WO2016138882A1 - 七元环小檗碱类似物及其药物组合物、制备方法和用途 - Google Patents

七元环小檗碱类似物及其药物组合物、制备方法和用途 Download PDF

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WO2016138882A1
WO2016138882A1 PCT/CN2016/076083 CN2016076083W WO2016138882A1 WO 2016138882 A1 WO2016138882 A1 WO 2016138882A1 CN 2016076083 W CN2016076083 W CN 2016076083W WO 2016138882 A1 WO2016138882 A1 WO 2016138882A1
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group
aryl
ring
alkyl
hydrogen
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PCT/CN2016/076083
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French (fr)
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朱维良
缪泽鸿
李波
宋姗姗
徐志建
张勇
蔡婷婷
陈凯先
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中国科学院上海药物研究所
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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
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D223/00—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom
    • C07D223/14—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
    • C07D223/16—Benzazepines; Hydrogenated benzazepines

Definitions

  • the present invention relates to the field of medicinal chemistry and pharmacotherapeutics, and in particular to a seven-membered cyclic berberine analog which can be used as a therapeutic tumor, a pharmaceutical composition thereof, a preparation method and a medical use.
  • the traditional Chinese medicine berberine mainly includes the dried rhizomes of Ranunculaceae, Coptis chinensis Franch, Coptis deltoidea C.Y.cheng et Hsiao, and Coptis teetaw all.
  • berberine is also found in the plants of the family Polygonaceae, Papaveraceae, Rutaceae, Anti-family, and Rhamnaceae.
  • studies have found that berberine and its derivatives are rich in pharmacological effects, and have anti-tumor, diabetes, cardiovascular disease, hypertension, inflammation, bacterial and viral infections, cerebral ischemic injury, mental illness, Alzheimer. Alzheimer disease, osteoporosis and other activities, the potential development and application prospects are very broad.
  • berberine Malignant tumors are one of the diseases that endanger human health and are among the highest in the mortality rate of various diseases. Pharmacological experiments show that berberine has obvious growth inhibition effects on lung cancer cells, gastric cancer cells, liver cancer cells, colon cancer cells, breast cancer cells, cervical cancer cells, prostate cancer cells, melanoma cells and leukemia cells. Its anti-tumor mechanism includes inhibition of tumor cell proliferation, promotion of tumor cell apoptosis, inhibition of tumor metastasis, and induction of tumor cell differentiation. The related molecular mechanisms are very complex and involve a wide range of signaling pathways. Therefore, the anti-tumor activity of berberine has the characteristics of multiple effects and multiple targets.
  • berberine is the key to this problem.
  • the parent can be regarded as a four-membered ring system composed of two basic units of isoquinoline. The whole system is in an approximately planar state.
  • the Shanghai Institute of Materia Medica, Chinese Academy of Sciences has used the drug design and total synthesis method to design and modify the B-ring of berberine for the first time, and obtained a series of novel berberine analogues, which significantly improved the solubility and Antitumor activity in vitro.
  • the compound sm38 in which the B ring is an unsaturated seven-membered ring has an inhibition rate of more than 50% on most test tumor cells (including colon cancer and melanoma cells) at a concentration of 10.0 uM.
  • a seven-membered cyclic berberine analog of the formula A a pharmaceutically acceptable salt thereof, a crystalline form thereof, a hydrate thereof, or a solvate thereof, or a prodrug thereof,
  • R 1 is located 1 to 4 (preferably 1-3 or 1-2) substituents the same or different, and R 2 is located 1-4 (preferably 1-3 or 1-2) identical or different substituents, and each of R 1 and R 2 is independently selected from the group consisting of hydrogen, hydroxy, amino, thiol, alkyl , aryl, alkoxy, alkylamino, alkylthio, aryl ether, arylamino, arylthio or halogen; or two R 1 may be adjacent to a carbon atom or two R 2 may be adjacent The carbon atoms together form a 5-7 membered ring containing 0-3 heteroatoms selected from O, S, N (eg, two R 1 and Or two R 2 and Together to form structures such as strontium and azapine;
  • R 3 is selected from the group consisting of hydrogen, oxygen, sulfur, hydroxy, amino, decyl, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylamino, alkylthio, aryl ether, arylamino , arylthio, aralkyl, C1-C6 aldehyde or halogen;
  • R 4 is selected from the group consisting of hydrogen, oxygen, sulfur, hydroxy, amino, decyl, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylamino, alkylthio, aryl ether, arylamino An arylthio group, an aralkyl group, a C1-C6 aldehyde group or a halogen.
  • R2 when R4 represents hydrogen and R3 represents hydrogen or oxygen, and D is a benzene ring, R2 is not hydrogen.
  • R 4 when R 4 is hydrogen and R 3 is hydrogen or oxygen, The ring is not a benzene ring; or when R 4 is hydrogen, R 3 is hydrogen or oxygen and When the ring is a benzene ring, R 2 is not hydrogen.
  • said R 1 or R 2 are each independently selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkynyl, hydroxy, substituted Or an unsubstituted alkoxy group, a substituted or unsubstituted -(CH 2 ) m -O-(CH 2 ) n -Ra, a substituted or unsubstituted -O-(CH 2 ) m -Ra; wherein Ra is C1-C3 alkyl, C1-C3 alkoxy or -(CH 2 ) m -CO-Rb, and Rb is hydroxy, C1-C6 alkoxy, m and n are each an integer of 0-3;
  • Rc is H, C1-C3 alkyl or C1-C3 haloalkyl.
  • substituted means that one or more of the groups H is substituted with a substituent selected from the group consisting of halogen, C1-C4 alkyl, C1-C4 haloalkyl, C2-C4 olefin.
  • a substituent selected from the group consisting of halogen, C1-C4 alkyl, C1-C4 haloalkyl, C2-C4 olefin.
  • the R 4 is -R 8 -R 9 or -R 8 -CO-R 10 , wherein R 8 is a divalent linking group, and R 9 is selected from the group consisting of C1-C8 esters.
  • Base C1-C8 carboxyl group, -O-C1-C6 alkyl group, -O-C2-C6 alkenyl group, hydroxyl group,
  • R 10 is selected from the group consisting of H, hydroxyl,
  • Rc is H, C1-C3 alkyl or C1-C3 haloalkyl.
  • the compound has the structure of formula A1:
  • Q and W each independently represent one or more of C, N, O or S atoms, and when each is independently N, the position and number of each in the ring may vary; when each independently contains O Or S, the carbon atoms in the ring are reduced, maintaining a five-membered aromatic ring system; Q or W may be bonded to each other or dispersed in different positions on the ring;
  • the position and number can be changed means that the position where Q or W appears on the respective ring is not limited; the number of Q or W may be 1-3. When the number is greater than or equal to ( ⁇ ) 2, in order to maintain the six-membered ring system, the number of carbon atoms on the corresponding ring is decreased;
  • R 1 and R 2 each independently represent hydrogen, hydroxy, amino, thiol, alkyl, aryl, alkoxy, alkylamino, alkylthio, aryl ether, arylamino, arylthio or halogen; or two R 1 may be bonded to an adjacent carbon atom or two R 2 together with an adjacent carbon atom to form a 5-7 membered ring containing 0-3 hetero atoms selected from O, S, N;
  • R 3 and R 4 each independently represent hydrogen, oxygen, sulfur, hydroxy, amino, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylamino, alkylthio, aryl ether, An arylamino group, an arylthio group, an aralkyl group, a C1-C6 aldehyde group or a halogen;
  • R 4 represents hydrogen and R 3 represents hydrogen or oxygen, and W is carbon, R 2 is not hydrogen.
  • the compound has the structure of formula A1:
  • Q and W each independently represent C or N, and when each is independently N, the position and number of each in the ring may vary; wherein said "when Q or W are each independently N, position and number "changeable” means that the position where Q or W appears on the respective ring is not limited; the number of Q or W may be 1-3, and when the number is greater than or equal to ( ⁇ ) 2, in order to maintain the six-membered ring system, The number of carbon atoms in the corresponding ring is reduced, and Q or W may be bonded to each other or dispersed in different positions on the ring;
  • R 1 , R 2 and R 3 each independently represent hydrogen, hydroxy, amino, thiol, alkyl, aryl, alkoxy, alkylamino, alkylthio, aryl ether, arylamino, arylthio or halogen;
  • R 4 represents oxygen, nitrogen, sulfur, hydroxy, amino, decyl, alkyl, aryl, alkoxy, alkylamino, alkylthio, aryl ether, arylamino, arylthio or halogen;
  • R 4 represents hydrogen and R 3 represents hydrogen or oxygen, W is not carbon or R 2 is not hydrogen.
  • R 2 when R 4 represents hydrogen and R 3 represents a hydrogen or oxygen, R 2 is not a two methoxy group (i.e., exclude the case where: R 2 is 2 and the number of simultaneously methoxy).
  • the halogen means fluorine, chlorine, bromine or iodine.
  • the aldehyde group comprises a C1-C6 -CHO, -CH 2 -CHO or the like.
  • the pharmaceutically acceptable salt includes a salt formed with an acid ion; preferably, the salt-forming acid ion includes a mineral acid ion, an organic acid ion, and/or a halogen ion. .
  • the acid ion is selected from the group consisting of nitrate ion, sulfate ion, phosphate ion, mesylate ion, benzenesulfonate ion, acetate ion, tartrate ion, and citrate Ions, maleate ions, succinate ions, citrate ions, salicylate ions, glycerate ions, Ascorbate ion, fluoride ion, chloride ion, bromide ion, iodide ion, or a combination thereof.
  • middle For a single button, Is a double bond
  • R 3 represents an oxygen, nitrogen, sulfur, alkyl or alkylamino group.
  • middle For double keys Is a single button.
  • R 4 represents a hydroxyl group, an amino group, a decyl group, an alkyl group, an aryl group, an alkoxy group, an alkylamino group, an alkyl fluorenyl group, an aryl ether group, an arylamino group, an aryl fluorenyl group or a halogen.
  • R 1 and R 2 each independently represent a hydroxyl group, an amino group, a decyl group, an alkyl group, an aryl group, an alkoxy group, an alkylamino group, an alkyl fluorenyl group, an aryl ether group, an arylamino group, an aryl fluorenyl group or a halogen.
  • R 1 , R 2 , R 3 , R 4 , ring, Rings, W, and/or Q are the corresponding groups in the specific compounds prepared in the examples, respectively.
  • the seven-membered ring berberine analog is selected from the group consisting of:
  • the compound is compounded by each of the specific formulas A prepared in Examples 2-18. Things.
  • a pharmaceutical composition comprising a therapeutically effective amount of one or more seven-membered cyclic berberine analogs of the first aspect of the invention, or a pharmaceutically acceptable thereof, is provided Salt and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition is an injection, a sachet, a tablet, a pill, a powder or a granule.
  • a third aspect of the invention there is provided the use of the seven-membered ring berberine analog of the first aspect of the invention, and a pharmaceutically acceptable salt thereof, for use in the preparation of a medicament for preventing or treating a tumor or cancer .
  • a method of preparing a pharmaceutical composition comprising the steps of: pharmaceutically acceptable carrier and a seven-membered cyclic berberine analog of the first aspect of the invention, or a crystal thereof A type, pharmaceutically acceptable salt, hydrate, solvate or prodrug is mixed to form a pharmaceutical composition.
  • a method of treatment comprising the steps of:
  • a safe and effective amount of the seven-membered cyclic berberine analog of the first aspect of the invention, and a pharmaceutically acceptable salt thereof, are administered to a subject in need of treatment.
  • the method of treatment is for treating a tumor.
  • the tumor comprises: lung cancer, gastric cancer, liver cancer, colorectal cancer, breast cancer, cervical cancer, prostate cancer, melanoma, leukemia cells.
  • a method for non-therapeutic inhibition of tumor cells in vitro comprising the steps of: the seven-membered cyclic berberine analog of the first aspect of the invention, and pharmaceutically acceptable thereof
  • tumor cells are cultured to inhibit the growth of the tumor cells.
  • the tumor cells are selected from the group consisting of lung cancer cells, gastric cancer cells, liver cancer cells, colorectal cancer cells, breast cancer cells, cervical cancer cells, prostate cancer cells, melanoma cells, and leukemia cells.
  • the inventors of the present application have extensively and intensively studied, and by first screening and testing a large number of compounds, a first class of compounds with novel structures and significantly improved antitumor activity have been unexpectedly discovered.
  • the structure of such compounds is shown in Formula A.
  • the in vitro antitumor activity of these compounds of formula A has a very significant increase (up to about 10 fold or more) than the natural product berberine.
  • the present invention has been completed on this basis.
  • the present invention conducts activity tests on a series of novel compounds having a novel saturated seven-membered ring structure as a core skeleton, and the results show that these newly synthesized compounds exhibit extremes on various tumor cells.
  • Significant anti-tumor activity there is a great potential for development.
  • the unsaturated double bond of the seven-membered ring of the compound sm38 is reduced to a saturated single bond, the in vitro antitumor activity of the formed compound (ie, compound 2 in Example 1) has a very significant improvement over the compound sm38 ( That is, the compound 1) in Example 1 was increased by 10 times.
  • the seven-membered ring berberine analog of the present invention and a pharmaceutically acceptable salt thereof can inhibit the growth of various tumor cells, and thus can be developed as a new drug for treating various tumors or cancers.
  • halogen refers to F, Cl, Br, and I. More preferably, the halogen atom is selected from the group consisting of F, Cl and Br.
  • C1-C6 alkyl refers to a straight or branched alkyl group comprising from 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl. , tert-butyl, or the like.
  • alkoxy refers to an alkoxy group having from 1 to 10 carbon atoms, preferably a C1-C6 alkoxy group.
  • C1-C6 alkoxy includes a straight or branched alkoxy group of 1 to 6 carbon atoms.
  • the following terms include unsubstituted forms as well as substituted forms: amino, mercapto, alkyl, aryl, alkoxy, alkylamino, alkylthio, aryl ether, arylamino, arylthio.
  • the number of carbon atoms in the alkyl group, the alkoxy group, the alkylamino group, and the alkylthio group is from 1 to 10, preferably from 1 to 6, more preferably from 1 to 3 .
  • the aryl group, the aryl ether group, the arylamino group, and the arylthio group have 6 to 30 carbon atoms, preferably 6 to 20, more preferably 6 -10.
  • aralkyl refers to a group formed by the substitution of one or more H atoms of a C1-C10 alkyl group by a C6-10 aryl group, and representative examples include a benzyl group.
  • alkyl includes saturated or unsaturated, linear, branched, cyclic 1-10 carbon atoms (C1-C10) all-carbon alkyl groups or 1-3 thereof An alkyl group in which a carbon atom is substituted with a hetero atom such as oxygen, nitrogen or sulfur, and an aralkyl group bonded through one or more carbon atoms. Further, the alkyl group is unsubstituted or substituted.
  • aryl includes fused or non-fused aryl groups, usually containing from 6 to 30 carbon atoms, and representative aryl groups include phenyl, naphthyl, or oxygen, nitrogen, sulfur, etc. Aromatic group of atoms.
  • 5-6 membered ring refers to a ring structure having 5 or 6 ring atoms, including C atoms and/or heteroatoms, which may be saturated or unsaturated, may be aromatic Or non-aromatic, with or without heteroatoms.
  • heteroatom refers to N, O or S.
  • the term "compound of the invention” refers to a compound of formula A, ie a seven-membered cyclic berberine analog.
  • the term also encompasses various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula A.
  • Ring and The ring may be saturated or unsaturated, and may be an aromatic ring or a non-aromatic ring, with or without a hetero atom.
  • a representative R1 or R2 group is the corresponding group of the following structural formula (exemplified by the R1 group):
  • representative R3 or R4 groups are the corresponding groups of the formula (exemplified by the R3 group):
  • a representative Q or W group is the corresponding group of the following structural formula (exemplified by a Q group):
  • R is selected from the group consisting of:
  • R is selected from the group consisting of a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C2-C6 alkenyl group, a substituted or unsubstituted benzyl group, a substituted or unsubstituted C2-C6 ester group. , substituted or unsubstituted C2-C8, wherein said substitution is substituted by one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy , hydroxyl.
  • one or more of the compounds of the formula A or the formula A1 or a pharmaceutically acceptable salt thereof may be deuterated, that is, the cerium isotope content of the cerium at the hydrazine-substituted position is at least greater than the natural strontium isotope.
  • the content (0.015%), preferably more than 30%, more preferably more than 50%, more preferably more than 75%, more preferably more than 95%, more preferably more than 99%.
  • the compound of formula A contains at least one deuterium atom, more preferably two deuterium atoms, more preferably four deuterium atoms, more preferably six deuterium atoms.
  • the term "pharmaceutically acceptable salt” refers to a salt of the compound of the invention formed with an acid or base suitable for use as a medicament.
  • Pharmaceutically acceptable salts include inorganic and organic salts.
  • a preferred class of salts are the salts of the compounds of the invention with acids.
  • Suitable acids for forming salts include, but are not limited to, mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid; formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, Organic acids such as fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid; Amino acids such as amino acid, phenylalanine, aspartic acid, and glutamic acid.
  • mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid
  • formic acid
  • salts of the compounds of the invention with bases such as alkali metal salts (for example sodium or potassium salts), alkaline earth metal salts (for example magnesium or calcium salts), ammonium salts (for example lower alkanolammonium).
  • bases such as alkali metal salts (for example sodium or potassium salts), alkaline earth metal salts (for example magnesium or calcium salts), ammonium salts (for example lower alkanolammonium).
  • Salts and other pharmaceutically acceptable amine salts such as methylamine, ethylamine, propylamine, dimethylamine, trimethylamine, diethylamine, triethylamine, tert-butyl
  • a base amine salt an ethylenediamine salt, a hydroxyethylamine salt, a dihydroxyethylamine salt, a trihydroxyethylamine salt, and an amine salt formed of morpholine, piperazine, and lysine, respectively.
  • solvate refers to a complex of a compound of the invention that is coordinated to a solvent molecule to form a specific ratio.
  • Hydrophilate means a complex formed by the coordination of a compound of the invention with water.
  • the compound of the present invention further includes a prodrug of a seven-membered cyclic berberine compound represented by Formula A.
  • prodrug includes a class of compounds which, by themselves, may be biologically active or inactive, which, upon administration in an appropriate manner, are metabolized or chemically reacted in the human body to form a compound of formula A, or formula A a salt or solution of a compound.
  • the prodrug includes, but is not limited to, a carboxylic acid ester, a carbonate, a phosphate, a nitrate, a sulfate, a sulfone ester, a sulfoxide, an amino compound, a carbamate, an azo compound of the compound. , phosphoramide, glucoside, ether, acetal and the like.
  • the preparation method of the structural compound of the formula A of the present invention is more specifically described below, but these specific methods are not correct.
  • the invention constitutes any limitation.
  • the compounds of the present invention may also be conveniently prepared by combining various synthetic methods described in the specification or known in the art, and such combinations are readily made by those skilled in the art to which the present invention pertains.
  • each reaction is usually carried out in an inert solvent at room temperature to reflux temperature (e.g., 0 ° C to 80 ° C, preferably 0 ° C to 50 ° C).
  • the reaction time is usually from 0.1 to 60 hours, preferably from 0.5 to 48 hours.
  • R 1 , R 2 , R 3 , W, and Q are as defined above.
  • the seven-membered ring double bond of Compound I is reduced by catalytic hydrogenation (e.g., palladium on carbon/hydrogen or palladium on carbon/ammonium formate, etc.) to give Compound II, which is then obtained from Compound II by a method similar to that in Example 2-18.
  • catalytic hydrogenation e.g., palladium on carbon/hydrogen or palladium on carbon/ammonium formate, etc.
  • the compound I it can be produced or purchased by, for example, the method of Chinese Patent Application No. 201010537594.1 or the like, for example, the compound 1 as a raw material in Example 1.
  • the compound of the present invention has excellent antitumor activity, the compound of the present invention and various crystal forms thereof, a pharmaceutically acceptable inorganic or organic salt, hydrate or solvate, and a drug containing the compound of the present invention as a main active ingredient
  • the composition can be used to treat, prevent, and alleviate tumor-related diseases.
  • compositions of the present invention comprise a safe or effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier.
  • safe and effective amount it is meant that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects.
  • the pharmaceutical compositions contain from 1 to 2000 mg of the compound of the invention per agent, more preferably from 10 to 1000 mg of the compound of the invention per agent.
  • the "one dose" is a capsule or tablet.
  • “Pharmaceutically acceptable carrier” means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By “compatibility” it is meant herein that the components of the composition are capable of intermingling with the compounds of the invention and with each other without significantly reducing the efficacy of the compound.
  • pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid).
  • magnesium stearate magnesium stearate
  • calcium sulfate vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as Tween ), a wetting agent (such as sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, an antioxidant, a preservative, a pyrogen-free water, and the like.
  • vegetable oil such as soybean oil, sesame oil, peanut oil, olive oil, etc.
  • polyol such as propylene glycol, glycerin, mannitol, sorbitol, etc.
  • emulsifier such as Tween
  • a wetting agent such as sodium lauryl sulfate
  • a coloring agent such as a flavoring agent, a stabilizer, an antioxidant, a preservative
  • the mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration. .
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) Absorbing accelerators, for example, quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and
  • Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs.
  • the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.
  • inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethyl
  • compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
  • the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.
  • suspending agents for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.
  • compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion.
  • Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.
  • Dosage forms for the compounds of the invention for topical administration include ointments, powders, patches, propellants and inhalants.
  • the active ingredient is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or, if necessary, propellants.
  • the compounds of the invention may be administered alone or in combination with other pharmaceutically acceptable other compounds, such as anti-neoplastic agents.
  • a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight
  • the dose to be administered is usually from 1 to 2000 mg, preferably from 50 to 1000 mg.
  • specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.
  • the compound of the present invention has excellent activity for inhibiting tumor cells, and not only significantly inhibits tumor cells in vitro, but also preferably has an IC 50 of ⁇ 2 nM.
  • the compounds of the present invention have a broad spectrum of antitumor activity.
  • the compounds of the present invention also have selective tumor cell inhibitory activity against certain tumor cells.
  • Method 1 Compound 1 (specifically prepared according to the method of Chinese Patent Application No. 201010537594.1) is dissolved in methanol, a catalytic amount of 10% palladium carbon is added, hydrogen gas is introduced, and the reaction is carried out at room temperature until the reaction of the starting material is complete. The reaction mixture was filtered over Celite, and evaporated.
  • tumor cells 2 liver cancer cells (BEL-7402, SMMC-7721); 2 gastric cancer cells (SGC-7901, MKN-45); 2 lung cancer cells (A549, SPC-A4); colon Two cancer cells (HT29, SW-620, HCT116) and melanoma cells (A375) were treated with compound for 72h, and the proliferation inhibition effect of the compound was detected by Sulforhodamine B (SRB) or MTT assay. The extent of it.
  • SRB Sulforhodamine B
  • Activity Experiment 1 tested the inhibitory activity of some of the compounds of the present invention on lung cancer cell lines (SPC-A4), colon cancer cell lines (SW-620, HCT116, HT29) and melanoma cell lines (A375), indicating that the compounds of the present invention have Strong anti-tumor activity, not only better than the natural product Berberine, the activity of some compounds is even better than the positive control drug etoposide (VP-16) or sorafenib (Sorafenib).
  • VP-16 positive control drug etoposide
  • Sorafenib sorafenib
  • Tumor cell inhibitory activity of the compound of the present invention (% inhibition rate of 5 ⁇ M)
  • the compound of the formula A prepared in the embodiments 1-18 of the present invention showed a significant inhibition rate of BEL-7402, SMMC-7721, SPC-A4 and other tumor cells at a concentration of 5 ⁇ M, and the inhibition rate was mostly 50. %-90%.
  • compound 2 had 2 liver cancer cells (BEL-7402, SMMC-7721); 2 gastric cancer cells (SGC-7901, MKN-45); 2 lung cancer cells (A549, SPC-A4); colon cancer cells 2
  • the strain (HT29, SW-620) has excellent tumor suppressing activity.

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Abstract

七元环小檗碱类似物及其药学上可接受的盐，其具有式A结构，其中各取代基的定义如说明书中所述；还提供了式A所示的七元环小檗碱类似物的制备方法，及其在制备预防或治疗肿瘤或癌症的药物中的用途。

Description

七元环小檗碱类似物及其药物组合物、制备方法和用途 技术领域
本发明涉及药物化学和药物治疗学领域，具体涉及可作为治疗肿瘤的七元环小檗碱类似物及其药物组合物、制备方法和医药用途。
背景技术
中医中药是我中华民族的瑰宝，因此，开展以活性天然产物为基础的现代创新药物研究是我国新药研发工作者应该予以高度关注的研究领域。已经广泛使用的从中草药中提取得到的活性天然产物很多，小檗碱是其中之一。小檗碱(Berberine)，属异喹啉类生物碱，商品名黄连素，主要用于治疗肠道细菌感染，是中药黄连中的主要活性成分。
Figure PCTCN2016076083-appb-000001
中药黄连主要包括毛茛科(Ranunculaceae)黄连(Coptis chinensis Franch)、三角叶黄连(Coptis deltoidea C.Y.cheng et Hsiao)、云连(Coptis teetaw all)的干燥根茎。除此之外，小檗碱还存在于小檗科、罂粟科、芸香科、防己科、鼠李科植物中。近年来研究发现，小檗碱及其衍生物药理作用十分丰富，具有抗肿瘤、糖尿病、心血管疾病、高血压、炎症、细菌和病毒感染、脑缺血性损伤、精神疾病、阿尔茨海默病(Alzheimer disease)、骨质疏松等多方面活性，潜在的开发应用前景非常广阔。
恶性肿瘤是危害人类健康的疾病之一，位居各类疾病死亡率的前列。药理实验研究表明，小檗碱对肺癌细胞、胃癌细胞、肝癌细胞、结肠癌细胞、乳腺癌细胞、宫颈癌细胞、前列腺癌细胞、黑色素瘤细胞、白血病细胞等均有明显的生长抑制作用。其抗肿瘤作用机制包括抑制肿瘤细胞增殖、促进肿瘤细胞凋亡、抑制肿瘤转移、以及诱导肿瘤细胞分化等，相关的分子机制非常复杂，涉及的信号通路也很广泛。因此，小檗碱抗肿瘤活性具有多效应、多靶点的作用特点。
已有的药理研究表明，小檗碱极具开发成多种抗肿瘤药物的潜力。但迄今为止，尚缺乏相关抗肿瘤新药上市。仔细比较文献报道的小檗碱抗肿瘤活性数据发现，其体内外活性测试时的有效剂量与目前临床应用的抗肿瘤药物相比，给药剂量太大。而造成这一问题的主要原因则是因为小檗碱口服生物利用度低。为了解决这一问题，制剂学家们想出了很多办法，比如脂质体、环糊精包合物以及与促吸收剂联合给药等，但都难以从根本上加以解决。据推测，小檗碱独特的结构特征是造成这一 问题的关键，其母体可以看作是两个异喹啉基本单元组合而成的四个六元环体系，整个体系处于一个近似平面的状态，再加上分子中拥有极性的C＝N+键，从而导致该化合物脂溶性和水溶性都很差，进而影响到生物利用度。因此，对小檗碱的结构进行改造，改善其溶解度和生物利用度并且增强抗肿瘤活性有助于加快该类化合物从天然活性成分到临床候选药物的转变，对于传统药物的深度开发也极具借鉴意义。
为了改善小檗碱的溶解度和生物利用度以及增强药理活性，国内外许多同行对小檗碱的结构进行了改造研究，取得了一些较好的实验结果。这些结构改造研究概括起来大致可分为以下几类：小檗碱A、C、D环上取代基的变化(改造位点多集中于2、3、8、9、12、13等位置，有单位点集中修饰和多位点同时修饰)；小檗碱C环的氢化(主要是C环的二氢化和四氢化，以及氢化后8、13位的取代)；小檗碱N上的季铵化(主要是C环氢化后氮上的不同基团季铵化)；小檗碱四元环骨架的改变(主要是小檗碱在碱性条件下开环变成苄基异喹啉类化合物)，绝大多数研究主要集中在小檗碱的A、C、D环上。
Figure PCTCN2016076083-appb-000002
近年来，中国科学院上海药物研究所综合采用药物设计及全合成的办法首次对小檗碱的B环进行了设计改造，得到一系列结构新颖的小檗碱类似物，明显地改善了溶解性和体外抗肿瘤活性。其中，B环为不饱和七元环的化合物sm38在10.0uM浓度时，对大多数测试肿瘤细胞(包括结肠癌和黑色素瘤细胞)的抑制率超过了50％。
迄今为止，本领域尚需研发更有效和/或更安全的小檗碱类似物，以便用于肿瘤等疾病的治疗。
发明内容
本发明的目的在于提供一种式A所示七元环小檗碱类似物及其药学上可接受的盐及其制法和应用。
本发明的第一方面，提供一种式A所示的七元环小檗碱类似物、其药学上可接受的盐、其晶型、其水合物、或其溶剂化物、或其前药，
Figure PCTCN2016076083-appb-000003
式中，
Figure PCTCN2016076083-appb-000004
代表双键或单键，并且
Figure PCTCN2016076083-appb-000005
中的两个
Figure PCTCN2016076083-appb-000006
不同时为双键，或者
Figure PCTCN2016076083-appb-000007
中的两个
Figure PCTCN2016076083-appb-000008
不同时为双键；
Figure PCTCN2016076083-appb-000009
和
Figure PCTCN2016076083-appb-000010
分别表示5-6元环，优选为芳基或杂芳基，其中所述的杂芳基具有1-3个N原子；
或者
Figure PCTCN2016076083-appb-000011
和
Figure PCTCN2016076083-appb-000012
分别表示芳基或5-6元杂芳基，其中当所述的杂芳基为6元环时，具有1-3个N原子；为5元环时，具有1-3个N或O或S杂原子；
R1为位于
Figure PCTCN2016076083-appb-000013
的1-4个(较佳地1-3个或1-2个)相同或不同的取代基，而R2为位于
Figure PCTCN2016076083-appb-000014
的1-4个(较佳地1-3个或1-2个)相同或不同的取代基，并且各R1和R2独立地选自下组：氢、羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基或卤素；或者两个R1可与相邻的碳原子或两个R2可与相邻的碳原子共同形成含有0-3个选自O、S、N的杂原子的5-7元环(例如，二个R1与
Figure PCTCN2016076083-appb-000015
或两个R2与
Figure PCTCN2016076083-appb-000016
共同形成吲哚、氮杂吲哚等结构)；
R3选自下组：氢、氧、硫、羟基、氨基、巯基、烷基、链烯基、链炔基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基、芳烷基、C1-C6醛基或卤素；
R4选自下组：氢、氧、硫、羟基、氨基、巯基、烷基、链烯基、链炔基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基、芳烷基、C1-C6醛基或卤素。
在另一优选例中，当R4代表氢且R3代表氢或氧，且D为苯环时，R2不为氢。
在另一优选例中，当R4为氢且R3为氢或氧时，
Figure PCTCN2016076083-appb-000017
环不为苯环；或者当R4 为氢，R3为氢或氧且当
Figure PCTCN2016076083-appb-000018
环为苯环时，R2不为氢。
在另一优选例中，所述的R1或R2各自独立地选自下组：取代或未取代的C1-C6烷基、取代或未取代的C2-C6链炔基、羟基、取代的或未取代的烷氧基、取代或未取代的-(CH2)m-O-(CH2)n-Ra、取代或未取代的-O-(CH2)m-Ra；其中，Ra为C1-C3烷基、C1-C3烷氧基或-(CH2)m-CO-Rb，而Rb为羟基、C1-C6烷氧基、
Figure PCTCN2016076083-appb-000019
m和n各自为0-3的整数；
其中，Rc为H、C1-C3烷基或C1-C3卤代烷基。
其中，所述的“取代的”指所述基团中一个或多个H被选自下组的取代基所取代：卤素、C1-C4烷基、C1-C4卤代烷基、C2-C4链烯基、C2-C4链炔基、C3-C6环烷基、苄基、卤代苄基、羟基、氨基、胺基。
在另一优选例中，所述的R4为-R8-R9或-R8-CO-R10，其中R8为二价连接基团，R9选自下组：C1-C8酯基、C1-C8羧基、-O-C1-C6烷基、-O-C2-C6链烯基、羟基、
Figure PCTCN2016076083-appb-000020
而R10选自下组：H、羟基、
Figure PCTCN2016076083-appb-000021
其中，Rc为H、C1-C3烷基或C1-C3卤代烷基。
在另一优选例中，R8为选自下组的二价连接基团：-CH2-、-CH2-CH2-、-CH＝CH-。
在另一优选例中，所述的化合物具有式A1结构：
Figure PCTCN2016076083-appb-000022
其中，
Figure PCTCN2016076083-appb-000023
代表双键或单键，并且
Figure PCTCN2016076083-appb-000024
中的两个
Figure PCTCN2016076083-appb-000025
不同时为双键，或者
Figure PCTCN2016076083-appb-000026
中的两个
Figure PCTCN2016076083-appb-000027
不同时为双键；
Q和W各自独立地代表C、N、O或S原子中的一种或2种以上，且当各自独立为N时，各自在环内所处的位置和数目可变化；当各自独立含有O或S时，环上碳原子减少，保持五元芳环体系；Q或W可以各自互相成键连接在一起或者各自分散在环上不同位置；
其中所述的“当Q或W各自独立为N时，位置和数目可变化”是指：Q或W出现在各自所在环上的位置不受限定；Q或W的数目可为1-3，当数目大于或等于(≥)2时，为保持六元环体系，相应环上碳原子数目减少；
R1和R2各自独立地代表氢、羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基或卤素；或者两个R1可与相邻的碳原子或两个R2可与相邻的碳原子共同形成含有0-3个选自O、S、N的杂原子的5-7元环；
R3和R4各自独立地代表氢、氧、硫、羟基、氨基、巯基、烷基、链烯基、链炔基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基、芳烷基、C1-C6醛基或卤素；
并且当R4代表氢且R3代表氢或氧，且W为碳时，R2不为氢。
在另一优选例中，所述的化合物具有式A1结构：
Figure PCTCN2016076083-appb-000028
其中，
Figure PCTCN2016076083-appb-000029
代表双键或单键，并且
Figure PCTCN2016076083-appb-000030
中的两个
Figure PCTCN2016076083-appb-000031
不同时为双键，或者
Figure PCTCN2016076083-appb-000032
中的两个
Figure PCTCN2016076083-appb-000033
不同时为双键。
Q和W各自独立地代表C或N，且当各自独立为N时，各自在环内所处的位置和数目可变化；其中所述的“当Q或W各自独立为N时，位置和数目可变化”是指：Q或W出现在各自所在环上的位置不受限定；Q或W的数目可为1-3，当数目大于或等于(≥)2时，为保持六元环体系，相应环上碳原子数目减少，Q或W可以各自互相成键连接在一起或者各自分散在环上不同位置；
R1，R2和R3各自独立地代表氢、羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基或卤素；
R4代表氧、氮、硫、羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基或卤素；
并且当R4代表氢且R3代表氢或氧时，W不为碳或者R2不为氢。
在另一优选例中，当R4代表氢且R3代表氢或氧时，R2不为二个甲氧基(即排除以下情况：R2数量为2且同时为甲氧基)。
在另一优选例中，所述的卤素是指氟、氯、溴或碘。
在另一优选例中，所述的C1-C6醛基包括-CHO、-CH2-CHO或类似基团。
在另一优选例中，所述的药学上可接受的盐包括与酸根离子形成的盐；较佳地，所述的成盐的酸根离子包括无机酸根离子、有机酸根离子、和/或卤素离子。
在另一优选例中，所述的酸根离子选自下组：硝酸根离子、硫酸根离子、磷酸根离子、甲磺酸根离子、苯磺酸根离子、醋酸根离子、酒石酸根离子、枸橼酸根离子、马来酸根离子、琥珀酸根离子、柠檬酸根离子、水杨酸根离子、甘油酸根离子、 抗坏血酸根离子、氟离子、氯离子、溴离子、碘离子、或其组合。
在另一优选例中，
Figure PCTCN2016076083-appb-000034
中的两个
Figure PCTCN2016076083-appb-000035
不同时为双键，且
Figure PCTCN2016076083-appb-000036
中的
Figure PCTCN2016076083-appb-000037
为单键，
Figure PCTCN2016076083-appb-000038
为双键；
Figure PCTCN2016076083-appb-000039
中的两个
Figure PCTCN2016076083-appb-000040
不同时为双键，且
Figure PCTCN2016076083-appb-000041
中的
Figure PCTCN2016076083-appb-000042
为双键，
Figure PCTCN2016076083-appb-000043
为单键；或
Figure PCTCN2016076083-appb-000044
中的两个
Figure PCTCN2016076083-appb-000045
不同时为双键，且
Figure PCTCN2016076083-appb-000046
中的为单键，
Figure PCTCN2016076083-appb-000048
为单键。
在另一优选例中，
Figure PCTCN2016076083-appb-000049
中的两个
Figure PCTCN2016076083-appb-000050
不同时为双键，且
Figure PCTCN2016076083-appb-000051
中的
Figure PCTCN2016076083-appb-000052
为双键，
Figure PCTCN2016076083-appb-000053
为单键；
Figure PCTCN2016076083-appb-000054
中的两个
Figure PCTCN2016076083-appb-000055
不同时为双键，且
Figure PCTCN2016076083-appb-000056
中的
Figure PCTCN2016076083-appb-000057
为单键，
Figure PCTCN2016076083-appb-000058
为双键；
Figure PCTCN2016076083-appb-000059
中的两个
Figure PCTCN2016076083-appb-000060
不同时为双键，且
Figure PCTCN2016076083-appb-000061
中的
Figure PCTCN2016076083-appb-000062
为单键，
Figure PCTCN2016076083-appb-000063
为单键。
在另一优选例中，
Figure PCTCN2016076083-appb-000064
中的
Figure PCTCN2016076083-appb-000065
为单键，
Figure PCTCN2016076083-appb-000066
为双键；和/或
R3代表氧、氮、硫、烷基或烷氨基。
在另一优选例中，
Figure PCTCN2016076083-appb-000067
中的
Figure PCTCN2016076083-appb-000068
为双键，
Figure PCTCN2016076083-appb-000069
为单键。
在另一优选例中，R4代表羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷巯基、芳醚基、芳氨基、芳巯基或卤素。
在另一优选例中，R1和R2各自独立地代表羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷巯基、芳醚基、芳氨基、芳巯基或卤素。
在另一优选例中，所述的R1、R2、R3、R4、
Figure PCTCN2016076083-appb-000070
环、
Figure PCTCN2016076083-appb-000071
环、W、和/或Q分别为实施例中所制备的具体化合物中相应基团。
在另一优选例中，所述七元环小檗碱类似物选自下组：
Figure PCTCN2016076083-appb-000072
Figure PCTCN2016076083-appb-000073
在另一优选例中，所述的化合物由实施例2-18中所制备的各具体的式A化合 物。
在本发明的第二方面，提供了一种药物组合物，它包含治疗有效量的一种或多种本发明第一方面所述的七元环小檗碱类似物或其药学上可接受的盐以及药学上可接受的载体。
在另一优选例中，所述的药物组合物为注射剂、囊剂、片剂、丸剂、散剂或颗粒剂。
在本发明的第三方面，提供了本发明第一方面所述的七元环小檗碱类似物及其药学上可接受的盐的用途，它被用于制备预防或治疗肿瘤或癌症的药物。
在本发明的第四方面，提供了一种制备药物组合物的方法，包括步骤：将药学上可接受的载体与本发明第一方面所述的七元环小檗碱类似物、或其晶型、药学上可接受的盐、水合物、溶剂合物或前药进行混合，从而形成药物组合物。
在本发明的第五方面，提供了一种治疗方法，包括步骤：
给需要治疗的对象施用安全有效量的本发明第一方面所述的七元环小檗碱类似物及其药学上可接受的盐。
在另一优选例中，所述的治疗方法用于治疗肿瘤。
在另一优选例中，所述的肿瘤包括：肺癌、胃癌、肝癌、结直肠癌、乳腺癌、宫颈癌、前列腺癌、黑色素瘤、白血病细胞。
在本发明的第六方面，提供了一种体外非治疗性地抑制肿瘤细胞的方法，包括步骤：在本发明第一方面所述的七元环小檗碱类似物及其药学上可接受的盐存在下，培养肿瘤细胞，从而抑制所述肿瘤细胞的生长。
在另一优选例中，所述的肿瘤细胞选自下组：肺癌细胞、胃癌细胞、肝癌细胞、结直肠癌细胞、乳腺癌细胞、宫颈癌细胞、前列腺癌细胞、黑色素瘤细胞、白血病细胞。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
具体实施方式
本申请的发明人经过广泛而深入地研究，通过对化合物的大量筛选和测试，首次意外发现了一类结构新颖且抗肿瘤活性显著提高的化合物。这类化合物的结构如式A所示。这些式A化合物的体外抗肿瘤活性较天然产物小檗碱具有非常显著的提高(可提高达约10倍或更多)。在此基础上完成了本发明。
具体地，本发明对于这类以新的饱和七元环结构为核心骨架的一系列新颖化合物进行了活性测试，结果显示，这些新合成的化合物在多种肿瘤细胞上表现出了极 显著的抗肿瘤活性，存在很大的潜在开发价值。例如，当化合物sm38七元环的不饱和双键被还原成饱和单键后，所形成的化合物(即实施例1中的化合物2)的体外抗肿瘤活性具有非常显著的提高，比化合物sm38(即实施例1中的化合物1)提高达10倍。
本发明的七元环小檗碱类似物及其药学上可接受的盐可抑制多种肿瘤细胞生长，因此可供开发成为新的治疗多种肿瘤或癌症的药物。
定义
如本文所用，“卤素”指F、Cl、Br、和I。更佳地，卤原子选自F、Cl和Br。
如本文所用，“C1-C6烷基”是指包括1-6个碳原子的直链或支链的烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基、或类似基团。
如本文所用，“烷氧基”指具有1-10个碳原子的烷氧基，优选C1-C6烷氧基。
如本文所用，“C1-C6烷氧基”包括1-6个碳原子的直链或支链的烷氧基。例如甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、异丁氧基、叔丁氧基、或类似基团。
在本发明中，以下术语包括未取代形式以及取代形式：氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基。
在本发明的优选例中，烷基、烷氧基、烷氨基、和烷硫基中的碳原子数为1-10个，较佳地为1-6个，更佳地为1-3个。
在本发明的优选例中，所述的芳基、芳醚基、芳氨基、和芳硫基中的碳原子数为6-30个，较佳地为6-20个，更佳地为6-10个。
如本文所用，术语“芳烷基”指C1-C10烷基中的1个或多个H原子被C6-10芳基取代所形成的基团，代表性的例子包括苄基。
此外，在本发明中，术语“烷基”包括饱和或不饱和、直链、支链、环状的1-10个碳原子(C1-C10)的全碳烷基或其中的1-3个碳原子被氧、氮、硫等杂原子取代的烷基，以及通过1个或1个以上碳原子连接的芳烷基。此外，所述的烷基是未取代的或取代的。
如本文所用，术语“芳基”包括稠合或非稠合的芳基，通常含有6-30个碳原子，代表性的芳基包括苯基、萘基，或含氧、氮、硫等杂原子的芳香基团。
如本文所用，“5-6元环”指具有5或6个环原子(包括C原子和/或杂原子)的环结构，所述的环结构可以是饱和的或不饱和的，可以是芳香的或非芳香的，可以含有或不含有杂原子。
如本文所用，“杂原子”指N、O或S。
活性成分
如本文所用，术语“本发明化合物”指式A所示的化合物，即七元环小檗碱类似物。该术语还包括及式A化合物的各种晶型形式、药学上可接受的盐、水合物或溶剂合物。
在本发明中，
Figure PCTCN2016076083-appb-000074
环和
Figure PCTCN2016076083-appb-000075
环可以是饱和的或不饱和的，可以是芳环或非芳环，可以含有或不含有杂原子。
在本发明的某些优选例中，代表性的R1或R2基团为下列结构式的相应基团(以R1基团为例)：
Figure PCTCN2016076083-appb-000076
在本发明某些优选例中，代表性的R3或R4基团为下列结构式的相应基团(以R3基团为例)：
Figure PCTCN2016076083-appb-000077
式中，“O or S”表示该位置可以是O或S。
在本发明某些优选例中，代表性的Q或W基团为下列结构式的相应基团(以Q基团为例)：
Figure PCTCN2016076083-appb-000078
此外，在本发明的一些优选例(包括上述各结构式)中，R为选自下组基团：
Figure PCTCN2016076083-appb-000079
在另一优选例中，R选自：取代或未取代C1-C6烷基、取代或未取代C2-C6链烯基、取代或未取代的苄基、取代或未取代的C2-C6酯基、取代或未取代的C2-C8，其中所述的取代指被一个或多个选自下组的取代基取代：卤素、C1-C3烷基、C1-C3卤代烷基、C1-C3烷氧基、羟基。
此外，在本发明中，式A或式A1化合物或其药学上可接受的盐中，一个或多个H可以为氘代的，即氘在氘取代位置的氘同位素含量至少是大于天然氘同位素含量(0.015％)，较佳地大于30％，更佳地大于50％，更佳地大于75％，更佳地大于95％，更佳地大于99％。
在另一优选例中，式A化合物至少含有1个氘原子,更佳地2个氘原子，更佳地4个氘原子，更佳地6个氘原子。
如本文所用，术语“药学上可接受的盐”指本发明化合物与酸或碱所形成的适合用作药物的盐。药学上可接受的盐包括无机盐和有机盐。一类优选的盐是本发明化合物与酸形成的盐。适合形成盐的酸包括但并不限于：盐酸、氢溴酸、氢氟酸、硫酸、硝酸、磷酸等无机酸；甲酸、乙酸、三氟乙酸、丙酸、草酸、丙二酸、琥珀酸、富马酸、马来酸、乳酸、苹果酸、酒石酸、柠檬酸、苦味酸、苯甲酸、甲磺酸、乙磺酸、对甲苯磺酸、苯磺酸、萘磺酸等有机酸；以及脯氨酸、苯丙氨酸、天冬氨酸、谷氨酸等氨基酸。
另一类优选的盐是本发明化合物与碱形成的盐，例如碱金属盐(例如钠盐或钾盐)、碱土金属盐(例如镁盐或钙盐)、铵盐(如低级的烷醇铵盐以及其它药学上可接受的胺盐)，例如甲胺盐、乙胺盐、丙胺盐、二甲基胺盐、三甲基胺盐、二乙基胺盐、三乙基胺盐、叔丁基胺盐、乙二胺盐、羟乙胺盐、二羟乙胺盐、三羟乙胺盐，以及分别由吗啉、哌嗪、赖氨酸形成的胺盐。
术语“溶剂合物”指本发明化合物与溶剂分子配位形成特定比例的配合物。“水合物”是指本发明化合物与水进行配位形成的配合物。
此外，本发明化合物还包括式A所示的七元环小檗碱类化合物的前药。术语“前药”包括其本身可以是具有生物学活性的或非活性的，当用适当的方法服用后，其在人体内进行代谢或化学反应而转变成式A的一类化合物，或式A的一个化合物所组成的盐或溶液。所述的前药包括(但不局限于)所述化合物的羧酸酯、碳酸酯、磷酸酯、硝酸酯、硫酸酯、砜酯、亚砜酯、氨基化合物、氨基甲酸盐、偶氮化合物、磷酰胺、葡萄糖苷、醚、乙缩醛等形式。
制备方法
下面更具体地描述本发明式A结构化合物的制备方法，但这些具体方法不对 本发明构成任何限制。本发明化合物还可以任选将在本说明书中描述的或本领域已知的各种合成方法组合起来而方便地制得，这样的组合可由本发明所属领域的技术人员容易地进行。
通常，在制备流程中，各反应通常在惰性溶剂中，在室温至回流温度(如0℃～80℃，优选0℃～50℃)下进行。反应时间通常为0.1小时-60小时，较佳地为0.5-48小时。
以式II和式III化合物为例，下面的通用制备路线可以用于合成本发明式A1结构的化合物：
Figure PCTCN2016076083-appb-000080
式中，R1、R2、R3、W、Q的定义如上所述。
具体地，化合物I的七元环双键被催化氢化(如钯碳/氢气或钯碳/甲酸铵等)还原得到化合物II，然后再参照与实施例2-18中类似的方法由化合物II得到不同化合物III。
对于化合物I，可用例如中国专利申请号201010537594.1的方法或类似方法进行制备或购买获得，例如实施例1中作为原料的化合物1。
药物组合物和施用方法
由于本发明化合物具有优异的抗肿瘤活性，因此本发明化合物及其各种晶型，药学上可接受的无机或有机盐，水合物或溶剂合物，以及含有本发明化合物为主要活性成分的药物组合物可用于治疗、预防以及缓解与肿瘤相关的疾病。
本发明的药物组合物包含安全有效量范围内的本发明化合物或其药理上可接受的盐及药理上可以接受的赋形剂或载体。其中“安全有效量”指的是：化合物的量足以明显改善病情，而不至于产生严重的副作用。通常，药物组合物含有1-2000mg本发明化合物/剂，更佳地，含有10-1000mg本发明化合物/剂。较佳地，所述的“一剂”为一个胶囊或药片。
“药学上可以接受的载体”指的是：一种或多种相容性固体或液体填料或凝胶物质，它们适合于人使用，而且必须有足够的纯度和足够低的毒性。“相容性”在此指的是组合物中各组份能和本发明的化合物以及它们之间相互掺和，而不明显降低化合物的药效。药学上可以接受的载体部分例子有纤维素及其衍生物(如羧甲基纤维素钠、乙基纤维素钠、纤维素乙酸酯等)、明胶、滑石、固体润滑剂(如硬脂酸、硬脂酸镁)、硫酸钙、植物油(如豆油、芝麻油、花生油、橄榄油等)、多元醇(如丙二 醇、甘油、甘露醇、山梨醇等)、乳化剂(如吐温
Figure PCTCN2016076083-appb-000081
)、润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。
本发明化合物或药物组合物的施用方式没有特别限制，代表性的施用方式包括(但并不限于)：口服、瘤内、直肠、肠胃外(静脉内、肌肉内或皮下)、和局部给药。
用于口服给药的固体剂型包括胶囊剂、片剂、丸剂、散剂和颗粒剂。在这些固体剂型中，活性化合物与至少一种常规惰性赋形剂(或载体)混合，如柠檬酸钠或磷酸二钙，或与下述成分混合：(a)填料或增容剂，例如，淀粉、乳糖、蔗糖、葡萄糖、甘露醇和硅酸；(b)粘合剂，例如，羟甲基纤维素、藻酸盐、明胶、聚乙烯基吡咯烷酮、蔗糖和阿拉伯胶；(c)保湿剂，例如，甘油；(d)崩解剂，例如，琼脂、碳酸钙、马铃薯淀粉或木薯淀粉、藻酸、某些复合硅酸盐、和碳酸钠；(e)缓溶剂，例如石蜡；(f)吸收加速剂，例如，季胺化合物；(g)润湿剂，例如鲸蜡醇和单硬脂酸甘油酯；(h)吸附剂，例如，高岭土；和(i)润滑剂，例如，滑石、硬脂酸钙、硬脂酸镁、固体聚乙二醇、十二烷基硫酸钠，或其混合物。胶囊剂、片剂和丸剂中，剂型也可包含缓冲剂。
固体剂型如片剂、糖丸、胶囊剂、丸剂和颗粒剂可采用包衣和壳材制备，如肠衣和其它本领域公知的材料。它们可包含不透明剂，并且，这种组合物中活性化合物或化合物的释放可以延迟的方式在消化道内的某一部分中释放。可采用的包埋组分的实例是聚合物质和蜡类物质。必要时，活性化合物也可与上述赋形剂中的一种或多种形成微胶囊形式。
用于口服给药的液体剂型包括药学上可接受的乳液、溶液、悬浮液、糖浆或酊剂。除了活性化合物外，液体剂型可包含本领域中常规采用的惰性稀释剂，如水或其它溶剂，增溶剂和乳化剂，例知，乙醇、异丙醇、碳酸乙酯、乙酸乙酯、丙二醇、1,3-丁二醇、二甲基甲酰胺以及油，特别是棉籽油、花生油、玉米胚油、橄榄油、蓖麻油和芝麻油或这些物质的混合物等。
除了这些惰性稀释剂外，组合物也可包含助剂，如润湿剂、乳化剂和悬浮剂、甜味剂、矫味剂和香料。
除了活性化合物外，悬浮液可包含悬浮剂，例如，乙氧基化异十八烷醇、聚氧乙烯山梨醇和脱水山梨醇酯、微晶纤维素、甲醇铝和琼脂或这些物质的混合物等。
用于肠胃外注射的组合物可包含生理上可接受的无菌含水或无水溶液、分散液、悬浮液或乳液，和用于重新溶解成无菌的可注射溶液或分散液的无菌粉末。适宜的含水和非水载体、稀释剂、溶剂或赋形剂包括水、乙醇、多元醇及其适宜的混合物。
用于局部给药的本发明化合物的剂型包括软膏剂、散剂、贴剂、喷射剂和吸入剂。活性成分在无菌条件下与生理上可接受的载体及任何防腐剂、缓冲剂，或必要时可能需要的推进剂一起混合。
本发明化合物可以单独给药，或者与其他药学上可接受的其他化合物(如抗肿瘤药物)联合给药。
使用药物组合物时，是将安全有效量的本发明化合物适用于需要治疗的哺乳动物(如人)，其中施用时剂量为药学上认为的有效给药剂量，对于60kg体重的人而言，日给药剂量通常为1～2000mg，优选50～1000mg。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。
本发明的主要优点在于：
(1)本发明化合物具有优异的抑制肿瘤细胞的活性，不仅可在体外显著抑制肿瘤细胞，而且优选化合物的IC50甚至<2nM。
(2)本发明化合物具有广谱的抗肿瘤活性。
(3)本发明化合物对于某些肿瘤细胞还具有选择性的肿瘤细胞抑制活性。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件如Sambrook等人，分子克隆：实验室手册(New York:Cold Spring Harbor Laboratory Press,1989)中所述的条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。
除非另行定义，文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。此外，任何与所记载内容相似或均等的方法及材料皆可应用于本发明方法中。文中所述的较佳实施方法与材料仅作示范之用。
实施例1
化合物2的制备
Figure PCTCN2016076083-appb-000082
方法一：化合物1(具体制备参考中国专利申请号201010537594.1的方法)溶于甲醇，加入催化量的10％钯碳，通入氢气，室温反应，至原料反应完全。反应液硅藻土过滤，滤液减压蒸干，得到白色固体2，产率95％。
方法二：化合物1(1.0eq.)溶于甲醇，加入催化量的10％钯碳，再加入甲酸铵(10当量)，加热回流反应，至原料反应完全。反应液硅藻土过滤，滤液减压蒸干，硅胶柱分离(PE/EA＝2:1)，得到白色固体2，产率90％。
1H NMR(400MHz,CDCl3)δ7.39–7.32(m,1H),7.28(d,J＝3.1Hz,1H),7.02 (s,1H),6.74(s,1H),6.40(s,1H),6.03(d,J＝1.4Hz,2H),5.14–5.01(m,1H),4.04(s,3H),3.97(s,3H),3.24–3.08(m,1H),2.68–2.51(m,2H),2.48–2.33(m,1H),1.97–1.82(m,1H).13C NMR(126MHz,CDCl3)δ160.10,151.50,149.47,148.37,146.44,142.41,132.38,128.74,122.24,119.47,118.90,109.18,108.30,105.57,101.37,61.65,56.90,41.60,30.00,28.77.HRMS(EI)计算值C21H19NO5365.1263[M]+,测量值365.1262.
实施例2
化合物3a-3d的制备
Figure PCTCN2016076083-appb-000083
3a的制备：化合物2(1.0eq.)溶于四氯化碳，氮气保护，加入NBS(氮溴代丁二酰亚胺，1.2eq.)，升温至50℃反应5小时，过滤，四氯化碳洗涤，滤液减压蒸干，硅胶柱分离(PE/EA＝2:1)，得到白色固体3a，产率80％。
1H NMR(400MHz,CDCl3)δ7.94–7.82(d,J＝9.1Hz,1H),7.47–7.38(d,J＝9.0Hz,1H),7.11(s,1H),6.76(s,1H),6.08(s,1H),6.05(s,1H),5.17–5.03(m,1H),4.03(s,3H),4.00(s,3H),3.05–2.94(m,1H),2.67–2.49(m,2H),2.39–2.27(m,1H),1.84–1.72(m,1H).13C NMR(126MHz,CDCl3)δ159.09,152.24,149.23,148.72,145.52,139.44,133.28,130.74,127.08,123.21,120.06,118.32,111.96,108.53,101.47,99.65,77.24,61.60,56.65,42.41,29.47,28.66.MS(ESI)m/z[M+H]+444.1.
3b的制备：化合物2(1.0eq.)溶于四氯化碳，氮气保护，加入NCS(氮氯代丁二酰亚胺，1.2eq.)，升温至80℃反应5小时，过滤，四氯化碳洗涤，滤液减压蒸干，硅胶柱分离(PE/EA＝2:1)，得到白色固体3b，产率83％。
1H NMR(400MHz,CDCl3)δ7.89–7.79(d,J＝9.0Hz,1H),7.50–7.39(d,J＝9.1Hz,1H),7.12(s,1H),6.77(s,1H),6.14–6.01(m,2H),5.13–5.01(m,1H),4.03(s,3H),4.00(s,3H),3.09–2.93(m,1H),2.72–2.46(m,2H),2.43–2.26(m,1H),1.85–1.75(m,1H).13C NMR(126MHz,CDCl3)δ158.77,152.30,149.39,148.64,145.60,137.69,133.40,129.81,125.26,120.19,120.01,118.34,111.47,108.66,101.42,61.55,56.63,42.09,29.50,28.60.HRMS(EI)计算值C21H18ClNO5399.0874[M]+,测量值399.0882.
3c的制备：化合物2(1.0eq.)溶于四氯化碳，氮气保护，加入NIS(氮碘代丁二酰亚胺，1.2eq.)，升温至80℃反应5小时，过滤，四氯化碳洗涤，滤液减压蒸干，硅胶柱分离(PE/EA＝2:1)，得到棕色固体3c，产率30％。
1H NMR(400MHz,CDCl3)δ7.86–7.78(d,J＝9.0Hz,1H),7.42–7.35(d,J＝9.1Hz,1H),7.04(s,1H),6.73(s,1H),6.07(s,1H),6.03(s,1H),5.20–5.10(m,1H),4.02(s,3H),3.98(s,3H),3.05–2.92(m,1H),2.65–2.47(m,2H),2.36–2.21(m,1H),1.82–1.68(m,1H).13C NMR(126MHz,CDCl3)δ159.41,152.00,148.84,145.38,143.00,133.00,132.58,130.16,128.63,119.45,118.27,112.57,108.30,101.43,77.25,75.75,61.52,56.59,42.91,29.32.HRMS(EI)计算值C21H18INO5491.0230[M]+,测量值491.0229.
3d的制备：化合物2(1.0eq.)溶于四氯化碳，氮气保护，加入NFSI(氮氟代双苯磺酰胺，1.2eq.)，升温至80℃反应5小时，过滤，四氯化碳洗涤，滤液减压蒸干，硅胶柱分离(PE/EA＝2:1)，得到白色固体3c，产率61％。
1H NMR(400MHz,DMSO)δ8.32(s,1H),7.39(m,3H),7.16(s,1H),6.84(s,1H),6.05(s,2H),4.13(s,1H),3.95(s,3H),3.85(s,3H),2.96(s,2H),2.23–1.93(m,1H),1.66–1.40(m,1H).13C NMR(126MHz,DMSO)δ160.20,156.45,149.20,148.12,146.07,133.37,128.25,120.53,116.13,110.57,110.36,101.73,79.64,61.37,56.58,30.07,25.79.HRMS(EI)计算值C21H18FNO5383.1169[M]+,测量值383.1172.
实施例3
化合物4的制备
Figure PCTCN2016076083-appb-000084
化合物3a(1.0eq.)溶于二氯甲烷，加入10％溶液体积的浓盐酸，搅拌反应2小时，反应液加水稀释，二氯甲烷萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，得到白色固体4，产率93％。
1H NMR(400MHz,CDCl3)δ13.48(s,1H),7.52–7.41(m,1H),7.41–7.32(m,1H),7.12(s,1H),6.75(s,1H),6.12(s,1H),6.05(s,1H),5.11–4.94(m,1H),4.01(s,3H),3.12–2.96(m,1H),2.73–2.58(m,1H),2.59–2.47(m,1H),2.36–2.18(m,1H),1.88–1.74(m,1H).13C NMR(126MHz,CDCl3)δ164.60,150.46,148.95,145.99, 145.69,138.14,133.14,129.22,126.35,118.31,116.41,112.09,110.97,108.57,102.76,101.57,77.24,56.53,41.90,29.34,28.55.MS(ESI)m/z[M+H]+430.2.
实施例4
化合物5a-5f的制备
Figure PCTCN2016076083-appb-000085
5a的制备：化合物4(1.0eq.)溶于丙酮，加入无水碳酸钾(2.0eq.)，溴乙酸甲酯(2.0eq.)，氮气保护，升温至60℃反应5小时，反应液加水稀释，过滤，水洗，干燥，得到微黄白色固体5a，产率77％。
1H NMR(400MHz,CDCl3)δ7.93–7.86(d,J＝9.1Hz,1H),7.47–7.35(d,J＝9.0Hz,1H),7.10(s,1H),6.75(s,1H),6.15–5.96(m,2H),5.13–5.01(m,1H),4.89–4.68(m,2H),3.97(s,3H),3.88(s,3H),3.08–2.87(m,1H),2.68–2.45(m,2H),2.41–2.24(m,1H),1.86–1.70(m,1H).13C NMR(126MHz,CDCl3)δ169.73,158.92,151.92,148.76,146.45,145.54,139.63,133.26,130.65,126.98,123.95,120.05,118.22,111.93,108.50,101.47,99.43,70.01,56.66,52.15,42.43,29.44,28.69.HRMS(EI)计算值C23H20BrNO7501.0423[M]+,测量值501.0408.
5b的制备：化合物4(1.0eq.)溶于DMF(二甲基甲酰胺)，冰浴降温，氮气保护，加入氢化钠(2.0eq.)，搅拌反应10分钟，再加入溴乙烷(2.0eq.)，四丁基碘化铵(0.1eq.)，然后升温至60℃反应5小时，反应液加水稀释，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(PE/EA＝1:1)，得到微黄白色固体5b，产率33％。
1H NMR(400MHz,CDCl3)δ7.89–7.83(d,J＝9.0Hz,1H),7.46–7.37(d,J＝9.0Hz,1H),7.11(s,1H),6.75(s,1H),6.12–6.01(m,2H),5.14–5.04(m,1H),4.28–4.10(m,2H),3.98(s,3H),3.05–2.92(m,1H),2.67–2.47(m,2H),2.40–2.27(m,1H),1.84–1.70(m,1H),1.58–1.53(m,3H).13C NMR(126MHz,CDCl3)δ159.11,152.49,148.63,145.51,139.39,133.27,130.83,127.18,122.98,120.27,118.49,111.96,108.48,101.44,99.63,77.22,70.06,56.77,42.35,29.47,28.71,15.77.HRMS (EI)计算值C22H20BrNO5457.0525[M]+,测量值457.0533.
5c的制备：化合物4(1.0eq.)溶于DMF，冰浴降温，氮气保护，加入氢化钠(2.0eq.)，搅拌反应10分钟，再加入烯丙基溴(2.0eq.)，搅拌反应5小时。反应液加水稀释，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(PE/EA＝5:1)，得到白色固体5c，产率54％。
1H NMR(400MHz,CDCl3)δ7.93–7.83(d,J＝9.1Hz,1H),7.47–7.38(d,J＝9.0Hz,1H),7.11(s,1H),6.75(s,1H),6.41–6.24(m,1H),6.14–6.01(m,2H),5.51–5.39(d,J＝17.2Hz,1H),5.32–5.23(m,1H),5.16–5.05(m,1H),4.75–4.61(d,J＝6.0Hz,2H),3.98(s,3H),3.05–2.92(m,1H),2.67–2.49(m,2H),2.40–2.24(m,1H),1.84–1.71(m,1H).13C NMR(126MHz,CDCl3)δ159.05,152.39,148.70,148.05,145.51,139.45,134.65,133.28,130.80,127.14,123.20,120.29,118.40,117.66,111.95,108.50,101.45,99.58,75.21,56.73,42.37,29.48,28.72.HRMS(EI)计算值C23H20BrNO5469.0525[M]+,测量值469.0506.
5d的制备：化合物4(1.0eq.)溶于DMF，冰浴降温，氮气保护，加入氢化钠(2.0eq.)，搅拌反应10分钟，再加入异戊烯基溴(2.0eq.)，搅拌反应5小时。反应液加水稀释，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(PE/EA＝4:1)，得到白色固体5d，产率50％。
1H NMR(400MHz,CDCl3)δ7.91–7.79(d,J＝9.0Hz,1H),7.45–7.35(d,J＝9.3Hz,1H),7.16–7.06(s,1H),6.77–6.71(s,1H),6.13–5.97(m,2H),5.85–5.72(m,1H),5.18–5.00(m,1H),4.73–4.58(m,2H),3.98(s,3H),3.07–2.93(m,1H),2.69–2.49(m,2H),2.41–2.21(m,1H),1.81(s,3H),1.78(s,3H).13C NMR(126MHz,CDCl3)δ159.05,152.56,148.66,148.37,145.49,139.39,137.68,133.28,130.77,127.21,122.98,121.04,120.43,118.29,111.96,108.47,101.43,99.53,70.74,56.69,42.29,29.48,28.74,25.88,18.10.HRMS(EI)计算值C25H24BrNO5497.0838[M]+,测量值497.0840.
5e的制备：化合物4(1.0eq.)溶于DMF，冰浴降温，氮气保护，加入氢化钠(2.0eq.)，搅拌反应10分钟，再加入二氟苄溴(2.0eq.)，搅拌反应5小时。反应液加水稀释，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(PE/EA＝5:1)，得到白色固体5e，产率54％。
1H NMR(400MHz,CDCl3)δ8.01–7.83(m,1H),7.76–7.56(m,1H),7.53–7.34(m,2H),7.24–7.07(m,2H),6.76(s,1H),6.16–5.97(m,2H),5.21–4.98(m,3H),3.97(s,3H),3.11–2.90(m,1H),2.71–2.48(m,2H),2.44–2.21(m,1H),1.89–1.70(m,1H).13C NMR(126MHz,CDCl3)δ159.01,152.28,148.76,147.42,145.55,139.59,133.26,130.78,127.07,124.50,123.69,120.37,118.22,117.79,116.82,111.94,108.51,101.48,99.54,74.19,56.61,42.40,29.47,28.76.HRMS(EI)计算值 C27H20BrF2NO5555.0493[M]+,测量值555.0491.
5f的制备：化合物4(1.0eq.)溶于DMF，冰浴降温，氮气保护，加入氢化钠(2.0eq.)，搅拌反应10分钟，再加入1-溴-2-(2-甲氧基乙氧基)乙烷(2.0eq.)，四丁基碘化铵(0.1eq.)，然后升温至60℃反应5小时，反应液加水稀释，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(PE/EA＝3:1)，得到微黄白色固体5f，产率19％。
1H NMR(400MHz,CDCl3)δ7.92–7.82(d,J＝9.0Hz,1H),7.45–7.36(d,J＝9.1Hz,1H),7.10(s,1H),6.75(s,1H),6.14–6.00(m,2H),5.14–5.02(m,1H),4.37–4.26(t,J＝5.3Hz,1H),4.07–4.00(t,J＝5.3Hz,2H),3.98(s,3H),3.84–3.76(dd,J＝5.6,3.8Hz,2H),3.64–3.56(dd,J＝5.8,3.7Hz,2H),3.41(s,3H),3.07–2.91(m,1H),2.69–2.46(m,2H),2.39–2.27(m,1H),1.84–1.72(m,1H).13C NMR(126MHz,CDCl3)δ158.55,151.84,148.24,147.89,145.06,138.93,132.79,130.29,126.68,122.71,119.62,117.96,111.48,108.02,100.97,99.14,72.67,71.54,70.19,69.98,58.57,56.24,41.88,29.24,28.99,28.23.HRMS(EI)计算值C25H26BrNO7531.0893[M]+,测量值531.0895.
实施例5
化合物6的制备
Figure PCTCN2016076083-appb-000086
化合物5a(1.0eq.)溶于甲醇，加入氢氧化钠(4.0eq.)水溶液，升温至80℃反应3小时。反应液减压蒸除甲醇，加水稀释，乙酸乙酯萃取除杂，水相再用稀盐酸调PH至3左右，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，得到微黄白色固体6，产率98％。
1H NMR(400MHz,CDCl3)δ8.00–7.91(d,J＝9.1Hz,1H),7.56–7.46(d,J＝9.1Hz,1H),7.10(s,1H),6.77(s,1H),6.12–6.02(m,2H),5.21–5.11(m,1H),5.02–4.79(m,2H),4.02(s,3H),3.16–2.99(m,1H),2.72–2.61(m,1H),2.56–2.43(m,1H),2.40–2.26(m,1H),1.90–1.75(m,1H).13C NMR(126MHz,CDCl3)δ171.27,160.32,150.62,149.10,146.99,145.72,139.53,133.24,130.64,126.36,123.80,118.94,117.14,111.88,108.64,101.56,72.23,56.51,43.44,29.35,28.56.HRMS(EI)计算值C22H18BrNO7487.0267[M]+,测量值487.0285.
实施例6
化合物7的制备
Figure PCTCN2016076083-appb-000087
化合物6(1.0eq.)溶于二氯甲烷，氮气保护，加入EDCI(1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐，1.2eq.)，DIPEA(二异丙基乙基胺，1.2eq.)，HOBt(1-羟基苯并三唑，0.1eq.)，N-甲基哌嗪(1.2eq.)，搅拌反应10小时。反应液加水稀释，二氯甲烷萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(CH2Cl2/MeOH＝15:1)，得到白色固体7，产率35％。
1H NMR(400MHz,CDCl3)δ8.00–7.85(d,J＝9.0Hz,1H),7.48–7.36(d,J＝9.1Hz,1H),7.09(s,1H),6.75(s,1H),6.16–5.96(d,J＝11.7Hz,2H),5.12–5.00(m,1H),4.89–4.67(m,2H),3.96(s,5H),3.76(s,2H),3.06–2.91(m,1H),2.68–2.45(m,6H),2.38(s,3H),2.32–2.25(m,1H),1.89–1.68(m,1H).13C NMR(126MHz,CDCl3)δ167.05,158.84,152.03,148.76,147.11,145.55,139.54,133.23,130.65,127.01,123.96,119.99,118.15,111.93,108.51,101.47,99.56,73.33,56.61,55.24,54.62,46.01,45.28,42.41,41.74,29.44,28.7.HRMS(EI)计算值C27H28BrN3O6569.1161[M]+,测量值569.1135.
实施例7
化合物8的制备
Figure PCTCN2016076083-appb-000088
化合物6(1.0eq.)溶于四氢呋喃，氮气保护，冰浴降温，加入硼烷四氢呋喃溶液(2.0eq.)，搅拌反应4小时。反应液加饱和氯化铵水溶液淬灭，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(CH2Cl2/MeOH＝20:1)，得到白色固体8，产率60％。
1H NMR(400MHz,CDCl3)δ7.96–7.85(d,J＝9.0Hz,1H),7.50–7.39(d,J＝9.1Hz,1H),7.10(s,1H),6.75(s,1H),6.15–6.01(d,J＝12.2Hz,2H),5.79(s,1H), 5.21–5.04(m,1H),4.42(brs,2H),3.99(s,3H),3.92(s,2H),3.09–2.93m,1H),2.69–2.57(m,1H),2.57–2.41(m,1H),2.40–2.22m,1H),1.90–1.70(m,1H).13C NMR(101MHz,CDCl3)δ159.61,151.43,148.39,147.97,145.12,138.99,132.85,130.43,126.41,122.57,118.63,118.17,111.47,108.10,101.05,100.06,61.10,56.07,42.42,28.98,28.14.HRMS(EI)计算值C22H20BrNO6473.0474[M]+,测量值473.0447.
实施例8
化合物9的制备
Figure PCTCN2016076083-appb-000089
化合物8(1.0eq.)溶于二氯甲烷，氮气保护，加入对甲苯磺酰氯(1.2eq.)，DMAP(二甲氨基吡啶，0.2eq.)，三乙胺(1.2eq.)，搅拌反应3小时。反应液加水稀释，二氯甲烷萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(PE/EA＝3:1)，得到微黄色油状物9，产率72％。
1H NMR(400MHz,CDCl3)δ7.91–7.86(d,J＝9.1Hz,1H),7.86–7.80(d,J＝7.9Hz,2H),7.43–7.38(d,J＝9.0Hz,1H),7.38–7.31(d,J＝7.9Hz,2H),7.10(s,1H),6.75(s,1H),6.10–6.02(m,2H),5.08–4.94(m,1H),4.56–4.47(t,J＝5.3Hz,2H),4.424.33(t,J＝5.1Hz,2H),3.96s,3H),3.022.92(m,1H),2.66–2.57(m,1H),2.57–2.49(m,1H),2.34–2.20(m,1H),1.84–1.71(m,1H).13C NMR(101MHz,CDCl3)δ158.48,151.74,148.31,147.02,145.10,144.14,139.00,132.74,130.26,129.29,127.57,126.56,123.24,119.46,118.07,111.48,108.05,101.02,99.19,70.89,68.99,56.28,41.94,28.98,28.20,21.20.HRMS(EI)计算值C29H26BrNO8S 627.0563[M]+,测量值627.0541.
实施例9
化合物10a-10c的制备
Figure PCTCN2016076083-appb-000090
10a的制备：化合物9(1.0eq.)溶于四氢呋喃，氮气保护，加入N-甲基哌嗪(1.2eq.)，搅拌反应10小时。反应液减压蒸除四氢呋喃，加水稀释，二氯甲烷萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(CH2Cl2/MeOH＝20:1)，得到微黄色固体10a，产率30％。
1H NMR(400MHz,CDCl3)δ7.98–7.80(d,J＝9.0Hz,1H),7.52–7.35(d,J＝9.1Hz,1H),7.07(s,1H),6.73(s,1H),6.19–5.97(m,2H),5.10–4.96(m,1H),4.31(s,2H),3.97(s,3H),3.55–2.85(m,11H),2.82–2.42(m,5H),2.34–2.19(m,1H),1.86–1.69(m,1H).13C NMR(101MHz,CDCl3)δ158.60,151.63,148.34,145.11,138.99,132.69,130.20,126.47,123.30,119.44,117.65,111.45,108.06,101.04,99.36,68.95,56.83,56.14,52.70,50.14,43.71,41.99,28.94,28.21.HRMS(ESI)计算值C27H31BrN3O5556.1442[M+H]+,测量值556.1458.
10b的制备：化合物9(1.0eq.)溶于四氢呋喃，氮气保护，加入二甲胺盐酸盐(1.2eq.)，三乙胺(1.2eq.)，搅拌反应10小时。反应液减压蒸除四氢呋喃，加水稀释，二氯甲烷萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(CH2Cl2/MeOH＝20:1)，得到微黄色固体10b，产率33％。
1H NMR(400MHz,CDCl3)δ7.97–7.87(d,J＝9.0Hz,1H),7.48–7.39(d,J＝9.0Hz,1H),7.07(s,1H),6.73(s,1H),6.12–5.97(m,2H),5.08–4.94(dd,J＝13.4,5.4Hz,1H),4.63–4.44(t,J＝4.0Hz,2H),4.00(s,3H),3.69(s,2H),3.14(s,6H),3.07–2.93(m,1H),2.68–2.56(m,1H),2.57–2.43(m,1H),2.31–2.21(m,1H),1.87–1.69(m,1H).13C NMR(101MHz,CDCl3)δ158.72,151.43,148.44,145.47,145.17,139.04,132.65,130.13,126.27,123.94,118.96,117.56,111.43,108.08,101.08,99.73,67.25,56.95,56.01,43.02,42.18,28.90,28.19.HRMS(ESI)计算值C24H26BrN2O5501.1020[M+H]+,测量值501.1034.
10c的制备：化合物9(1.0eq.)溶于四氢呋喃，氮气保护，加入甲胺醇溶液(1.2eq.)，搅拌反应10小时。反应液减压蒸除四氢呋喃，加水稀释，二氯甲烷萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(CH2Cl2/MeOH＝20:1)，得到微黄色固体10c，产率69％。
1H NMR(400MHz,CDCl3)δ10.57–10.21(m,1H),8.00–7.89(d,J＝9.0Hz,1H),7.53–7.42(d,J＝9.0Hz,1H),7.09(s,1H),6.75(s,1H),6.15–5.99(d,J＝11.6Hz,2H),5.09–4.96(m,1H),4.71–4.52(m,2H),3.99(s,3H),3.42(s,2H),3.12–2.97(m,1H),2.88(s,3H),2.70–2.57(m,1H),2.56–2.42(m,1H),2.34–2.20(m,1H),1.85–1.77(m,1H).13C NMR(101MHz,CDCl3)δ159.88,151.15,148.60,145.97,145.25,138.94,132.72,130.34,126.00,123.63,118.21,117.67,111.45,108.14,101.13,68.94,55.94,49.17,42.82,33.14,28.88,28.13.HRMS(ESI)计算值C23H24BrN2O5487.0863[M+H]+,测量值487.0854.
实施例10
化合物11的制备
Figure PCTCN2016076083-appb-000091
化合物3a(1.0eq.)溶于DMF，加入三苯基磷(0.1eq.)，无水碳酸钾(2.0eq.)，丙烯酸甲酯(2.0eq.)，氮气保护，加入醋酸钯(0.05eq.)，升温至100℃反应10小时。反应液加水稀释，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(PE/EA＝2:1)，得到微黄色固体11，产率78％。
1H NMR(400MHz,CDCl3)δ7.78–7.71(d,J＝9.1Hz,1H),7.65–7.58(d,J＝16.2Hz,1H),7.40–7.34(d,J＝9.1Hz,1H),6.81–6.71(d,J＝3.2Hz,2H),6.11–5.97(m,3H),5.17–5.06(m,1H),4.04(s,3H),3.99(s,3H),3.78(s,3H),3.08–2.92(m,1H),2.70–2.48(m,2H),2.38–2.27(m,1H),1.90–1.76(m,1H).13C NMR(126MHz,CDCl3)δ167.23,159.11,151.81,149.68,148.86,145.90,141.63,133.48,130.13,125.69,122.68,120.23,120.05,118.28,111.64,109.93,108.77,101.46,77.19,61.57,56.67,51.63,41.27,29.54,29.00.HRMS(EI)计算值C25H23NO7449.1475[M]+,测量值449.1476.
实施例11
化合物12的制备
Figure PCTCN2016076083-appb-000092
化合物11(1.0eq.)溶于甲醇，加入催化量的10％钯碳，再加入甲酸铵(10当量)，加热回流反应，至原料反应完全。反应液硅藻土过滤，滤液减压蒸干，硅胶柱分离(PE/EA＝2:1)，得到白色固体12，产率98％。
1H NMR(400MHz,CDCl3)δ7.55–7.45(m,1H),7.45–7.36(m,1H),6.83(s,1H),6.76(s,1H),6.14–5.98(m,2H),5.12–4.98(m,1H),4.04(s,3H),3.98(s,3H),3.64(s,3H),3.23–3.10(m,1H),3.04–2.84(m,2H),2.65–2.56(m,1H),2.55–2.36(m,3H),2.36–2.22(m,1H),1.80–1.72(m,1H).13C NMR(126MHz,CDCl3)δ173.10,159.28,151.52,149.92,148.43,145.93,139.09,133.49,131.43,126.40,120.69,119.08,118.38,111.07,109.85,108.78,101.38,77.19,61.48,56.73,51.67,41.18,34.31,29.54,28.62,23.79.HRMS(EI)计算值C25H25NO7451.1631[M]+,测量值451.1637.
实施例12
化合物13的制备
Figure PCTCN2016076083-appb-000093
化合物11(1.0eq.)溶于甲醇，加入氢氧化钠(4.0eq.)水溶液，升温至80℃反应3小时。反应液减压蒸除甲醇，加水稀释，乙酸乙酯萃取除杂，水相再用稀盐酸调PH至3左右，过滤，水洗，干燥，得到微黄白色固体13，产率69％。
1H NMR(400MHz,CDCl3)δ7.80–7.74(d,J＝9.1Hz,1H),7.74–7.66(m,1H),7.41–7.35(d,J＝9.1Hz,1H),6.80–6.73(m,2H),6.11–6.00(m,3H),5.22–5.03(m,1H),4.04(s,3H),3.99(s,3H),3.07–2.95(m,1H),2.69–2.50(m,2H),2.40–2.27(m,1H),1.90–1.78(m,1H).13C NMR(126MHz,CDCl3)δ170.92,159.13,151.88,149.70,149.00,145.95,143.80,142.31,133.55,129.89,125.49,121.56,120.11,118.34,111.60,109.73,108.85,101.52,77.19,61.58,56.65,41.37,29.54,29.05.HRMS(EI)计算值C24H21NO7435.1318[M]+,测量值435.1292.
实施例13
化合物14的制备
Figure PCTCN2016076083-appb-000094
化合物12(1.0eq.)溶于甲醇，加入氢氧化钠(4.0eq.)水溶液，升温至80℃反应3小时。反应液减压蒸除甲醇，加水稀释，乙酸乙酯萃取除杂，水相再用稀盐酸调PH至3左右，过滤，水洗，干燥，得到白色固体14，产率83％。
1H NMR(400MHz,CDCl3)δ7.55–7.44(m,1H),7.44–7.36(m,1H),6.83(s,1H),6.74(s,1H),6.11–5.99(m,2H),5.12–4.98(m,1H),4.03(s,3H),3.98(s,3H),3.25–3.11(m,1H),3.05–2.84(m,2H),2.65–2.37(m,4H),2.37–2.20(m,1H),1.82–1.66(m,1H).13C NMR(126MHz,CDCl3)δ177.22,159.33,151.54,149.91,148.47,145.94,139.18,133.54,131.33,126.29,120.62,119.00,118.44,110.98,109.78,108.81,101.40,61.49,56.71,41.28,34.08,29.53,28.62,23.60.HRMS(EI)计算值C24H23NO7437.1475[M]+,测量值437.1465.
实施例14
化合物15的制备
Figure PCTCN2016076083-appb-000095
化合物13(1.0eq.)溶于二氯甲烷，氮气保护，加入EDCI(1.2eq.)，DIPEA(1.2eq.)，HOBt(0.1eq.)，N-甲基哌嗪(1.2eq.)，搅拌反应10小时。反应液加水稀释，二氯甲烷萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(CH2Cl2/MeOH＝30:1)，得到黄色固体15，产率52％。
1H NMR(400MHz,CDCl3)δ7.73–7.54(m,2H),7.37–7.24(m,1H),6.73(s,2H),6.21–5.86(m,3H),5.21–4.99(m,1H),4.07–3.91(m,6H),3.87–3.68(m,1H),3.67–3.48(m,1H),3.32–3.05(m,2H),3.04–2.91(m,1H),2.90–2.68(m,1H),2.68–2.53(m,2H),2.54–2.09(m,9H),1.94–1.64(m,1H).13C NMR(101MHz,CDCl3)δ165.24,159.21,151.84,149.53,148.53,145.96,140.07,138.26,133.35,130.78, 126.71,124.63,120.23,119.86,118.30,111.69,111.01,108.46,101.40,77.30,61.54,56.66,45.83,41.17,29.60,28.79.HRMS(EI)计算值C29H31N3O6517.2213[M]+,测量值517.2206.
实施例15
化合物16的制备
Figure PCTCN2016076083-appb-000096
化合物14(1.0eq.)溶于二氯甲烷，氮气保护，加入EDCI(1.2eq.)，DIPEA(1.2eq.)，HOBt(0.1eq.)，N-甲基哌嗪(1.2eq.)，搅拌反应10小时。反应液加水稀释，二氯甲烷萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(CH2Cl2/MeOH＝20:1)，得到微黄白色固体16，产率77％。
1H NMR(400MHz,CDCl3)δ7.61–7.48(d,J＝9.0Hz,1H),7.44–7.35(d,J＝9.0Hz,1H),6.84(s,1H),6.74(s,1H),6.14–5.92(m,2H),5.12–4.95(m,1H),4.01(s,3H),3.96(s,3H),3.71–3.50(m,2H),3.29–3.15(m,2H),3.15–3.06(m,1H),3.03–2.83(m,2H),2.64–2.52(m,1H),2.52–2.41(m,2H),2.40–2.33(m,2H),2.33–2.19(m,7H),1.82–1.63(m,1H).13C NMR(101MHz,CDCl3)δ170.36,159.22,151.55,149.81,148.43,145.99,138.74,133.39,131.41,126.51,120.64,119.32,118.42,111.66,110.02,108.75,101.44,77.28,76.87,61.47,56.69,54.91,54.47,45.82,45.05,41.17,33.60,29.58,28.61,24.24.HRMS(EI)计算值C29H33N3O6519.2369[M]+,测量值519.2370.
实施例16
化合物17的制备
Figure PCTCN2016076083-appb-000097
N,N-二甲基甲酰胺溶液，氮气保护，冰浴降温，加入三氯氧磷(10.0eq.)，搅拌反应2小时，加入化合物2(1.0eq.)的DMF溶液，搅拌反应10小时。反应液加水稀释，10％氢氧化钠溶液调PH至碱性，水稀释，过滤，固体水洗，干燥，得到 黄色固体产物17，产率95％。
1H NMR(400MHz,CDCl3)δ9.62(s,1H),9.11–9.00(d,J＝9.2Hz,1H),7.49–7.37(d,J＝9.2Hz,1H),6.88(s,1H),6.83(s,1H),6.09(s,2H),5.11–5.00(dd,J＝13.3,5.9Hz,1H),4.02(s,3H),4.00(s,3H),3.15–2.99(td,J＝13.2,4.9Hz,1H),2.79–2.57(m,2H),2.51–2.37(dt,J＝13.2,6.4Hz,1H),2.03–1.89(tt,J＝12.6,6.2Hz,1H).13C NMR(101MHz,CDCl3)δ190.63,165.17,153.38,149.77,145.80,145.57,133.67,125.84,121.93,118.61,114.67,113.47,112.61,110.31,108.79,101.53,76.53,55.97,40.57,29.00.HRMS(EI)计算值C22H19NO6393.1212[M]+,测量值393.1220.
实施例17
化合物18的制备
Figure PCTCN2016076083-appb-000098
化合物17(1.0eq.)溶于乙醇，冰浴降温，加入硼氢化钠(2.0eq.)，搅拌反应5小时。反应液加少量稀盐酸淬灭，减压蒸干甲醇，水稀释，过滤，稀盐酸洗，水洗，干燥，得到黄色固体18，产率78％。
1H NMR(400MHz,CDCl3)δ7.81–7.70(d,J＝9.0Hz,1H),7.45–7.34(d,J＝9.0Hz,1H),7.13(s,1H),6.75(s,1H),6.05(s,2H),5.12–4.96(m,1H),4.90–4.77(d,J＝11.8Hz,1H),4.55–4.41(d,J＝11.8Hz,1H),4.02(s,3H),3.97(s,3H),3.05–2.87(td,J＝13.1,4.8Hz,1H),2.67–2.56(m,1H),2.56–2.43(td,J＝12.9,7.2Hz,1H),2.41–2.24(dt,J＝13.3,6.7Hz,1H),1.83–1.75(m,1H).13C NMR(126MHz,CDCl3)δ150.96,134.81,121.20,120.07,111.77,110.17,102.84,78.71,78.46,78.20,62.92,61.34,58.17,42.93,31.12,30.17.HRMS(EI)计算值C22H21NO6395.1369[M]+,测量值395.1377.
实施例18
化合物19a-19b的制备
Figure PCTCN2016076083-appb-000099
19a的制备：化合物18(1.0eq.)溶于DMF，冰浴降温，氮气保护，加入氢化钠(2.0eq.)，搅拌反应10分钟，再加入碘甲烷(2.0eq.)，搅拌反应5小时。反应液加水淬灭，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(PE/EA＝1:1)，得到微黄白色固体19a，产率38％。
1H NMR(400MHz,CDCl3)δ7.63–7.56(d,J＝8.9Hz,1H),7.45–7.38(d,J＝9.0Hz,1H),7.08(s,1H),6.76(s,1H),6.13–5.99(m,2H),5.10–4.98(m,1H),4.51–4.42(m,1H),4.23–4.11(m,1H),4.01(s,3H),3.97(s,3H),3.40(s,3H),3.07–2.92(m,1H),2.68–2.46(m,2H),2.44–2.28(m,1H),1.90–1.75(m,1H).13C NMR(126MHz,CDCl3)δ159.78,151.59,149.47,148.60,145.84,142.14,133.43,132.27,126.16,120.18,119.86,118.80,110.40,109.63,108.68,101.39,69.57,61.53,57.84,56.83,41.61,29.72,28.81.HRMS(EI)计算值C23H23NO6409.1525[M]+,测量值409.1533.
19b的制备：化合物18(1.0eq.)溶于DMF，冰浴降温，氮气保护，加入氢化钠(2.0eq.)，搅拌反应10分钟，再加入异戊烯基溴(2.0eq.)，搅拌反应5小时。反应液加水淬灭，乙酸乙酯萃取，分离有机相，水洗，饱和氯化钠洗，无水硫酸钠干燥，减压蒸干，硅胶柱分离(PE/EA＝3:1)，得到微黄色固体19b，产率22％。
1H NMR(400MHz,CDCl3)δ7.69–7.58(d,J＝8.9Hz,1H),7.47–7.36(d,J＝8.9Hz,1H),7.16(s,1H),6.76(s,1H),6.17–5.98(m,2H),5.52–5.40(m,1H),5.12–4.98(m,1H),4.58–4.42(d,J＝10.4Hz,1H),4.27–4.16(d,J＝10.6Hz,1H),4.12–3.91(m,8H),3.06–2.92(m,1H),2.70–2.46(m,2H),2.43–2.27(m,1H),1.87–1.78(m,1H),1.77(s,3H),1.73(s,3H).13C NMR(101MHz,CDCl3)δ159.35,151.10,149.01,148.10,145.31,141.57,137.50,132.89,132.00,125.78,120.26,119.64,118.37,110.14,109.35,108.16,100.90,76.78,66.47,66.09,61.08,56.40,41.15,29.28,28.34,25.33,17.56.HRMS(EI)计算值C27H29NO6463.1995[M]+,测量值463.1999.
实验例19
本发明化合物对肿瘤细胞的活性影响。
实验方法：
采用常规的或常用的肿瘤细胞：肝癌细胞2株(BEL-7402，SMMC-7721)；胃癌细胞2株(SGC-7901，MKN-45)；肺癌细胞2株(A549，SPC-A4)；结肠癌细胞2株(HT29，SW-620，HCT116)，黑色素瘤细胞(A375)分别加入化合物处理72h，用磺酰罗丹明B(Sulforhodamine B，SRB)法或MTT法检测化合物的增殖生长抑制作用及其程度。
实验结果：
1、部分化合物的肿瘤细胞抑制活性(IC50，μM)
化合物 SPC-A4 SW-620 A375 HCT116 HT29
小檗碱(Berberine) 5.42 2.66 3.57 10 -
依托铂苷(VP-16) 0.94 1.01 - - -
索拉菲尼(Sorafenib) - - 12.7 8.6 3.67
1(即SM38) 0.54 0.73 0.59 4.87 -
2 0.01 0.06 0.04 0.67 0.69
3a <0.006 <0.006 0.02 0.4 0.08
3b <0.006 <0.006 0.08 0.99 0.25
3c <0.006 <0.006 0.097 1.17 -
5b 0.06 0.44 0.025 0.12 0.03
5c 0.07 0.19 0.126 0.71 0.09
8 0.03 0.21 0.284 2.36 0.62
活性实验1测试了本发明部分化合物对肺癌细胞株(SPC-A4)、结肠癌细胞株(SW-620，HCT116，HT29)和黑色素瘤细胞株(A375)的抑制活性，说明本发明化合物具有较强抑瘤活性，不仅优于天然产物小檗碱(Berberine)，部分化合物的活性甚至优于阳性对照药依托泊苷(VP-16)或索拉菲尼(Sorafenib)。
2、本发明化合物的肿瘤细胞抑制活性(5μM浓度抑制率％)
本发明实施例1-18中所制备的式A化合物，在5μM浓度下，对于BEL-7402、SMMC-7721、SPC-A4等多种肿瘤细胞表现出明显的抑制率％，抑制率大多为50％-90％。
以化合物2为例，其对一些常见肿瘤细胞株的抑制活性列于下表：
化合物 BEL-7402 SMMC-7721 SGC-7901 A549 SPC-A4 HT29 SW-620
2 57.608 51.479 38.613 51.099 88.846 68.634 77.073
结果表明，化合物2对肝癌细胞2株(BEL-7402，SMMC-7721)；胃癌细胞2株(SGC-7901，MKN-45)；肺癌细胞2株(A549，SPC-A4)；结肠癌细胞2株(HT29，SW-620)有优良的肿瘤抑制活性。
上述实施例说明，本发明的化合物对各种肿瘤细胞有不同程度的抑制活性，证明本发明的化合物有潜在的抗肿瘤活性。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (13)

  1. 一种式A所示的七元环小檗碱类似物、其药学上可接受的盐，其晶型、其水合物、其溶剂化物、或其前药，
    Figure PCTCN2016076083-appb-100001
    式中，
    Figure PCTCN2016076083-appb-100002
    代表双键或单键，并且
    Figure PCTCN2016076083-appb-100003
    中的两个
    Figure PCTCN2016076083-appb-100004
    不同时为双键，或者
    Figure PCTCN2016076083-appb-100005
    中的两个
    Figure PCTCN2016076083-appb-100006
    不同时为双键；
    Figure PCTCN2016076083-appb-100007
    分别表示芳基或5-6元杂芳基，其中当所述的杂芳基为6元环时，具有1-3个N原子；为5元环时，具有1-3个N或O或S杂原子；
    R1为位于
    Figure PCTCN2016076083-appb-100008
    的1-4个相同或不同的取代基，而R2为位于
    Figure PCTCN2016076083-appb-100009
    的1-4个相同或不同的取代基，并且各R1和R2独立地选自下组：氢、羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基或卤素；或者两个R1可与相邻的碳原子或两个R2可与相邻的碳原子共同形成含有0-3个选自O、S、N的杂原子的5-7元环；
    R3选自下组：氢、氧、硫、羟基、氨基、巯基、烷基、链烯基、链炔基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基、芳烷基、C1-C6醛基或卤素；
    R4选自下组：氢、氧、硫、羟基、氨基、巯基、烷基、链烯基、链炔基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基、芳烷基、C1-C6醛基或卤素；
    并且当R4代表氢且R3代表氢或氧，且D为苯环时，R2不为氢。2.如权利要求1所述的七元环小檗碱类似物及其药学上可接受的盐，其特征在于，所述的化合物具有式A1结构：
    Figure PCTCN2016076083-appb-100010
    其中，
    Figure PCTCN2016076083-appb-100011
    代表双键或单键，并且
    Figure PCTCN2016076083-appb-100012
    中的两个
    Figure PCTCN2016076083-appb-100013
    不同时为双键，或者
    Figure PCTCN2016076083-appb-100014
    中的两个
    Figure PCTCN2016076083-appb-100015
    不同时为双键；
    Q和W各自独立地代表C、N、O或S原子中的一种或2种以上，且当各自独立为N时，各自在环内所处的位置和数目可变化；当各自独立含有O或S时，环上碳原子减少，保持五元芳环体系；Q或W可以各自互相成键连接在一起或者各自分散在环上不同位置；
    其中所述的“当Q或W各自独立为N时，位置和数目可变化”是指：Q或W出现在各自所在环上的位置不受限定；Q或W的数目可为1-3，当数目大于或等于(≥)2时，为保持六元环体系，相应环上碳原子数目减少；
    R1和R2各自独立地代表氢、羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基或卤素；或者两个R1可与相邻的碳原子或两个R2可与相邻的碳原子共同形成含有0-3个选自O、S、N的杂原子的5-7元环；
    R3和R4各自独立地代表氢、氧、硫、羟基、氨基、巯基、烷基、链烯基、链炔基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基、芳烷基、C1-C6醛基或卤素；
    并且当R4代表氢且R3代表氢或氧，且W为碳时，R2不为氢。
  2. 一种式A所示的七元环小檗碱类似物、其药学上可接受的盐、其晶型、其水合物、或其溶剂化物、或其前药，
    Figure PCTCN2016076083-appb-100016
    式中，
    Figure PCTCN2016076083-appb-100017
    代表双键或单键，并且
    Figure PCTCN2016076083-appb-100018
    中的两个
    Figure PCTCN2016076083-appb-100019
    不同时为双键，或者
    Figure PCTCN2016076083-appb-100020
    中的两个
    Figure PCTCN2016076083-appb-100021
    不同时为双键；
    Figure PCTCN2016076083-appb-100022
    分别表示5-6元环；
    R1为位于
    Figure PCTCN2016076083-appb-100023
    的1-4个(较佳地1-3个或1-2个)相同或不同的取代基，而R2为位于
    Figure PCTCN2016076083-appb-100024
    的1-4个(较佳地1-3个或1-2个)相同或不同的取代基，并且各R1和R2独立地选自下组：氢、羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基或卤素；或者两个R1可与相邻的碳原子或两个R2可与相邻的碳原子共同形成含有0-3个选自O、S、N的杂原子的5-7元环(例如，二个R1与
    Figure PCTCN2016076083-appb-100025
    或两个R2与
    Figure PCTCN2016076083-appb-100026
    共同形成吲哚、氮杂吲哚等结构)；
    R3选自下组：氢、氧、硫、羟基、氨基、巯基、烷基、链烯基、链炔基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基、芳烷基、C1-C6醛基或卤素；
    R4选自下组：氢、氧、硫、羟基、氨基、巯基、烷基、链烯基、链炔基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基、芳烷基、C1-C6醛基或卤素。
  3. 如权利要求3所述的七元环小檗碱类似物及其药学上可接受的盐，其特征在于，
    Figure PCTCN2016076083-appb-100027
    分别为芳基或杂芳基，其中所述的杂芳基具有1-3个N原子；
  4. 如权利要求3所述的七元环小檗碱类似物及其药学上可接受的盐，其特征在于，所述化合物具有式A1结构：
    Figure PCTCN2016076083-appb-100028
    其中，
    Figure PCTCN2016076083-appb-100029
    代表双键或单键，并且
    Figure PCTCN2016076083-appb-100030
    中的两个
    Figure PCTCN2016076083-appb-100031
    不同时为双键，或者
    Figure PCTCN2016076083-appb-100032
    中的两个
    Figure PCTCN2016076083-appb-100033
    不同时为双键；
    Q和W各自独立地代表C、N、O或S原子中的一种或2种以上，且当各自独立为N时，各自在环内所处的位置和数目可变化；当各自独立含有O或S时，环上碳原子减少，保持五元芳环体系；Q或W可以各自互相成键连接在一起或者各自分散在环上不同位置；
    其中所述的“当Q或W各自独立为N时，位置和数目可变化”是指：Q或W出现在各自所在环上的位置不受限定；Q或W的数目可为1-3，当数目大于或等于(≥)2时，为保持六元环体系，相应环上碳原子数目减少；
    R1和R2各自独立地代表氢、羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基或卤素；或者两个R1可与相邻的碳原子或两个R2可与相邻的碳原子共同形成含有0-3个选自O、S、N的杂原子的5-7元环；
    R3和R4各自独立地代表氢、氧、硫、羟基、氨基、巯基、烷基、链烯基、链炔基、芳基、烷氧基、烷氨基、烷硫基、芳醚基、芳氨基、芳硫基、芳烷基、C1-C6醛基或卤素；
    并且当R4代表氢且R3代表氢或氧，且W为碳时，R2不为氢。
  5. 如权利要求1-5中任一所述的七元环小檗碱类似物及其药学上可接受的盐，其特征在于，所述的药学上可接受的盐包括与酸根离子形成的盐；
    较佳地，所述的成盐的酸根离子包括无机酸根离子、有机酸根离子、和/或卤素离子。
  6. 根据权利要求1-5中任一所述的七元环小檗碱类似物或其药学上可接受的盐，其特征在于，
    Figure PCTCN2016076083-appb-100034
    中的
    Figure PCTCN2016076083-appb-100035
    为单键，
    Figure PCTCN2016076083-appb-100036
    为双键；和/或
    R3代表氧、氮、硫、烷基或烷氨基。
  7. 根据权利要求1-5中任一所述的七元环小檗碱类似物或其药学上可接受的盐，其特征在于，
    Figure PCTCN2016076083-appb-100037
    中的
    Figure PCTCN2016076083-appb-100038
    为双键，
    Figure PCTCN2016076083-appb-100039
    为单键。
  8. 根据权利要求1-5中任一所述的七元环小檗碱类似物或其药学上可接受的盐，其特征在于，R4代表羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷巯基、芳醚基、芳氨基、芳巯基或卤素；和/或
    R1和R2各自独立地代表羟基、氨基、巯基、烷基、芳基、烷氧基、烷氨基、烷巯基、芳醚基、芳氨基、芳巯基或卤素。
  9. 根据权利要求1-5中任一所述的七元环小檗碱类似物及其药学上可接受的盐，其中，所述七元环小檗碱类似物选自下组：
    Figure PCTCN2016076083-appb-100040
    Figure PCTCN2016076083-appb-100041
  10. 一种药物组合物，其特征在于，包含治疗有效量的一种或多种权利要求1～10所述的七元环小檗碱类似物或其药学上可接受的盐以及药学上可接受的载体。
  11. 根据权利要求1～10中任一所述的七元环小檗碱类似物及其药学上可接受的盐在制备预防或治疗肿瘤或癌症的药物中的用途。
  12. 一种制备药物组合物的方法，其特征在于，包括步骤：将药学上可接受的载体与权利要求1-10中任一所述的七元环小檗碱类似物、或其晶型、药学上可接受的盐、水合物或溶剂合物进行混合，从而形成药物组合物。
  13. 一种体外非治疗性地抑制肿瘤细胞的方法，其特征在于，包括步骤：在权利要求1-10中任一所述的七元环小檗碱类似物及其药学上可接受的盐存在下，培养肿瘤细胞，从而抑制所述肿瘤细胞的生长。
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