.

[0124] (4) After incubation, the absorbance (OD value) of the fully automated enzyme-linked immunosorbent assay (ELISA) is detected at a wavelength of 450 nm.

[0125] (5) The experiment was repeated 3 times, and the cell inhibition rate was calculated according to the following formula:

[0126]

[0127] 3. Experimental Results

[0128] The activity test results of the test compounds are shown in Tables 1 and 2 below:

[0129] Table 1

[0130]

[0131]

[0132] Table 2

[0133]

[0134] The data above show that some of the tested compounds exhibited good inhibitory activity against human prostate cancer cells PC-3 and human colon cancer cells SW620. Compound 8c showed better inhibitory activity against all cell lines than compounds 8a and 8b, compound 10c showed better inhibitory activity against all cell lines than compounds 10a and 10b, and compound 9c showed better inhibitory activity against HepG2, PC-3, KYSE-150, and SW620 than compounds 9a and 9b.

[0135] The foregoing has described specific exemplary embodiments of the present invention. It should be understood that the scope of protection of the present invention is not limited to the exemplary embodiments described above. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the scope of protection of the claims of this application.

Claims

1. Use of a compound of formula (I-4) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of colon cancer or prostate cancer: in, X is O; Y is OH; R1 is selected from H and C. 1-6 alkyl; R4 is selected from C 1-6 alkyl; R5 is H; Z is selected from O and OH, when When it is a double bond, Z is selected from O; when When it is a single bond, Z is selected from OH.

2. The use as described in claim 1, wherein the compound represented by formula (I-4) is selected from the following compounds:

3. Use of the compound represented by formula (I-4) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for inhibiting cancer cells selected from: colon cancer cells, prostate cancer cells, wherein, The compound shown in formula (I-4) has the definition as claimed in claim 1 or 2.

4. The use of the compound represented by formula (I-4) in the preparation of a medicament for the prevention or treatment of cancer or for the inhibition of cancer cells, wherein, The compound shown in formula (I-4) has the definition of claim 1 or 2; The cancer is selected from prostate cancer or colon cancer; The cancer cells are selected from prostate cancer cells or colon cancer cells.

5. The use as described in claim 4, wherein the compound is selected from at least one of compounds 8c, 9c, and 10c of claim 2.

6. The use as described in claim 1, wherein the drug is a pharmaceutical composition.

7. The use as described in claim 6, wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

8. The use as described in claim 6, wherein the pharmaceutical composition contains one or more additional therapeutic agents.

Images