阅读说明：本技术 带桥环的环己烷二羧酸衍生物及其药物组合物和应用 (Cyclohexane dicarboxylic acid derivative with bridged ring, and pharmaceutical composition and application thereof ) 是由 汤磊 陈文章 彭严 彭金刚 崔杏 于 2020-05-07 设计创作，主要内容包括：本发明公开了一种具有通式(I)所示的带桥环的环己烷二羧酸衍生物、立体异构体和药学上可接受的盐在制备抗肿瘤药物中的应用,特别是针对血癌、肝癌、肺癌症、胃癌和卵巢癌具有明显的抑制作用。本发明的化合物具有较高的抗肿瘤活性、抗肿瘤谱宽且毒性低,适用于制备抗癌药物。(The invention discloses application of cyclohexane dicarboxylic acid derivatives with bridged rings, stereoisomers and pharmaceutically acceptable salts shown in a general formula (I) in preparation of antitumor drugs, and particularly has an obvious inhibiting effect on leukemia, liver cancer, lung cancer, gastric cancer and ovarian cancer. The compound of the invention has higher antitumor activity, wide antitumor spectrum and low toxicity, and is suitable for preparing anticancer drugs.)

1. A compound of formula (I), a stereoisomer thereof, a pharmaceutically acceptable salt thereof or a solvate thereof,

wherein, X₁And X₂Identical or different, independently of one another, from NH and O; or X₁And X₂Together with the carbon atom to which they are attached form a five-or six-membered ring, and when forming a five-membered ring, X₁And X₂Is an atom;

R₁、R₂identical or different, independently of one another, from hydrogen, OH, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-8Cycloalkyl and C_3-30Cycloalkenyl radical, whereinC_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-8Cycloalkyl and C_3-30The cycloalkenyl is optionally substituted with one, two or more of the following groups: hydroxy, carboxy, sulfonic acid, oxo (═ O), C_1-6Alkyl radical, C_2-6Alkenyl radicals, substituted by one, two or more hydroxy radicals, C_1-6Alkyl or C_2-6Alkenyl-substituted C_3-8Cycloalkyl or C_3-8A cycloalkenyl group;

R₃、R₄identical or different, independently of one another, from hydrogen, hydroxy, C_1-6Alkyl and C_1-6An alkoxy group;

or, R₃、R₄Together with the carbon atom to which they are attached form C_3-6Cycloalkyl or 3-6 membered heterocyclyl, wherein said C is_3-6Cycloalkyl or 3-6 membered heterocyclyl is optionally substituted by one, two or more oxo (═ O), hydroxy or C_1-6Alkyl substituted and the 3-6 membered heterocyclyl contains one or two heteroatoms selected from O, N and S;

y is selected from O, carbonyl and methylene, wherein the methylene is optionally substituted with one, two or more OH, C_1-6Alkyl radical, C_1-6Alkoxy or C_2-6And (3) alkenyl substitution.

2. The compound according to claim 1, wherein, in the compound of formula (I),

X₁and X₂Is an O atom or is an N atom, in which case X₁And X₂Together with the carbon atoms to which they are attached form a succinic anhydride or optionally substituted succinimide five-membered ring;

R₁、R₂identical or different, independently of one another, from hydrogen, C_1-6Alkyl radical, C_3-8Cycloalkyl and C_3-30Cycloalkenyl radical, wherein said C_1-6Alkyl radical, C_3-8Cycloalkyl and C_3-30The cycloalkenyl is optionally substituted with one, two or more of the following groups: hydroxy, carboxy, sulfonic acid, oxo (═ O), C_1-6Alkyl radical, C_2-6Alkenyl radicals, substituted by one, two or more hydroxy radicals, C_1-6Alkyl or C_2-6Alkenyl-substituted C_3-8Cycloalkyl or C_3-8A cycloalkenyl group.

3. The compound according to claim 1 or 2, wherein, in the compound of formula (I),

X₁and X₂Are all O;

R₁、R₂identical or different, independently of one another, from hydrogen, or a monovalent radical formed by a monocyclic monoterpene compound or a pentacyclic triterpene compound: perillyl alcohol, menthol, isopulegol, alpha-terpineol, beta-terpineol, gamma-terpineol, carveol, oleanolic acid, ursolic acid, maslinic acid, corosolic acid, glycyrrhetinic acid, asiatic acid, saikosaponin E, saikosaponin F, saikosaponin G, secondary saikosaponin A, secondary saikosaponin B, secondary saikosaponin C, secondary saikosaponin D, saikosaponin, phytolaccagenic acid-30-methyl ester, 2-hydroxyphytolaccacid, 2-hydroxyphytolaccagenic acid-30-methyl ester, betulinic acid, 23-hydroxybetulinic acid, triptonide;

preferably, the monovalent group formed by the monocyclic monoterpene compound or the pentacyclic triterpene compound may be 4- (1-methylvinyl) -1-cyclohexene-1-methoxy group, (1R,2S,5R) -2-isopropyl-5-methylcyclohexyloxy group, 5-methyl-2- (1-methylvinyl) cyclohexyloxy group, 2- (4-methyl-3-cyclohexenyl) -2-propoxy group, 1-methyl-4- (1-methylvinyl) cyclohexyloxy group, 1-methyl-4-isopropylidene-cyclohexyloxy group, 2-methyl-5- (1-methylvinyl) -2-cyclohexenyloxy group, 12-ene-28-carboxyl-oleanane-3-oxyl, 12-ene-28-carboxyl-ursane-3 beta-oxyl, 12-ene-28-acid-2 alpha-hydroxyl oleanane-3 beta-oxyl, 12-ene-28-acid-3 beta-hydroxyl oleanane-2 alpha-oxyl, 12-ene-28-oic acid-2 alpha-hydroxy ursane-3 beta-oxyl, 12-ene-28-oic acid-3 beta-hydroxy ursane-2 alpha-oxyl and (3 beta, 20 beta) -11-oxo-olean-12-ene-29-oic acid-3-oxyl.

4. A compound according to any one of claims 1 to 3, wherein, in the compound of formula (I),

R₃、R₄identical or different, independently of one another, from hydrogen, C_1-3Alkyl and C_1-3An alkoxy group;

or, R₃、R₄Together with the carbon atom to which they are attached form a cyclopropane, cyclobutyl, cyclopentyl, cyclohexyl, oxirane, oxetane, oxolanyl, oxocyclohexyl, azetidinyl, azacyclopentyl, azacyclohexyl or thiacyclopentyl group;

y is selected from O, carbonyl, or by OH or C_1-3An alkoxy-substituted methylene group.

5. The compound according to any one of claims 1-4, wherein the compound of formula (I) is selected from the following compounds (1) to (13):

6. a process for the preparation of a compound according to any one of claims 1 to 5, characterized in that it comprises: reacting compound M1 with a compound capable of providing R₁And/or R₂Reacting the compound of the group to obtain the compound shown in the formula (I):

wherein R is₁、R₂、R₃、R₄、Y、X₁And X₂Each having the definitions given in any one of claims 1 to 5;

preferably, said can provide R₁And/or R₂The group is selected from R₁OH、R₁NH₂、R₂OH and R₂NH₂；

Preferably, said can provide R₁And/or R₂The group is selected from methanol, taurine, monocyclic monoterpene compound or pentacyclic triterpene compound;

preferably, said can provide R₁And/or R₂The group is selected from perillyl alcohol, menthol, isopulegol, alpha-terpineol, beta-terpineol, gamma-terpineol, and carveol;

preferably, said can provide R₁And/or R₂The group is selected from oleanolic acid, ursolic acid, crataegolic acid, corosolic acid, glycyrrhetinic acid, asiatic acid, saikosaponin E, saikosaponin F, saikosaponin G, secondary saikosaponin A, secondary saikosaponin B, secondary saikosaponin C, secondary saikosaponin D, radix bupleuri sapogenin, phytolaccagenic acid-30-methyl ester, 2-hydroxy phytolaccagenic acid-30-methyl ester, betulinic acid, 23-hydroxy betulinic acid, and triptonide.

7. The production method according to claim 6, wherein the reaction is carried out in the presence of a solvent such as an organic solvent.

8. The production method according to claim 6 or 7, wherein the reaction may be carried out in the presence of an acid or a base;

the acid is organic acid or inorganic acid; for example, at least one selected from formic acid, acetic acid, propionic acid, trifluoroacetic acid, HCl, sulfuric acid;

the alkali is organic alkali or inorganic alkali; the inorganic base may be selected from at least one of: hydrides, hydroxides, alkoxides, acetates, fluorides, phosphates, carbonates and bicarbonates of alkali metals or alkaline earth metals. Preferred bases are sodium amide, sodium hydride, lithium diisopropylamide, sodium methoxide, potassium tert-butoxide, sodium hydroxide, potassium hydroxide, sodium acetate, sodium phosphate, potassium fluoride, cesium fluoride, sodium carbonate, potassium bicarbonate, sodium bicarbonate, and cesium carbonate; the organic base may be selected from at least one of the following: tertiary amines, substituted or unsubstituted pyridines and substituted or unsubstituted triethylamine, trimethylamine, N, N-diisopropylethylamine, tri-N-propylamine, tri-N-butylamine, tri-N-hexylamine, tricyclohexylamine, N-methylcyclohexylamine, N-methylpyrrolidine, N-methylpiperidine, N-ethylpiperidine, N, N-dimethylaniline, N-methylmorpholine, pyridine, 2, 3-or 4-methylpyridine, 2-methyl-5-ethylpyridine, 2, 6-dimethylpyridine, 2,4, 6-trimethylpyridine, 4-dimethylaminopyridine, quinoline, methylquinoline, N, N, N-tetramethylethylenediamine, N, N-dimethyl-1, 4-diazacyclohexane, N, N-diethyl-1, 4-diazabicyclocyclohexane, 1, 8-bis (dimethylamino) naphthalene, Diazabicyclooctane (DABCO), Diazabicyclononane (DBN), Diazabicycloundecane (DBU), butylimidazole and methylimidazole.

9. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds of formula (I), racemates, stereoisomers, pharmaceutically acceptable salts or solvates thereof as claimed in any one of claims 1 to 5.

10. Use of at least one compound of formula (I), a stereoisomer, a pharmaceutically acceptable salt or a solvate according to any one of claims 1 to 5 in the preparation of an anti-tumor medicament;

the antitumor drug is preferably used for treating leukemia, liver cancer, lung cancer, gastric cancer or ovarian cancer.

Technical Field

The invention belongs to the field of antitumor drugs, and particularly relates to a cyclohexane dicarboxylic acid derivative with a bridged ring, a pharmaceutical composition and an application thereof.

Background

Tumors are a complex disease with a multi-etiology network, and the mortality rate is second worldwide. Currently, antitumor drugs mainly aim at a single target or route, and the drugs have good treatment effects (nat. Rev. drug Discv.2006,5, 993-. However, most drugs have damage to the hematopoietic system, so that they show serious toxic and side effects clinically. In addition, most tumor drugs used in clinic belong to immunosuppressants, which inhibit the immune response of the body while controlling the proliferation of cancer cells, thereby reducing the defense capability of the body against tumors, and possibly increasing the risk of other diseases, such as infectious diseases (bioorg. med. chem.2007,15, 6126-6134). On the other hand, with a single target anti-tumor drug, after the drug blocks a target or a pathway, tumor cells may compensate the change through other potential pathways, so that the effect is weakened or eliminated (curr. med.2008,15, 422-. Therefore, the research and development of the anti-tumor drugs with better activity and multiple functions, such as drugs without immune suppression and with multiple targets and targeting, is one of the key points and difficulties in the research and development of the anti-tumor drugs.

Norcantharidin (NCTD), a bridged ring cyclohexanedicarboxylic acid compound with the chemical name of exo-7-oxabicyclo [2,2,1] heptane-2, 3-dicarboxylic anhydride, has the following structural formula:

norcantharidin is clinically used for treating primary liver cancer, and has unique leukocyte increasing function while inhibiting liver cancer cells. However, the norcantharidin still has the problems of high toxicity, narrow antitumor spectrum and the like, and the clinical application of the norcantharidin is limited.

Terpenoids are widely found in nature and possess many biological activities. Terpenes can be classified into hemiterpenes (containing one isoprene unit), monoterpenes (containing two isoprene units), sesquiterpenes (containing three isoprene units), diterpenes (containing four isoprene units), triterpenes (containing six isoprene units), and the like, according to the number of isoprene units in their molecular skeleton. Wherein the monoterpene compounds such as perillyl alcohol (4- (1-methylethenyl) -1-cyclohexene-1-methanol) are natural products with terpineol smell, have a tumor inhibition mechanism different from norcantharidin, have prevention and treatment effects on various tumors, have stronger anti-colon cancer, breast cancer and prostate cancer effects, and have entered the 2 nd-phase clinical research (NCS-641066) abroad at present. But the perilla alcohol has weak inhibition effect on liver tumor, blood tumor and lung tumor, has pungent smell and is not beneficial to being prepared into oral preparations.

Triterpenes, such as oleanolic acid (3-hydroxy-oleanane-12-ene-28-acid), have an antioxidation effect, are often used as liver protection preparations clinically, and have certain antitumor activity in recent years.

There is a need in the art for more norcantharidin derivative compounds with anti-tumor activity.

Disclosure of Invention

In order to improve the technical problems, the invention provides a cyclohexane dicarboxylic acid derivative with a bridged ring or norcantharidin derivative, namely a compound shown as a formula (I), a stereoisomer thereof, a pharmaceutically acceptable salt thereof or a solvate thereof,

wherein, X₁And X₂Identical or different, independently of one another, from NH and O; or X₁And X₂Together with the carbon atom to which they are attached form a five-or six-membered ring, and when forming a five-membered ring, X₁And X₂Is an atom;

R₁、R₂identical or different, independently of one another, from hydrogen, OH, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-8Cycloalkyl and C_3-30Cycloalkenyl radical, wherein said C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-8Cycloalkyl and C_3-30The cycloalkenyl is optionally substituted with one, two or more of the following groups: hydroxy, carboxy, sulfonic acid, oxo (═ O), C_1-6Alkyl radical, C_2-6Alkenyl radicals, substituted by one, two or more hydroxy radicals, C_1-6Alkyl or C_2-6Alkenyl-substituted C_3-8Cycloalkyl or C_3-8A cycloalkenyl group;

R₃、R₄identical or different, independently of one another, from hydrogen, hydroxy, C_1-6Alkyl and C_1-6An alkoxy group;

or, R₃、R₄Together with the carbon atom to which they are attached form C_3-6Cycloalkyl or 3-6 membered heterocyclyl, wherein said C is_3-6Cycloalkyl or 3-6 membered heterocyclyl optionally substituted by one, two or moreOxo (═ O), hydroxy or C_1-6Alkyl substituted and the 3-6 membered heterocyclyl contains one or two heteroatoms selected from O, N and S;

y is selected from O, carbonyl and methylene, wherein the methylene is optionally substituted with one, two or more OH, C_1-6Alkyl radical, C_1-6Alkoxy or C_2-6And (3) alkenyl substitution.

According to an embodiment of the invention, in the compound of formula (I),

X₁and X₂Is an O atom or is an N atom, in which case X₁And X₂Together with the carbon atoms to which they are attached form a succinic anhydride or optionally substituted succinimide five-membered ring;

R₁、R₂identical or different, independently of one another, from hydrogen, C_1-6Alkyl radical, C_3-8Cycloalkyl and C_3-30Cycloalkenyl radical, wherein said C_1-6Alkyl radical, C_3-8Cycloalkyl and C_3-30The cycloalkenyl is optionally substituted with one, two or more of the following groups: hydroxy, carboxy, sulfonic acid, oxo (═ O), C_1-6Alkyl radical, C_2-6Alkenyl radicals, substituted by one, two or more hydroxy radicals, C_1-6Alkyl or C_2-6Alkenyl-substituted C_3-8Cycloalkyl or C_3-8A cycloalkenyl group.

According to an embodiment of the invention, in the compound of formula (I),

X₁and X₂Are all O;

R₁、R₂identical or different, independently of one another, from hydrogen, or a monovalent radical formed by a monocyclic monoterpene compound or a pentacyclic triterpene compound: perillyl alcohol, menthol, isopulegol, alpha-terpineol, beta-terpineol, gamma-terpineol, carveol, oleanolic acid, ursolic acid, crataegolic acid, corosolic acid, glycyrrhetinic acid, asiatic acid, saikogenin E, saikogenin F, saikogenin G, secondary saikogenin A, secondary saikogenin B, secondary saikogenin C, secondary saikogenin D, radix bupleuri sapogenin, phytolaccagenic acid-30-methyl ester, 2-hydroxy phytolaccagenic acid, phytolaccaine, and other compoundsPhytolaccic acid-30-methyl ester, betulinic acid, 23-hydroxy betulinic acid, and triptonide;

for example, the monovalent group formed by the monocyclic monoterpene compound or the pentacyclic triterpene compound may be 4- (1-methylvinyl) -1-cyclohexene-1-methoxy group, (1R,2S,5R) -2-isopropyl-5-methylcyclohexyloxy group, 5-methyl-2- (1-methylvinyl) cyclohexyloxy group, 2- (4-methyl-3-cyclohexenyl) -2-propoxy group, 1-methyl-4- (1-methylvinyl) cyclohexyloxy group, 1-methyl-4-isopropylidene-cyclohexyloxy group, 2-methyl-5- (1-methylvinyl) -2-cyclohexenyloxy group, 12-ene-28-carboxyl-oleanane-3-oxyl, 12-ene-28-carboxyl-ursane-3 beta-oxyl, 12-ene-28-acid-2 alpha-hydroxyl oleanane-3 beta-oxyl, 12-ene-28-acid-3 beta-hydroxyl oleanane-2 alpha-oxyl, 12-ene-28-oic acid-2 alpha-hydroxy ursane-3 beta-oxyl, 12-ene-28-oic acid-3 beta-hydroxy ursane-2 alpha-oxyl and (3 beta, 20 beta) -11-oxo-olean-12-ene-29-oic acid-3-oxyl.

According to an embodiment of the invention, in the compound of formula (I),

R₃、R₄identical or different, independently of one another, from hydrogen, C_1-3Alkyl and C_1-3An alkoxy group;

or, R₃、R₄Together with the carbon atom to which they are attached form a cyclopropane, cyclobutyl, cyclopentyl, cyclohexyl, oxirane, oxetane, oxolanyl, oxocyclohexyl, azetidinyl, azacyclopentyl, azacyclohexyl or thiacyclopentyl group;

y is selected from O, carbonyl, or by OH or C_1-3An alkoxy-substituted methylene group.

According to an embodiment of the present invention, the compound of formula (I) is selected from the following compounds (1) to (13):

the invention also provides a preparation method of the compound shown in the formula (I), which comprises the following steps: reacting compound M1 with a compound capable of providing R₁And/or R₂Conversion of radicalsReacting the compound to obtain the compound shown in the formula (I):

wherein R is₁、R₂、R₃、R₄、Y、X₁And X₂Each having the definitions set forth above;

according to an embodiment of the invention, said can provide R₁And/or R₂The group may be selected from R₁OH、R₁NH₂、R₂OH and R₂NH₂；

According to an embodiment of the invention, said can provide R₁And/or R₂The group can be selected from methanol, taurine, monocyclic monoterpene compounds or pentacyclic triterpenoid compounds.

Said can provide R₁And/or R₂The group can be selected from perillyl alcohol, menthol, isopulegol, alpha-terpineol, beta-terpineol, gamma-terpineol, and carveol.

Said can provide R₁And/or R₂The group is selected from oleanolic acid, ursolic acid, maslinic acid, corosolic acid, glycyrrhetinic acid, asiatic acid, saikosaponin E, saikosaponin F, saikosaponin G, secondary saikosaponin A, secondary saikosaponin B, secondary saikosaponin C, secondary saikosaponin D, saikosaponin F, phytolaccagenic acid-30-methyl ester, 2-hydroxy phytolaccagenic acid-30-methyl ester, betulinic acid, 23-hydroxy betulinic acid, and triptonide.

According to an embodiment of the present invention, the preparation method may be performed in the presence of a solvent such as an organic solvent. For example, the organic solvent may be selected from at least one of the following: ethers such as ethyl propyl ether, n-butyl ether, anisole, phenetole, cyclohexylmethyl ether, dimethyl ether, diethyl ether, dimethyl glycol, diphenyl ether, dipropyl ether, diisopropyl ether, di-n-butyl ether, diisobutyl ether, diisoamyl ether, ethylene glycol dimethyl ether, isopropyl ethyl ether, methyl tert-butyl ether, tetrahydrofuran, methyl tetrahydrofuran, dioxane, dichlorodiethyl ether, and polyethers of ethylene oxide and/or propylene oxide; aliphatic, cycloaliphatic or aromatic hydrocarbons, such as pentane, hexane, heptane, octane, nonane, and possibly substituted by fluorine and chlorine atoms, such as methylene chloride, dichloromethane, trichloromethane, carbon tetrachloride, fluorobenzene, chlorobenzene or dichlorobenzene; cyclohexane, methylcyclohexane, petroleum ether, octane, benzene, toluene, chlorobenzene, bromobenzene, xylene; esters such as methyl acetate, ethyl acetate, butyl acetate, isobutyl acetate and dimethyl carbonate, dibutyl carbonate or ethylene carbonate.

According to an embodiment of the present invention, the preparation process may be carried out, for example, in the presence of an acid or a base.

The acid may be an organic acid or an inorganic acid, for example, at least one selected from formic acid, acetic acid, propionic acid, trifluoroacetic acid, HCl, sulfuric acid.

The base may be an organic base or an inorganic base. For example, the inorganic base may be selected from at least one of the following: hydrides, hydroxides, alkoxides, acetates, fluorides, phosphates, carbonates and bicarbonates of alkali metals or alkaline earth metals. Preferred bases are sodium amide, sodium hydride, lithium diisopropylamide, sodium methoxide, potassium tert-butoxide, sodium hydroxide, potassium hydroxide, sodium acetate, sodium phosphate, potassium fluoride, cesium fluoride, sodium carbonate, potassium bicarbonate, sodium bicarbonate, and cesium carbonate; the organic base may be selected from at least one of the following: tertiary amines, substituted or unsubstituted pyridines and substituted or unsubstituted triethylamine, trimethylamine, N, N-diisopropylethylamine, tri-N-propylamine, tri-N-butylamine, tri-N-hexylamine, tricyclohexylamine, N-methylcyclohexylamine, N-methylpyrrolidine, N-methylpiperidine, N-ethylpiperidine, N, N-dimethylaniline, N-methylmorpholine, pyridine, 2, 3-or 4-methylpyridine, 2-methyl-5-ethylpyridine, 2, 6-dimethylpyridine, 2,4, 6-trimethylpyridine, 4-dimethylaminopyridine, quinoline, methylquinoline, N, N, N-tetramethylethylenediamine, N, N-dimethyl-1, 4-diazacyclohexane, N, N-diethyl-1, 4-diazabicyclocyclohexane, 1, 8-bis (dimethylamino) naphthalene, Diazabicyclooctane (DABCO), Diazabicyclononane (DBN), Diazabicycloundecane (DBU), butylimidazole and methylimidazole.

The invention also provides a pharmaceutical composition, which comprises a therapeutically effective amount of at least one of the compounds shown in the formula (I), and racemate, stereoisomer, pharmaceutically acceptable salt or solvate thereof.

According to an embodiment of the invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

According to an embodiment of the invention, the pharmaceutical composition may further comprise one or more additional therapeutic agents.

The present invention also provides a method of treating cancer comprising administering to a patient a prophylactically or therapeutically effective amount of at least one compound, stereoisomer, pharmaceutically acceptable salt, or solvate of formula (I);

the cancer may be leukemia, liver cancer, lung cancer, gastric cancer and ovarian cancer.

In some embodiments, the patient is a human.

The invention also provides at least one of a compound, stereoisomer, pharmaceutically acceptable salt or solvate shown as the formula (I), or a pharmaceutical composition thereof for treating cancer.

The invention also provides application of at least one of the compound shown in the formula (I), stereoisomer, pharmaceutically acceptable salt or solvate in preparing antitumor drugs.

According to the present invention, the antitumor drug can be used for treating the following cancers: leukemia, liver cancer, lung cancer, gastric cancer or ovarian cancer.

As a medicament, the compounds of the present invention may be administered in the form of a pharmaceutical composition. These compositions may be prepared in a manner well known in the pharmaceutical arts and may be administered by a variety of routes depending on whether local or systemic treatment is desired and the area to be treated. Can be administered topically (e.g., transdermal, dermal, ocular, and mucosal including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal), orally, or parenterally. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intracerebroventricular, administration. The administration may be parenteral in a single bolus form, or may be by, for example, a continuous infusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, water, powder or oily bases, thickeners and the like may be necessary or desirable.

In preparing the compositions of the invention, the active ingredient is typically mixed with an excipient, diluted by an excipient or enclosed within such a carrier, for example, in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it may be a solid, semi-solid, or liquid material that serves as a vehicle, carrier, or medium for the active ingredient. Thus, the composition may be in the form of: tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or dissolved in a liquid vehicle); ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders containing, for example, up to 10% by weight of the active compound.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. The formulation may also contain: lubricants such as talc, magnesium stearate and mineral oil; a humectant; emulsifying and suspending agents; preservatives such as methyl benzoate and hydroxypropyl benzoate; sweetening agents and flavoring agents. The compositions of the present invention may be formulated so as to provide immediate, sustained or delayed release of the active ingredient after administration to the patient by employing methods known in the art.

The compositions may be formulated in unit dosage forms, each dosage containing from about 5 to about 1000mg, more usually from about 100 to about 500mg, of the active ingredient. The term "unit dosage form" refers to physically discrete single dosage units suitable as unitary dosages for human patients and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in admixture with suitable pharmaceutical excipients.

The effective dose of the active compound can vary widely and is generally administered in a pharmaceutically effective amount. However, it will be understood that the amount of the compound actually administered will generally be determined by a physician, in the light of the relevant circumstances, and will include the condition to be treated, the chosen route of administration, the actual compound administered; age, weight and response of the individual patient; severity of patient symptoms, etc.

For preparing solid compositions such as tablets, the principal active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogeneous mixture of the compound of the invention. When referring to these preformulation compositions as homogeneous, it is meant that the active ingredient is generally uniformly distributed throughout the composition such that the composition may be readily divided into equally effective unit dosage forms such as tablets, pills and capsules. The solid pre-formulations are then divided into unit dosage forms of the type described above containing, for example, from about 0.1 to 1000mg of the active ingredient of the invention.

The tablets or pills of the present invention may be coated or compounded to provide a dosage form affording the advantage of prolonged action. For example, a tablet or pill contains an inner dose and an outer dose component, the latter being in the form of a capsule of the former. The two components may be separated by an enteric layer which serves to resist disintegration in the stomach, leaving the inner component intact through the duodenum or delayed in release. A variety of materials may be used for such enteric layers or coatings, such materials including polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol and cellulose acetate.

Liquid forms in which the compounds and compositions of the present invention may be incorporated for oral or injectable administration include aqueous solutions, suitably flavoured syrups, aqueous or oil suspensions; and flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil; as well as elixirs and similar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions, suspensions, and powders dissolved in pharmaceutically acceptable water or organic solvents or mixtures thereof. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In certain embodiments, the composition is administered by the oral or nasal respiratory route to achieve a local or systemic effect. The composition may be atomized by the use of an inert gas. The nebulized solution may be inhaled directly from the nebulizing device, or the nebulizing device may be connected to a mask or intermittent positive pressure ventilator. The solution, suspension or powder composition may be administered orally or nasally by means of a device that delivers the formulation in a suitable manner.

The amount of compound or composition administered to a patient is not fixed and depends on the drug administered, the purpose of the administration such as prevention or treatment; the condition of the patient, the mode of administration, etc. In therapeutic applications, compositions can be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dosage will depend on the disease state being treated and the judgment of the attending clinician, which will depend on factors such as the severity of the disease, the age, weight and general condition of the patient.

The composition administered to the patient may be in the form of a pharmaceutical composition as described above. These compositions may be sterilized by conventional sterilization techniques or may be sterilized by filtration. The aqueous solutions may be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the compound preparation is usually 3 to 11, more preferably 5 to 9, and most preferably 7 to 8. It will be appreciated that the use of certain of the aforementioned excipients, carriers or stabilizers may result in the formation of a pharmaceutical salt.

Therapeutic dosages of the compounds of the invention may be determined, for example, by: the particular use of the treatment, the mode of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compound of the invention in the pharmaceutical composition may not be fixed and will depend on a variety of factors including dosage, chemical properties (e.g., hydrophobicity), and the route of administration. For example, the compounds of the present invention can be provided for parenteral administration by a physiological buffered aqueous solution containing about 0.1-10% w/v of the compound. Some typical dosage ranges are from about 1. mu.g/kg to about 1g/kg body weight/day. In certain embodiments, the dosage range is from about 0.01mg/kg to about 100mg/kg body weight/day. The dosage will likely depend on such variables as the type and extent of progression of the disease or disorder, the general health status of the particular patient, the relative biological efficacy of the selected compound, the excipient formulation and its route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

Advantageous effects

The compound shown in the formula (I) has obvious inhibition effect on cancer cells, particularly liver cancer cells HepG2, stomach cancer cells BGC803, human lung cancer cells H460, blood cancer cells HL60 and ovarian cancer cells HO 8901. The compound of the invention has obviously stronger inhibiting effect on cancer cells than positive control perilla alcohol, oleanolic acid and norcantharidin, and has obviously lower toxicity on cells than norcantharidin. In addition, the compounds of the present invention, such as compound 6, are non-irritating and pharmaceutically acceptable. The compound of the invention has good antitumor activity, wide antitumor spectrum and low toxicity, and is suitable for preparing anticancer drugs.

Definition and description of terms

Unless otherwise indicated, the definitions of groups and terms described in the specification and claims of the present application, including definitions thereof as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. The definitions of the groups and the structures of the compounds in such combinations and after the combination are within the scope of the present specification.

Unless otherwise indicated, the numerical ranges set forth in the specification and claims are equivalent to at least each and every specific integer numerical value set forth therein. For example, a numerical range of "1-30" is equivalent to reciting each of the integer values in the numerical range of "1-10," i.e., 1,2, 3, 4,5, 6, 7, 8, 9, 10, and each of the integer values in the numerical range of "11-40," i.e., 11, 12, 13, 14, 15, 25, 26, 27, 28, 29, 30. It is to be understood that "more" in one, two, or more of the substituents used herein when describing substituents shall mean an integer ≧ 3, such as 3, 4,5, 6, 7, 8, 9, or 10. Further, when certain numerical ranges are defined as "numbers," it should be understood that the two endpoints of the range, each integer within the range, and each decimal within the range are recited. For example, "a number of 0 to 10" should be understood to not only recite each integer of 0, 1,2, 3, 4,5, 6, 7, 8, 9, and 10, but also to recite at least the sum of each integer and 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, respectively.

The term "C_1-6Alkyl "denotes straight-chain and branched alkyl groups having 1,2, 3, 4,5 or 6 carbon atoms. The alkyl group is, for example, a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, an isopropyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, an isopentyl group, a 2-methylbutyl group, a 1-ethylpropyl group, a 1, 2-dimethylpropyl group, a neopentyl group, a 1, 1-dimethylpropyl group, a 4-methylpentyl group, a 3-methylpentyl group, a 2-ethylbutyl group, a 1-ethylbutyl group, a 3, 3-dimethylbutyl group, a 2, 2-dimethylbutyl group, a 1, 1-dimethylbutyl group, a 2, 3-dimethylbutyl group, a 1, 3-dimethylbutyl group or a 1, 2-dimethylbutyl group, or the like, or isomers thereof.

The term "C_2-6Alkenyl "is understood to preferably mean a straight-chain or branched monovalent hydrocarbon radical which contains one or more double bonds and has 2,3, 4,5 or 6 carbon atoms, for example 2,3, 4,5 or 6 carbon atoms (i.e. C)_2-6Alkenyl) having 2 or 3 carbon atoms (i.e., C)_2-3Alkenyl). It is understood that where the alkenyl group contains more than one double bond, the double bonds may be separated from each other 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) -pent-1-enyl, (Z) -pent-1-enyl, hex-5-enyl, (E) -hex-4-enyl, (Z) -hex-4-enyl, m-n-2-enyl, m-n-1-enyl, m-n-E-4-enyl, m-n-2-enyl, m-n-enyl, m-E-4-enyl, m-2-enyl, m-pent-1-enyl, m-2-methyl-enyl, m-2-methylvinyl, m-2-methyl-2-methylvinyl, m-but-2-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-eneA group, (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-methylbut-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-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.

The term "C_3-8Cycloalkyl "is understood to mean a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane having 3, 4,5, 6, 7 or 8 carbon atoms. Said "C_3-8Cycloalkyl "may be, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl.

The term "C_3-30Cycloalkenyl is understood to mean an unsaturated monovalent monocyclic, bicyclic or polycyclic hydrocarbon ring having 3 to 30 carbon atoms and one, two or more olefinic bonds. Said "C_3-10The cycloalkyl group "may be a monocyclic hydrocarbon group such as a cyclobutenyl group, cyclopentenyl group, cyclopentadienyl group, cyclohexenyl group, cycloheptenyl group, cyclooctenyl group, cyclooctadienyl group or cyclooctatrienyl group, or a bicyclic hydrocarbon group such as a decahydronaphthalene ring; or is a fused ring of said single ring.

The term "3-8 membered heterocyclyl" means a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane, which contains from 1 to 5 heteroatoms independently selected from N, O and S, and a total number of ring atoms of from 3 to 8 (e.g., numbers of 3, 4,5, 6, 7, 8, etc.) non-aromatic cyclic group, preferably "3-6 membered heterocyclyl". The term "3-6 membered heterocyclyl" means a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane comprising 1-5, preferably 1-3 heteroatoms selected from N, O and S. The heterocyclic group may be attached to the rest of the molecule through any of the carbon atoms or nitrogen atom (if present). In particular, the heterocyclic group may include, but is not limited to: 4-membered rings such as azetidinyl, oxetanyl; 5-membered rings such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl; according to the invention, the heterocyclic radical is non-aromatic. When the 3-to 8-membered heterocyclic group is linked to another group to form the compound of the present invention, the carbon atom of the 3-to 8-membered heterocyclic group may be linked to another group, or the heterocyclic atom of the 3-to 8-membered heterocyclic ring may be linked to another group. For example, when the 3-to 8-membered heterocyclic group is selected from piperazinyl, it may be such that the nitrogen atom on the piperazinyl group is linked to another group. Or when the 3-8 membered heterocyclyl group is selected from piperidinyl, it may be that the nitrogen atom on the piperidinyl ring and the carbon atom in the para position are attached to other groups.

Unless otherwise indicated, heterocyclyl includes all possible isomeric forms thereof, e.g. positional isomers thereof. Thus, for some illustrative non-limiting examples, forms may be included that are substituted at one, two or more of their 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-positions, etc. (if present) or bonded to other groups, including pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, and pyridin-4-yl; thienyl or thienylene includes thien-2-yl, thien-3-yl and thien-3-yl; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

The term "oxo" refers to an oxy substitution (═ O) formed by oxidation of a carbon atom, a nitrogen atom or a sulfur atom in a substituent.

It will be appreciated by those skilled in the art that the compounds of formula (I) may 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; these compounds may also form inner salts if they contain both an acidic centre (e.g. carboxyl) and a basic centre (e.g. amino). Acid addition salts include, but are not limited to: hydrochloride, hydrofluoride, hydrobromide, hydroiodide, sulfate, pyrosulfate, phosphate, nitrate, methanesulfonate, ethanesulfonate2-hydroxyethanesulfonate, benzenesulfonate, tosylate, sulfamate, 2-naphthalenesulfonate, formate, acetoacetate, pyruvate, laurate, cinnamate, benzoate, acetate, glyoxylate, trifluoroacetate, pivalate, propionate, butyrate, hexanoate, heptanoate, undecanoate, stearate, ascorbate, camphorate, camphorsulfonate, citrate, fumarate, malate, maleate, hydroxymaleate, oxalate, salicylate, succinate, gluconate, quinic acid, pamoate, glycolate, tartrate, lactate, 2- (4-hydroxybenzoyl) benzoate, cyclopentanepropionate, digluconate, 3-hydroxy-2-naphthoate, nicotinate, and the like, Pamoate, pectinate, 3-phenylpropionate, picrate, pivalate, itaconate, trifluoromethanesulfonate, dodecylsulfate, p-toluenesulfonate, napadisylate, malonate, adipate, alginate, mandelate, glucoheptonate, glycerophosphate, sulfosalicylate, hemisulfate or thiocyanate, aspartate and the like; base addition salts such as alkali metal salts, alkaline earth metal salts, ammonium salts and the like, specifically include but are not limited to: sodium salt, lithium salt, potassium salt, ammonium salt, aluminum salt, magnesium salt, calcium salt, barium salt, ferric salt, ferrous salt, manganese salt, manganous salt, zinc salt and ammonium salt (including NH)₃Salts with organic amines (NH)₄Salts), methylammonium salts, trimethylammonium salts, diethylammonium salts, triethylammonium salts, propylammonium salts, tripropylammonium salts, isopropylammonium salts, tertiarybutylammonium salts, N' -dibenzylethylenediamine salts, dicyclohexylammonium salts, 1, 6-hexadimethrine salts, benzylammonium salts, ethanolamine salts, N-dimethylethanolamine salts, N-diethylethanolamine salts, triethanolamine salts, tromethamine salts, lysine salts, arginine salts, histidine salts, glucammonium salts, N-methylglucammonium salts, dimethylglucammonium salts, ethylglucammonium salts, meglumine salts, betaine salts, caffeine salts, chloroprocaine salts, procaine salts, lidocaine salts, pyridine salts, picoline salts, piperidine salts, morpholine salts, piperazine salts, purine salts, cacao salt, choline salts), and the like.

The compounds of the invention may be present in the form of solvates, such as hydrates, wherein the compounds of the invention comprise as structural element of the crystal lattice of the compound a polar solvent, such as in particular water, methanol or ethanol. The amount of polar solvent, particularly water, may be present in stoichiometric or non-stoichiometric proportions.

Depending on their molecular structure, the compounds of the invention may be chiral and may therefore exist in various enantiomeric forms. These compounds may thus be present in racemic or optically active form. The compounds of the invention or intermediates thereof may be separated into enantiomeric compounds by chemical or physical methods well known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from mixtures by reaction with optically active resolving agents. Examples of suitable resolving agents are optically active acids such as the R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g. N-benzoylproline or N-benzenesulfonylproline) or various optically active camphorsulphonic acids. The chromatographic enantiomeric resolution can also advantageously be carried out with the aid of optically active resolving agents, such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers, which are immobilized on silica gel. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example hexane/isopropanol/acetonitrile.

The corresponding stable isomers can be isolated according to known methods, for example by extraction, filtration or column chromatography.

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

The term "therapeutically effective amount" means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response that is being sought by a researcher, veterinarian, medical doctor or other clinician in a tissue, system, animal, individual, or human, which includes one or more of the following: (1) prevention of diseases: for example, preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not experienced or developed disease pathology or symptomatology. (2) Inhibiting the disease: for example, inhibiting the disease, disorder or condition (i.e., arresting the further development of the pathology and/or condition) in an individual who is experiencing or presenting the pathology or condition of the disease, disorder or condition. (3) And (3) relieving the diseases: for example, relieving the disease, disorder or condition (i.e., reversing the pathology and/or symptomatology) in an individual who is experiencing or presenting with the pathology or symptomatology of the disease, disorder or condition.

Detailed Description

The technical solution of the present invention will be further described in detail with reference to specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.

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

Example 1:

the cyclohexane dicarboxylic acid derivatives with bridged rings of the present invention can be prepared by the following route:

(1) preparation of nordehydrocantharidin

20g (204mmol) of maleic anhydride are weighed out and added into a 250mL round-bottomed flask, 120mL of dry ether are added for dissolution, 19g (279mmol) of furan are slowly added dropwise, and stirring is carried out at room temperature for 24h after the addition is finished. Suction filtration is carried out, and the filter cake is evaporated to dryness to obtain 30g (181mmol) of white solid with the yield of 89%. Mp 120-122 deg.C; IR (KBr, cm)^-1):3050,1860,1795；¹H NMR(CDCl₃,400MHz)δ:3.19(s,2H),5.44-5.47(m,2H),6.57-6.59(m,2H).

(2) Preparation of Demethyl dehydrocantharidin dimethyl ester

1.7g (10mmol) of nordehydrocantharidin is weighed and added toIn a 25mL round-bottom flask, 8mL of saturated hydrogen chloride methanol solution is added for reflux reaction overnight, the solvent is distilled off under normal pressure, 10mL of distilled water and ethyl acetate are added for extraction (15mL multiplied by 3), organic phases are combined, the organic phase is washed by sodium bicarbonate, washed by water and saturated sodium chloride, dried by anhydrous sodium sulfate and evaporated to dryness under reduced pressure to obtain 2.0g of white solid with the yield of 94%. Mp:119-120 deg.C; IR (KBr, cm)^-1):2998,1768,1434,1363,1302,1254,1194,1157,932,737；¹H NMR(CDCl₃,400MHz)δ:2.81(s,2H),3.69(s,6H),5.26(s,2H),6.44(s,2H).

(3) Preparation of 4, 5-cyclopropyl norcantharidin bis-methyl ester

650mg (10mmol) of zinc powder and 99mg (1.0mmol) of cuprous chloride are weighed and added into a 25mL two-necked bottle with a nitrogen protection device, stirred for 5min at room temperature under the protection of nitrogen, then added with a diiodomethane solution dissolved with 320mg (1.2mmol) of diiodomethane, reacted for 20min in an ultrasonic way, the reaction solution turns into purple brown, and then added with 4mL of a dried glycol dimethyl ether solution dissolved with 212mg (1.0mmol) of nordehydrocantharidin, and reacted at room temperature overnight. After the reaction, adding saturated ammonium chloride for quenching, performing suction filtration, washing a filter cake with diethyl ether, separating an organic layer from a filtrate, washing the organic layer with saturated ammonium chloride, washing with water, washing with saturated sodium chloride, drying with anhydrous sodium sulfate, and evaporating under reduced pressure to obtain 151mg of white solid with the yield of 67%. Mp is 61-62 ℃; IR (KBr, cm)^-1):2966,2850,1739,1436,1202,803,640；¹H NMR(CDCl₃,400MHz)δ:0.18-0.22(m,1H),0.61-0.62(m,1H),1.10-1.13(m,2H),3.08(s,2H),3.67(s,6H),4.67(s,2H)；¹³C NMR(CDCl₃,100MHz)δ:15.3,48.87,52.0,53.0,77.9,171.42；ESI-MS:249.1[M+Na]⁺,475[2M+Na]⁺.

(4) Preparation of sodium 4, 5-cyclopropyl norcantharidate

Taking 113mg (0.5mmol) of 5, 6-cyclopropyl norcantharidin dimethyl ester, adding 3mL of water and 20mg (0.5mmol) of sodium hydroxide, stirring at room temperature for 2h, adding a certain amount of ethanol, precipitating a solid, performing suction filtration, and evaporating to dryness to obtain 106mg of white solid with the yield of 88%. IR (KBr, cm-1):2987,2947,1559,1436,1404,1086,862,803,634;¹H NMR(D₂O,400MHz)δ:0.04-0.11(m,1H),0.28-0.31(m,1H),1.07-1.06(m,2H),4.37(s,2H).

(5) preparation of 4, 5-cyclopropyl norcantharidin (Compound 1)

57mg (0.25mmol) of sodium 4, 5-cyclopropyl norcantharidate is dissolved in 0.3mL of water, the pH is adjusted to about 3 by 0.1N of dilute hydrochloric acid, 4mL of trifluoroacetic anhydride is added after evaporation under reduced pressure, the mixture reacts at 40 ℃ overnight, and 38mg (0.21mmol) of white solid is obtained after evaporation under reduced pressure, namely the title compound, and the yield is 84%. IR (KBr, cm)^-1):2992,1880,1780；¹H NMR(CDCl₃,400MHz)δ:0.04-0.11(m,1H),0.28-0.31(m,1H),1.07-1.06(m,2H),4.37(s,2H)4.98-5.02(m,2H)；ESI-MS:181.1[M+H]⁺.

(6) Preparation of 4, 5-cyclopropyl norcantharidin perillyl alcohol ester (Compound 2)

Adding 180mg (1mmol) of 4, 5-cyclopropyl norcantharidin, 152mg (1mmol) of perillyl alcohol and 56mg (1mmol) of potassium hydroxide into a 25mL round-bottomed bottle, adding 10mL of dried dichloromethane, refluxing for 5h, evaporating to dryness under reduced pressure, adding a proper amount of water, adjusting the pH to about 3 with 1N hydrochloric acid, extracting with ethyl acetate (10mL multiplied by 3), combining organic phases, washing the organic phases with water, saturated sodium chloride, drying with anhydrous sodium sulfate, evaporating to dryness under reduced pressure, and performing column chromatography (ethyl acetate: petroleum ether: 1:5) to obtain 173mg of the title compound with the yield of 52%. IR (KBr, cm)^-1):3322,3003,2922,1730；¹H NMR(CDCl₃,400MHz)δ:0.05-0.11(m,1H),0.28-0.32(m,1H),1.07-1.09(m,2H),1.71(s,3H),1.80-1.87(m,3H),1.89-1.98(m,1H),2.02-2.16(m,4H),3.03(s,2H),4.39-4.51(m,2H),4.68-4.70(m,2H),4.88-4.94(m,2H),5.74(s,1H)；ESI-MS:355.5[M+Na]⁺.

(7) Preparation of 4, 5-epoxy norcantharidin

0.5g (3mmol) of dehydromethylcantharidin is taken and dissolved in 2mL of glacial acetic acid, 0.7mL of hydrogen peroxide is added dropwise in ice bath, the temperature is slowly returned to room temperature and stirred overnight, the mixture is filtered by suction, and the solid is dissolved in 6mL of acetyl chloride and refluxed for 12 h. The temperature is reduced to room temperature, 0.19g of white solid is obtained by suction filtration, and the yield is 35%. ESI-MS 183.14[ M + H ]]⁺.

(8) Preparation of 4, 5-epoxy norcantharidin perillyl alcohol ester (Compound 3)

Dissolving 91mg (0.5mmol) of 4, 5-epoxy norcantharidin and 608mg (4mmol) of perillyl alcohol in 6mL of dry dichloromethane, and adding 0.7mL (0.7 mL)4mmol) DIPEA was stirred at room temperature for 48h, and the column chromatography was evaporated under reduced pressure (ethyl acetate: petroleum ether-1: 4) gave 72mg of the title compound in 43% yield.¹H NMR(CDCl₃,400MHz):1.70(s,3H),1.80-1.87(m,3H),1.89-1.98(m,1H),2.02-2.16(m,4H),3.03(s,2H),4.39-4.51(m,4H),4.68-4.71(m,2H),4.88-4.92(m,2H),5.75(s,1H)；ESI-MS:335.38[M+H]⁺.

(9) Preparation of norcantharidin

Adding 8g (48mmol) of dehydromethylcantharidin into a 100mL round-bottom bottle, dissolving with 50mL of dry acetone, adding 5mg (10%) of palladium on carbon under hydrogen at room temperature for 12h, adding a little activated carbon for assisting filtration, performing suction filtration, washing a filter cake with ethyl acetate, evaporating the filtrate to dryness to obtain 7.7g, wherein the yield is 95%. Mp 109-111 ℃; IR (KBr, cm)^-1):2991,1867,1791,1600,1219,1021,636；¹H NMR(CDCl₃,400MHz)δ:1.59-1.64(m,2H),1.86-1.91(m,2H),3.17(s,2H),5.02-5.03(m,2H).

(10) Preparation of norcantharidin oleanolic acid ester (Compound 4 and Compound 5)

228mg (0.5mmol) of oleanolic acid, 168mg (1.0mmol) of norcantharidin and 122mg (1.0mmol) of DMAP are added into a 50mL round-bottomed bottle, dissolved by 15mL of dry ethylene glycol dimethyl ether, refluxed for 12h, after the reaction is finished, the solvent is evaporated under reduced pressure, a small amount of tetrahydrofuran is added for dissolution, silica gel is stirred, and column chromatography (petroleum ether: dichloromethane: tetrahydrofuran: 5:9:2) is carried out to obtain 69mg of compound 4, wherein the yield is 22%. Mp is 265 ℃ and 266 ℃; IR (KBr, cm)^-1):2947,1733,1701,1466,1240,1206,912,734；¹H NMR(CDCl₃,400MHz)δ:0.66(s,3H),0.74(s,3H),0.83(s,3H),0.86(s,3H),0.90(s,3H),0.91(s,3H),0.93(s,3H),2.80-2.84(m,1H),3.02-3.05(m,1H),3.21-3.27(m,1H),4.40-4.53(m,1H),4.79-4.83(m,1H),5.05-5.06(m,1H),5.25-5.26(m,1H)；¹³C NMR(CDCl₃,400MHz)δ:15.65,17.38,17.46,17.62,18.32,22.47,23.36,23.71,25.54,26.07,26.12,28.51,28.85,30.74,33.14,36.60,37.10,37.16,37.65,37.85,39.24,40.84,41.17,45.63,46.57,47.10,50.69,53.76,54.87,54.91,80.75,81.32,81.91,122.88,143.62,171.44,177.32,185.62；ESI-MS:647.5[M+Na]⁺,1271.7[2M+Na]⁺.

To obtain 72mg of the compound 5,the yield thereof was found to be 23%. Mp 268-269 deg.C; IR (KBr, cm)^-1):2948,1732,1704,1464,1365,1255,1206,910,649；¹H NMR(CDCl₃,400MHz)δ:0.71(s,3H),0.79(s,3H),0.82(s,3H),0.85(s,3H),0.87(s,3H),0.90(s,3H),0.92(s,3H),0.93-0.94(m,2H),1.05-1.13(m,6H),1.22-1.36(m,4H),1.54-1.62(m,10H),1.72-1.77(m,4H),1.87-1.89(m,3H),2.79-2.83(m,1H),2.98-3.01(m,1H),3.48-3.53(m,1H),4.51-4.55(m,1H),4.81-4.87(m,2H),5.26(s,1H)；¹³C NMR(CDCl₃,400MHz)δ:15.47,15.62,17.08,17.28,18.29,22.67,24.24,23.40,22.67,25.23,26.02,27.71,28.44,30.74,31.04,33.14,37.07,37.11,37.69,37.99,39.29,40.89,41.37,45.78,46.56,47.351,51.08,51.2,54.98,55.21,80.75,81.32,80.91,122.68,143.72,170.49 176.68,184.77；ESI-MS:647.5[M+Na]⁺,1271.7[2M+Na]⁺.

(11) Preparation of norcantharidin perillyl alcohol ester (Compound 6)

Adding 168mg (1mmol) of norcantharidin, 152mg (1mmol) of perillyl alcohol and 56mg (1mmol) of potassium hydroxide into a 25mL round-bottomed bottle, adding 10mL of dried dichloromethane solution, refluxing for 5h, evaporating to dryness under reduced pressure, adding an appropriate amount of water, adjusting the pH to 3 with 1N hydrochloric acid, extracting with ethyl acetate (10mL multiplied by 3), combining organic phases, washing the organic phases with water, saturated sodium chloride, drying with anhydrous sodium sulfate, evaporating to dryness under reduced pressure, and recrystallizing to obtain 198mg of the title compound with the yield of 62%. IR (KBr, cm)^-1):3006,2922,1725,1706,1352,1305,1146,560；¹H NMR(CDCl₃,400MHz)δ:1.40-1.46(m,1H),1.49-1.54(m,2H),1.71(s,3H),1.80-1.87(m,3H),1.89-1.97(m,1H),2.02-2.15(m,4H),3.01(s,2H),4.39-4.51(m,2H),4.68-4.70(m,2H),4.88-4.94(m,2H),5.74(s,1H)；¹³C NMR(CDCl₃,400MHz)δ:20.73,26.33,27.27,28.94,29.66,30.45,40.74,52.17,69.24,78.43,78.59,80.14,126.29,132.2,149.59,170.75,175.67；ESI-MS:343.1[M+Na]⁺,663.2[2M+Na]⁺.

(12) Norcantharidin taurine (Compound 11)

125mg (1mmol) of taurine and 136mg (1mmol) of sodium acetate are weighed and added into a 10mL round-bottomed bottle, 6mL of glacial acetic acid is added, the mixture is stirred for 10min at room temperature, then 168mg (1mmol) of norcantharidin is added,then refluxing reaction is carried out overnight, suction filtration is carried out the next day, a filter cake is recrystallized by methanol, dissolved by water, added with Amberlite IR-120, stirred for 10min, and evaporated to dryness by suction filtration to obtain 214mg of white solid, namely the title compound, with the yield of 78%. IR (KBr, cm)^-1):2988,1720,1449,1416,1355,1197,1053,661；¹H NMR(D₂O,400MHz)δ:1.63-1.73(m,4H),3.02-3.08(m,4H),3.76(s,2H),4.81(s,2H)；¹³C NMR(CD₃OD,100MHz)δ:29.3,35.8,48.8,51.2,80.40,179.0.

(13) Preparation of cyclopentadiene

13g of dicyclopentadiene is taken and added into a three-necked bottle provided with a thermometer, a nitrogen protection device and a normal pressure distillation device, then the reaction device is placed in an oil bath at 170 ℃, meanwhile, a receiving bottle is placed in an ice salt bath at-20 ℃, the distillate at the front section is discarded, 5.8g of distillate at the middle section is reserved, and the yield is 22%.

(14) Preparation of 6, 6-dimethylfulvene

Adding 2.0g (30mmol) of cyclopentadiene, 1.7g (30mmol) of acetone and 18mL of methanol into a 50mL round-bottomed bottle, slowly adding a pyrrolidine (1.5mL, 18mmol) solution diluted with 4mL of methanol under the protection of nitrogen and in the dark, reacting for 1h at room temperature after the addition is finished, adding 1.1mL (19mmol) of acetic acid after the reaction is finished, stirring for about 10min at room temperature, pouring the reaction solution into ice water, extracting with diethyl ether (20mL multiplied by 3), combining organic phases, washing with water, washing with saturated sodium chloride, drying with anhydrous magnesium sulfate, and evaporating the solvent by suspending under reduced pressure to obtain 2.4g of pale yellow oily matter (used for steaming and stored at-20 ℃) with the yield of 75%.¹H NMR(CDCl₃,400MHz)δ:2.19(s,6H),6.53-6.45(m,4H).

(15) Preparation of 7-isopropylidene nordehydrocantharidin

Weighing 1.8g (18mmol) of maleic anhydride, adding the maleic anhydride into a 100mL round-bottom bottle, taking 20mL of dry toluene, heating the toluene to reflux, adding 2.4g (23mmol) of 6-dimethyl fulvene, carrying out reflux reaction for 1h, after the reaction is finished, evaporating the solvent under reduced pressure, and recrystallizing twice with ethyl acetate to obtain 3.4g of colorless blocky crystals with the yield of 93%. Mp 128-129 ℃; IR (KBr, cm)^-1):3076,2963,1854,1772,1449,1227,1072,931,632；¹H NMR(CDCl₃,400MHz)δ:1.59(s,6H),3.04(s,2H),3.87-3.88(m,2H),6.45-6.46(m,2H).

(16) Preparation of 7-isopropylidene norcantharidin

Taking 0.52g (2.5mmol) of 7-isopropylidene dehydronorcantharidin, adding the 7-isopropylidene dehydrocantharidin into a 50mL round-bottomed bottle, adding 25mL of ethyl acetate for dissolving, adding 10mg (10%) of palladium carbon, reacting at room temperature for 15min under normal pressure and hydrogen, after the reaction is finished, carrying out suction filtration, and evaporating the filtrate to dryness to obtain 4.9g of white solid with the yield of 97%. Mp:130-132 deg.C; IR (KBr, cm)^-1):2979,2879,1771,1452,1735,1083,909,589；¹H NMR(CDCl₃,400MHz)δ:1.43-1.48(m,2H),1.66(s,6H),1.72-1.75(m,2H),2.96(s,2H),3.19(t,J＝2.0Hz,2H)；ESI-MS:229.1[M+Na]⁺,425.0[2M+Na]⁺.

(17) Preparation of 7-carbonyl norcantharidin (Compound 7)

206mg (1mmol) of 7-isopropylidene norcantharidin is dissolved in 10mL of carbon tetrachloride, ozone gas is introduced at the temperature of minus 25 ℃ until the reaction solution turns blue, the stirring is continued for 10min, 89 mu L of dimethyl sulfide is slowly added, the temperature is slowly raised to the room temperature, the evaporation is carried out, the ethyl acetate/petroleum ether recrystallization is carried out to obtain 85mg of the title compound, and the yield is 47%. IR (KBr, cm)^-1):2995,1825,1785,1770；¹H NMR(CDCl₃,400MHz)δ:1.63-1.69(m,2H),2.02-2.04(m,2H),2.99(s,2H),3.24(t,J＝2.0Hz,2H)；ESI-MS:181.15[M+H]⁺.

(18) Preparation of 7-isopropylidene norcantharidin dimethyl ester

Taking 0.4g (1.9mmol) of 7-isopropylidene norcantharidin, adding the 7-isopropylidene norcantharidin into a 10mL round-bottomed bottle, adding 5mL of saturated hydrogen chloride methanol for reflux reaction for 6 hours, distilling the solvent out under reduced pressure, adding 8mL of ethyl acetate for dissolution, washing with saturated sodium bicarbonate, washing with water, drying with anhydrous sodium sulfate, and evaporating to dryness under reduced pressure to obtain 0.39g of white solid with the yield of 81%. Mp 104-; IR (KBr, cm)^-1):2996,2958,2916,1745,1332,1200,1155；¹H NMR(CDCl₃,400MHz)δ:1.14-1.63(m,4H),1.69(s,6H),2.95(m,2H),3.56(s,6H).

(19) Preparation of 7-carbonyl norcantharidin dimethyl ester

252mg (1mmol) of 7-isopropylidene norcantharidin dimethyl ester are dissolved in 10mL of carbon tetrachloride, and ozone gas is introduced at-25 ℃ until the reaction solution turns into blueAnd (3) continuously stirring for 10min, then slowly adding 89 mu L of dimethyl sulfide, slowly raising the temperature to room temperature, evaporating to dryness, and recrystallizing twice in isopropanol to obtain 122mg of white solid with the yield of 54%. IR (KBr, cm)^-1):2993,1786,1744；¹H NMR(CDCI₃):δ:1.64-1.65(m,2H),2.00-2.03(m,2H),2.33(dd,J＝2.0Hz,J＝2.8Hz,2H),3.08(s,2H),3.65(s,6H)。ESI-MS:227.3[M+H]⁺.

(20) Preparation of 7-carbonyl norcantharidinic acid (Compound 8)

57mg (0.25mmol) of 7-carbonyl norcantharidin bis-methyl ester and 10mg (0.25mmol) of sodium hydroxide are taken, 3mL of water is added for dissolving at room temperature and stirring for 2h, the pH value is adjusted to about 3 by 1N hydrochloric acid, ethyl acetate is used for extraction, and 48mg of the title compound is obtained by evaporation to dryness, wherein the yield is 97%. ESI-MS 199.2[ M + H ]]⁺。

Example 2:

in vitro antitumor Activity test of norcantharidin derivative

The test adopts MTT method to test the inhibitory activity of the tested compound on different tumor strains (human liver cancer cell HepG2, gastric cancer BGC803, human lung cancer cell H460, blood cancer cell HL60 and ovarian cancer cell HO8901), and the test is divided into a positive control group, a blank control group and an experimental group. Adding perillyl alcohol, oleanolic acid and norcantharidin with corresponding concentrations into the positive group respectively to serve as positive controls; blank control group added DMSO (1%); test groups were dosed with the corresponding concentrations of test compound.

Taking the above five tumor cells in exponential growth phase, adjusting cell suspension concentration to 1 × 10 with RPMI-1640 culture medium containing 10% calf serum⁴Per mL; inoculating into 96-well plate, adding 100 μ L cell suspension per well, and placing in 5% CO₂And culturing in an incubator at 37 ℃ for 24 hours to ensure that the cells are completely attached to the wall. Adding corresponding drugs (concentration 5.0 × 10) into the positive control group and the experimental group respectively^-4，1.0×10^-4，2.0×10^-5，4.0×10^-6，8.0×10^-7，1.6×10^-7，3.2×10^-8mol/L) medicine, adding equal volume of DMSO (1%) into blank control group, setting 3 multiple wells in each group, placing at 37 deg.C and 5% CO₂The culture in the incubator is respectively carried out for 24 h. MTT (5mg/mL), 10. mu.L/well was added, and the mixture was incubated at 37 ℃ with 5% CO₂Culture boxContinuously culturing for 4 h; removing upper layer liquid after finishing culturing, adding 100 μ L DMSO solution into each multiple hole, shaking on shaking table for 10min to dissolve crystal completely, detecting absorbance A value of each hole at 490nm with microplate reader, and calculating IC with graphpad prism software₅₀The values, experimental results are shown in table 1.

TABLE 1 inhibitory Activity of some of the compounds on five tumor cells (compound numbers same as example 1)

The inhibitory activity of compounds 4,6 and 11 and norcantharidin on normal human lung fibroblasts WI-38 cells was determined using the same assay as described above. The results are shown in Table 2.

TABLE 2 IC of Compounds 4,6 and 11 on Normal human embryonic lung fibroblasts WI-38₅₀(μM)(n＝3)

From the above, the cyclohexane dicarboxylic acid derivative with bridged ring provided by the invention has different degrees of inhibition effects on liver cancer cell HepG2, gastric cancer cell BGC803, human lung cancer cell H460, blood cancer cell HL60 and ovarian cancer cell HO 8901. Wherein the inhibitory activity of the compound 1 on the lung cancer cell H460 is equivalent to that of norcantharidin as a positive control; the compound 2 has stronger inhibition on liver cancer cells HepG2 and stomach cancer cells BGC803, and particularly has the inhibition activity on ovarian cancer cells HO8901 which is improved by about 3 times compared with norcantharidin; the compound 7 shows equivalent or stronger inhibitory activity to the positive control on four tumor strains, such as the inhibitory activity to human lung cancer cell H460 and ovarian cancer HO8901, which are respectively improved by about 2 times and 3 times. The compound 9 has the same inhibiting effect on liver cancer cell HepG2, blood cancer cell HL60 and human lung cancer cell H460 as norcantharidin. It is appreciated that compounds 4,6 and 11 all show very significant inhibition of five tumor strains. The inhibitory activity of the compound 11 on HepG2, H460 and HL60 cells is about 9 times that of norcantharidin, and the inhibitory activity on BGC803 and HO8901 cells is about 18 times that of norcantharidin. The inhibitory activity of compound 4 on HL60 and HO8901 cells was about 18 and 8 times that of norcantharidin as a positive control. The inhibitory activity of compound 6 on H460 and HO8901 cells was about 60 and 16 times that of the positive control norcantharidin. More importantly, the toxicity of the compound 11 to WI-38 cells is equivalent to that of norcantharidin, which means that the compound 11 has wider treatment window and higher safety compared with norcantharidin. While the toxicity of compound 4 and compound 6 to HO8901 cells was much lower than that of norcantharidin, suggesting that it has a larger therapeutic index and a wider therapeutic window. Therefore, the compound provided by the invention can be used for preparing candidate medicines for resisting liver cancer, gastric cancer, lung cancer, leukemia and ovarian cancer, wherein the leukemia and ovarian cancer are preferred.

The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.