WO2018196581A1 - 双氢青蒿素-甾体结合物及其制备方法和应用 - Google Patents

双氢青蒿素-甾体结合物及其制备方法和应用 Download PDF

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WO2018196581A1
WO2018196581A1 PCT/CN2018/082120 CN2018082120W WO2018196581A1 WO 2018196581 A1 WO2018196581 A1 WO 2018196581A1 CN 2018082120 W CN2018082120 W CN 2018082120W WO 2018196581 A1 WO2018196581 A1 WO 2018196581A1
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carbon atoms
cancer
group
dihydroartemisinin
acid
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PCT/CN2018/082120
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English (en)
French (fr)
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沈征武
张菁华
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沈征武
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Priority to EP18790816.5A priority Critical patent/EP3617218B1/en
Publication of WO2018196581A1 publication Critical patent/WO2018196581A1/zh
Priority to US16/657,908 priority patent/US10881636B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/554—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J17/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, having an oxygen-containing hetero ring not condensed with the cyclopenta(a)hydrophenanthrene skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J31/00—Normal steroids containing one or more sulfur atoms not belonging to a hetero ring
    • C07J31/006—Normal steroids containing one or more sulfur atoms not belonging to a hetero ring not covered by C07J31/003
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J41/00—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
    • C07J41/0033—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005
    • C07J41/0055—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 the 17-beta position being substituted by an uninterrupted chain of at least three carbon atoms which may or may not be branched, e.g. cholane or cholestane derivatives, optionally cyclised, e.g. 17-beta-phenyl or 17-beta-furyl derivatives
    • C07J41/0061—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 the 17-beta position being substituted by an uninterrupted chain of at least three carbon atoms which may or may not be branched, e.g. cholane or cholestane derivatives, optionally cyclised, e.g. 17-beta-phenyl or 17-beta-furyl derivatives one of the carbon atoms being part of an amide group
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J71/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton is condensed with a heterocyclic ring
    • C07J71/0005—Oxygen-containing hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J71/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton is condensed with a heterocyclic ring
    • C07J71/0036—Nitrogen-containing hetero ring
    • C07J71/0042—Nitrogen only
    • C07J71/0052—Nitrogen only at position 16(17)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
    • C07J9/005—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane containing a carboxylic function directly attached or attached by a chain containing only carbon atoms to the cyclopenta[a]hydrophenanthrene skeleton

Definitions

  • the invention relates to the technical field of medicinal chemistry, in particular to a dihydroartemisinin-steroidal conjugate and a preparation method and application thereof.
  • artemisinin Since artemisinin has been found to have a strong anti-malarial effect in the 1970s, artemisinin, artesunate, artemether, dihydroartemisinin and its combination have been approved as antimalarial drugs in China. And it is widely used around the world. At the same time, such compounds have also been widely reported to have anticancer effects. Among them, the 11,13-dihydroartemisinin showed more obvious anticancer activity. In recent years, the anticancer activities of various artemisinin derivatives, including dihydroartemisinin and its dimers, artesunate, artemether and other compounds have been extensively and intensively studied. Such compounds are effective not only for leukemia and colon cancer, but also for other cancers such as melanoma, breast cancer, ovarian cancer, prostate cancer, and kidney cancer.
  • Lung cancer is one of the most malignant tumors with the fastest growth in morbidity and mortality and the greatest threat to population health and life. In the past 50 years, the incidence and mortality of lung cancer have been significantly increased in many countries. The incidence and mortality of lung cancer in males accounted for the first place among all malignant tumors. The incidence rate of females ranked second and the mortality rate ranked second. Lung cancer is divided into two categories: small cell lung cancer and non-small cell lung cancer.
  • Small cell lung cancer is mainly characterized by neuroendocrine characteristics, high degree of malignancy, rapid growth, early lymphatic metastasis and hematogenous dissemination, sensitive to chemotherapy and radiotherapy; most non-small cell lung cancer lacks neuroendocrine characteristics, including histologically squamous cell carcinoma
  • the three major categories of adenocarcinoma and large cell carcinoma are significantly less sensitive to chemotherapy and radiotherapy than small cell lung cancer.
  • Non-small cell lung cancer accounts for 80%-85% of all lung cancer cases, and small cell lung cancer accounts for 15%-20%.
  • Chemotherapy is the main treatment for lung cancer, and more than 90% of lung cancers require chemotherapy. Chemotherapy is also the main method for the treatment of non-small cell lung cancer.
  • the tumor remission rate of chemotherapy for non-small cell lung cancer is 40% to 50%.
  • Chemotherapy generally does not cure non-small cell lung cancer, only prolonging patient survival and improving quality of life.
  • molecular biology technology a variety of molecular targeted drugs have been put into clinical treatment of non-small cell lung cancer.
  • 16 targeted drugs for the treatment of non-small cell lung cancer which is the most targeted cancer drug among all cancer types.
  • CFDA has approved five types of drugs listed in China, including four small molecule targeted drugs and one macromolecular targeted drug.
  • Gliomas are the most common primary brain tumors produced by carcinogenesis of brain and spinal glial cells. Gliomas are high in children and adolescents, especially in children aged 5-9 years. Childhood patients often have poorly differentiated polar glioblastoma, medulloblastoma, and ependymoma, and adult patients have astrocytoma. Children with short course of disease and rapid progress often cause severe brainstem symptoms in a short period of time; adult patients have long course of disease and slow progress, and severe brain stem symptoms can occur in several months or even more than one year. In the past 30 years, the incidence of primary malignant brain tumors has increased year by year, with an annual growth rate of about 1.2%, especially in the middle-aged and elderly population.
  • Glioma is an invasive growth product. It has no obvious boundary with normal brain tissue. It is difficult to completely remove it. It is not sensitive to radiotherapy and chemotherapy. It is very easy to relapse. It grows in benign and malignant tumors in important parts such as the brain. It is difficult to remove or fundamentally. Can not be operated. Chemical drugs and general anti-tumor Chinese medicines are not ideal due to factors such as the blood-brain barrier. Therefore, glioma is still one of the worst prognosis in systemic tumors.
  • temozolomide is the only chemotherapeutic drug for the treatment of glioma with clear efficacy.
  • a chemotherapeutic drug for the treatment of glioma with clear efficacy.
  • the use of temozolomide can significantly prolong the survival of patients.
  • Other anti-tumor drugs have no obvious therapeutic effect on brain tumors.
  • the technical problem to be solved by the present invention is to provide a dihydroartemisinin-steroidal conjugate which has stronger anticancer activity than dihydroartemisinin and can be applied to non-small cell lung cancer, brain cancer and Treatment of other malignant tumors.
  • a dihydroartemisinin-steroidal complex having the general formula (I), or an isomer thereof, or a pharmaceutically acceptable salt thereof, or a prodrug molecule thereof,
  • the dihydroartemisinin molecule is linked to the steroid nucleus via the linker X at its 10 position; X can be attached to the 2, 3, 4, 6, 7, 11, 12, 16 or 17 position of the steroid .
  • Linker X is an atom or group having a molecular function linking the two ends, including CH 2 , O, S, N, -CO 2 -, SO 2 -, a diol or a triol of 2 to 10 carbon atoms, benzene Phenol or benzenetriol derivative, aminophenol derivative, aminobenzoic acid derivative, hydroxybenzoic acid derivative, polysubstituted benzene ring or heterocyclic ring containing two or more alkyl acid groups, 2-10 carbon atoms a diacid or polybasic acid, a diindenyl compound of 2 to 10 carbon atoms, a diamine of 2 to 10 carbon atoms, a hydroxy acid of 2 to 10 carbon atoms, an amino acid of 3 to 10 carbon atoms or an amino alcohol;
  • R 1 is H, OH, NH 2 , halogen, fluorenyl, alkoxy group of 1 to 10 carbon atoms, haloalkoxy group of 2 to 10 carbon atoms, alkoxy alcohol or alkoxy group of 1 to 10 carbon atoms a polyhydric alcohol, an alkoxy alcohol of 1 to 10 carbon atoms or a sulfonic acid ester of an alkoxy polyol, a salt thereof, an alkoxy ether of 1 to 10 carbon atoms, an alkoxy group of 1 to 10 carbon atoms a carboxylic acid or alkoxy polycarboxylic acid and a salt thereof, an alkoxysulfonic acid of 1 to 10 carbon atoms and a salt thereof, an alkoxyamine of 1 to 10 carbon atoms, an alkane of 1 to 10 carbon atoms Oxyamide, oxy olefin having 2 to 10 carbon atoms, oxyalkyne having 2 to 10 carbon atoms, cycloalkyl
  • Heterocyclic or a 3-6 carbon monosaccharides, galactose, ribose and deoxyribose, etc., which are connected via a glycosidic bond to a sugar steroid nucleus; R 1 may be attached to the steroid 3,4,6, 7th place.
  • R 2 is H, OH, NH 2 , halogen, halogen, decyl, sulfonate and its salts, sulfinate and its salts, amide group, alkoxy group of 1 to 10 carbon atoms, 2-10 carbons a halogenated alkoxy group of an atom, an alkoxy alcohol or alkoxy polyol of 1 to 10 carbon atoms, an alkoxy alcohol of 1 to 10 carbon atoms or a sulfonic acid ester of an alkoxy polyol, and a salt thereof, 1 An alkoxy ether of 10 carbon atoms, an alkoxycarboxylic acid or alkoxy polycarboxylic acid of 1 to 10 carbon atoms and a salt thereof, an alkoxysulfonic acid of 1 to 10 carbon atoms and a salt thereof, Alkoxyamine of 1 to 10 carbon atoms, alkoxyamide of 1 to 10 carbon atoms, oxyalkylene having 2 to 10 carbon atoms,
  • R 3 is H, OH, NH 2 , halogen, decyl, sulfonate and its salts, sulfinate and its salts, amide group, alkyl group of 1 to 12 carbon atoms, halogenated alkane of 2 to 12 carbon atoms a group, an alcohol or a polyol of 1 to 12 carbon atoms, a sulfonate of an alcohol or a polyol of 1 to 12 carbon atoms and a salt thereof, an ether of 1 to 12 carbon atoms or a monoether of a polyhydric alcohol, 1- a carboxylic acid or a polycarboxylic acid having 12 carbon atoms and a salt thereof, an alkylsulfonic acid having 1 to 12 carbon atoms and a salt thereof, an amine having 1 to 12 carbon atoms, an amide having 1 to 12 carbon atoms, and 2 Olefins of -12 carbon atoms, alkynes of 2 to 12 carbon atoms,
  • the C-17 side chain of cholic acid and its derivatizing group Connected), the C-17 side chain of cholic acid and its derivatizing group; the C-17 side chain of a plant sterol such as ergosterol and soy sterol or a derivative thereof; the C-17 side chain of a steroidal steroid, such as an estrogen Ketone, estradiol, estriol, hydrocortisone, cortisone, dexamethasone, prednisone, prednisone, aldosterone, etc.
  • a plant sterol such as ergosterol and soy sterol or a derivative thereof
  • the C-17 side chain of a steroidal steroid such as an estrogen Ketone, estradiol, estriol, hydrocortisone, cortisone, dexamethasone, prednisone, prednisone, aldosterone, etc.
  • C-17 side chain; and 16-position hydroxyl group becomes a spiro ring side chain
  • a spiro ring side chain Such as diosgenin, saponin, sisal, saponin, side chain of saponin, saponin, etc.
  • the stereo configuration of R 3 may be either an alpha configuration or a beta configuration ;
  • the linker X comprises O, S, N, -CH 2 -, -NR-, -BH-, -BR-, -P(O)-, -NH-NH-, -OSO 2 -, -O-CO-O-,-OCO-(CH 2 )n-COO-,-O(CH 2 )nO-,-O(CH 2 OCH 2 )nO-,-O-SO 2 -O-,- O-PO 2 -O-,-O-BO-O-,-(CH 2 )nO-,-O-Ph-O-,-O(CH 2 )nNR(CH 2 )mO-,-OOC(CH 2 ) nNR(CH 2 )m-COO-, -O(CH 2 ) 2 N(CH 2 ) 2 N(CH 2 ) 2 O-, -OPh(Y)-O-,-O-Ph(Y)-COO-,-OOC-(CH 2 ) )
  • R is H or a saturated or unsaturated hydrocarbon of 1 to 10 carbon atoms
  • Y is OR', NHR', NR' 2 , halogen, -CF 3 , -COOH, -COOR', -CONH 2 , -CONR' 2 , -CN, -NO 2 , -SO 3 -, or -SO 2 R', Y may be substituted at different positions of the benzene ring;
  • R' is a saturated or unsaturated hydrocarbon group of H or 1 to 10 carbon atoms ;n and m are 0-10, respectively.
  • the linker X is O, -CH 2 O- or -O-CH 2 CH 2 -O-.
  • linker X One end of the linker X is connected to the 10 position of the dihydroartemisinin, and the other end is connected to the 2, 3, 4, 6, 7 position, 11 position, 12 position, 15 position of the steroid body. 16-bit, or 17-bit, or attached to the side chain of the carcass. X is preferably attached to the 3, 6 or 7 position of the steroid. Its stereo configuration may be either an alpha configuration or a beta configuration.
  • R 1 may be located at the 2-position, or the 3-position, or the 4-position, or the 6-position, or the 7-position of the steroid core.
  • R 1 may also be a double bond located at 4, 5 or 6 positions.
  • R 1 is preferably OH, NH 2 , OSO 3 H, a dialkylamino group such as a dimethylamino group or the like.
  • the preferred connection position of R 1 is 4 bits, 6 bits or 7 bits. Its stereo configuration may be either an alpha configuration or a beta configuration.
  • R 2 may be located at the 11th, or 12th, or 15th, or 16th position of the corpus callosum.
  • R 2 is preferably OH, NH 2 , OSO 3 H, a dialkylamine group such as a dimethylamine group or the like.
  • the preferred connection position of R 2 is 12 bits or 16 bits. Its stereo configuration may be either an alpha configuration or a beta configuration.
  • the steroidal nucleus refers to all structural units having cyclopentane oxime polyhydrophenanthrene, which include natural and non-natural steroids and derivatives thereof, which include decane, androstane, and estradiol. , the structure type of pregnane, cholane, ergosterol, myristane and isoindole.
  • bile acid including natural or unnatural amino acid derivatives such as taurocholic acid, glycocholic acid), deoxycholic acid (including taurodeoxycholic acid, glycodeoxycholic acid, etc.)
  • Non-natural amino acid derivatives include hyodeoxycholic acid (including natural or unnatural amino acid derivatives such as tauroporine deoxycholic acid, glycine hyodeoxycholic acid), ursodeoxycholic acid (including tauroursodeoxycholic acid) , natural or unnatural amino acid derivatives such as glycosubicin deoxycholic acid), chenodeoxycholic acid (including natural or unnatural amino acid derivatives such as taurochenodeoxycholic acid, glyco-deoxycholic acid), Obecholic acid and its analogues or derivatives, cholesteryl, ergosterol, soybean sterol, plant sterols such as 5 ⁇ or 5 ⁇ sitosterol, steroid hormones and derivatives thereof such as estrone, estradi
  • glucocorticoids and their derivatives such as hydrocortisone, cortisone, dexamethasone, prednisone, prednisone and its derivatives; mineralocorticoids and their derivatives such as aldosterone, steroids Saponins such as diosgenin, saponin, sisal saponin, saponins, saponins and other plant soaps ⁇ or these plant steroidal saponins such as diosgenin, saponin, sisal saponin, saponin, saponin, steroidal alkaloids such as solanine, beta-citronine, spicy solanine , checkerboard alkaloids, cucurbitamine, mestreamine, Fritillaria alkaloids, cyclopamine and the like and derivatives thereof.
  • mineralocorticoids and their derivatives such as aldosterone, steroids Saponins such as diosgenin, saponin, sisal saponin, saponins, saponins and
  • the conjugates of the formula (I) of the present invention include all of their isomers such as positional isomers, stereoisomers and optical isomers.
  • the conjugate of the formula (I) comprises a compound of the following specific structure:
  • the present invention provides the following compounds:
  • the pharmaceutically acceptable salt of the dihydroartemisinin-steroidal combination includes a potassium salt, a sodium salt, a calcium salt, a magnesium salt, or an organic salt formed with natural and non-natural organic nitrogen-containing compounds. .
  • composition comprising one or more of the above-described dihydroartemisinin-steroidal conjugates or isomers thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof A drug molecule, and a pharmaceutically acceptable carrier.
  • the above acceptable carriers are non-toxic, can aid in administration and have no adverse effect on the therapeutic effect of the combination.
  • Such carriers can be any solid excipient, liquid excipient, semi-solid excipient or a gaseous excipient in an aerosol composition which is generally available to those skilled in the art.
  • Solid pharmaceutical excipients include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, stearyl glyceryl ester, sodium chloride , anhydrous skim milk, etc.
  • the liquid and semi-solid excipients may be selected from the group consisting of glycerin, propylene glycol, water, ethanol, and various oils, including those derived from petroleum, animal, vegetable, or synthetic oils, for example, peanut oil, soybean oil, mineral oil, sesame oil, and the like, preferably Liquid carriers, particularly for injectable solutions, include water, saline, aqueous dextrose, and glycol. It is also possible to add other adjuvants such as flavoring agents, sweeteners and the like to the composition.
  • the pharmaceutical composition of the present invention can be administered orally or by other administration means such as injection, transdermal administration, spray administration, rectal administration, vaginal administration and the like.
  • administration means such as injection, transdermal administration, spray administration, rectal administration, vaginal administration and the like.
  • the preferred mode of administration is oral, which can be adjusted depending on the extent of the disease.
  • the pharmaceutical composition of the invention can be combined with other anticancer drugs, including traditional Chinese medicine anticancer drugs, and other anticancer drugs include doxorubicin, bleomycin, vinblastine, taxanes, etoposide, 5- Fluorouracil, cyclophosphamide, methotrexate, cisplatin, retinoic acid, temozolomide, actinomycin, and targeted drugs such as imatinib, gefitinib, sorafenib, erlotinib, sulphate Nitinib, rituximab, cetuximab, trastuzumab, etc., PD-1 inhibitors such as nivolumab and pembrolizumab, PD-L1 inhibitors Such as atetizumab (atezolizumab), divavalumab (durvalumab) and avumab (avelumab).
  • doxorubicin bleo
  • the pharmaceutical composition of the present invention can be administered in combination with a heme biosynthesis precursor such as 5-aminolevulinic acid (ALA) and its homologues.
  • a heme biosynthesis precursor such as 5-aminolevulinic acid (ALA) and its homologues.
  • the pharmaceutical composition of the present invention can also be used in combination with an amino acid compound such as ⁇ -aminolevulinic acid to treat cancer.
  • the pharmaceutical composition of the present invention can also be used in combination with other cancer treatment methods such as surgery, radiation therapy, immunotherapy, bone marrow transplantation and the like.
  • the various dosage forms of the pharmaceutical compositions of the present invention can be prepared according to conventional methods in the pharmaceutical arts.
  • the compound is mixed with one or more carriers and then formulated into a desired dosage form such as a tablet, a pill, a capsule, a semisolid, a powder, a sustained release dosage form, a solution, a suspension, a formulation, a gas Aerosol and the like.
  • Dihydroartemisinin is linked at its position by an ether bond and a 2-position, a 3-position, a 5-position, a 6-position, a 7-position, an 11-position, a 12-position, a 15-position, a 16-position, or a 17-position hydroxyl group on the steroid.
  • a dihydroartemisinin-steroidal complex is obtained.
  • the specific preparation method is as follows:
  • Route 2 In the case where the steroid compound has a plurality of hydroxyl groups, in particular, a hydroxy group having a relatively small steric hindrance, such as a 3-position hydroxyl group and a hydroxyl group at another position, in order to connect the dihydroartemisinin to For the specific position of the steroid, it is necessary to first use a protecting group such as acetyl, benzoyl, Piv, chloroacetyl, benzyl, MOM, TMS, TES, TMBS for the hydroxyl group having a lower steric hindrance. , TIPS, TBDPS, etc.
  • a protecting group such as acetyl, benzoyl, Piv, chloroacetyl, benzyl, MOM, TMS, TES, TMBS
  • dihydroartemisininin is protected, and then these protected steroids are reacted with dihydroartemisinin in the presence of a Lewis acid, such that the dihydroartemisinin is linked by an ether bond to a hydroxyl group at a specific position of the steroid.
  • the protective group is removed under acidic, basic or neutral conditions to obtain a compound in which the steroid and the dihydroartemisinin are linked.
  • Dihydroartemisinin or its derivative is first linked to the linker X via an ester bond, an ether bond, an amide bond, a carbon-carbon single bond or a carbon-nitrogen, carbon-sulfur single bond, and the intermediate is passed through a total of The valence bond is linked to the 2, 3, 5, 6, 7, 7 or 12, 15th, 16th, or 17th positions of the steroid, forming a dihydroartemisinin-steroidal conjugate.
  • the linker X generally contains two to three reactive groups which can be bonded, such as a halogenated alcohol, a halogenated acid, an amino acid, a hydroxy acid, an amino alcohol, a polyhydric alcohol such as propylene glycol, glycerin, etc., a polybasic acid such as oxalic acid.
  • Preferred linkers X are polyhydric alcohols or halohydrins such as bromoethanol, ethylene glycol, glycerol and the like.
  • Polyhydric phenol derivatives such as m-phenol, phloroglucinol, dihydroxybenzoic acid, and the like.
  • the specific preparation method is as follows:
  • Route 1 The formation of dihydroartemisinin and a linker such as a polyol or a halohydrin (such as bromoethanol, ethylene glycol, glycerol, m-phenol, phloroglucin, etc.) in the presence of a Lewis acid Intermediate Z with a hydroxyl group, a phenolic hydroxyl group or a bromine, and then the intermediate Z in the presence of a Lewis acid or a base and at the 2-position, 3-position, 5-position, 6-position, 7-position, 11-position, 12-position of the steroid The 15th, 16th, or 17th hydroxyl group is linked to an ether.
  • a linker such as a polyol or a halohydrin (such as bromoethanol, ethylene glycol, glycerol, m-phenol, phloroglucin, etc.) in the presence of a Lewis acid Intermediate Z with a hydroxyl group, a phenolic hydroxyl group or
  • Route 2 In the case where the steroid compound has a plurality of hydroxyl groups, particularly a hydroxy group having a relatively small steric hindrance such as a 3-position hydroxyl group and a hydroxyl group at another position, in order to pass the dihydroartemisinin through the linker Linking to a specific position of the steroid, it is necessary to first use a protecting group such as acetyl, benzoyl, Piv, chloroacetyl, benzyl, MOM, TMS, for a hydroxyl group having a lower steric hindrance such as a 3-hydroxyl group.
  • the protecting group is reacted with the intermediate Z in the presence of a Lewis acid, so that the dihydroartemisinin is linked to the specific position of the steroid by an ether bond. Finally, the protecting group is removed under acidic, basic or neutral conditions to give a compound in which the steroid and a dihydroartemisinin are linked.
  • the acid used in the acidic condition may be a protic acid or a Lewis acid, preferably boron trifluoride etherate, p-toluenesulfonic acid, trifluoroacetic acid, sulfuric acid or hydrochloric acid.
  • the base used in the basic condition may be an organic base or an inorganic base, preferably triethylamine, decrolidine, sodium carbonate or sodium hydrogencarbonate.
  • the solvent used in each reaction may be a protic solvent or an aprotic solvent. Preferred are: diethyl ether, tetrahydrofuran, acetonitrile, DMF, DMSO, alcohol, water.
  • the reaction temperature is usually from 10 to 80 degrees Celsius.
  • the crude product of the above reaction can be further purified by chromatography, generally by column chromatography, using silica gel or alumina as a filler, and the eluent can be mixed in different ratios of petroleum ether-acetone or petroleum ether-ethyl acetate.
  • the cancer includes brain cancer, glioma, endometrial cancer, ovarian cancer, cervical cancer, breast cancer, colon cancer, lung cancer, prostate cancer, liver cancer, leukemia, lymphoma, skin cancer, basal cell tumor, hemangioma , uterine cancer, laryngeal cancer, stomach cancer, lip cancer, esophageal cancer, nasopharyngeal cancer, gallbladder cancer, pancreatic cancer, kidney cancer, tongue cancer, bladder cancer, melanoma, lipoma, thyroid cancer, thymic cancer, bone cancer, etc. .
  • lung cancer includes non-small cell lung cancer, EGFR-TKI resistant non-small cell lung cancer, and the like.
  • the dihydroartemisinin-steroidal conjugate of the invention has simple synthesis, good chemical stability and low toxicity, and exhibits strong inhibitory activity against a plurality of tumor cell lines in an in vitro test, especially for a very malignant degree. High medulloblastoma cells and EGFR-TKI resistant non-small cell lung cancer cells show high inhibitory activity.
  • the dihydroartemisinin-steroidal conjugate of the invention has a significantly higher inhibition rate on tumor cells than dihydroartemisinin, and the toxicity is much smaller than that of paclitaxel, and has broad application prospects in the treatment of cancer.
  • Figure 1 is a graph showing the tumor growth curve of the anti-cancer effect of the JZ-2 compound of the present invention on the human lung cancer cell A549 nude mouse transplantation model;
  • Fig. 2 is a graph showing the body weight of experimental animals in which the JZ-2 compound of Example 15 of the present invention has an anticancer effect on a human lung cancer cell A549 nude mouse transplantation model.
  • boron trifluoride etherate (1 mL) was slowly added dropwise to dihydroartemisinin (569 mg, 2.0 mmol) and ursodeoxycholic acid (785 mg, 2.0 mmol) in diethyl ether solution (50 mL), the reaction system It was naturally stirred to room temperature and stirred overnight.
  • Potassium carbonate (276 mg, 2.0 mmol) and a catalytic amount of potassium iodide were added to ethyl bromide dihydroartemisinin (391 mg, 1.0 mmol) and ursodeoxycholic acid (392 mg, 1.0 mmol) in DMF (20 mL) at room temperature. In the solution, the reaction solution was heated to 70 ° C for 2 hours.
  • the boron trifluoride etherate complex (180 mg) was added to a solution of dihydroartemisinin deoxycholate (compound JZ-08, 130 mg, 0.18 mmol) in pyridine (2 mL) at 0 ° C, and the reaction was allowed to react at room temperature.
  • the tumor cells used were: human prostate cancer cell-PC-3 cell (CRL-1435), human cervical cancer cell line-HeLa cell (CCL-2), human acute T cell leukemia cell-Jurkat cell (TIB-152), Human renal clear cell adenocarcinoma-786-O cells (CRL-1932) and human breast cancer cells-MCF7 cells (HTB-22) were tested for the effect of the above-mentioned compound samples of the present invention on the growth of these tumor cell lines; Renal cell-Vero cells are normal cells (CCL-81) and the cytotoxic effects of the compound samples are examined.
  • the living cells can be stained by the cell proliferation-toxicity test kit Cell Counting Kit-8 (CCK-8 solution), and the OD value of the cells can be measured to reflect the survival and growth state of the cells, that is, the cell survival rate ( %) and cell growth inhibition (%).
  • the compound concentration CC 50 at half cell survival and the compound concentration IC 50 at which half of the cell growth was inhibited were calculated.
  • Vero cells (CCL-81), PC-3 cells (CRL-1435), HeLa cells (CCL-2), Jurkat cells (TIB-152), 786-O cells (CRL-1932), MCF7 cells (HTB-22) ) were purchased from ATCC; DMEM medium, F12 medium, MEM medium, RPMI 1640 medium were purchased from Gibco (Life Technologies); fetal bovine serum (FBS) was purchased from Hyclone (Thermo Scientific); From Gibco (Life Technologies); CCK-8 solution was purchased from Shanghai Dongren Chemical Technology Co., Ltd.; 96-well cell culture plate (transparent flat bottom) was purchased from Corning; SpectraMax 190 microplate reader was purchased from Molecular Devices Corporation.
  • DMEM medium, F12 medium, MEM medium, RPMI 1640 medium were purchased from Gibco (Life Technologies); fetal bovine serum (FBS) was purchased from Hyclone (Thermo Scientific); From Gibco (Life Technologies); CCK-8 solution was purchased from Shanghai Dongren Chemical Technology Co., Ltd
  • the culture solution containing 10% FBS is a complete culture solution.
  • Vero cells were cultured in DMEM complete medium; PC-3 cells were cultured in F12 complete medium; Hela cells were cultured in MEM complete medium; Jurkat cells were cultured in RPMI1640 complete medium; 786-O cells were completely treated with RPMI 1640 The culture solution was cultured; MCF-7 cells were cultured in MEM complete medium. After all the cells were cultured in logarithmic growth, the following tests were performed.
  • the adherent cells were collected by trypsinization; the suspended cells were collected directly.
  • the cells were centrifuged, and after counting, the cells were prepared into 5 ⁇ 103 cells/100 ⁇ l/well with respective complete culture solutions, and seeded in a transparent 96-well culture plate; at the same time, each cell was set without cells or samples. Blank wells; and control wells with or without cells.
  • the 96-well culture plates after inoculation were cultured overnight at 37 ° C in a 5% CO 2 incubator. A test compound sample of 50 ⁇ l/well was added the next day, and 50 ⁇ l/well of the complete culture solution was added.
  • the final test concentrations of the compound samples were 100 ⁇ M, 25 ⁇ M, 6.25 ⁇ M, 1.56 ⁇ M, 0.39 ⁇ M, 0.1 ⁇ M, 0.02 ⁇ M, 0.006 ⁇ M, and the test concentrations were double duplicate wells; the control wells and blank wells were replaced with complete culture solutions, ie, 100 ⁇ l/well of complete culture solution was added. Incubation was continued for 48 hours at 37 ° C in a 5% CO 2 incubator. Two hours before the end of the culture, the culture solution was removed, and 100 ⁇ l/well of a complete culture solution containing 10% CCK-8 solution was added until the end of the culture, and the OD value was measured at 450 nM (reference 650 nM).
  • the half cell death concentration CC 50 was calculated to reflect the toxic effect of the compound sample; the cell growth inhibition (%) of the tumor cell line was detected, and the concentration of the cell growth number was determined at half time.
  • IC 50 which reflects the antitumor activity of the compound sample.
  • the IC 50 results are shown in Table 1 and Table 2.
  • the results show that the dihydroartemisinin-steroidal conjugate has different degrees of inhibition on the growth of various tumor cells, and this series of compounds have a far lower inhibition rate on the tested tumor cells. Far stronger than dihydroartemisinin.
  • the inhibition rate of individual compounds on test tumor cells is similar to that of paclitaxel, but the toxicity is much smaller than that of paclitaxel.
  • Group 3 was administered once every three days, and paclitaxel 10 mg/kg was intraperitoneally injected. The entire treatment process lasted for 28 days. Among them, three died in group one and one died in group three. There was no animal death in group 2, and the death of group 1 mice was caused by uncontrolled growth of tumors. No death occurred in group 2, indicating that the tumors in this group were well controlled, and JZ-02 was less toxic than paclitaxel.
  • the tumor growth curve shown in Fig. 1 indicates that the compound JZ-2 of the present invention can significantly inhibit tumor growth, and the inhibitory effect is better than that of the positive control paclitaxel.
  • the body weight curve shown in Figure 2 indicates that there was no significant decrease in body weight of the mice after 28 days of continuous administration of JZ-2. It was proved that the compounds were less toxic and had no significant effect on the body weight and quality of life of the mice.
  • the tumor inhibition rate of the group 2 (the treatment group) was 58%.
  • the tumor inhibition rate of the positive control group was 52%.

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Abstract

本发明公开了一种具有下述通式(I)的双氢青蒿素-甾体结合物或其可药用盐，其中，双氢青蒿素分子在其10位通过连接体X与甾体母核相连接。本发明还公开了该双氢青蒿素-甾体结合物的制备方法及其在制备治疗癌症的药物中的应用。本发明的双氢青蒿素-甾体结合物，对多株肿瘤细胞显示出很强的抑制活性，且毒性小，可透过血脑屏障，应用前景非常广阔。

Description

双氢青蒿素-甾体结合物及其制备方法和应用 技术领域
本发明涉及药物化学技术领域，具体涉及一种双氢青蒿素-甾体结合物及其制备方法和应用。
背景技术
自从上世纪70年代发现青蒿素具有强大的抗疟作用以来，青蒿素，青蒿琥酯，蒿甲醚，双氢青蒿素及其复方制剂分别先后在中国被批准为抗疟药物，并在全球得到广泛应用。同时，该类化合物也被广泛报道具有抗癌作用。其中11，13-位双氢青蒿素显示较明显的抗癌活性。近年来各类青蒿素的衍生物，包括双氢青蒿素及其二聚物，青蒿琥酯，蒿甲醚等化合物的抗癌活性得到广泛和深入的研究。这类化合物在体外实验中不仅对白血病、结肠癌有效，对其他癌症如黑色素瘤、乳腺癌、卵巢癌、前列腺癌、肾癌等均具有良好的活性。
随着中国经济建设的发展，人民生活得到了很大的改善。但是同时，由于中国人口老龄化加剧、生态环境遭受破坏、不健康生活方式及食品安全问题凸现，肿瘤发病率多年持续上升，已成为一个必须高度重视的公共卫生问题乃至社会问题，根据国际癌症研究署预测，如不采取有效措施，中国癌症发病数和死亡数到2020年将上升至400万人和300万人；2030年将上升至500万人和350万人。国家癌症中心发布的《2012中国肿瘤登记年报》显示，全国肿瘤登记地区恶性肿瘤发病第一位的是肺癌，其次为胃癌、结直肠癌、肝癌和食管癌；死亡第一位的是肺癌，其次为肝癌、胃癌、食管癌和结直肠癌。
肺癌是发病率和死亡率增长最快，对人群健康和生命威胁最大的恶性肿瘤之一。近50年来许多国家都报道肺癌的发病率和死亡率均明显增高，男性肺癌发病率和死亡率均占所有恶性肿瘤的第一位，女性发病率占第二位，死亡率占第二位。肺癌分为两大类：小细胞肺癌和非小细胞肺癌。小细胞肺癌主要表现为神经内分泌特性，恶性程度高，生长快，较早出现淋巴转移和血行播散，对化疗和放疗敏感；大多数非小细胞肺癌缺乏神经内分泌特性，组织学上包括鳞癌、腺癌和大细胞癌三大类，对化疗和放疗的敏感性明显低于小细胞肺癌。非小细胞肺癌约占所有肺癌病例的80％-85％，小细胞肺癌占15％-20％。化疗是肺癌的主要治疗方法，90％以上的肺癌需要接受化疗治疗。化疗也是治疗非小细胞肺癌的主要手段，化疗治疗非小细胞肺癌的肿瘤缓解率为40％～50％。化疗一般不能治愈非小细胞肺癌，只能延长患者生存和改善生活质量。随着分子生物学技术的发展，多种分子靶向药物已经投入了临床治疗非小细胞肺癌。目前，治疗非小细胞肺癌的靶向药物有16种，是所有癌种中靶向药物最多的癌种。抗体类大分子靶向药物有6种，小分子靶向药物有10种。CFDA批准在中国上市的有5种， 包含4种小分子靶向药物，1种大分子靶向药物。从每年FDA批准上市的靶向药物的数量可以看出，从2011年开始几乎每年都会有针对肺癌的靶向药物上市，2014年非小细胞肺癌的靶向药物上市数量达到了历史峰值，有5种之多。这些靶向药物的上市，减轻了患者痛苦，延长了其生存期。但是目前而言，由于在大部分肿瘤中仍未检测到驱动突变，因而无法采用靶向药物进行治疗，同时获得性耐药是这类靶向药物最大的缺陷之一。因此发展新的低毒广谱类的肿瘤药物，仍是治疗非小细胞肺癌和耐药的非小细胞肺癌所面临的重大挑战。
脑胶质瘤是由于大脑和脊髓胶质细胞癌变所产生的、最常见的原发性颅脑肿瘤。脑胶质瘤在儿童及青少年高发，特别是5～9岁儿童发病率最高。儿童病人常以分化较差的极性成胶质细胞瘤、髓母细胞瘤和室管膜瘤为多，成年病人则以星形细胞瘤为多。儿童患者病程短、进展快；常在较短时间内即引起严重的脑干症状；成年患者病程长、进展慢，可数月甚至1年以上始出现严重的脑干症状。近30年来，原发性恶性脑肿瘤发生率逐年递增，年增长率约为1.2％,中老年人群尤为明显。据文献报道，中国脑胶质瘤年发病率为3-8人/10万人，年死亡人数达3万人。胶质瘤系浸润性生长物，它和正常脑组织没有明显界限，难以完全切除，对放疗化疗不甚敏感，非常容易复发，生长在大脑等重要部位的良、恶性肿瘤，手术难以切除或根本不能手术。化学药物和一般抗肿瘤的中药，因血脑屏障等因素的影响，疗效也不理想，因此脑胶质瘤至今仍是全身肿瘤中预后最差的肿瘤之一。
目前，替莫唑胺是治疗胶质瘤唯一有明确疗效的化疗药物。对于高度恶性的脑胶质瘤，替莫唑胺的应用，可以显著延长患者的生存预后。而其他抗肿瘤药物对脑瘤没有明显治疗作用。
发明内容
本发明要解决的技术问题是提供一种双氢青蒿素-甾体结合物，该结合物比双氢青蒿素具有更强的抗癌活性，可应用于非小细胞肺癌，脑癌及其他恶性肿瘤的治疗。
为了解决上述技术问题，本发明通过如下技术方案实现:
在本发明的一个方面，提供了一种具有通式(I)的双氢青蒿素-甾体结合物，或其异构体、或其可药用盐、或其前药分子，
Figure PCTCN2018082120-appb-000001
其中，双氢青蒿素分子在其10位通过连接体X与甾体母核相连接；X可以连接在甾体的2，3，4，6，7，11，12，16或17位上。
连接体X为具有连接两端分子功能的原子或基团，包括CH 2，O，S，N，-CO 2-，SO 2-，2-10个碳原子的二醇或三醇，苯二酚或苯三酚衍生物，氨基酚衍生物，氨基苯甲酸衍生物，羟基苯甲酸衍生物，多取代的含有2个以上烷基酸取代的苯环或杂环，2-10个碳原子的二酸或多元酸，2-10个碳原子的二巯基化合物，2-10个碳原子的二胺，2-10个碳原子的羟基酸，3-10个碳原子的氨基酸或氨基醇；
R 1为H，OH，NH 2，卤素，巯基，1-10个碳原子的烷氧基，2-10个碳原子的卤代烷氧基，1-10个碳原子的烷氧基醇或烷氧基多元醇，1-10个碳原子的烷氧基醇或烷氧基多元醇的磺酸酯及其盐，1-10个碳原子的烷氧基醚，1-10个碳原子的烷氧基羧酸或烷氧基多元羧酸及其盐，1-10个碳原子的烷氧基磺酸及其盐，1-10个碳原子的烷氧基胺，1-10个碳原子的烷氧基酰胺，含2-10个碳原子的氧基烯烃，含2-10个碳原子的氧基炔烃，3-8个碳的环烷基氧基，酮基，亚胺，1-10个碳原子的烷基亚胺，1-10个碳原子的烷氧基亚胺，羟胺，羟胺磺酸酯及其盐，亚磺酸酯，酰胺基，带有或不带有取代基的苯氧基，带有或不带有取代基的苄氧基，带有或不带有取代基的萘氧基，二甲基胺基、二乙基胺基等1-10个碳原子的二烷基胺基，如哌嗪，哌啶，吡咯，四氢吡咯等含有1-4个杂原子的五元-八元的杂环，或3-6个碳的单糖如果糖，半乳糖，核糖和脱氧核糖等，这些糖通过糖苷键与甾体母核相连；R 1可以连接在甾体的3，4，6，7位上。
R 2为H，OH，NH 2，卤素，卤素，巯基，磺酸酯及其盐，亚磺酸酯及其盐，酰胺基，1-10个碳原子的烷氧基，2-10个碳原子的卤代烷氧基，1-10个碳原子的烷氧基醇或烷氧基多元醇，1-10个碳原子的烷氧基醇或烷氧基多元醇的磺酸酯及其盐，1-10个碳原子的烷氧基醚，1-10个碳原子的烷氧基羧酸或烷氧基多元羧酸及其盐，1-10个碳原子的烷氧基磺酸及其盐，1-10 个碳原子的烷氧基胺，1-10个碳原子的烷氧基酰胺，含2-10个碳原子的氧基烯烃，含2-10个碳原子的氧基炔烃，3-8个碳的环烷基氧基，酮基，亚胺，1-10个碳原子的烷基亚胺，1-10个碳原子的烷氧基亚胺，羟胺，羟胺磺酸酯及其盐，带有或不带有取代基的苯氧基，带有或不带有取代基的苄氧基，带有或不带有取代基的萘氧基，二甲基胺基、二乙基胺基等1-10个碳原子的二烷基胺基，如哌嗪，哌啶，吡咯，四氢吡咯等含有1-4个杂原子的五元-八元的杂环，或3-6个碳的单糖，这些糖通过糖苷键与甾体母核相连；
R 3为H，OH，NH 2，卤素，巯基，磺酸酯及其盐，亚磺酸酯及其盐，酰胺基，1-12个碳原子的烷基，2-12个碳原子的卤代烷基，1-12个碳原子的醇或多元醇，1-12个碳原子的醇或多元醇的磺酸酯及其盐，1-12个碳原子的醚或多元醇的单醚，1-12个碳原子的羧酸或多元羧酸及其盐，1-12个碳原子的烷基磺酸及其盐，1-12个碳原子的胺，1-12个碳原子的酰胺，含2-12个碳原子的烯烃，含2-12个碳原子的炔烃，3-8个碳的环烷，1-12个碳原子的酮，1-12个碳原子的烷基亚胺，1-12个碳原子的烷氧基亚胺，羟胺，羟胺磺酸酯及其盐，1-12个碳原子的烷基磺酸酯及其盐，1-12个碳原子的烷基亚磺酸酯，1-12个碳原子的烷基酰胺，带有或不带有取代基的苯基或苯氧基，带有或不带有取代基的苄基，带有或不带有取代基的萘基，二甲基胺基、二乙基胺基等1-12个碳原子的二烷基胺基，如哌嗪，哌啶，吡咯，四氢吡咯等含有1-4个杂原子的五元-八元的杂环或苯并杂环，或3-6个碳的单糖(这些糖通过糖苷键与甾体母核相连)，胆酸的C-17侧链及其衍生基团；植物甾醇如麦角甾醇和大豆甾醇的C-17侧链或其衍生基团；激素类甾体的C-17侧链，如雌酮，雌二醇，雌三醇，氢化可的松，可的松，地塞米松，强的松，泼尼松，醛固酮等的C-17侧链；与16位羟基成为螺环的侧链，如薯蓣皂甙元，海柯皂甙元，剑麻皂甙元，替告皂甙元，叉蕊皂甙元等的侧链；甾体生物碱的C-17侧链，如茄碱，β-苦茄碱，辣茄碱，棋盘花碱，藜芦胺碱，介藜芦胺，贝母生物碱，环巴胺等的侧链；R 3的立体构型可以是α构型，也可以是β构型；
式(I)中，
Figure PCTCN2018082120-appb-000002
代表α-取代，
Figure PCTCN2018082120-appb-000003
代表β-取代。
优选的，所述连接体X包括O，S，N，-CH 2-，-NR-，-BH-，-BR-，-P(O)-，-NH-NH-，-OSO 2-，-O-CO-O-，-OCO-(CH 2)n-COO-，-O(CH 2)nO-，-O(CH 2OCH 2)nO-，-O-SO 2-O-，-O-PO 2-O-，-O-BO-O-，-(CH 2)n-O-，-O-Ph-O-，-O(CH 2)nNR(CH 2)mO-，-OOC(CH 2)nNR(CH 2)m-COO-，-O(CH 2) 2N(CH 2) 2N(CH 2) 2O-，
Figure PCTCN2018082120-appb-000004
-OPh(Y)-O-，-O-Ph(Y)-COO-，-OOC-(CH 2)nPh(Y)(CH 2)mCOO-，-OOCPh(Y)COO-，-OCH 2CH 2NHPh(Y)NHCH 2CH 2O-，含有1-4个杂原子的五元或六元杂环及其衍生物如呋喃， 嘧啶，噻唑，噻吩，噁唑，嘌呤等；
其中R为H或1-10个碳原子的饱和或不饱和烃；Y为OR’，NHR’，NR’ 2，卤素，-CF 3，-COOH，-COOR’，-CONH 2，-CONR’ 2，-CN，-NO 2，-SO 3-，或-SO 2R’，Y可在苯环不同的位置上取代；R’为H或1-10个碳原子的饱和或不饱和的烃基；n和m分别为0-10。
更优选的，所述连接体X为O，-CH 2O-或-O-CH 2CH 2-O-。
所述连接体X一端连接在双氢青蒿素的10位，另一端连接在甾体母核的2位，3位，4位，6位，7位，11位，12位，15位，16位，或者17位，或者连接在甾体的侧链上。X优选为连接在甾体3位，6位或者7位。其立体构型可以是α构型，也可以是β构型。
本发明式(I)中，R 1可以位于甾体母核的2位，或者3位，或者4位，或者6位，或者7位。R 1还可以是位于4位，5位或者6位的双键。R 1优选为OH，NH 2，OSO 3H，二烷基胺基如二甲基胺基等。R 1优选的连接位置为4位，6位或者7位。其立体构型可以是α构型，也可以是β构型。
R 2可以位于甾体母核的11位，或者12位，或者15位，或者16位。R 2优选为OH，NH 2，OSO 3H，二烷基胺基如二甲基胺基等。R 2优选的连接位置为12位或者16位。其立体构型可以是α构型，也可以是β构型。
在本发明中，甾体母核是指所有具有环戊烷骈多氢菲的结构单元，其包括天然和非天然的甾体及其衍生物，其包含甾烷，雄甾烷，雌甾烷，孕甾烷，胆烷，麦角甾烷，豆甾烷及异甾体的结构类型。具体来说，包含胆酸(包括牛磺胆酸，甘氨胆酸等天然或非天然氨基酸衍生物)，去氧胆酸(包括牛磺去氧胆酸，甘氨去氧胆酸等天然或非天然氨基酸衍生物)，猪去氧胆酸(包括牛磺猪去氧胆酸，甘氨猪去氧胆酸等天然或非天然氨基酸衍生物)，熊去氧胆酸(包括牛磺熊去氧胆酸，甘氨熊去氧胆酸等天然或非天然氨基酸衍生物)，鹅去氧胆酸(包括牛磺鹅去氧胆酸，甘氨鹅去氧胆酸等天然或非天然氨基酸衍生物)，奥贝胆酸及其类似物或衍生物，胆甾醇，麦角甾醇，大豆甾醇，5α或5β谷甾醇等植物甾醇，甾体激素及其衍生物如雌酮，雌二醇，雌三醇等及其衍生物，糖皮质激素及其衍生物如氢化可的松，可的松，地塞米松，强的松，泼尼松等及其衍生物；盐皮质激素及其衍生物如醛固酮，甾体皂甙如薯蓣皂甙，海柯皂甙，剑麻皂甙，替告皂甙，叉蕊皂甙等植物皂甙或这些植物甾体皂甙元如薯蓣皂甙元，海柯皂甙元，剑麻皂甙元，替告皂甙元，叉蕊皂甙元，甾体生物碱如茄碱，β-苦茄碱，辣茄碱，棋盘花碱，藜芦胺碱，介藜芦胺，贝母生物碱，环巴胺等及其上述甾体的衍生物。
本发明通式(I)的结合物包括其所有的异构体，如位置异构体，立体异构体和光学异构体。
优选的，所述通式(I)结合物包括下述具体结构的化合物：
Figure PCTCN2018082120-appb-000005
Figure PCTCN2018082120-appb-000006
Figure PCTCN2018082120-appb-000007
更优选的，本发明实施例提供了下述化合物：
Figure PCTCN2018082120-appb-000008
Figure PCTCN2018082120-appb-000009
在本发明中，双氢青蒿素-甾体结合物的可药用盐，包括钾盐，钠盐，钙盐，镁盐，或者与天然的和非天然的有机含氮化合物形成的有机盐。
在本发明的另一方面，还提供了一种药物组合物，包含一种或几种上述双氢青蒿素-甾体结合物或其异构体、或其可药用盐、或其前药分子，以及药学上可接受的载体。
上述可接受的载体是无毒的、能辅助施用并且对结合物的治疗效果没有不利影响。此类载体可以是本领域的技术人员通常能得到的任何固体赋形剂、液体赋形剂、半固体赋形剂或者在气雾剂组合物中的气体赋形剂。固体药物赋形剂包括淀粉、纤维素、滑石、葡萄糖、乳糖、蔗糖、明胶、麦芽、稻米、面粉、白垩、硅胶、硬脂酸镁、硬脂酸钠、甘油硬脂酰酯、氯化钠、无水脱脂乳等。液体和半固体赋形剂可以选自甘油、丙二醇、水、乙醇和各种油，包括那些源于石油、动物、植物或人工合成的油，例如，花生油、豆油、矿物油、芝麻油等、优选的液体载体，特别是用于可注射溶液的，包括水、盐水、葡萄糖水溶液和甘醇。另外还可以在组合物中加入其它辅剂如香味剂、甜味剂等。
本发明的药物组合物可以通过口服或其他给药方式给药，如注射，透皮给药，喷雾给药，直肠给药，阴道给药等。优选的给药方式是口服，它可根据疾病程度调节。
本发明的药物组合物可以和其他抗癌药物包括中药抗癌药物联合用药，其他抗癌药物包括阿霉素类，博莱霉素，长春碱类，紫杉烷类，依托泊苷，5-氟尿嘧啶，环磷酰胺，甲氨蝶呤，顺铂，维甲酸，替莫唑胺，放线菌素，以及靶向药物如伊马替尼、吉非替尼、索拉非尼、厄洛替尼、舒尼替尼、利妥昔单抗、西妥昔单抗、曲妥珠单抗等，PD-1抑制剂如尼伏单抗(nivolumab)和潘利珠单抗(pembrolizumab)等，PD-L1抑制剂如阿替珠单抗(atezolizumab)、度伐单抗(durvalumab)和阿维单抗(avelumab)等。
本发明的药物组合物可以和血红素生物合成前体如5-氨基乙酰丙酸(ALA)及其同系物联合用药。
本发明的药物组合物还可以和δ-氨基酮戊酸等氨基酸类化合物联合用药治疗癌症。
本发明的药物组合物也可以和其他癌症治疗方法联合使用，如手术治疗，放射治疗，免疫治疗，骨髓移植等。
本发明药物组合物的各种剂型可以按照药学领域的常规方法制备。例如使该化合物与一种或者多种载体混合，然后将其制成所需的剂型，如片剂、药丸、胶囊、半固体、粉末、缓释剂型、溶液、混悬液、配剂、气雾剂等。
在本发明的另一方面，还提供了上述双氢青蒿素-甾体结合物的制备方法，包括以下步骤：
双氢青蒿素在其10位以醚键和甾体上的2位，3位，5位，6位，7位，11位，12位，15位，16位，或者17位羟基连接，得双氢青蒿素-甾体结合物。
具体的制备方法如下：
路线一：双氢青蒿素在路易斯酸的存在下，直接和甾体的2位，3位，5位，6位，7位，11位，12位，15位，16位，或者17位羟基连接成醚，得双氢青蒿素-甾体结合物。
路线二：在甾体化合物具有多个羟基的情况下，特别是具有立体位阻相对比较小的羟基，如3-位羟基和其他位置的羟基共同存在下，为了使双氢青蒿素连接于甾体的特定位置，就需要先将甾体上位阻较小的羟基如3-位羟基用保护基如乙酰基，苯甲酰基，Piv，氯乙酰基，苄基，MOM，TMS，TES，TMBS，TIPS，TBDPS等进行保护，然后这些有保护基的甾体在路易斯酸存在下与双氢青蒿素进行反应，使得双氢青蒿素以醚键与甾体的特定位置的羟基连接，最后，在酸性、碱性或中性的条件下脱掉保护基，得到甾体和双氢青蒿素相连接的化合物。
在本发明的另一方面，还提供了上述双氢青蒿素-甾体结合物的第二种制备方法，包括以下步骤：
双氢青蒿素或其衍生物先和连接体X通过酯键，醚键，酰胺键，碳-碳单键或者碳-氮、碳-硫单键连接成为中间体，该中间体再通过共价键与甾体的2位，3位，5位，6位，7位，11位，12位，15位，16位，或17位连接，形成双氢青蒿素-甾体结合物。
其中的连接体X一般含有两到三个可以成键的活性基团，如卤代醇，卤代酸，氨基酸，羟基酸，氨基醇，多元醇如丙二醇、甘油等，多元酸如乙二酸、丙二酸、EDTA等，糖如果糖、核糖、赤藓糖、赤藓糖醇等，多元胺类如乙二胺、丙二胺、丁二胺、苯二胺、哌嗪等，多元酚如间苯酚、间苯三酚及其衍生物，杂环类如哌嗪、氨基噻唑乙酸类衍生物等。优选的连接体X为多元醇或卤代醇如溴代乙醇、乙二醇、丙三醇等。多元酚类衍生物如间苯酚、间苯三酚、二羟基苯甲酸等。
具体的制备方法如下：
路线一：采用双氢青蒿素和连接体如多元醇或卤代醇(如溴代乙醇，乙二醇，丙三醇，间苯酚，间苯三酚等)在路易斯酸的存在下，形成带有羟基、酚羟基或溴的中间体Z，然后该中间体Z在路易斯酸或碱的存在下和甾体的2位，3位，5位，6位，7位，11位，12位，15位，16位，或17位羟基连接成醚。
Figure PCTCN2018082120-appb-000010
路线二：在甾体化合物具有多个羟基的情况下，特别是具有立体位阻相对比较小的羟基如3-位羟基和其他位置的羟基共同存在下，为了使双氢青蒿素通过连接体连接于甾体的特定位置，就需要先将甾体上位阻较小的羟基如3-位羟基用保护基团如乙酰基，苯甲酰基，Piv，氯乙酰基，苄基，MOM，TMS，TES，TMBS，TIPS，TBDPS等进行保护，然后加上了保护基的甾体在路易斯酸的存在下与中间体Z进行反应，使得双氢青蒿素以醚键与甾体的特定位置连接，最后，在酸性、碱性或中性的条件下脱掉保护基，得到甾体和一个双氢青蒿素相连接的化合物。
上述反应中，所述酸性条件使用的酸可以是质子酸或路易斯酸，优选为三氟化硼乙醚、对甲苯磺酸、三氟乙酸、硫酸或盐酸。所述碱性条件使用的碱可以是有机碱或无机碱，优选为三乙胺、砒咯烷、碳酸钠或碳酸氢钠。各反应所使用的溶剂可以是质子溶剂或非质子溶剂。优选为：乙醚，四氢呋喃，乙腈，DMF，DMSO，醇，水。反应温度一般在摄氏温度10-80度。
上述反应的粗产物可以用色谱法进一步纯化，一般用柱层析法，填料用硅胶或氧化铝，洗脱剂可以用石油醚-丙酮或石油醚-乙酸乙酯的不同比例的混合。
在本发明的另一方面，还提供了上述双氢青蒿素-甾体结合物或其异构体、或其可药用盐、或其前药分子在制备治疗癌症的药物中的应用。
所述癌症包括脑癌、脑胶质瘤、子宫内膜癌、卵巢癌、宫颈癌、乳腺癌、结肠癌、肺癌、前列腺癌、肝癌、白血病、淋巴癌、皮肤癌、基底细胞瘤、血管瘤、子宫癌、喉癌、胃癌、唇癌、食道癌、鼻咽癌、胆囊癌、胰腺癌、肾癌、舌癌、膀胱癌、黑素瘤、脂肪瘤、甲状腺癌、胸腺癌、骨癌等。其中肺癌包括非小细胞肺癌、EGFR-TKI耐药的非小细胞肺癌等。
本发明的双氢青蒿素-甾体结合物，合成简单，化学稳定性好，毒性小，在体外试验中对多株肿瘤细胞株都显示出非常强的抑制活性，尤其是对恶性程度非常高的髓母细胞瘤株和EGFR-TKI耐药的非小细胞肺癌细胞显示出了高度的抑制活性。本发明双氢青蒿素-甾体结合物对肿瘤细胞的抑制率明显高于双氢青蒿素，而毒性却远小于紫杉醇，在癌症的治疗方面具有广阔的应用前景。
附图说明
下面结合附图和具体实施方式对本发明作进一步详细的说明。
图1是本发明实施例15的JZ-2化合物在人源肺癌细胞A549裸鼠移植模型上的抗癌作用的肿瘤生长曲线图；
图2是本发明实施例15的JZ-2化合物在人源肺癌细胞A549裸鼠移植模型上的抗癌作用的实验动物体重曲线图。
具体实施方式
实施例1 化合物JZ-01的制备
Figure PCTCN2018082120-appb-000011
-78℃下，三氟化硼乙醚(1mL)缓慢滴加至双氢青蒿素(569mg，2.0mmol)和胆酸(817mg，2.0mmol)的乙醚溶液(50mL)中，反应体系自然升至室温后搅拌过夜。TLC监测至反应结束后，用饱和碳酸氢钠水溶液(30mL)缓慢淬灭反应，乙酸乙酯萃取(30mL×3)，有机相合并后用水(50mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝3:1-1:1)得无色油状化合物(编号化合物JZ-01)726mg(产率：53.8％)。
1H NMR(400MHz,CDCl 3)：δ5.47(s,1H),4.93(d,J＝3.3Hz,1H),4.00(s,1H),3.88(s,1H),3.51(t,J＝11.0Hz,1H),2.64–2.56(m,1H),2.47–2.21(m,4H),2.10–2.02(m,2H),1.99–1.85(m,5H),1.72(m,8H),1.53(m,5H),1.45(m,6H),1.40–1.30(m,5H),1.31–1.23(m,4H),1.16(m,2H),0.99(m,7H),0.89(m,8H),0.71(s,3H)。
13C NMR(100MHz,CDCl 3)：δ104.03(s),100.14(s),88.08(s),81.29(s),77.24(s),68.67(s),52.65(s),46.86(s),46.51(s),44.56(s),42.21(s),41.33(s),39.32(s),37.47(s),36.51(s),35.58(s),35.29(s),35.22(s),34.80(d),30.94–30.35(m),30.25(d),29.72(s),28.71(d),27.58(s),26.70(s),26.30(s),24.70(s),23.22(s),22.60(s),20.41(s),17.24(s),13.08(s),12.46(s)。
实施例2 化合物JZ-02的制备
Figure PCTCN2018082120-appb-000012
-78℃下，三氟化硼乙醚(1mL)缓慢滴加至双氢青蒿素(569mg，2.0mmol)和熊去氧胆酸(785mg，2.0mmol)的乙醚溶液(50mL)中，反应体系自然升至室温后搅拌过夜。TLC监测至反应结束后，用饱和碳酸氢钠水溶液(30mL)缓慢淬灭反应，乙酸乙酯萃取(30mL×3)， 有机相合并后用水(50mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝4:1-2:1)得无色油状化合物(编号化合物JZ-02)748mg(产率：56.8％)。
1H NMR(400MHz,CDCl 3)：δ5.46(s,1H),4.92(d,J＝3.3Hz,1H),3.61(m,2H),2.67–2.55(m,1H),2.47–2.34(m,2H),2.28(ddd,J＝15.8,9.6,6.5Hz,1H),2.04(m,2H),1.94–1.71(m,9H),1.71–1.55(m,4H),1.54–1.41(m,11H),1.40–1.21(m,9H),1.19–1.01(m,3H),0.98–0.89(m,12H),0.70(s,3H)。
13C NMR(100MHz,CDCl 3)：δ178.71(s),104.07(s),100.21(s),88.08(s),81.22(s),77.24(s),71.41(s),55.61(s),54.92(s),52.61(s),44.51(s),43.79(s),42.37(s),40.07(s),39.19(s),37.49(s),36.99(s),36.48(s),35.21(s),35.01(s),34.71(s),34.25(s),33.60(s),30.78(d),28.74(d),26.90(s),26.27(s),24.63(d),23.53(s),21.24(s),20.41(s),18.42(s),13.16(s),12.15(s)。
实施例3 化合物JZ-03的制备
Figure PCTCN2018082120-appb-000013
-78℃下，三氟化硼乙醚(1mL)缓慢滴加至双氢青蒿素(569mg，2.0mmol)和猪去氧胆酸(785mg，2.0mmol)的乙醚溶液(50mL)中，反应体系自然升至室温后搅拌过夜。TLC监测至反应结束后，用饱和碳酸氢钠水溶液(30mL)缓慢淬灭反应，乙酸乙酯萃取(30mL×3)，有机相合并后用水(50mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝8:1-1:1)得无色油状化合物(编号化合物JZ-03)653mg(产率：49.5％)。
1H NMR(400MHz,CDCl 3)；δ5.41(s,1H),4.96(d,J＝3.5Hz,1H),4.14–4.02(m,1H),3.64(dt,J＝15.5,5.5Hz,1H),2.68–2.58(m,1H),2.35(m,3H),2.12–1.95(m,2H),1.95–1.56(m,12H),1.56–1.21(m,16H),1.12(m,6H),1.02–0.89(m,10H),0.86(d,J＝7.3Hz,3H),0.65(s,3H)。
13C NMR(100MHz,CDCl 3)；δ179.13(s),104.09(s),98.78(s),88.20(s),81.19(s),77.24(s),72.49(s),71.73(s),56.21(s),55.93(s),52.64(s),47.88(s),44.45(s),42.88(s),39.99(d),37.55(s),36.51(s),36.00(s),35.39(d),34.60(d),31.64(s),30.24(s),30.36(s),30.44(s),30.75(s), 30.77(s),30.87(s),29.67(s),28.11(s),26.26(s),24.64(d),24.22(s),23.77(s),20.75(s),20.41(s),18.25(s),13.16(s),12.02(s)。
实施例4 化合物JZ-04的制备
Figure PCTCN2018082120-appb-000014
-78℃下，三氟化硼乙醚(1mL)缓慢滴加至双氢青蒿素(569mg，2.0mmol)和鹅去氧胆酸(785mg，2.0mmol)的乙醚溶液(50mL)中，反应体系自然升至室温后搅拌过夜。TLC监测至反应结束后，用饱和碳酸氢钠水溶液(30mL)缓慢淬灭反应，乙酸乙酯萃取(30mL×3)，有机相合并后用水(50mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝10:1-1:1)得无色油状化合物(编号化合物JZ-04)723mg(产率：54.8％)。
1H NMR(400MHz,CDCl 3)；δ5.46(s,1H),4.93(d,J＝3.1Hz,1H),3.88(s,1H),3.48(t,J＝10.8Hz,1H),2.65–2.55(m,1H),2.46–2.33(m,2H),2.27(m,1H),1.87(m,16H),1.52–1.13(m,21H),0.95(m,14H),0.66(s,3H)。
13C NMR(100MHz,CDCl 3)；δ179.74(s),104.02(s),100.40(s),88.09(s),81.30(s),77.49(s),77.25(s),68.70(s),55.79(s),52.63(s),50.53(s),44.59(s),42.73(s),41.33(s),39.56(s),39.33(s),37.47(s),36.51(s),35.86(s),35.30(d),34.80(d),32.88(s),30.86(d),29.12(s),28.18(s),26.29(s),24.63(d),23.70(s),22.90(s),20.64(s),20.41(s),18.27(s),13.20(s),11.78(s)。
实施例5 化合物JZ-05的制备
Figure PCTCN2018082120-appb-000015
-78℃下，三氟化硼乙醚(1mL)缓慢滴加至双氢青蒿素(569mg，2.0mmol)和薯蓣皂苷元(829mg，2.0mmol)的乙醚溶液(50mL)中，反应体系自然升至室温后搅拌过夜。TLC 监测至反应结束后，用饱和碳酸氢钠水溶液(30mL)缓慢淬灭反应，乙酸乙酯萃取(30mL×3)，有机相合并后用水(50mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝40:1-15:1)得白色固体化合物(编号化合物JZ-05)975mg(产率：71.6％)。
1H NMR(400MHz,CDCl 3)；δ5.48(s,1H),5.36(d,J＝4.9Hz,1H),4.94(d,J＝3.4Hz,1H),4.43(dd,J＝15.0,7.5Hz,1H),3.60(dt,J＝9.6,4.7Hz,1H),3.50(dd,J＝10.3,3.5Hz,1H),3.40(t,J＝10.9Hz,1H),2.67–2.57(m,1H),2.44–2.26(m,3H),2.02(m,3H),1.93–1.80(m,5H),1.78–1.72(m,2H),1.71–1.57(m,7H),1.56–1.44(m,8H),1.40–1.09(m,7H),1.06–0.93(m,11H),0.90(d,J＝7.3Hz,3H),0.81(m,6H)。
13C NMR(100MHz,CDCl 3)；δ140.74(s),121.39(s),109.32(s),104.05(s),99.45(s),88.09(s),81.27(s),80.87(s),77.25(s),76.13(s),66.87(s),62.05(s),56.46(s),52.62(s),50.01(s),44.52(s),41.61(s),40.27(s),39.78(s),37.47(s),36.90(d),36.49(s),34.71(s),31.97(d),31.40(d),30.76(s),30.32(s),28.81(s),27.59(s),26.28(s),24.61(d),20.87(s),20.42(s),19.52(s),17.18(s),16.32(s),14.57(s),13.12(s)。
实施例6 化合物JZ-06的制备
Figure PCTCN2018082120-appb-000016
-78℃下，三氟化硼乙醚(1mL)缓慢滴加至双氢青蒿素(569mg，2.0mmol)和麦角甾醇(793mg，2.0mmol)的乙醚溶液(50mL)中，反应体系自然升至室温后搅拌过夜。TLC监测至反应结束后，用饱和碳酸氢钠水溶液(30mL)缓慢淬灭反应，乙酸乙酯萃取(30mL×3)，有机相合并后用水(50mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝30:1-15:1)得白色固体(编号化合物JZ-06)895mg(产率：67.5％)。
1H NMR(400MHz,CDCl 3)；δ5.56(dd,J＝5.6,2.2Hz,1H),5.49(s,1H),5.39(dt,J＝5.3,2.5Hz,1H),5.33–5.14(m,2H),4.97(d,J＝3.4Hz,1H),3.70(ddd,J＝11.2,7.8,4.4Hz,1H),2.69–2.58(m,1H),2.56–2.46(m,1H),2.38(m,2H),2.11–2.01(m,3H),2.00–1.82(m,7H), 1.79–1.65(m,4H),1.65–1.56(m,2H),1.53–1.45(m,5H),1.42–1.21(m,10H),1.04(d,J＝6.7Hz,3H),0.89-0.98(m,12H),0.85(t,J＝6.4Hz,6H),0.64(s,3H)。
13C NMR(100MHz,CDCl 3)：δ140.99(s),139.97(s),135.63(s),131.93(s),119.47(s),116.47(s),104.05(s),99.60(s),88.10(s),81.24(s),77.24(s),75.07(s),55.69(s),54.50(s),52.63(s),46.05(s),44.52(s),42.81(d),40.50(s),39.07(s),38.81(s),37.97(s),37.48(s),37.26(s),36.49(s),34.72(s),33.10(s),30.77(s),29.72(s),28.35(s),27.98(s),26.25(s),24.52(s),24.70(s)23.03(s),21.09(d),20.40(s),19.98(s),19.67(s),17.62(s),16.21(s),13.11(s),12.07(s)。
实施例7 化合物JZ-07的制备
Figure PCTCN2018082120-appb-000017
-78℃下，三氟化硼乙醚(1mL)缓慢滴加至双氢青蒿素(569mg，2.0mmol)和石胆酸(753mg，2.0mmol)的乙醚溶液(50mL)中，反应体系自然升至室温后搅拌过夜。TLC监测至反应结束后，用饱和碳酸氢钠水溶液(30mL)缓慢淬灭反应，乙酸乙酯萃取(30mL×3)，有机相合并后用水(50mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝12:1-2:1)得无色油状化合物(编号化合物JZ-07)843mg(产率：65.5％)。
1H NMR(400MHz,CDCl 3)：δ5.47(s,1H),4.92(d,J＝3.3Hz,1H),3.73–3.53(m,1H),2.66–2.56(m,1H),2.39(m,2H),2.27(ddd,J＝15.8,9.7,6.4Hz,1H),2.05(dt,J＝14.5,3.7Hz,1H),1.99–1.94(m,1H),1.93–1.70(m,8H),1.68–1.55(m,3H),1.51–1.21(m,21H),1.08(m,4H),1.00–0.89(m,13H),0.66(s,3H)。
13C NMR(100MHz,CDCl 3)δ180.33(s),104.04(s),100.16(s),88.07(s),81.26(s),76.26(s),56.42(s),55.98(s),52.63(s),44.57(s),42.77(s),41.97(s),40.42(s),40.12(s),37.48(s),36.50(s),35.85(s),35.32(s),35.42(s),34.74(s),32.59(s),31.00(s),30.85(s),30.78(s),29.72(s),29.01(s),28.20(s),27.33(s),26.40(s),26.27(s),24.63(d),24.21(s),23.51(s),20.89(s),20.42(s),18.30(s),13.18(s),12.07(s)。
实施例8 化合物JZ-08的制备
Figure PCTCN2018082120-appb-000018
-78℃下，三氟化硼乙醚(1mL)缓慢滴加至双氢青蒿素(569mg，2.0mmol)和脱氧胆酸(785mg，2.0mmol)的乙醚溶液(50mL)中，反应体系自然升至室温后搅拌过夜。TLC监测至反应结束后，用饱和碳酸氢钠水溶液(30mL)缓慢淬灭反应，乙酸乙酯萃取(30mL×3)，有机相合并后用水(50mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝12:1-3:1)得无色油状化合物(编号化合物JZ-08)723mg(产率：54.8％)。
1H NMR(400MHz,CDCl 3)：δ5.47(s,1H),4.91(d,J＝3.4Hz,1H),4.02(s,1H),3.67–3.55(m,1H),2.65–2.56(m,1H),2.48–2.25(m,3H),2.07–2.01(m,1H),1.92–1.81(m,5H),1.77–1.70(m,3H),1.68–1.58(m,4H),1.54(m,2H),1.50–1.34(m,12H),1.27(m,8H),1.18–1.06(m,2H),1.01(d,J＝6.2Hz,3H),0.96(d,J＝6.3Hz,3H),0.93–0.85(m,7H),0.71(s,3H)。
13C NMR(100MHz,CDCl 3)：δ179.27(s),104.03(s),100.49(s),88.10(s),81.30(s),77.24(s),73.32(s),52.62(s),48.36(s),47.33(s),46.46(s),44.55(s),42.00(s),37.43(s),36.51(s),36.06(s),35.26(s),35.04(s),34.66(s),34.30(s),33.79(s),32.64(s),31.05–30.54(m),29.86–29.32(m),28.98(s),27.35(d),26.29(s),26.06(s),24.64(d),23.62(s),23.34(s),20.38(s),17.33(s),13.20(s),12.77(s)。
实施例9 溴乙烷双氢青蒿素醚的制备
0℃下，三氟化硼乙醚(50μL)缓慢滴加至双氢青蒿素(284mg，1.0mmol)和2-溴乙醇(125mg，1.0mmol)的乙醚溶液(20mL)中，反应体系在0℃下搅拌过夜。TLC监测至反应结束后，用饱和碳酸氢钠水溶液(10mL)缓慢淬灭反应，乙酸乙酯萃取(10mL×3)，有机相合并后用水(20mL)洗涤1次，饱和食盐水(20mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝30:1～8:1)得白色固体目标产物203mg(产率：51.9％)。
1H NMR(400MHz,CDCl3)δ5.48(s,1H),4.84(d,J＝3.3Hz,1H),4.18–4.07(m,1H),3.81–3.74(m,1H),3.51(t,J＝5.2Hz,2H),2.69–2.59(m,1H),2.40–2.33(m,1H),2.06–1.99(m, 1H),1.92–1.84(m,2H),1.76(m,1H),1.68–1.61(m,1H),1.55–1.43(m,5H),1.37–1.21(m,3H),0.95–0.90(m,6H).
13C NMR(101MHz,CDCl3)δ104.10(s),102.02(s),88.13(s),81.08(s),77.42(s),77.11(s),76.79(s),68.16(s),52.56(s),44.35(s),37.38(s),36.39(s),34.66(s),31.47(s),30.89(s),26.16(s),24.65(s),24.36(s),20.39(s),13.00(s).
实施例10 化合物JZ-09的制备
Figure PCTCN2018082120-appb-000019
室温下，碳酸钾(276mg，2.0mmol)和催化量的碘化钾分别加入至溴乙烷双氢青蒿素醚(391mg，1.0mmol)和熊去氧胆酸(392mg，1.0mmol)的DMF(20mL)溶液中，该反应液升至70℃反应2小时。TLC监测至反应结束后，用水(20mL)缓慢淬灭反应，乙酸乙酯萃取(20mL×3)，有机相合并后用水(30mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝4:1-1:1)得无色油状化合物(编号化合物JZ-09)536mg(产率：76.2％)。
1H NMR(400MHz,CDCl 3)：δ5.45(s,1H),4.84(d,J＝3.4Hz,1H),4.28–4.21(m,2H),4.08–3.98(m,1H),3.68–3.57(m,3H),2.72–2.58(m,1H),2.43–2.34(m,2H),2.28–2.19(m,1H),2.04(m,2H),1.91(m,2H),1.85–1.77(m,5H),1.71–1.60(m,5H),1.53–1.44(m,11H),1.39–1.25(m,12H),1.19–1.03(m,3H),0.94(m,12H),0.69(s,3H)。
13C NMR(100MHz,CDCl 3)δ174.08(s),104.13(s),102.08(s),87.91(s),81.09(s),77.25(s),71.44(d),66.00(s),63.38(s),55.72(s),54.85(s),52.55(s),44.40(s),43.78(d),42.43(s),40.14(s),39.17(s),37.58(s),37.30(s),36.85(s),36.43(s),35.28(s),34.93(s),34.67(s),34.09(s),31.20(s),30.91(d),30.35(s),29.72(s),28.63(s),26.91(s),26.21(s),24.75(s),24.41(s),23.40(s),21.18(s),20.43(s),18.40(s),12.96(s),12.15(s)。
实施例11 化合物JZ-10的制备
Figure PCTCN2018082120-appb-000020
室温下，碳酸钾(276mg，2.0mmol)和催化量的碘化钾分别加入至溴乙烷双氢青蒿素醚(391mg，1.0mmol)和猪去氧胆酸(392mg，1.0mmol)的DMF(20mL)溶液中，该反应液升至70℃反应2小时。TLC监测至反应结束后，用水(20mL)缓慢淬灭反应，乙酸乙酯萃取(20mL×3)，有机相合并后用水(30mL)洗涤1次，饱和食盐水(50mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过硅胶柱纯化(石油醚：乙酸乙酯＝4:1-1:1)得无色油状化合物(编号化合物JZ-10)589mg(产率：83.7％)。
1H NMR(400MHz,CDCl 3)：δ5.45(s,1H),4.84(d,J＝3.4Hz,1H),4.28–4.22(m,2H),4.10–4.00(m,2H),3.67–3.62m,2H),2.71–2.58(m,1H),2.43–2.35(m,2H),2.22(s,1H),2.07(m,1H),1.98–1.87(m,4H),1.82–1.74(m,4H),1.67–1.59(m,8H),1.46–1.28(m,15H),1.17–1.08(m,6H),0.98–0.89(m,12H),0.65(s,3H)。
13C NMR(100MHz,CDCl 3)：δ174.10(s),104.12(s),102.06(s),87.90(s),81.09(s),77.26(s),71.60(s),68.07(s),66.00(s),63.38(s),56.14(s),55.86(s),52.55(s),48.39(s),44.40(s),42.85(s),39.95(s),39.80(s),37.58(s),36.43(s),35.96(s),35.35(s),35.55(s),35.03(s),34.83(s),34.66(s),31.19(s),30.92(s),30.83(s),30.24(s),29.20(s),28.13(s),26.20(s),24.74(s),24.41(s),24.21(s),23.50(s),20.75(s),20.42(s),18.27(s),12.95(s),12.05(s)。
实施例12 化合物JZ-11的制备
Figure PCTCN2018082120-appb-000021
0℃下，三氟化硼乙醚复合物(180mg)加入至熊去氧胆酸双氢青蒿素醚(化合物JZ-02，130mg，0.18mmol)的吡啶(2mL)溶液中，反应体系在室温下反应6小时，TLC监测至反应结束后，用1N稀盐酸(5mL)缓慢淬灭反应，二氯甲烷萃取(10mL×3)，有机相合并后用1N稀盐酸(20mL)洗涤2次，饱和食盐水(30mL)洗涤1次，无水硫酸镁干燥，过滤， 滤液减压旋干即得粗产物，粗产物过柱纯化(二氯甲烷：甲醇＝20:1～10:1)得白色固体目标产物(编号化合物JZ-11)103mg(产率：70.6％)。
1H NMR(400MHz,DMSO)δ11.97(s,1H),5.34(s,1H),4.80(d,J＝3.3Hz,1H),3.94(s,1H),3.39(s,1H),2.42–2.30(m,1H),2.29–2.04(m,5H),2.01–1.89(m,2H),1.84–1.54(m,10H),1.38(m,8H),1.27(m,7H),1.21–1.10(m,6H),1.00(m,2H),0.92–0.81(m,12H),0.61(s,3H).
13C NMR(100MHz,DMSO)δ175.89(s),103.71(s),100.13(s),87.51(s),80.98(s),77.27(s),76.33(s),55.29(s),55.08(s),43.66(s),41.43(s),35.28(s),35.01(s),34.75(s),34.60(s),34.15(s),31.76(s),31.62(s),31.44(s),31.27(s),30.94(s),30.28(s),29.49(s),28.97(s),26.13(s),25.87(s),24.67(d,J＝13.2Hz),23.74(s),20.65(s),18.78(s),13.40(s),12.42(s).
实施例13 化合物JZ-12的制备
Figure PCTCN2018082120-appb-000022
0℃下，三氟化硼乙醚复合物(180mg)加入至脱氧胆酸双氢青蒿素醚(化合物JZ-08，130mg，0.18mmol)的吡啶(2mL)溶液中，反应体系在室温下反应6小时，TLC监测至反应结束后，用1N稀盐酸(5mL)缓慢淬灭反应，二氯甲烷萃取(10mL×3)，有机相合并后用1N稀盐酸(20mL)洗涤2次，饱和食盐水(30mL)洗涤1次，无水硫酸镁干燥，过滤，滤液减压旋干即得粗产物，粗产物过柱纯化(二氯甲烷：甲醇＝20:1～12:1)得白色固体目标产物(编号化合物JZ-12)112mg(产率：76.8％)。
1H NMR(400MHz,DMSO)δ11.75(s,1H),5.33(s,1H),4.79(d,J＝3.3Hz,1H),4.34(s,1H),3.46(d,J＝10.7Hz,1H),2.37(m,1H),2.27–2.11(m,3H),2.08–1.92(m,2H),1.83–1.57(m,10H),1.53–1.43(m,5H),1.40–0.97(m,18H),0.96–0.78(m,14H),0.64(s,3H).
13C NMR(100MHz,DMSO)δ177.08(s),103.70(s),98.99(s),87.45(s),81.01(s),78.29(s),76.61(s),52.56(s),48.62(s),46.45(s),46.07(s),44.41(s),42.18(s),37.21(s),36.54(s),36.03(s),35.70(s),34.68(s),34.50(s),33.89(s),32.51(s),31.61(s),30.89(s),28.68(s),27.70(s),26.46(s),26.12(s),25.12(s),24.73(s),24.02(s),23.79(s),20.63(s),17.77(s),13.41(s),12.73(s).
实施例14 双氢青蒿素-甾体结合物对体外培养的各种肿瘤细胞的抑制作用
1.实验原理
采用的肿瘤细胞为：人前列腺癌细胞-PC-3细胞(CRL-1435)、人宫颈癌细胞系-HeLa细胞(CCL-2)、人急性T细胞白血病细胞-Jurkat细胞(TIB-152)、人肾透明细胞腺癌-786-O细胞(CRL-1932)和人乳腺癌细胞-MCF7细胞(HTB-22)，检测本发明上述化合物样品对这些肿瘤细胞株生长的影响作用；以非洲绿猴肾细胞-Vero细胞为正常细胞(CCL-81)，检测化合物样品的细胞毒性作用。在化合物样品干预下，活细胞可利用细胞增殖-毒性检测试剂盒Cell Counting Kit-8(CCK-8溶液)进行染色，通过测定其OD值来反映细胞的存活和生长状态，即细胞存活率(％)和细胞生长抑制(％)。并计算出半数细胞存活时的化合物浓度CC 50，以及抑制细胞生长为半数时的化合物浓度IC 50。
2.实验材料
Vero细胞(CCL-81)、PC-3细胞(CRL-1435)、HeLa细胞(CCL-2)、Jurkat细胞(TIB-152)、786-O细胞(CRL-1932)、MCF7细胞(HTB-22)均购自ATCC；DMEM培养液、F12培养液、MEM培养液、RPMI 1640培养液均购自Gibco公司(Life Technologies)；胎牛血清(FBS)购自Hyclone公司(Thermo Scientific)；胰酶购自Gibco公司(Life Technologies)；CCK-8溶液购自上海东仁化学科技公司；96孔细胞培养板(透明平底)购自Corning公司；SpectraMax 190microplate reader酶标仪购自Molecular Devices Corporation。
3.实验方法
含10％FBS的培养液为完全培养液。Vero细胞用DMEM完全培养液进行培养；PC-3细胞用F12完全培养液进行培养；Hela细胞用MEM完全培养液进行培养；Jurkat细胞用RPMI1640完全培养液进行培养；786-O细胞用RPMI 1640完全培养液进行培养；MCF-7细胞用MEM完全培养液进行培养。所有细胞培养成对数生长后，进行以下试验。
贴壁细胞以胰酶消化收集细胞；悬浮细胞直接收集。细胞离心，计数后，用各自完全培养液将细胞调制至5*103个/100μl/孔，接种于透明96孔培养板中；同时每块培养液板中都设置不加细胞、不加样品的空白孔；以及加细胞、不加样品的对照孔。接种后的96孔培养板在37℃，5％CO 2培养箱中培养过夜。次日将加入测试化合物样品50μl/孔，并加入完全培养液50μl/孔。化合物样品最终试验浓度为100μM、25μM、6.25μM、1.56μM、0.39μM、0.1μM、0.02μM、0.006μM，各个试验浓度为双复孔；对照孔和空白孔用完全培养液替代化合物样品，即加入完全培养液100μl/孔。37℃，5％CO 2培养箱中继续培养48小时。在结束培养前2小时，去培养液，加入含10％CCK-8溶液的完全培养液100μl/孔，至培养结束，酶标仪于450nM(参比650nM)处测定OD值。
根据检测Vero细胞存活率(％)，计算出半数细胞死亡浓度CC 50，反应出化合物样品的毒性作用；检测肿瘤细胞株的细胞生长抑制(％)，计算出细胞生长数抑制在半数时的浓度IC 50，反应出化合物样品的抗肿瘤活性。
计算公式如下：
细胞存活率(％)＝(化合物样品孔的OD值-空白孔的OD值)/(对照孔的OD值-空白孔的OD值)×100％
细胞生长抑制(％)＝[1-(化合物样品孔的OD值-空白孔的OD值)/(对照孔的OD值-空白孔的OD值)]×100％
4.实验结果
IC 50结果见表1和表2，结果显示，双氢青蒿素-甾体结合物对各类肿瘤细胞生长均具有不同程度的抑制，这一系列化合物对受试肿瘤细胞的抑制率均远远强于双氢青蒿素。个别化合物对受试肿瘤细胞的抑制率与紫杉醇相近，而毒性却远远小于紫杉醇。
表1 双氢青蒿素-甾体结合物对各类肿瘤细胞的抑制作用
Figure PCTCN2018082120-appb-000023
表2 化合物JZ-2和JZ-8对各类肿瘤细胞的抑制作用
Figure PCTCN2018082120-appb-000024
实施例15 JZ-2在人源肺癌细胞A549裸鼠移植模型上的抗癌作用
实验用2-4周龄的裸鼠(18-22g)23只，每只裸鼠皮下注射人非小细胞肺癌细胞A549(1-5x10 6),待肿瘤长至1-2cm 3(约40天),将23只裸鼠随机分为三组，组一(9只)为对照组，组二(9只)为治疗组，组三(5只)为紫杉醇阳性对照组。组一每天腹腔注射生理盐水20ul,组二每天腹腔注射给药JZ-02 10mg/kg，每七天给药休息2天。组三每三天给药一次，腹腔注射紫杉醇10mg/kg。整个治疗过程持续了28天。其中组一死亡三只，组三死亡一只。组二无动物死亡，组一小鼠的死亡是由于肿瘤无控制的生长所造成，组二无一死亡，说明该组的肿瘤得到了较好的控制，同时JZ-02的毒性小于紫杉醇。
图1所示的肿瘤生长曲线表明，本发明化合物JZ-2能明显抑制肿瘤生长，抑制效果比阳性对照紫杉醇好。
图2所示的体重曲线表明，JZ-2连续给药28天后，小鼠的体重没有显著下降。证明该类化合物毒性较小，对小鼠的体重和生活质量没有显著影响。
将各组动物处死后，对肿瘤组织进行称重，计算抑瘤率，得出组二(治疗组)的抑瘤率为58％。阳性对照组的抑瘤率为52％。
以上所述实施例仅表达了本发明的实施方式，其描述较为具体和详细，但并不能因此而理解为对本发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。

Claims (16)

  1. 具有通式(I)的双氢青蒿素-甾体结合物，或其异构体、或其可药用盐、或其前药分子，
    Figure PCTCN2018082120-appb-100001
    其中，双氢青蒿素分子在其10位通过连接体X与甾体母核相连接；
    连接体X为具有连接两端分子功能的原子或基团，包括CH 2，O，S，N，-CO 2-，-SO 2-，2-10个碳原子的二醇或三醇，苯二酚或苯三酚衍生物，氨基酚衍生物，氨基苯甲酸衍生物，羟基苯甲酸衍生物，多取代的含有2个以上烷基酸取代的苯环或杂环，2-10个碳原子的二酸或多元酸，2-10个碳原子的二巯基化合物，2-10个碳原子的二胺，2-10个碳原子的羟基酸，3-10个碳原子的氨基酸或氨基醇；X可以连接在甾体的2，3，4，6，7，11，12，16或17位上。
    R 1为H，OH，NH 2，NR’ 2(R’为1-10个碳原子的烷烃)，卤素，羧基，酰胺基，巯基，磺酸酯及其盐，亚磺酸酯及其盐，4-位或5-位或6-位的双键，1-10个碳原子的烷，环烷，卤代烷，多卤代烷或卤代环烷烃，含有或不含有卤素的烯基或炔烃，烷胺基或者烷基氨基。5-15个碳原子的含有环烷烃取代或含有芳香环取代或含有杂环取代的烷烃，卤代烷烃，含有或不含有卤素的烯烃或炔烃。1-10个碳原子的烷氧基，2-10个碳原子的卤代烷氧基，1-10个碳原子的烷氧基醇或烷氧基多元醇，1-10个碳原子的烷氧基醇或烷氧基多元醇的磺酸酯及其盐，1-10个碳原子的烷氧基醚，1-10个碳原子的烷氧基羧酸或烷氧基多元羧酸及其盐，1-10个碳原子的烷氧基磺酸及其盐，1-10个碳原子的烷氧基胺，1-10个碳原子的烷氧基酰胺，含2-10个碳原子的带有或不带有卤素的氧基烯烃，含2-10个碳原子的带有或不带有卤素的氧基炔烃，3-8个碳的环烷基氧基，酮基，亚胺，1-10个碳原子的烷基亚胺，1-10个碳原子的烷氧基亚胺，羟胺，羟胺磺酸酯及其盐，酰胺基，带有或不带有取代基的苯氧基，带有或不带有取代基的苄氧基，带有或不带有取代基的萘氧基，1-10个碳原子的二烷基胺基，含有1-4个杂原 子的五元-八元的杂环或苯并杂环，或3-6个碳的单糖；R 1取代的位置可以在甾体的3-位，4-位，6位或者在7位。
    R 2为H，OH，NH 2，NR’ 2(R’为1-10个碳原子的烷烃)，卤素，巯基，羧基，酮基，磺酸酯及其盐，亚磺酸酯及其盐，酰胺基，11-位双键，1-10个碳原子的烷烃，环烷烃，卤代烷，多卤代烷或卤代环烷，含有或不含有卤素的烯烃或炔烃，烷胺基或者烷基氨基。5-15个碳原子的含有环烷烃取代或含有芳香环取代或含有杂环取代的烷烃，卤代烷烃，含有或不含有卤素的烯烃或炔烃。1-10个碳原子的烷氧基，2-10个碳原子的卤代烷氧基，1-10个碳原子的烷氧基醇或烷氧基多元醇，1-10个碳原子的烷氧基醇或烷氧基多元醇的磺酸酯及其盐，1-10个碳原子的烷氧基醚，1-10个碳原子的烷氧基羧酸或烷氧基多元羧酸及其盐，1-10个碳原子的烷氧基磺酸及其盐，1-10个碳原子的烷氧基胺，1-10个碳原子的烷氧基酰胺，含2-10个碳原子的带有或不带有卤素的氧基烯烃，含2-10个碳原子的带有或不带有卤素的氧基炔烃，3-8个碳的带有或不带有卤素的环烷基氧基，酮基，亚胺，1-10个碳原子的烷基亚胺，1-10个碳原子的烷氧基亚胺，羟胺，羟胺磺酸酯及其盐，带有或不带有取代基的苯氧基，带有或不带有取代基的苄氧基，带有或不带有取代基的萘氧基，1-10个碳原子的二烷基胺基，含有1-4个杂原子的五元-八元的杂环或苯并杂环，或3-6个碳的单糖；R 1取代的位置可以在甾体的11-位或12-位。
    R 3为H，OH，NH 2，NR’ 2(R’为1-10个碳原子的烷烃)，卤素，巯基，磺酸酯及其盐，亚磺酸酯及其盐，酰胺基，1-12个碳原子的烷基，2-12个碳原子的卤代烷基或多卤代烷基，1-12个碳原子的醇或多元醇，1-12个碳原子的醇或多元醇的磺酸酯及其盐，1-12个碳原子的醚或多元醇的单醚，1-12个碳原子的羧酸或多元羧酸及其盐，1-12个碳原子的烷基磺酸及其盐，1-12个碳原子的胺，1-12个碳原子的酰胺，含2-12个碳原子的带有或不带有卤素的烯烃，含2-12个碳原子的带有或不带有卤素的炔烃，3-8个碳的环烷，1-10个碳原子的酮，1-12个碳原子的烷基亚胺，1-12个碳原子的烷氧基亚胺，羟胺，羟胺磺酸酯及其盐，1-12个碳原子的烷基磺酸酯及其盐，1-12个碳原子的烷基亚磺酸酯，1-12个碳原子的烷基酰胺，带有或不带有取代基的苯基或苯氧基，带有或不带有取代基的苄基，带有或不带有取代基的萘基，1-10个碳原子的二烷基胺基，含有1-4个杂原子的五元-八元的杂环或苯并杂环，3-6个碳的单糖，胆酸的C-17侧链及其衍生基团，植物甾醇的C-17侧链或其衍生基团，激素类甾体的C-17侧链，与16位羟基成为螺环的侧链，或甾体生物碱的C-17侧链以及上述侧链的衍生物。
    式(I)中，
    Figure PCTCN2018082120-appb-100002
    代表α-取代，
    Figure PCTCN2018082120-appb-100003
    代表β-取代。
  2. 根据权利要求1所述的双氢青蒿素-甾体结合物，其特征在于，所述连接体X包括O，S，N，-CH 2-，-NR-，-BH-，-BR-，-P(O)-，-NH-NH-，-OSO 2-，-O-CO-O-，-OCO-(CH 2)n-COO-， -O(CH 2)nO-，-O(CH 2OCH 2)nO-，-O-SO 2-O-，-O-PO 2-O-，-O-BO-O-，-(CH 2)n-O-，-O-Ph-O-，-O(CH 2)nNR(CH 2)mO-，-OOC(CH 2)nNR(CH 2)m-COO-，-O(CH 2) 2N(CH 2) 2N(CH 2) 2O-，
    Figure PCTCN2018082120-appb-100004
    -OPh(Y)-O-，-O-Ph(Y)-COO-，-OOC-(CH 2)nPh(Y)(CH 2)mCOO-，-OOCPh(Y)COO-，-OCH 2CH 2NHPh(Y)NHCH 2CH 2O-，含有1-4个杂原子的五元或六元杂环及其衍生物；
    其中R为H或1-10个碳原子的饱和或不饱和烃；Y为OR’，NHR’，NR’ 2，卤素，-CF 3，-COOH，-COOR’，-CONH 2，-CONR’ 2，-CN，-NO 2，-SO 3-，或-SO 2R’，Y可在苯环不同的位置上取代；R’为H或1-10个碳原子的饱和或不饱和的烃基；n和m分别为0-10。
  3. 根据权利要求2所述的双氢青蒿素-甾体结合物，其特征在于，所述连接体X为氧，-CH 2O-或-O-CH 2CH 2-O-。
  4. 根据权利要求1所述的双氢青蒿素-甾体结合物，其特征在于，所述连接体X一端连接在双氢青蒿素的10位，另一端连接在甾体母核的2位，3位，5位，6位，7位，11位，12位，15位，16位，17位，或者连接在甾体的侧链上。
  5. 根据权利要求4所述的双氢青蒿素-甾体结合物，其特征在于，所述连接体X的另一端连接在甾体母核的3位，6位，或者7位。
  6. 根据权利要求1所述的双氢青蒿素-甾体结合物，其特征在于，所述甾体母核为具有环戊烷骈多氢菲的结构单元，其包括天然和非天然的甾体及其衍生物，该甾体及其衍生物包含甾烷，雄甾烷，雌甾烷，孕甾烷，胆烷，麦角甾烷，豆甾烷及异甾体的结构类型。
  7. 根据权利要求6所述的双氢青蒿素-甾体结合物，其特征在于，所述天然和非天然的甾体及其衍生物包含胆酸，去氧胆酸，猪去氧胆酸，熊去氧胆酸，鹅去氧胆酸，奥贝胆酸及其类似物或衍生物，胆甾醇，麦角甾醇，大豆甾醇，5α或5β谷甾醇及其衍生物，甾体激素及其衍生物，糖皮质激素及其衍生物，盐皮质激素及其衍生物，甾体皂甙，甾体皂甙元，甾体生物碱。
  8. 根据权利要求1所述的双氢青蒿素-甾体结合物，其特征在于，所述式(I)结合物包括：
    Figure PCTCN2018082120-appb-100005
    Figure PCTCN2018082120-appb-100006
    Figure PCTCN2018082120-appb-100007
    Figure PCTCN2018082120-appb-100008
  9. 一种药物组合物，其特征在于，包含一种或几种权利要求1所述双氢青蒿素-甾体结合物或其异构体、或其可药用盐、或其前药分子，以及药学上可接受的载体。
  10. 权利要求1所述双氢青蒿素-甾体结合物的制备方法，其特征在于，包括以下步骤：
    双氢青蒿素在其10位以醚键和甾体上的2位，3位，5位，6位，7位，11位，12位，15位，16位，17位，或甾体侧链羟基连接，得双氢青蒿素-甾体结合物。
  11. 权利要求1所述双氢青蒿素-甾体结合物的制备方法，其特征在于，包括以下步骤：
    双氢青蒿素或其衍生物先和连接体X通过酯键，醚键，酰胺键，碳-碳单键或者碳-氮、碳-硫单键连接成为中间体，该中间体再通过共价键与甾体的2位，3位，5位，6位，7位，11位，12位，15位，16位，17位，或甾体侧链连接，形成双氢青蒿素-甾体结合物。
  12. 权利要求1-8中任一项所述双氢青蒿素-甾体结合物或其异构体、或其可药用盐、或 其前药分子，或权利要求9所述的药物组合物在制备治疗癌症的药物中的应用。
  13. 根据权利要求12所述的应用，其特征在于，所述癌症包括脑癌、脑胶质瘤、子宫内膜癌、卵巢癌、宫颈癌、乳腺癌、结肠癌、肺癌、前列腺癌、肝癌、白血病、淋巴癌、皮肤癌、基底细胞瘤、血管瘤、子宫癌、喉癌、胃癌、唇癌、食道癌、鼻咽癌、胆囊癌、胰腺癌、肾癌、舌癌、膀胱癌、黑素瘤、脂肪瘤、甲状腺癌、胸腺癌、骨癌。
  14. 权利要求1-8中任一项所述双氢青蒿素-甾体结合物或其异构体、或其可药用盐、或其前药分子，与至少一种另外的抗癌剂或氨基酸类化合物联用在制备治疗癌症的药物中的应用。
  15. 根据权利要求14所述的应用，其特征在于，所述另外的抗癌剂包括阿霉素类、博莱霉素、长春碱类、紫杉烷类、依托泊苷、5-氟尿嘧啶、环磷酰胺、甲氨蝶呤、顺铂、维甲酸、替莫唑胺、放线菌素、伊马替尼、吉非替尼、索拉非尼、厄洛替尼、舒尼替尼、利妥昔单抗、西妥昔单抗、曲妥珠单抗、尼伏单抗、潘利珠单抗、阿替珠单抗、度伐单抗、和/或阿维单抗。
  16. 权利要求1-8中任一项所述双氢青蒿素-甾体结合物或其异构体、或其可药用盐、或其前药分子，与血红素生物合成前体联用在制备治疗癌症的药物中的应用。
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