WO2017167183A1 - 二芳基-β-内酰胺类化合物及其制备方法和在制药中的用途 - Google Patents

二芳基-β-内酰胺类化合物及其制备方法和在制药中的用途 Download PDF

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WO2017167183A1
WO2017167183A1 PCT/CN2017/078444 CN2017078444W WO2017167183A1 WO 2017167183 A1 WO2017167183 A1 WO 2017167183A1 CN 2017078444 W CN2017078444 W CN 2017078444W WO 2017167183 A1 WO2017167183 A1 WO 2017167183A1
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mmol
compound
group
nmr
cdcl
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PCT/CN2017/078444
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French (fr)
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王洋
刘明明
周鹏飞
冯克昌
丁奎岭
王晓明
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复旦大学
中国科学院上海有机化学研究所
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Priority claimed from CN201710162725.4A external-priority patent/CN107235883B/zh
Application filed by 复旦大学, 中国科学院上海有机化学研究所 filed Critical 复旦大学
Priority to US16/090,568 priority Critical patent/US20200392150A1/en
Priority to EP17773213.8A priority patent/EP3438105B1/en
Publication of WO2017167183A1 publication Critical patent/WO2017167183A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D205/00—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom
    • C07D205/02—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings
    • C07D205/06—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
    • C07D205/08—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with one oxygen atom directly attached in position 2, e.g. beta-lactams
    • 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
    • A61K31/397—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having four-membered rings, e.g. azetidine
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4192—1,2,3-Triazoles
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • 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
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • 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
    • A61K31/695—Silicon compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D205/00—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom
    • C07D205/02—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings
    • C07D205/06—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
    • C07D205/08—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with one oxygen atom directly attached in position 2, e.g. beta-lactams
    • C07D205/085—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with one oxygen atom directly attached in position 2, e.g. beta-lactams with a nitrogen atom directly attached in position 3
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C07D491/113—Spiro-condensed systems with two or more oxygen atoms as ring hetero atoms in the oxygen-containing ring

Definitions

  • the present invention belongs to the field of chemical pharmacy, and relates to a diaryl- ⁇ -lactam compound having significant antitumor activity, a preparation method thereof, and an antitumor activity in vitro and in vivo, and such compounds and pharmaceutically acceptable salts thereof or The use of a pharmaceutical compound whose medicinal salt is a component in the field of prevention and treatment of tumor-related diseases.
  • Cancer is one of the major diseases that threaten human health, and its mortality rate is second only to cardiovascular and cerebrovascular diseases. It is estimated that there will be 15 million new cases by 2020, and the death toll will reach 10 million;
  • the treatment of cancer includes surgery, radiation therapy, chemotherapy (drug therapy) and biological therapy.
  • Chemotherapy is the most common method, which is to treat cancer patients with one or more cytotoxic anti-tumor drugs. treatment.
  • cytotoxic anti-tumor drugs Although a large number of drugs are currently in clinical use, due to the long-term use of chemotherapy drugs or mutations in tumor cells themselves, many malignant tumors are resistant to chemotherapy drugs, resulting in weakened or disappeared chemotherapy effects, as well as traditional anti-tumor drugs due to Lack of selectivity-induced toxicity makes chemotherapy drugs still unable to meet clinical needs. Therefore, the search for new anti-tumor drugs with higher efficiency and low toxicity has always been a hot spot in the field of research and development and an important problem that needs to be solved urgently.
  • anti-tumor drugs approved for marketing including the following types: (1) anti-tumor drugs acting on DNA: such as alkylating agents, metal platinum complexes, DNA topoisomerase inhibitors And anti-metabolite anti-tumor drugs; (2) anti-tumor drugs acting on kinases: such as tyrosine kinase inhibitors and serine/threonine kinase inhibitors; (3) anti-tumor drugs acting on microtubules: Microtubule aggregation inhibitors and microtubule stabilizers, and the like.
  • DNA such as alkylating agents, metal platinum complexes, DNA topoisomerase inhibitors And anti-metabolite anti-tumor drugs
  • anti-tumor drugs acting on kinases such as tyrosine kinase inhibitors and serine/threonine kinase inhibitors
  • anti-tumor drugs acting on microtubules Microtubule aggregation inhibitors and microtubule stabilizer
  • the anti-tumor drugs acting on microtubules are currently the most effective chemotherapeutic drugs for treating prostate cancer, breast cancer, ovarian cancer and other solid tumors, and are one of the hotspots of anti-tumor drug research in recent years.
  • tubulin aggregation inhibitors Much of the important results and progress have been made in the inhibition of tubulin aggregation inhibitors, especially in the structural modification of CompAir. These drugs are effective in inhibiting tumor growth, but the disadvantage is that although many inhibitors have entered clinical trials, there are not many compounds that are approved for marketing due to certain toxicity. Therefore, the search for novel tubulin aggregation inhibitors and angiogenesis inhibitors with more active and less side effects is still a clinically urgent need.
  • the object of the present invention is to disclose a novel diaryl- ⁇ -lactam compound having a compound of the formula I or a pharmaceutical salt thereof.
  • diaryl- ⁇ -lactam compound having the structure of Formula I, and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof:
  • R 1 is one or more groups selected from the group consisting of a substituted or unsubstituted C1-C4 alkoxy group, a C1-C4 alkyl group, a halogen, an amino group, a hydroxyl group, a carboxyl group, a substituted or unsubstituted group.
  • R is selected from the group consisting of vinyl, halogen, amino, hydroxy, carboxy, fluorosulfonyloxy, methylsulfonyl (Ms), substituted or unsubstituted C1-C4 alkoxy, substituted or Unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C1-C4 alkylamino, substituted or unsubstituted C2-C10 ester, substituted or unsubstituted C1- C6 alkyl-hydroxy, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heteroaryl;
  • substitution means that one or more hydrogen atoms on the group are substituted with a substituent selected from the group consisting of C1-C4 alkoxy, C1-C4 alkyl, halogen, C2-C10 acyloxy, C2-C10.
  • the substituted phenyl means that the phenyl ring is substituted with from 1 to 5 substituents selected from the group consisting of a nitro group, a fluorine atom or a methoxy group.
  • the compound has the structure shown in the following formula I-1:
  • the compound has the structure shown in the following formula I-2:
  • the compound has the structure shown by the following formula I-3, I-4, I-5, I-6 or I-7:
  • the compound is selected from the group consisting of:
  • a pharmaceutical composition according to the first aspect of the present invention for the preparation of a pharmaceutical composition for treating or preventing a disease selected from the group consisting of microtubule-associated protein aggregation Mammalian diseases, mammalian diseases associated with angiogenesis.
  • the mammalian disease associated with aggregation of microtubule-associated proteins is a tumor.
  • the tumor is selected from the group consisting of thyroid cancer, head and neck squamous cell carcinoma, cervical cancer, ovarian cancer, breast cancer, colorectal cancer, pancreatic cancer, esophageal cancer, osteosarcoma, and kidney.
  • a pharmaceutical composition comprising: (i) an effective amount of a compound of formula I, and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof And (ii) a pharmaceutically acceptable carrier.
  • the compound of formula I is prepared using the compound of formula Ig as described.
  • Figure 1 is a graph showing the inhibition of in vitro microtubule aggregation by compounds 69, 70 and 97.
  • Figure 2 shows immunoblot analysis experiments showing that Compounds 1, 69, 70 and 97 significantly inhibited microtubule aggregation and maintained tubulin in a depolymerized state.
  • Figure 4 shows that inhibition of angiogenesis experiments showed that compounds 69, 70 and 97 significantly inhibited the formation of capillary-like structures in HUVEC cells.
  • Figure 5 is a Matrigel plug assay showing that compounds 69, 70 and 97 significantly inhibit VEGF-mediated neovascularization.
  • Figure 6 is a graph of colony inhibition assay showing that compounds 69, 70 and 97 significantly inhibited the formation of colonies of tumor cells.
  • Figure 7 is an in vitro cell cycle assay showing that compounds 69, 70 and 97 significantly arrest cells in the G2/M phase.
  • Figure 8 shows the results of an in vitro cell cycle-associated protein assay showing that compounds 69, 70 and 97 can be significantly promoted. Phospho-histone H3, cyclin B1, mitotic checkpoint protein BuBR1 expression.
  • Figure 9 is a result of an in vitro apoptosis assay showing that compounds 69, 70 and 97 significantly promote apoptosis.
  • Figure 10 shows the results of in vitro apoptosis-related protein detection experiments, showing that compounds 69, 70 and 97 can significantly promote the expression of the pro-apoptotic protein Bax, the tumor suppressor gene p53, and the spliced DNA repair enzyme.
  • Figure 11 is a tumor treatment and mechanism study experiment at the animal level and tissue level, showing that compounds 69, 70 and 97 can significantly inhibit tumor growth in vivo and have no significant effect on mouse body weight; tissue staining results show that compounds 69, 70 and 97 can cause tumor tissue to produce necrotic areas (arrows in the figure), and no abnormal areas were observed in the liver, kidney, and spleen tissues.
  • the inventors have unexpectedly discovered a new class of diaryl- ⁇ -lactam compounds after a long and intensive study.
  • the compound has excellent tubulin aggregation inhibitory activity and thus can be used as an angiogenesis inhibitor for treating cancer. Based on the above findings, the inventors completed the present invention.
  • C1-C4 alkyl refers to a straight or branched alkyl group having from 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, Tert-butyl, or a similar group.
  • C3-C6 cycloalkyl refers to a cycloalkyl group having from 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, or the like.
  • C1-C4 alkoxy refers to a straight or branched alkoxy group having from 1 to 4 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso Butoxy, sec-butoxy, tert-butoxy, or the like.
  • halogen refers to F, Cl, Br and I.
  • C1-C4 alkylamino refers to an amine group substituted by a C1-C4 alkyl group, for example, having "C1-C4 alkyl-NH-" or "(alkyl) 2 -N- (total number of carbon atoms is 1- 4)", “-C1-C4 alkylene-NH 2 ", “alkyl-N-alkylene- (total number of carbon atoms 1-12)", or "(alkyl) 2 -N-alkylene a group of a radical - (the total number of carbon atoms is 1-12)” structure, such as CH 3 NH-, C 2 H 5 NH-, C 3 H 7 NH-, (CH 3 ) 2 N-, -CH 2 NH 2 , -C 2 H 5 NH 2 , -C 3 H 7 NH 2 , -C 2 H 4 N(CH 3 ) 2 , or the like.
  • the definition of C 1-12 alkyl is as
  • C2-C10 ester group refers to a substituent having the structure "linear or branched alkyl/cycloalkyl/aryl/heteroaryl-carbonyl-oxy-" having 1 to 9 carbon atoms, For example, an ethyl ester group, a propyl ester group, a butyl ester group, or the like.
  • C1-C6 amido refers to a substituent of the form “linear or branched alkyl/cycloalkyl/aryl/heteroaryl-carbonyl-amino-" having 0 to 5 carbon atoms, Such as acetamido, propionamide, butanamide, or the like.
  • C6-C10 aryl refers to an aryl group having 6 to 10 carbon atoms, such as phenyl, naphthyl and the like, which may be substituted or unsubstituted.
  • 5-12 membered heteroaryl refers to a heteroaryl group having 5 to 12 ring atoms, wherein the ring atom includes a carbon atom and one or more (preferably 1 to 3) are selected from O, S and/or N.
  • a heteroatom heteroaryl group preferably a 5-8 membered heteroaryl group.
  • the heteroaryl group can be substituted or unsubstituted.
  • C3 -C6 heterocyclyl refers to a non-aromatic cyclic group having 3 to 6 carbon atoms and one or more (preferably 1 to 3) heteroatoms selected from O, S and/or N. Preferably, a 5-6 membered heterocyclic group is used.
  • the heterocyclic group may be substituted or unsubstituted.
  • the term "pharmaceutically acceptable” ingredient means a substance which is suitable for use in humans and/or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reaction), that is, a reasonable benefit/risk ratio.
  • the term "effective amount" means an amount of a therapeutic agent that treats, alleviates or prevents a target disease or condition, or an amount that exhibits a detectable therapeutic or prophylactic effect.
  • the precise effective amount for a subject will depend on the size and health of the subject, the nature and extent of the condition, and the combination of therapeutic and/or therapeutic agents selected for administration. Therefore, it is useless to specify an accurate effective amount in advance. However, for a given condition, routine experimentation can be used to determine the effective amount that the clinician can determine.
  • each of the chiral carbon atoms may be optionally in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.
  • the "pharmaceutically acceptable salt” as described in the present invention may specifically be an inorganic acid such as hydrohalic acid, sulfuric acid, phosphoric acid or nitric acid, and capric acid, fumaric acid, oxalic acid, malic acid, lactic acid, a salt formed by an organic acid such as camphorsulfonic acid.
  • an inorganic acid such as hydrohalic acid, sulfuric acid, phosphoric acid or nitric acid, and capric acid, fumaric acid, oxalic acid, malic acid, lactic acid, a salt formed by an organic acid such as camphorsulfonic acid.
  • Another object of the present invention is to provide a use of the above compound or a pharmaceutically acceptable salt thereof and a composition comprising the same or a salt thereof for the preparation of a medicament for preventing or treating a tumor-related disease.
  • the cancer-related diseases may be specifically thyroid cancer, head and neck squamous cell carcinoma, cervical cancer, ovarian cancer, breast cancer, colorectal cancer, pancreatic cancer, esophageal cancer, osteosarcoma, renal cancer, stomach cancer, lung cancer.
  • the present invention provides and demonstrates a diaryl- ⁇ -lactam compound having a significant antitumor effect or a pharmaceutically acceptable salt thereof, which inhibits the growth of tumor cells by inhibiting tubulin aggregation, in vitro and in vivo.
  • the anti-tumor experiment has a good inhibitory effect on tumor growth.
  • a preferred class of compounds in the present invention are diaryl- ⁇ -lactam compounds having the structure of Formula I:
  • N-morpholinyl 2-(1,3-dioxoisoindol-2-yl)ethyl
  • 4-acetamidophenyl benzyl, n-butyl, 3,4-dimethoxy
  • Phenylphenyl R 6 is taken from vinyl, 4-nitrophenyl, cyclopropyl.
  • a more preferred class of compounds are diaryl- ⁇ -lactam compounds having the structure of Formula II:
  • R is taken from the methoxy group, and the number is 1 or 2, which may be substituted at the 3, 4, and 5 positions.
  • R is derived from methyl, trimethylsilyl, phenyl or tert-butylphenyl, and the configuration of the ethylenic bond is Z or E.
  • R 1 is independently selected from methyl, ethyl, hydroxymethyl, hydrogen atom, alkoxy, acyloxy, hydroxy, halogen, amino, phenylamino, benzylamino, acetylamino, p-toluenesulfonic Amido, methanesulfonylamino, benzoylamino, 3-fluorobenzoylamino, methanesulfonyloxy, methoxymethyl, N,N-dimethylaminomethyl, 4-hydroxybenzyl, trimethyl Silylethyl, ethoxycarbonylmethyl, carboxypropionyloxy, R 2 is independently taken from hydroxy, amino, halo, methoxy, methyl, fluorosulfonyloxy, hydrogen or acyloxy.
  • the dominant compounds are:
  • R 1 is derived from a hydrogen atom, a methyl group, an acetyl group or an acryloyl group
  • R 2 is derived from a hydrogen atom, a benzyl group, an acyl group or an acryloyl group.
  • a particularly preferred class of compounds has the structure shown below:
  • the compound of the present invention has excellent inhibitory activity against tubulin, the compound of the present invention and various crystal forms thereof, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and compounds containing the present invention are mainly
  • the pharmaceutical composition of the active ingredient can be used for the treatment, prevention, and alleviation of diseases associated with tubulin activity or expression levels, particularly for diseases in which tubulin activity or expression levels are all related.
  • the compounds of the invention are useful in the treatment of diseases such as cancer, neurodegenerative diseases, malaria, AIDS, gout, diabetes and the like.
  • 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 5 to 200 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 Wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.
  • 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.
  • An example of an embedding component that can be used is poly Compound and waxy substances. 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 compounds.
  • 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 5 to 500 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.
  • a novel structure of a compound having a tubulin inhibitory activity is provided.
  • Direct silica gel column chromatography first remove the ethyl acrylate with PE, then carry out gradient elution, collect the corresponding eluent, recover 1.55 g of the raw material, the recovery rate is 58%, and obtain the colorless oily product (1c) 1.12 g, deducted and recovered.
  • the yield of the raw material was 73%.
  • the compound 1 (20 mg, 0.054 mmol) was dissolved in 1.5 mL of dry DCM, and then stirred under stirring at 0 ° C, then EtOAc (15 mg, 0.162 mmol), 1-butylsulfonyl chloride (21 mg, 0.081 mmol).
  • the compound 1 (20 mg, 0.054 mmol) was dissolved in 1.5 mL of dry DCM, and then, under stirring at 0 ° C, TEA (15 mg, 0.162 mmol), benzylsulfonyl chloride (16 mg, 0.081 mmol), and then allowed to warm to room temperature overnight.
  • the synthesis method of compound 38 is similar to the synthesis method of compound 1:
  • Tris(dibenzylideneacetone)dipalladium (4.3 mg, 0.0047 mmol) and 1e (8.5 mg, 0.0118 mmol) were separately added to a schlenk tube under nitrogen atmosphere, and anhydrous CH 2 Cl 2 (5 mL) was added at room temperature.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • Tris(dibenzylideneacetone)dipalladium (4.3 mg, 0.0047 mmol) and 1e (8.5 mg, 0.0118 mmol) were separately added to a schlenk tube under nitrogen atmosphere, and anhydrous CH 2 Cl 2 (5 mL) was added at room temperature.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • Tris(dibenzylideneacetone)dipalladium (9.2 mg, 0.01 mmol) and 1e (18 mg, 0.025 mmol) were separately added to a schlenk tube under nitrogen atmosphere, and anhydrous CH 2 Cl 2 (5 mL) was added and stirred at room temperature.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • Tris(dibenzylideneacetone)dipalladium (3.8 mg, 0.0041 mmol) and 1e (7.2 mg, 0.01 mmol) were separately added to a Schlenk tube under nitrogen atmosphere, and anhydrous CH 2 Cl 2 (5 mL) was added at room temperature.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • Tris(dibenzylideneacetone)dipalladium (5.7 mg, 0.0062 mmol) and 1e (11.1 mg, 0.016 mmol) were separately added to a schlenk tube under nitrogen atmosphere, and anhydrous CH 2 Cl 2 (5 mL) was added at room temperature.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • Tris(dibenzylideneacetone)dipalladium (5.7 mg, 0.0062 mmol) and 1e (11.1 mg, 0.016 mmol) were separately added to a schlenk tube under nitrogen atmosphere, and anhydrous CH 2 Cl 2 (5 mL) was added at room temperature.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • Tris(dibenzylideneacetone)dipalladium (3.8 mg, 0.0042 mmol) and 1e (7.56 mg, 0.0105 mmol) were separately added to a schlenk tube under nitrogen atmosphere, and anhydrous CH 2 Cl 2 (5 mL) was added at room temperature.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • Tris(dibenzylideneacetone)dipalladium (3.8 mg, 0.0042 mmol) and 1e (7.56 mg, 0.0105 mmol) were separately added to a schlenk tube under nitrogen atmosphere, and anhydrous CH 2 Cl 2 (5 mL) was added at room temperature.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • Tris(dibenzylideneacetone)dipalladium (4.7 mg, 0.0052 mmol) and 1e (9.3 mg, 0.0129 mmol) were separately added to a schlenk tube under nitrogen atmosphere, and anhydrous CH 2 Cl 2 (5 mL) was added at room temperature.
  • the ligand is replaced by 0.005 equivalent of triphenylphosphine, and the other operations are identical, and the corresponding racemate can be obtained.
  • the tumor cells were seeded in a 96-well plate at 37 ° C, 5% CO 2 for 24 hours, then 6 different concentrations of samples were added, and the positive samples paclitaxel, CA-4 as a positive control. After 48 hours of incubation, MTT was added, and after 4 hours, the supernatant was discarded, purple crystals were dissolved by adding DMSO, and the OD value was measured at 540 nm on a microplate reader, and the inhibition rate was calculated. The half-inhibitory concentration IC 50 value of the compound was calculated from the inhibition rates of the six concentrations.
  • Diaryl- ⁇ -lactam target compounds inhibit tumor cell proliferation activity (IC 50 , ⁇ M)
  • Antitumor activity was determined by the MTT method and the data were the average of three measurements.
  • A2780 and SKOV-3 are human ovarian cancer cell lines; MDA-MB-231 is a human breast cancer cell line; Hela is a human cervical cancer cell line.
  • the inhibition of in vitro microtubule aggregation by test compounds 69, 70 and 97 was tested by turbidity method.
  • the test kit was purchased from Cytoskeleton, Inc., USA. The specific steps are as follows.
  • the reaction solution was pre-incubated on ice, and various concentrations of the test compound were added, and the DMSO (4%, v/v) group was set as the negative control group, and the Colchicine-treated group was used as the positive control.
  • Hela cells were treated with different concentrations of test compounds for 6 hours, and the DMSO group was set as a negative control group.
  • the cells were collected and washed twice with PBS and then digested with cell lysate containing microtubule stabilizer (containing 100 mM PIPES, pH 6.8, 1 mM). MgCl 2 , 2 mM EGTA, 0.5% NP-40, 2M glycerol, 5 ⁇ M paclitaxel and protein kinase inhibitor).
  • the digestive juice was centrifuged at 15,000 rpm for 15 minutes. The supernatant was aspirated and the pellet was dissolved in SDS digestion buffer.
  • the tumor cells were cultured, and the cells were seeded on a cover glass pretreated (2M NaOH soaked for 2 hours, 75% ethanol soaked for 30 minutes). After the cells were attached, the coverslips were placed in a 24-well plate, and the medium was added. After 24 hours of culture, different concentrations of the test compound were added for 24 h, and the DMSO-treated group was set as a negative control. The medium was discarded, the cells were washed twice with PBS, fixed with methanol for 15 min, washed three times with PBS, permeabilized with 0.1% triton for 15 min, and washed three times with PBS.
  • the cells were blocked with 5% BSA for 1 h at room temperature, added with primary antibody at 4 ° C overnight, and rinsed 3 times with PBST for 5 min each time. Fluorescent secondary antibody was added, incubated at room temperature for 1 h in the dark, and rinsed 3 times with PBST for 5 min each time.
  • One drop (including DAPI) of the sealer was added dropwise, and the morphology of tubulin was observed by confocal microscopy. The effect of the test compound on the structure of the microtubule was examined, and related photographs were taken. The results showed that Compounds 1, 69, 70 and 97 significantly inhibited the aggregation of microtubules.
  • the extracellular matrix gel MatriGel (BD Biosciences, USA) was mixed with PBS in a 1:1 dilution and added to a 24-well plate. Incubate for 1 hour at 37 ° C in a 5% CO 2 incubator. After gel formation, HUVEC cells were seeded onto the gel at a concentration of 3 ⁇ 10 4 /well, and DMEM medium containing 10% PBS was added. The test sample was added, and CA-4 was set as a positive control, and DMSO was used as a negative control. The culture was incubated at 37 ° C in a 5% CO 2 incubator for 12 hours. The capillary formation was observed under an inverted phase contrast microscope, and related photographs were taken. The results showed that compounds 69, 70 and 97 significantly inhibited the formation of capillary-like structures in HUVEC cells (Fig. 4).
  • test compound was added at 4 ° C in Matrigel containing 100 ng/mL human recombinant VEGF-A165, while the DMSO treated group was set as a negative control.
  • the 6-well plate 1000 cells per well were used. After the cells were attached, the cells were treated with different concentrations of the test sample for 48 hours, and the positive control group (CA-4) and the negative control group (blank solvent) were replaced. After the fresh medium was further cultured for 7-10 days, the medium was discarded, the cells were fixed with methanol, and stained with Giemsa or crystal violet dye, and the number of colonies formed by more than 50 cells was counted under a microscope. The results showed that compounds 69, 70 and 97 significantly inhibited the formation of colonies of tumor cells (Fig. 6).
  • the cells were seeded at 2 ⁇ 10 5 cells/well. After the cells were attached, the cells were treated with different concentrations of the test samples for 24 hours, and the positive control group (CA-4) and the negative control group were set at the same time. (blank solvent). The cells were collected, washed twice with PBS, and fixed overnight with 75% ethanol at -20 ° C, stained with PI and tested by flow cytometry. As a result, as shown in Fig. 7, compounds 69, 70 and 97 were able to significantly arrest cells in the G2/M phase.
  • the cells were seeded at 2 ⁇ 10 5 cells/well. After the cells were attached, the cells were treated with different concentrations of the test samples for 24 hours, and the positive control group (CA-4) and the negative control group were set at the same time. (blank solvent). The cells were collected, washed twice with PBS, fixed overnight with 75% ethanol at -20 ° C, and double stained with PI and Annexin V. The cells were stained and tested by flow cytometry. The results showed that compounds 69, 70 and 97 significantly promoted apoptosis (Fig. 9).
  • A. Western blot After treatment of tumor cells with different concentrations of compounds (while designing DMSO as a negative control), the cells were collected and lysed with lysate. After the protein sample is denatured by heating, it is separated by electrophoresis, transferred, blocked, and then subjected to primary antibody reaction and secondary antibody reaction, and then exposed to color.
  • mice were housed individually in cages without pathogens. Each group of mice (10, 5 female, 5 male) was intraperitoneally injected with test drugs 69 (95, 70, 50, 35 and 25 mg/kg), 70 (500, 425, 350, 275 and 200 mg/kg) or 97 (275, 200, 150, 125 and 100 mg/kg). , or placebo (8.3% castor oil with 8.3% ethanol in PBS). 70, 69 was dissolved in castor oil ethanol (1:1, v/v) and then diluted with PBS (1:5, v/v). Mouse deaths were recorded daily for 14 days. The results showed that the LD 50 of Compound 69 was 61.5 mg/kg, the LD 50 of Compound 70 was more than 500 mg/kg, and the LD 50 of Compound 97 was 136.5 mg/kg.
  • LD 50 61.5 mg / kg
  • 70 LD 50 > 500 mg / kg
  • 97: LD50 136.5 mg / kg.
  • the ovarian cancer cell line A2780 was cultured, and the cells were inoculated during the vigorous growth period. Each 2 ⁇ 10 6 cells were intraperitoneally injected into 6-week old Balb/C nude mice to establish a tumor metastasis model of nude mice, which was grown under SPF conditions.
  • the subcutaneous xenografts of nude mice were grown to a volume of about 100 mm 3 , and the established tumor-bearing mice were randomly divided into 4 groups, 10 in each group, and intraperitoneally injected with different concentrations of 7.5 mg/kg of 69 and 12.5 mg/ The tumor growth size was recorded at 70, 4 mg/kg and 8 mg/kg of 97, 5 mg/kg or 10 mg/kg paclitaxel and a blank control (same as the acute toxicity test).
  • Tumor inhibition rate (1 - average tumor mass of the experimental group / average tumor mass of the control group) ⁇ 100%

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Abstract

本发明属合成药物化学领域，涉及一类新型具有显著抗肿瘤活性的二芳基-β-内酰胺类化合物及其在制药中的应用。本发明还包括这类化合物、其药用盐及其药物组合物在制备预防或治疗与肿瘤相关疾病的药物中的应用。所述的二芳基-β-内酰胺类化合物具有如下通式结构:（I）.

Description

二芳基-β-内酰胺类化合物及其制备方法和在制药中的用途 技术领域
本发明属化学制药领域,涉及具有显著抗肿瘤活性的二芳基-β-内酰胺类化合物、制备方法及其体外、体内抗肿瘤活性，以及这类化合物及其药学盐或以这类化合物或其药学盐为成分的复方药物在预防和治疗肿瘤相关疾病领域的应用。
背景技术
癌症是威胁人类健康的主要疾病之一，其死亡率仅次于心脑血管疾病、位列第二；据估计，到2020年将有1500万的新增病例，死亡人数将达到1000万；目前癌症的治疗方法有手术治疗、放射治疗、化学治疗(药物治疗)和生物治疗等，其中化疗是最为常见的一种手段，即用一种或多种具有细胞毒性的抗肿瘤药物对癌症患者进行治疗。尽管目前已有大量的药物在临床应用，然而由于化疗药物的长期使用或肿瘤细胞自身的突变、许多恶性肿瘤对化疗药物产生耐药性、导致化疗效果减弱或消失，以及传统的抗肿瘤药物由于缺乏选择性导致的毒性,使得化疗药物仍不能满足临床的需要。因此，寻找更为高效、低毒的新型抗肿瘤药物始终是医药领域的研发热点和急需解决的重要问题。
目前,被批准上市的抗肿瘤药物已有近百种，主要包括以下几种类型：(1)作用于DNA的抗肿瘤药物：如烷化剂、金属铂配合物、DNA拓扑异构酶抑制剂和抗代谢抗肿瘤药等；(2)作用于激酶的抗肿瘤药物：如酪氨酸激酶抑制剂和丝氨酸/苏氨酸激酶抑制剂等；(3)作用于微管的抗肿瘤药物：如微管聚集抑制剂和微管稳定剂等。其中，作用于微管的抗肿瘤药物是目前治疗前列腺癌、乳腺癌、卵巢癌和其它实体瘤的最有效的化疗药物，是近年来抗肿瘤药物研究的热点之一。
在微管蛋白聚集抑制剂方面、尤其是针对康普立停的结构改造方面已经取得了许多重要成果和进展。这些药物都可以有效地抑制肿瘤增长，但缺点是虽然有许多抑制剂进入了临床试验，由于存在一定毒性、最终被批准上市的化合物却不多。因此，寻找活性更强且副作用更小的新型微管蛋白聚集抑制剂以及血管生成抑制剂仍是临床的迫切需求。
发明内容
本发明的目的是公开新型的二芳基-β-内酰胺类化合物，具有通式Ⅰ结构的化合物或其药学盐。
本发明的第一方面，提供了一种具有通式Ⅰ结构的二芳基-β-内酰胺类化合物，及其药学上可接受的盐，水合物，溶剂合物或前药：
Figure PCTCN2017078444-appb-000001
其特征在于，
R1为一个或多个位于环上的选自下组的基团：取代或未取代的C1-C4烷氧基、C1-C4 烷基、卤素、氨基、羟基、羧基、取代或未取代的C2-C10酰氧基、C2-C10酯基、甲氧甲酰基、烯丙氧基、炔丙氧基、磺酰氧基、烷氨基、酰氨基、磺酰氨基或者2-3个上述相同或不同基团的组合；
R2为一个或多个位于环上的选自下组的基团：取代或未取代的C1-C6烷氧基、取代或未取代的C1-C6烷基、卤素、氨基、羟基、羧基、氟磺酰氧基、烯丙氧基、炔丙氧基、C1-C4烷氨基、C2-C10酯基、取代或未取代的C1-C6烷基-羟基、取代或未取代的C6-C10的芳基、取代或未取代的5-12元杂芳基、-OTBS、-CH2-R、-OR、-O(C＝O)R、-O-(SO2)-R、-O(PO)-R2、-NH(C＝O)R、-NH-(SO2)-R；
R3和R4各自独立地选自下组：取代或未取代的C1-C6烷基、氢原子、酰氧基、羟基、羧基、环丙基、氨基、取代或未取代的C1-C4烷氨基、磺酰氧基、取代或未取代的C1-C4烷氧基、取代或未取代的C1-C6烷基-羟基、取代或未取代的芳基、取代或未取代的吗啉基、-CH2-R、-OR、-O(C＝O)R、-O-(SO2)-R、-O(PO)-R2、-NH(C＝O)R、-NH-(SO2)-R；
或者R3和R4共同构成＝CHR、-OC(＝O)OCH2-、＝O、C3-C6环烷烃、C3-C6杂环或取代或未取代的-(CH2)n-，其中n选自下组：1、2、3、4、5或6；
R5和R6各自独立地为H；或者R5和R6共同构成＝CHR、-OC(＝O)OCH2-、＝O、＝S；
其中，所述的R选自下组：乙烯基、卤素、氨基、羟基、羧基、氟磺酰氧基、甲基磺酰基(Ms)、取代或未取代的C1-C4烷氧基、取代或未取代的C1-C6烷基、取代或未取代的C3-C6环烷基、取代或未取代的C1-C4烷氨基、取代或未取代的C2-C10酯基、取代或未取代的C1-C6烷基-羟基、取代或未取代的C6-C10的芳基、取代或未取代的5-12元杂芳基；
所述的取代指基团上的一个或多个氢原子被选自下组的取代基取代：C1-C4烷氧基、C1-C4烷基、卤素、C2-C10酰氧基、C2-C10酯基、羟基、环丙基、乙烯基、氨基、氧基(＝O)、吗啉基、磺酰氧基、酰胺基、-NO2、-NHBoc、-NHCbz、-NHC(＝O)Me、-OBn、-NHBn、-SiMe3、未取代或被1-3个选自下组的取代基取代的苯基或吡啶基：C1-C4烷氧基、C1-C4烷基、卤素、羟基。
在另一优选例中，所述的取代苯基是指苯环被1-5个选自下组的取代基取代：硝基、氟原子或甲氧基。
在另一优选例中，所述的化合物具有如下式I-1所示的结构：
Figure PCTCN2017078444-appb-000002
其中，各基团的定义如上文中所述。
在另一优选例中，所述的化合物具有如下式I-2所示的结构：
Figure PCTCN2017078444-appb-000003
其中，各基团的定义如上文中所述。
在另一优选例中，所述的化合物具有如下式I-3、I-4、I-5、I-6或I-7所示的结构：
Figure PCTCN2017078444-appb-000004
其中，各基团的定义如上文中所述。
本发明的第二方面，提供了一种如下式所示的化合物：
Figure PCTCN2017078444-appb-000005
在另一优选例中，所述的化合物选自下组：
Figure PCTCN2017078444-appb-000006
Figure PCTCN2017078444-appb-000007
Figure PCTCN2017078444-appb-000008
Figure PCTCN2017078444-appb-000009
Figure PCTCN2017078444-appb-000010
本发明的第三方面，提供了一种如本发明第一方面所述的药物组合物用于制备治疗或预防选自下组的疾病的药物组合物的用途：与微管相关蛋白聚集有关的哺乳动物疾病、与血管生成相关的哺乳动物疾病。
在另一优选例中，所述的与微管相关蛋白聚集有关的哺乳动物疾病为肿瘤。
在另一优选例中，所述的肿瘤选自下组：甲状腺癌、头颈部鳞状细胞癌、宫颈癌、卵巢癌、乳腺癌、结直肠癌、胰腺癌、食管癌、骨肉瘤、肾癌、胃癌、肺癌、肝癌、黑色素瘤、淋巴瘤、前列腺癌、肾癌、膀胱癌、脑胶质瘤、鼻咽癌，神经内分泌癌、未分化癌、间质肉瘤、绒癌、恶性葡萄胎、恶性畸胎瘤、良性肿瘤。
本发明的第四方面，提供一种药物组合物，所述的药物组合物包括：(i)有效量的式I化合物，及其药学上可接受的盐，水合物，溶剂合物或前药；和(ii)药学上可接受的载体。
本发明的第五方面，提供一种式I化合物的制备方法，所述方法包括步骤：
Figure PCTCN2017078444-appb-000011
在惰性溶剂中，用式If化合物进行反应，得到式Ig化合物；
和任选的步骤：
Figure PCTCN2017078444-appb-000012
用所述的式Ig化合物制备得到式I化合物。
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。
附图说明
图1是化合物69、70和97对体外微管聚集的抑制作用的曲线图。
图2免疫印迹分析实验显示了化合物1、69、70和97能明显抑制微管聚集，使微管蛋白维持解聚状态。
图3免疫荧光检测微管蛋白形态实验显示了化合物1、69、70和97能明显抑制微管的聚集。
图4抑制血管生成实验显示了化合物69、70和97能明显抑制HUVEC细胞生成毛细血管样结构。
图5是基质胶塞实验，显示化合物69、70和97能明显抑制由VEGF介导的新生血管生成。
图6为菌落抑制试验图，其中显示了化合物69、70和97能明显抑制肿瘤细胞菌落的形成。
图7是体外细胞周期实验，显示了化合物69、70和97能明显将细胞阻滞于G2/M期。
图8是体外细胞周期相关蛋白检测实验结果，显示化合物69、70和97能明显促进 磷酸化组蛋白H3、细胞周期蛋白B1、有丝分裂检验点蛋白BuBR1表达。
图9是体外细胞凋亡实验结果，显示了化合物69、70和97能明显促进细胞凋亡。
图10是体外凋亡相关蛋白检测实验结果，显示化合物69、70和97能明显促进促凋亡蛋白Bax、抑癌基因p53、剪切的DNA修复酶的表达。
图11是动物水平和组织水平的肿瘤治疗作用和机理研究实验，显示化合物69、70和97在体内能明显抑制肿瘤生长，并且对小鼠体重无明显影响；组织染色结果显示化合物69、70和97能使肿瘤组织产生坏死区(图中箭头标示)，并且对肝、肾、脾的组织染色并未观察到非正常区域。
具体实施方法：
本发明人经过长期而深入的研究，意外地发现了一类新型的二芳基-β-内酰胺类化合物。所述的化合物具有优异的微管蛋白聚集抑制活性，因此可以被用作为血管生成抑制剂用于治疗癌症。基于上述发现，发明人完成了本发明。
术语
在本文中，除特别说明之处，术语“取代”指基团上的一个或多个氢原子被选自下组的取代基取代：C1-C4烷氧基、C1-C4烷基、卤素、C2-C10酰氧基、C2-C10酯基、羟基、环丙基、乙烯基、氨基、氧基(＝O)、吗啉基、磺酰氧基、酰胺基、-NO2、-NHBoc、-NHCbz、-NHC(＝O)Me、-OBn、-NHBn、-SiMe3、未取代或被1-3个选自下组的取代基取代的苯基或吡啶基：C1-C4烷氧基、C1-C4烷基、卤素、羟基。
术语“C1-C4烷基”指具有1～4个碳原子的直链或支链烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、或类似基团。
术语“C3-C6环烷基”指具有3-6个碳原子的环烷基，例如环丙基、环丁基、环戊基、或类似基团。
术语“C1-C4烷氧基”指具有1-4个碳原子的直链或支链烷氧基，例如甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、异丁氧基、仲丁氧基、叔丁氧基、或类似基团。
术语“卤素”指F、Cl、Br和I。
术语“C1-C4烷胺基”指被C1-C4烷基取代的胺基，例如具有“C1-C4烷基-NH-”或“(烷基)2-N-(碳原子总数为1-4)”、“-C1-C4亚烷基-NH2”、“烷基-N-亚烷基-(碳原子总数为1-12)”、或“(烷基)2-N-亚烷基-(碳原子总数为1-12)”结构的基团，例如CH3NH-、C2H5NH-、C3H7NH-、(CH3)2N-、-CH2NH2、-C2H5NH2、-C3H7NH2、-C2H4N(CH3)2，或类似基团。其中，C1-12烷基的定义如前所述。
术语“C2-C10酯基”指形如“具有1-9个碳原子的直链或支链烷基/环烷基/芳基/杂芳基-羰基-氧基-”结构的取代基，如乙酯基、丙酯基、丁酯基，或类似基团。
术语“C1-C6酰胺基”指形如“具有0-5个碳原子的直链或支链烷基/环烷基/芳基/杂芳基-羰基-胺基-”结构的取代基，如乙酰胺基、丙酰胺基、丁酰胺基，或类似基团。
术语“C6-C10芳基”指具有6-10个碳原子的芳基，例如苯基、萘基等，所述的芳基可以是取代或未取代的。
术语“5-12元杂芳基”指具有5-12个环原子的杂芳基，其中环原子包括碳原子和一个或多个(优选1-3个)选自O、S和/或N的杂原子的杂芳基，优选5-8元杂芳基。所述的杂芳基可以是取代或未取代的。
术语“C3-C6杂环基”指具有3-6个碳原子和一个或多个(优选1-3个)选自O、S和/或N的杂 原子的非芳香性环状基团，优选5-6元杂环基。所述的杂环基可以是取代或未取代的。
本发明中，术语“药学上可接受的”成分是指适用于人和/或动物而无过度不良副反应(如毒性、刺激和变态反应)，即有合理的效益/风险比的物质。
本发明中，术语“有效量”指治疗剂治疗、缓解或预防目标疾病或状况的量，或是表现出可检测的治疗或预防效果的量。对于某一对象的精确有效量取决于该对象的体型和健康状况、病症的性质和程度、以及选择给予的治疗剂和/或治疗剂的组合。因此，预先指定准确的有效量是没用的。然而，对于某给定的状况而言，可以用常规实验来确定该有效量，临床医师是能够判断出来的。
除非特别说明，本发明中，所有出现的化合物均意在包括所有可能的光学异构体，如单一手性的化合物，或各种不同手性化合物的混合物(即外消旋体)。本发明的所有化合物之中，各手性碳原子可以任选地为R构型或S构型，或R构型和S构型的混合物。
本发明中所述的“药学上可接受的盐”，具体地可列举为与氢卤酸、硫酸、磷酸、硝酸等无机酸，以及枸橼酸、富马酸、草酸、苹果酸、乳酸、樟脑磺酸等有机酸形成的盐。
本发明的另一个目的是提供上述化合物或这些化合物在药学上可接受的盐以及包含该化合物或其盐的组合物用于制备预防或治疗与肿瘤相关疾病的药物中的应用。所述的与肿瘤相关疾病具体可列举为甲状腺癌、头颈部鳞状细胞癌、宫颈癌、卵巢癌、乳腺癌、结直肠癌、胰腺癌、食管癌、骨肉瘤、肾癌、胃癌、肺癌、肝癌、黑色素瘤、淋巴瘤、前列腺癌、肾癌、膀胱癌、脑胶质瘤、鼻咽癌，神经内分泌癌、未分化癌、间质肉瘤、绒癌、恶性葡萄胎、恶性畸胎瘤等，以及良性肿瘤，但不受限于此。本发明提供并证明了具有显著抗肿瘤作用的二芳基-β-内酰胺类化合物或其在药学上可接受的盐，通过抑制微管蛋白聚集抑制肿瘤细胞生长的调控机制，在体外、体内的抗肿瘤实验中对肿瘤的生长具有良好的抑制作用。
本发明中一类优选的化合物是具有通式Ⅰ结构的二芳基-β-内酰胺类化合物：
Figure PCTCN2017078444-appb-000013
其特征在于，R1取自羟基、氨基、卤素、1-3个碳原子的烷氧基、甲氧甲酰基、烯丙氧基、炔丙氧基、O(CH2)nR2、OCH2COR3、OCOR4、OSO2R5、NHCOR6、NHSO2NH2，其中R2取自卤素、羟基、N,N-二甲基氨基、N-吗啉基，且n＝2或3，R3取自1-3个碳原子的烷氧基、氨基，R4取自1-3个碳原子的烷基、苯基、取代苯基、吡啶基、环丙基、乙烯基、N-吗啉基，所述的取代苯基是指苯环的2，3或4位有且仅有1个硝基、氟原子或甲氧基取代，R5取自氨基、4-甲基苯基、N-吗啉基、2-(1,3-二氧代异吲哚-2-基)乙基、4-乙酰氨基苯基、苄基、正丁基、3,4-二甲氧基苯基，R6取自乙烯基、4-硝基苯基、环丙基。
一类更优选的化合物是具有通式Ⅱ结构的二芳基-β-内酰胺类化合物：
Figure PCTCN2017078444-appb-000014
其中R取自甲氧基，数目是1个或2个，可为3、4、5-位取代。
或具有通式Ⅲ结构的二芳基-β-内酰胺类化合物：
Figure PCTCN2017078444-appb-000015
其特征在于R取自甲基、三甲基硅基、苯基或者叔丁基苯基，烯键的构型为Z式或E式。
或具有通式Ⅳ和Ⅴ结构的二芳基-β-内酰胺类化合物：
其特征在于R1独立地选自甲基、乙基、羟甲基、氢原子、烷氧基、酰氧基、羟基、卤素、氨基、苯基氨基、苄基氨基、乙酰氨基、对甲苯磺酰氨基、甲磺酰氨基、苯甲酰氨基、3-氟苯甲酰氨基、甲磺酰氧基、甲氧甲基、N,N-二甲氨甲基、4-羟基苄基、三甲基硅乙基、乙氧羰甲基、羧丙酰氧基，R2独立地取自羟基、氨基、卤素、甲氧基、甲基、氟磺酰氧基、氢原子或酰氧基。其中优势化合物为：
Figure PCTCN2017078444-appb-000017
或具有通式Ⅵ结构的二芳基-β-内酰胺类化合物：
Figure PCTCN2017078444-appb-000018
其特征在于R1取自氢原子、甲基、乙酰基、丙烯酰基，R2取自氢原子、苄基、酰 基、丙烯酰基。
一类特别优选的化合物具有如下所示的结构：
Figure PCTCN2017078444-appb-000019
药物组合物和施用方法
由于本发明化合物具有优异的对微管蛋白的抑制活性，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、预防以及缓解由微管蛋白活性或表达量相关的疾病，尤其适用于微管蛋白活性或表达量均相关的疾病。根据现有技术，本发明化合物可用于治疗以下疾病：癌症、神经退行性疾病、疟疾、艾滋病、痛风、糖尿病等等。
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5-200mg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
Figure PCTCN2017078444-appb-000020
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化合物联合给药。
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5～500mg。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。
与现有技术相比，本发明的主要优点包括：
(1)提供了一类结构新颖的化合物，所述的化合物具有微管蛋白抑制活性。
(2)提供了一种具有肿瘤抑制活性的化合物，所述的化合物可以用于制备治疗肿瘤的药物。
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，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。本发明目标化合物的制备方法可以进一步用代表性化合物制备过程体现如下：
实施例1 化合物(S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(1)的合成
参照文献(Wang,X.；Meng,F.；Wang,Y.；Han,Z.；Chen,Y.-J.；Liu,L.；Wang,Z.；Ding,K.Angewandte Chemie Int.Ed.2012,124,9410-9416.)方法，我们设计如下路线合成目标化合物1：
Figure PCTCN2017078444-appb-000021
1.1 3-叔丁基二甲基硅氧基-4-甲氧基苯甲醛(1b)的合成
100mL三口瓶中加入3-羟基-4-甲氧基苯甲醛(1a)(1.58g,10.4mmol)、DMAP(25mg,0.2mmol)、无水DCM(50mL)和无水三乙胺(2mL,14.5mmol)，冷至冰浴下，TBSCl(1.88g,12.5mmol)溶于无水DCM(10mL)中，由滴液漏斗缓慢加入。滴完后撤去冰浴，室温搅拌两小时。加入饱和碳酸氢钠水溶液淬灭反应，分出有机相，水相用DCM萃取三遍，合并有机相，无水硫酸钠干燥。湿法上柱层析分离纯化，收集对应的洗脱液，蒸干溶剂得无色油状化合物(1b)2.66g，收率96％。
1.2 2-[1-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-1-羟基甲基]丙烯酸乙酯(1c)的合成
在50mL茄形瓶中，加入化合物4-甲氧基-3-叔丁基二甲基硅氧基苯甲醛(1b)(2.66g,10mmol)、丙烯酸乙酯(1.00g,10mmol)和DABCO(1.12g,10mmol)，室温搅拌反应2周后，TLC(展开剂：石油醚/乙酸乙酯＝9:1)显示原料(Rf＝0.4)明显减少，有大量新点生成(Rf＝0.3)，停止反应。直接硅胶柱层析，用PE先除去丙烯酸乙酯，继而进行梯度洗脱，收集对应的洗脱液，回收原料1.55g，回收率58％，得无色油状产物(1c)1.12g，扣除回收原料的收率为73％。1H NMR(400MHz,CDCl3)δ6.92(dd,J＝8.3,2.1Hz,1H),6.85(d,J＝2.1Hz,1H),6.81(d,J＝8.3Hz,1H),6.70-6.76(m,1H),6.31(s,1H),5.77(d,J＝1.1Hz,1H),5.47(d,J＝5.0Hz,1H),4.17(q,J＝7.1Hz,2H),3.79(s,3H),1.24(t,J＝7.1Hz,3H),0.98(s,9H),0.13(s,6H)；ESI-LRMS m/z(％):367.2[M+H]+.
1.3 2-[1-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-1-乙酰氧基甲基]丙烯酸乙酯(1d)的合成
化合物1c(1.12g,3.1mmol)、DMAP(45mg,0.31mmol)和TEA(0.63g,6.2mmol)溶于20mL无水DCM中，0℃搅拌条件下，缓慢滴加乙酸酐(0.63g,6.2mmol)，加完后在冰浴中继续搅拌5min后，TLC(展开剂：石油醚/乙酸乙酯＝9:1)显示原料(Rf＝0.3)完全消失，有新点(Rf＝0.5)生成，缓慢加入饱和碳酸氢钠水溶液10mL淬灭反应。分液，水层用DCM(3x 20mL)萃取，合并有机相，无水硫酸钠干燥。拌硅胶柱层析分离纯化，收集对应的洗脱液，减压蒸干溶剂得无色油状化合物(1d)1.2g,收率96％。1H NMR (400MHz,CDCl3)δ6.93(dd,J＝8.3,2.2Hz,1H),6.84(d,J＝2.2Hz,1H),6.79(d,J＝8.3Hz,1H),6.58(s,1H),6.36(s,1H),5.77(s,1H),4.14(q,J＝7.1Hz,2H),3.78(s,3H),2.09(s,3H),1.21(t,J＝7.1Hz,3H),0.98(s,9H),0.13(s,6H)；ESI-LRMS m/z(％):409.2[M+H]+.
1.4 (S)-2-[1-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-1-(3,4,5-三甲氧基苯基氨基)甲基]丙烯酸乙酯(1f)的合成
氮气氛围下,三(二亚苄基丙酮)二钯(27mg,0.029mmol)和1e(49mg,0.074mmol)分别加入一Schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入底物1d(1.2g,2.9mmol),K2CO3(1.0M水溶液,5mL,5mmol)和3,4,5-,三氧基苯胺(809mg,0.62mmol)；室温下搅拌2h后,TLC(展开剂：石油醚/乙酸乙酯＝9:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.2)生成,用二氯甲烷萃取(3×10mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得黄色油状物(1f)1.3g,收率84％；1H NMR(400MHz,CDCl3)δ6.91(dd,J＝8.3,2.1Hz,1H),6.83(d,J＝2.1Hz,1H),6.80(d,J＝8.3Hz,1H),6.34(s,1H),5.88(s,1H),5.81(s,2H),5.28(s,1H),4.15(q,J＝7.1Hz,2H),3.78(s,3H),3.76(s,6H),3.74(s,3H),1.22(t,J＝7.1Hz,3H),0.96(s,9H),0.11(s,6H).
1.5 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(1g)的合成
化合物1f(1.3g)和Sn[N(TMS)2]2(1.6g,3.7mmol)加入一Schlenk管中,加入无水甲苯(20mL),加热回流4.5小时后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.8)基本消失,有新点(Rf＝0.5)生成,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状化合物(1g)1.1g,收率92％；[α]D 20＝+37.3(c 1.00,CHCl3)；1H NMR(400MHz,CDCl3)δ6.91-6.88(m,1H),6.78-6.76(m,2H),6.52(s,2H),5.72(s,1H),5.20(s,1H),5.06(s,1H),3.71(s,3H),3.68(s,3H),3.64(s,6H),0.86(t,J＝2.8Hz,9H),0.01(d,J＝4.8Hz,6H)；13C NMR(100MHz,CDCl3)δ160.5,153.1,151.1,149.6,145.2,134.2,133.4,128.4,120.1,118.9,111.8,110.1,94.5,63.2,60.5,55.6,55.1,25.3,18.1,-5.01；ESI-LRMS m/z:486.2[M+H+].
1.6 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(1)的合成
化合物1g(1.1g,2.3mmol)溶于25mL THF，冷至冰浴下，TBAF(888mg,3.4mmol)溶于5mL THF，由分液漏斗缓慢滴加，滴完后继续搅拌15min后，TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料(Rf＝0.7)完全消失，有新点(Rf＝0.5)生成，减压旋干，20mL乙酸乙酯溶解，水洗(20mL x 2)，饱和食盐水洗(20mL)，无水硫酸钠干燥。过滤浓缩后，柱层析纯化，得白色固体(1)597mg,收率71％。Mp 115–117℃；[α]D 20＝+38.6(c1.0,CHCl3)；1H NMR(400MHz,CDCl3):δ6.93(d,J＝1.7Hz,1H),6.87(dd,J＝8.2,1.7Hz,1H),6.82(d,J＝8.2Hz,1H),6.58(s,2H),5.79(s,1H),5.26(s,1H),5.13(s,1H),3.84(s,3H),3.73(s,3H),3.71(s,6H).13C NMR(150MHz,CDCl3):δ160.3,152.9,149.2,146.5,145.6,134.1,133.2,128.9,118.1,112.3,110.3,110.0,94.31,63.0,60.3,55.5,55.4.ESI-MS(m/z):372.1(M+H+).ESI-HRMS(m/z):calcd for C20H21NO6+H+[M+H]+,372.1442；found,372.1441.
实施例2 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-甲氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(2)的合成
Figure PCTCN2017078444-appb-000022
20mL茄形瓶中加入无水DMF(1mL)、化合物1(22mg,0.059mmol)和碳酸钾(15mg,0.108mmol)，室温搅拌10min，此时溶液为黄色，加入碘甲烷(17mg,0.12mmol)，搅拌反应3h后，TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失，有新点生成(Rf＝0.5)。停止反应，加入10mL乙酸乙酯，以水(2x 10mL)洗涤，饱和食盐水(10mL)洗，无水硫酸钠干燥。拌硅胶柱层析分离纯化，收集对应的洗脱液，蒸干溶剂得白色固体(2)23mg，收率97％。Mp 138-139℃；[α]D 20＝+35.6(c 0.21,CHCl3)；1H NMR(400MHz,CDCl3)δ7.00(dd,J＝8.2,1.8Hz,1H),6.86(d,J＝8.2Hz,1H),6.83(d,J＝1.8Hz,1H),6.60(s,2H),5.84(s,1H),5.31(s,1H),5.17(s,1H),3.88(s,3H),3.83(s,3H),3.76(s,3H),3.73(s,6H)；13C NMR(100MHz,CDCl3)δ161.3,153.8,150.1,150.0,149.9,134.9,134.2,129.1,120.2,111.4,111.2,109.3,95.1,64.3,61.3,56.3；ESI-LRMS m/z(％):386.2[M+H]+；ESI-HRMS m/z(％):Calcd for C21H24NO6[M+H]+386.1598,found386.1598.
实施例3 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-乙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(3)的合成
Figure PCTCN2017078444-appb-000023
20mL茄形瓶中，加入无水DMF 1mL、化合物1(22mg,0.059mmol)和碳酸钾(15mg,0.108mmol)，室温搅拌10min，此时溶液为黄色，加入溴乙烷(12mg,0.12mmol)，过夜反应，TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失，有新点生成(Rf＝0.5)，停止反应，加入10mL乙酸乙酯，以水洗涤(3x 10mL)，无水硫酸钠干燥。直接拌硅胶柱层析分离纯化，收集对应的洗脱液，蒸干溶剂得白色固体(3)23mg，收率97％。Mp 92-93℃；[α]D 20＝+34.0(c 0.27,CHCl3).1H NMR(400MHz,CDCl3)δ6.98(dd,J＝8.2,2.0Hz,1H),6.86(d,J＝8.2Hz,1H),6.83(d,J＝1.9Hz,1H),6.60(s,2H),5.83(s,1H),5.29(s,1H),5.16(s,1H),4.02(q,J＝7.0Hz,2H),3.86(s,3H),3.75(s,3H),3.72(s,6H),1.41(t,J＝7.0Hz,3H)；13C NMR(100MHz,CDCl3)δ161.2,153.6,150.0,149.9,149.1,134.8,134.0,128.9,112.0,111.5,111.1,110.6,94.9,64.6,64.2,61.2,56.2,56.1,14.8；ESI-LRMS m/z(％):400.2[M+H]+；ESI-HRMS m/z(％):Calcd for C22H26NO6[M+H]+400.1755,found 400.1756.
实施例4 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-丙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(4)的合成
Figure PCTCN2017078444-appb-000024
20mL茄形瓶中加入无水DMF 1mL,化合物1(20mg,0.054mmol)和碳酸钾(15mg,0.108mmol),室温搅拌10min(此时溶液为黄色)后加入溴丙烷(13mg,0.108mmol),室温7h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)约40％剩余,有新点生成(Rf＝0.5),升温至50℃过夜,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.2),停止反应,冷至室温后加入10mL乙酸乙酯,以水洗涤(2x 10mL),饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(4)19mg,收率85％；Mp 90-91℃；[α]D 20＝+40.0(c 0.07,CHCl3)；1H NMR(400MHz,CDCl3)δ6.98(dd,J＝8.2Hz,J＝1.4Hz,1H),6.86(d,J＝8.2Hz,1H),6.84(d,J＝1.4Hz,1H),6.60(s,2H),5.84(s,1H),5.30(s,1H),5.17(s,1H),3.90(t,J＝6.8Hz,2H),3.86(s,3H),3.76(s,3H),3.73(s,6H),1.87–1.76(m,2H),1.00(t,J＝7.4Hz,3H)；13C NMR(100MHz,CDCl3)δ161.4,153.8,150.3,150.1,149.5,134.9,134.2,129.0,120.1,111.8,111.1,110.9,95.1,70.8,64.4,61.3,56.3,22.7,10.7；ESI-LRMS m/z(％):436.2[M+Na]+；ESI-HRMS m/z(％):Calcd for C23H28NO6[M+H]+414.1911,found 414.1912.
实施例5 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-异丙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(5)的合成
Figure PCTCN2017078444-appb-000025
20mL茄形瓶中加入无水THF/DMSO(3/1,1.5mL),化合物1(20mg,0.054mmol)和碳酸钾(37mg,0.27mmol),室温搅拌10min(此时溶液为黄色)后加入2-溴丙烷(15mg,0.12mmol),过夜反应后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.2),停止反应,加入10mL乙酸乙酯,以水洗涤(3x 10mL),无水硫酸钠干燥3h,直接拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(5)21mg,收率98％；Mp 146-147℃；[α]D 20＝+13.1(c 0.21,CHCl3)；1H NMR(400MHz,CDCl3)δ6.99(dd,J＝8.2,2.0Hz,1H),6.93–6.85(m,2H),6.61(s,2H),5.84(t,J＝1.7Hz,1H),5.31(s,1H),5.18(s,1H),4.55–4.42(m,1H),3.86(s,3H),3.77(s,3H),3.73(s,6H),1.33(d,J＝6.1Hz,3H),1.28(d,J＝6.1Hz,3H)；13C NMR(100MHz,CDCl3)δ160.4,152.9,150.4,149.3,147.2,134.1,133.2,128.0,119.5,133.3,111.4,110.1,70.1,63.3,60.3,55.4,55.3,21.3,21.2；ESI-LRMS m/z(％):436.2[M+Na]+；ESI-HRMS m/z(％):Calcd for C23H28NO6[M+H]+414.1911,found 414.1916.
实施例6 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-烯丙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(6)的合成
Figure PCTCN2017078444-appb-000026
20mL茄形瓶中加入DMF 2mL,化合物1(20mg,0.054mmol),碳酸铯(35mg,0.108mmol),搅拌0.5h后,加入3-溴丙烯(13mg,0.054mmol),过夜反应后,TLC(展开剂：石油醚/乙酸乙酯＝3:2)显示原料1(Rf＝0.6)完全消失,有新点生成(Rf＝0.3),停止反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(6)21mg,收率98％；M.p.92～93℃；[α]D 20＝+40.0(c 0.39,CHCl3)；1H NMR(400MHz,CDCl3)δ6.99(dd,J＝8.2,1.2Hz,1H),6.87(d,J＝8.2Hz,1H),6.85(s,1H),6.59(s,2H),6.00(ddd,J＝17.4,10.5,5.5Hz,1H),5.83(s,1H),5.33(d,J＝17.4Hz,1H),5.29(s,1H),5.22(d,J＝10.5Hz,1H),5.16(s,1H),4.55(d,J＝5.5Hz,2H),3.87(s,3H),3.76(s,3H),3.73(s,6H)；13C NMR(100MHz,CDCl3)δ161.0,153.5,150.1,149.8,148.6,134.7,133.8,132.8,128.7,120.1,118.3,111.7,111.5,110.7,94.9,69.9,64.0,61.0,56.1,56.0；ESI-LRMS m/z(％):412.2[M+H]+；ESI-HRMS m/z(％):Calcd for C23H26NO6[M+H]+412.1755,found412.1756.
实施例7 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-炔丙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(7)的合成
Figure PCTCN2017078444-appb-000027
20mL茄形瓶中加入DMF 2mL,化合物1(20mg,0.054mmol),碳酸铯(35mg,0.108mmol),搅拌0.5h后,加入3-溴丙炔(13mg,0.054mmol),过夜反应后,TLC(展开剂：石油醚/乙酸乙酯＝3:2)显示原料1(Rf＝0.6)完全消失,有新点生成(Rf＝0.3),停止反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(7)18mg,收率85％；M.p.106～107℃；[α]D 20＝+50.5(c 0.33,CHCl3)；1H NMR(400MHz,CDCl3)δ7.04(d,J＝8.2Hz,1H),7.02(s,1H),6.90(d,J＝8.2Hz,1H),6.60(s,2H),5.84(s,1H),5.33(s,1H),5.17(s,1H),4.73(d,J＝2.2Hz,2H),3.87(s,3H),3.76(s,3H),3.73(s,6H),2.34(t,J＝2.2Hz,1H)；13C NMR(100MHz,CDCl3)δ160.9,153.5,150.3,149.8,147.2,134.7,133.8,128.7,121.0,112.9,111.9,110.8,95.0,78.0,76.0,63.8,60.96,56.9,56.1,56.0；ESI-LRMS m/z(％):410.2[M+H]+；ESI-HRMS m/z(％):Calcd for C23H24NO6[M+H]+ 410.1598,found 410.1600.
实施例8 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟乙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(8)的合成
Figure PCTCN2017078444-appb-000028
20mL茄形瓶中加入无水DMSO 1mL,化合物1(20mg,0.054mmol)和碳酸钾(37mg,0.27mmol),室温搅拌10min(此时溶液为黄色)后加入2-氯乙醇(5mg,0.12mmol),70℃过夜反应,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.1),停止反应,加入10mL乙酸乙酯,以水洗涤(3x 10mL),无水硫酸钠干燥3h,直接拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(8)12mg,收率54％；[α]D 20＝+22.0(c 0.25,CHCl3)；1H NMR(400MHz,CDCl3)δ7.03(dd,J＝4.0Hz,J＝8.0Hz,1H),6.87(d,J＝8.2Hz,2H),6.58(s,2H),5.83(s,1H),5.29(s,1H),5.15(s,1H),4.05(t,J＝4.0Hz,2H),3.92(t,J＝4.0Hz,2H),3.85(s,3H),3.75(s,3H),3.72(s,6H),2.81(s,1H)；13C NMR(100MHz,CDCl3)δ160.3,152.9,149.7,149.1,148.2,134.1,133.2,128.4,120.2,111.6,111.1,110.3,94.2,70.7,63.2,60.5,60.3,55.4,55.3；ESI-LRMS m/z(％):416.2[M+H]+；ESI-HRMS m/z(％):Calcd for C22H26NO7[M+H]+416.1704,found416.1704.
实施例9 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟丙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(9)的合成
Figure PCTCN2017078444-appb-000029
20mL茄形瓶中加入无水乙腈2mL,化合物1(20mg,0.054mmol),碳酸钾(15mg,0.108mmol),碘化钾(1.5mg,0.0081mmol),搅拌条件下加入3-溴丙醇(9mg,0.065mmol),升温至50摄氏度过夜反应后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.1),停止反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(9)22mg,收率95％；[α]D 20＝+18.7(c 0.27,CHCl3)；1H NMR(400MHz,CDCl3)δ7.01(d,J＝8.2Hz,1H),6.91–6.79(m,2H),6.61(s,2H),5.85(s,1H),5.31(s,1H),5.18(s,1H),4.14(t,J＝5.8Hz,2H),3.86–3.84(m,5H),3.77(s,3H),3.74(s,6H),2.59(brd,1H),2.08–2.03(m,2H)；13C NMR(100MHz,CDCl3)δ160.4,152.9,149.4,149.2,148.3,134.1,133.2,128.2,119.8,110.7,110.2,110.1,94.2,67.7,63.3, 60.7,60.3,55.5,55.3,31.0,29.1；ESI-LRMS m/z(％):431.2[M+H]+；ESI-HRMS m/z(％):Calcd for C23H28NO7[M+H]+430.1860,found 430.1861.
实施例10 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氯乙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(10)的合成
Figure PCTCN2017078444-appb-000030
20mL茄形瓶中加入2-丁酮2mL,化合物1(20mg,0.054mmol),碳酸钾(15mg,0.108mmol),搅拌条件下加入2-溴氯乙烷(31mg,0.162mmol),回流反应14h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)基本消失,有新点生成(Rf＝0.4),停止反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得淡黄色固体(10)17mg,收率73％；M.p.91～92℃；[α]D 20＝+36.4(c 0.33,CHCl3)；1H NMR(400MHz,CDCl3)δ7.05(dd,J＝8.2,1.8Hz,1H),6.90–6.88(m,2H),6.59(s,2H),5.84(s,1H),5.30(s,1H),5.16(s,1H),4.22(t,J＝6.0Hz,2H),3.87(s,3H),3.80(t,J＝6.0Hz,2H),3.76(s,3H),3.73(s,6H)；13C NMR(100MHz,CDCl3)δ161.2,153.7,150.6,149.9,148.5,134.9,134.0,129.1,121.4,112.7,112.3,111.2,95.0,69.7,64.0,61.2,56.3,42.0；ESI-LRMS m/z(％):434.1[M+H]+；ESI-HRMS m/z(％):Calcd for C22H25ClNO6[M+H]+434.1365,found434.1366.
实施例11 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氯丙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(11)的合成
Figure PCTCN2017078444-appb-000031
20mL茄形瓶中加入丙酮2mL,化合物1(20mg,0.054mmol),碳酸钾(15mg,0.108mmol),搅拌条件下加入3-溴氯丙烷(34mg,0.216mmol),回流过夜反应后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.5),停止反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(11)18mg,收率77％；M.p.72～73℃；[α]D 20＝+20.1(c 0.15,CHCl3)；1H NMR(400MHz,CDCl3)δ7.01(d,J＝8.2Hz,1H),6.88–6.86(m,2H),6.60(s,2H),5.84(s,1H),5.30(s,1H),5.17(s,1H),4.18–4.01(m,2H),3.85(s,3H),3.76–3.73(m,11H),2.27–2.22(m,2H)；13C NMR(100MHz,CDCl3)δ161.2,153.7,150.3,150.0,149.0,134.8,134.0,129.0,120.6,112.0, 111.6,111.1,95.0,65.9,64.1,61.2,56.3,56.2,41.7,32.4；ESI-LRMS m/z(％):449.1[M+H]+；ESI-HRMS m/z(％):Calcd for C23H27ClNO6[M+H]+448.1521,found 448.1523.
实施例12 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-乙氧酰基甲氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(12)的合成
Figure PCTCN2017078444-appb-000032
20mL茄形瓶中加入无水DMF 1.5mL,化合物1(20mg,0.054mmol)和碳酸铯(35mg,0.108mmol),冰浴条件下搅拌0.5h(此时溶液为黄色,有不溶物)后加入溴乙酸乙酯(18mg,0.108mmol)(溶液由黄色变为无色,仍有不溶物),升温至90摄氏度反应6h,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.5),停止反应,冷却至室温后加入10mL乙酸乙酯,以水洗涤(3x 10mL),饱和食盐水(10mL)洗,无水硫酸钠干燥3h,直接拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得无色油状物(12)33mg,收率95％；1H NMR(400MHz,CDCl3)δ7.05(dd,J＝8.3,1.9Hz,1H),6.90(d,J＝8.3Hz,1H),6.81(d,J＝1.9Hz,1H),6.58(s,2H),5.82(s,1H),5.29(s,1H),5.14(s,1H),4.64(s,2H),4.17–4.11(m,2H),3.88(s,3H),3.76(s,3H),3.73(s,6H),1.22(t,J＝7.2Hz,3H)；ESI-LRMS m/z(％):458.0[M+H]+.
实施例13 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-叔丁氧酰基甲氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(13)的合成
Figure PCTCN2017078444-appb-000033
20mL茄形瓶中加入化合物1(20mg,0.054mmol)和碳酸铯(35mg,0.108mmol),溴乙酸叔丁酯(21mg,0.108mmol),无水DMF 1.5mL,室温搅拌反应5h,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.5),停止反应,冷却至室温后加入10mL乙酸乙酯,以水洗涤(3x 10mL),饱和食盐水(10mL)洗,无水硫酸钠干燥3h,直接拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得淡黄色油状物(13)24mg,收率92％；[α]D 20＝+37.8(c 0.12,CHCl3)；1H NMR(400MHz,CDCl3)δ7.03(d,J＝8.2Hz,1H),6.89(d,J＝8.2Hz,1H),6.76(s,1H),6.58(s,2H),5.81(s,1H),5.27(s,1H),5.13(s,1H),4.53(s,2H),3.88(s,3H),3.75(s,3H),3.72(s,6H),1.39(s,9H)；13C NMR(150MHz,CDCl3)δ160.9,153.5,150.2,149.9,148.9,134.8,133.8,128.9,120.4,111.7,111.6,110.7,94.9,67.3,63.9,60.9,58.0,56.1,55.9,45.9；ESI-LRMS m/z(％):486.2[M+H]+；ESI-HRMS m/z(％):Calcd for C26H31NNaO8[M+Na]+508.1942,found 508.1944.
实施例14 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-二甲氨基乙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(14)的合成
Figure PCTCN2017078444-appb-000034
20mL茄形瓶中加入无水乙腈2mL,化合物1(30mg,0.081mmol),碳酸钾(45mg,0.323mmol),2-氯乙基二甲胺盐酸盐(14mg,0.097mmol),回流反应8h后,TLC(展开剂：二氯甲烷/甲醇＝15:1)显示原料1(Rf＝0.8)完全消失,有新点生成(Rf＝0.5),停止反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(14)27mg,收率88％；[α]D 20＝+37.8(c 0.12,CHCl3)；1H NMR(400MHz,CDCl3)δ7.03(d,J＝8.2Hz,1H),6.89(d,J＝8.2Hz,1H),6.76(s,1H),6.58(s,2H),5.81(s,1H),5.27(s,1H),5.13(s,1H),4.53(s,2H),3.88(s,3H),3.75(s,3H),3.72(s,6H),1.39(s,9H)；13C NMR(150MHz,CDCl3)δ160.9,153.5,150.2,149.9,148.9,134.8,133.8,128.9,120.4,111.7,111.6,110.7,94.9,67.3,63.9,60.9,58.0,56.1,55.9,45.9；ESI-LRMS m/z(％):486.2[M+H]+；ESI-HRMS m/z(％):Calcd for C26H31NNaO8[M+Na]+508.1942,found 508.1944.
实施例15 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-吗啉基乙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(15)的合成
Figure PCTCN2017078444-appb-000035
20mL茄形瓶中加入无水乙腈2mL,化合物1(20mg,0.054mmol),碳酸钾(30mg,0.216mmol),N-(2-氯乙基)吗啉盐酸盐(12mg,0.065mmol),回流反应9h后,TLC(展开剂：石油醚/乙酸乙酯＝1:2)显示原料1(Rf＝0.7)完全消失,有新点生成(Rf＝0.2),停止反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(15)23mg,收率88％；Mp 86-87℃；[α]D 20＝+115.4(c 0.13,CHCl3)；1H NMR(400MHz,CDCl3)δ7.01(d,J＝8.2Hz,1H),6.86(d,J＝8.2Hz,1H),6.84(d,J＝0.9Hz,1H),6.59(s,2H),5.83(s,1H),5.29(s,1H),5.15(s,1H),4.12–3.97(m,2H),3.85(s,3H),3.76(s,3H),3.74–3.67(m,10H),2.78(t,J＝6.0Hz,2H),2.55(s,4H)；13C NMR(150MHz,CDCl3)δ160.4,152.9,149.5,149.2,148.3,134.1,133.2,128.3,119.9,111.0,110.4,110.2,94.2,66.2,66.0,63.3,60.3,56.8,55.4,55.3,53.4；ESI-LRMS m/z(％):485.2[M+H]+；ESI-HRMS m/z(％):Calcd for C26H33N2O7[M+H]+485.2282,found 485.2284.
实施例16 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氨甲酰甲氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(16)的合成
Figure PCTCN2017078444-appb-000036
20mL茄形瓶中加入丁酮2mL,化合物1(20mg,0.054mmol),碳酸钾(22mg,0.162mmol),碘化钾(1mg,0.0054mmol)和氯乙酰胺(7mg,0.068mmol),回流反应9h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.1),停止反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得无色油状物(16)22mg,收率95％；[α]D 20＝+20.9(c 0.29,CHCl3)；1H NMR(400MHz,CDCl3)δ7.08(d,J＝8.3Hz,1H),6.92–6.90(m,3H),6.57(s,2H),6.02(s,1H),5.84(s,1H),5.30(s,1H),5.15(s,1H),4.48(s,2H),3.87(s,3H),3.76(s,3H),3.74(s,6H)；13C NMR(150MHz,CDCl3)δ170.5,160.1,153.0,149.7,149.0,147.1,134.2,133.0,128.7,121.2,113.1,111.6,110.3,94.3,68.7,62.8,60.3,55.5,55.3；ESI-LRMS m/z(％):429.2[M+H]+；ESI-HRMS m/z(％):Calcd for C22H24NaO7[M+Na]+451.1476,found 451.1476.
实施例17 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-苄氧酰氨基丙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(17)的合成
Figure PCTCN2017078444-appb-000037
20mL茄形瓶中加入无水DMF 2mL,化合物1(20mg,0.054mmol),碳酸钾(15mg,0.108mmol),碘化钾(1.5mg,0.0081mmol),搅拌条件下加入N-苄氧羰基-3-溴丙胺(44mg,0.162mmol),升温至55摄氏度过夜反应后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.25),停止反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(17)22mg,收率73％；1H NMR(400MHz,CDCl3)δ7.39–7.27(m,5H),7.00(d,J＝8.0Hz,1H),6.83–6.81(m,2H),6.59(s,2H),6.00(s,1H),5.83(s,1H),5.28(s,1H),5.15(s,1H),5.09(s,2H),4.04(t,J＝5.5Hz,2H),3.75(s,3H),3.72(s,6H),3.71(s,3H),3.43(dd,J＝11.2,5.5Hz,2H),2.03–1.96(m,2H)；ESI-LRMS m/z(％):563.2[M+H]+.
实施例18 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-苯甲酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(18)的合成
Figure PCTCN2017078444-appb-000038
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(11mg,0.108mmol),苯甲酰氯(12mg,0.081mmol),后升至室温反应0.5h,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.4),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(18)23mg,收率90％；M.p.151～152℃；[α]D 20＝+22.2(c 0.34,CHCl3)；1H NMR(400MHz,CDCl3)δ8.18(d,J＝7.8Hz,2H),7.66–7.62(m,1H),7.53–7.49(m,2H),7.30(dd,J＝8.4,2.0Hz,1H),7.24(d,J＝2.0Hz,1H),7.02(d,J＝8.4Hz,1H),6.61(s,2H),5.86(s,1H),5.34(s,1H),5.22(s,1H),3.82(s,3H),3.78(s,3H),3.76(s,6H)；13C NMR(100MHz,CDCl3)δ164.5,160.9,153.6,151.9,149.6,140.5,134.8,133.8,133.7,130.3,129.1,129.0,128.6,125.3,121.9,113.0,111.1,94.9,63.4,61.0,56.1,56.0；ESI-LRMS m/z(％):476.2[M+H]+；ESI-HRMS m/z(％):Calcd for C27H26NO7[M+H]+476.1704,found476.1705.
实施例19 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-对硝基苯甲酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(19)的合成
Figure PCTCN2017078444-appb-000039
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(11mg,0.108mmol),对硝基苯甲酰氯(15mg,0.081mmol),后升至室温反应0.5h,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.4),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得淡黄色固体(19)27mg,收率96％；M.p.88～89℃；[α]D 20＝+45.2(c 0.31,CHCl3)；1H NMR(400MHz,CDCl3)δ8.35(s,4H),7.34(dd,J＝8.5,2.1Hz,1H),7.24(d,J＝2.1Hz,1H),7.05(d,J＝8.5Hz,1H),6.60(s,2H),5.87(s,1H),5.35(s,1H),5.22(s,1H),3.83(s,3H),3.77(s,3H),3.76(s,6H)；13C NMR(100MHz,CDCl3)δ162.6,160.7,153.6,151.6,151.0,149.6,140.0,134.9,134.5,133.7,131.4,129.2,125.9,123.7,121.4,113.0,111.2,94.9,63.3,61.0,56.2,56.1；ESI-LRMS m/z(％):521.1[M+H]+；ESI-HRMS m/z(％):Calcd for C27H25N2O9[M+H]+521.1555,found 521.1554.
实施例20 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-对氟苯甲酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(20)的合成
Figure PCTCN2017078444-appb-000040
化合物1(29mg,0.078mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入无水吡啶(130mg,1.6mmol),对氟苯甲酰氯(109mg,0.78mmol),升至室温反应0.5h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.4),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(20)35mg,收率91％；M.p.185～186℃；[α]D 20＝+38.5(c 0.15,CHCl3)；1H NMR(400MHz,CDCl3)δ8.22–8.14(m,2H),7.30(dd,J＝8.5,2.2Hz,1H),7.22(d,J＝2.2Hz,1H),7.19–7.15(m,2H),7.02(d,J＝8.5Hz,1H),6.60(s,2H),5.86(t,J＝1.7Hz,1H),5.34(s,1H),5.21(s,1H),3.81(s,3H),3.77(s,3H),3.75(s,6H)；13C NMR(100MHz,CDCl3)δ167.6,165.1,163.6,161.0,153.7,152.0,149.7,140.4,134.8,133.9,133.1,133.0,129.1,125.6,125.5,125.4,122.0,116.1,116.0,113.0,111.4,95.0,63.5,61.1,56.3,56.2；ESI-LRMS m/z(％):494.2[M+H]+；ESI-HRMS m/z(％):Calcd for C27H25FNO8[M+H]+494.1610,found 494.1611.
实施例21 (S)-1-(3,4,5-三甲氧基苯基)-4-[3-(2-甲氧基)苯甲酰氧基-4-甲氧基苯基]-3-亚甲基氮杂环丁烷-2-酮(21)的合成
Figure PCTCN2017078444-appb-000041
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(11mg,0.108mmol),邻甲氧基苯甲酰氯(14mg,0.081mmol),后升至室温反应0.5h,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.4),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得无色油状物(21)23mg,收率84％；[α]D 20＝+22.2(c 0.11,CHCl3)；1H NMR(400MHz,CDCl3)δ8.15–8.11(m,2H),7.35–7.32(m,1H),7.28(dd,J＝8.7,2.0Hz,1H),7.22(d,J＝2.0Hz,1H),7.02–6.95(m,3H),6.91–6.87(m,1H),6.61(s,2H),5.85(s,1H),5.33(s,1H),5.21(s,1H),3.88(s,3H),3.81(s,3H),3.77(s,3H),3.76(s,6H)；13C NMR(151MHz,CDCl3)δ171.3,164.2,164.0,160.9,160.3,153.6,152.0,149.6,140.6,134.8,133.8,132.4,129.5,128.9,128.2,127.2,125.2,122.0,121.4,113.9,113.7,112.9,111.1,94.9,92.8, 77.3,77.1,76.8,69.9,66.9,63.4,60.9,56.1,56.1,55.5,55.3,10.6；ESI-LRMS m/z(％):506.1[M+H]+；ESI-HRMS m/z(％):Calcd for C28H27NNaO8[M+Na]+528.1629,found528.1630.
实施例22 (S)-1-(3,4,5-三甲氧基苯基)-4-[3-(3-吡啶甲酰氧基-4-甲氧基苯基)]-3-亚甲基氮杂环丁烷-2-酮(22)的合成
Figure PCTCN2017078444-appb-000042
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(17mg,0.216mmol),2-吡啶甲酰氯(15mg,0.081mmol),升至室温反应0.5h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.2),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(22)16mg,收率62％；M.p.113～114℃；[α]D 20＝+44.9(c 0.35,CHCl3)；1H NMR(400MHz,CDCl3)δ9.37(s,1H),8.85(d,J＝4.4Hz,1H),8.43(d,J＝8.0Hz,1H),7.46(dd,J＝8.0,4.4Hz,1H),7.32(dd,J＝8.4,1.7Hz,1H),7.24(d,J＝1.7Hz,1H),7.03(d,J＝8.5Hz,1H),6.60(s,2H),5.86(s,1H),5.34(s,1H),5.22(s,1H),3.82(s,3H),3.77(s,3H),3.76(s,6H)；13C NMR(150MHz,CDCl3)δ163.2,160.8,154.0,153.6,151.7,151.5,149.6,140.0,137.7,134.9,133.7,129.1,125.7,125.3,123.5,121.7,113.0,111.2,94.9,63.3,61.0,56.1；ESI-LRMS m/z(％):477.2[M+H]+；ESI-HRMS m/z(％):Calcd for C26H25N2O7[M+H]+477.1656,found 477.1659.
实施例23 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-环丙酰甲氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(23)的合成
Figure PCTCN2017078444-appb-000043
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入无水吡啶(90mg,1.2mmol),环丙甲酰氯(40mg,0.54mmol),升至室温反应0.5h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.4),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(23)20mg,收率84％；M.p.68～69℃；[α]D 20＝+12.4(c 0.44,CHCl3)；1H NMR(400MHz,CDCl3)δ7.23(dd,J＝8.4,2.1Hz,1H),7.11(d,J＝2.1Hz,1H),6.95(d,J＝8.5Hz,1H),6.58(s,2H),5.84(s,1H),5.30(s,1H),5.18(s,1H),3.83(s,3H),3.76(s,3H), 3.73(s,6H),1.91–1.78(m,1H),1.20–1.12(m,2H),1.07–0.98(m,2H)；13C NMR(100MHz,CDCl3)δ161.0,153.6,150.0,149.8,148.9,133.8,128.9,120.4,111.4,110.9,110.8,94.9,68.3,64.0,61.3,61.0,56.1,56.0,31.7；ESI-LRMS m/z(％):440.2[M+H]+；ESI-HRMS m/z(％):Calcd for C24H25NNaO7[M+Na]+462.1523,found 462.1526.
实施例24 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-丙烯酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(24)的合成
Figure PCTCN2017078444-appb-000044
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(8.2mg,0.108mmol),丙烯酰氯(10mg,0.081mmol),升至室温反应0.5h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.4),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(24)17mg,收率74％；M.p.109～110℃；[α]D 20＝+52.1(c 0.26,CHCl3)；1H NMR(400MHz,CDCl3)δ7.26(dd,,J＝8.4Hz,2.0Hz,1H),7.15(d,J＝2.0Hz,1H),6.98(d,J＝8.4Hz,1H),6.61–6.57(m,3H),6.32(dd,J＝17.6,10.4Hz,1H),6.02(d,J＝10.4Hz,1H),5.85(s,1H),5.31(s,1H),5.19(s,1H),3.82(s,3H),3.76(s,3H),3.74(s,6H)；13C NMR(100MHz,CDCl3)δ163.8,160.8,153.6,151.7,149.6,140.0,134.8,133.8,132.9,128.9,127.4,125.3,121.8,112.9,111.1,94.8,63.3,61.0,56.1,56.0；ESI-LRMS m/z(％):426.1[M+H]+；ESI-HRMS m/z(％):Calcd for C23H23NNaO7[M+Na]+448.1367,found448.1369.
实施例25 (S)-1-(3,4,5-三甲氧基苯基)-4-[3-(4-吗啉碳酰氧基)-4-甲氧基苯基]-3-亚甲基氮杂环丁烷-2-酮(25)的合成
Figure PCTCN2017078444-appb-000045
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入无水吡啶(90mg,1.2mmol),N-吗啉酰氯(81mg,0.54mmol),升至室温反应9h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)基本消失,有新点生成(Rf＝0.2),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(25)20mg,收率84％；M.p.82～83℃；[α]D 20＝+30.0(c 0.16,CHCl3)；1H NMR(400MHz,CDCl3)δ7.23(dd,J＝8.4,2.1Hz,1H),7.15(d,J＝2.1Hz,1H),6.95(d,J＝8.4Hz,1H),6.59(s,2H),5.84(s,1H),5.31(s,1H),5.19(s,1H),3.84(s,3H),3.76(s,3H),3.76(s,3H),3.75–3.73(m,10H),3.67 (s,2H),3.54(s,2H)；13C NMR(150MHz,CDCl3)δ160.2,153.0,152.5,151.5,149.0,140.2,134.1,133.1,128.2,124.3,121.3,112.0,110.5,94.3,66.0,62.8,60.3,55.5,55.4,44.5,43.7；ESI-LRMS m/z(％):485.2[M+H]+；ESI-HRMS m/z(％):Calcd for C25H28N2NaO8[M+Na]+507.1738,found 507.1739.
实施例26 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氨基磺酰氧基-4-甲氧基苯基)-3-乙氧酰基甲基丁胺-2-酮(26)的合成
Figure PCTCN2017078444-appb-000046
氮气保护条件下,化合物1(40mg,0.108mmol)和氨基磺酰氯(62mg,0.54mmol)置于Schlenk中,加入2.5mL经分子筛除水的DMA,室温搅拌反应4.5h,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.2),停止反应(1mL水淬灭),加入10mL乙酸乙酯,以水洗涤(3x 10mL),饱和食盐(10mL)洗,无水硫酸钠干燥3h,直接拌硅胶柱层析分离纯化,收集对应的洗脱液并蒸干溶剂,得白色固体(26)46mg,收率95％；Mp 199-200℃；[α]D 20＝-14.3(c 0.10,DMSO)；1H NMR(400MHz,DMSO)δ7.98(d,J＝2.2Hz,1H),7.83(dd,J＝8.5,2.2Hz,1H),7.62(d,J＝8.5Hz,1H),7.12(s,2H),6.23(t,J＝1.7Hz,1H),6.09(s,1H),5.70(s,1H),4.30(s,3H),4.15(s,6H),4.07(s,3H)；13C NMR(100MHz,CDCl3)δ160.1,153.1,151.8,149.2,138.6,133.9,132.9,128.6,125.9,122.7,113.7,111.6,94.6,61.6,60.0,55.7,55.6；ESI-LRMS m/z(％):451.1[M+H]+；ESI-HRMS m/z(％):Calcd for C20H26N3O8S[M+NH4]+468.1435,found 468.1435.
实施例27 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-对甲苯磺酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(27)的合成
Figure PCTCN2017078444-appb-000047
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(15mg,0.162mmol),对甲基苯磺酰氯(16mg,0.081mmol),升至室温反应0.5h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.2),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(27)27mg,收率95％；Mp 162-163℃；[α]D 20＝+53.1(c 0.16,CHCl3)；1H NMR(400MHz,CDCl3)δ7.66(d,J＝8.3Hz,2H),7.30–7.20(m,3H),7.14(d,J＝2.1Hz,1H),6.88(d,J＝8.5Hz,1H),6.56(s,2H),5.84(t,J＝1.5Hz,1H),5.29(s,1H),5.16(s,1H),3.79(s,3H),3.77(s,6H),3.61(s,3H),2.44(s,3H)；13C NMR(150MHz,CDCl3)δ160.0,153.0,151.80,148.9,144.7,138.1,134.2,133.0,132.4,128.8,128.3,127.8,125.6,122.2, 112.7,110.5,94.2,62.3,60.3,55.5,55.2,21.0；ESI-LRMS m/z(％):548.1[M+Na]+；ESI-HRMS m/z(％):Calcd for C27H31N2O8S[M+NH4]+543.1796,found 543.1794.
实施例28 (S)-1-(3,4,5-三甲氧基苯基)-4-[3-(2-邻苯二甲酰亚氨基乙磺酰氧基)]-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(28)的合成
Figure PCTCN2017078444-appb-000048
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(14mg,0.162mmol),2-邻苯二甲酰亚氨基乙烷磺酰氯(16mg,0.081mmol),升至室温反应2d后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示有新点生成(Rf＝0.2),原料1(Rf＝0.3)不再有减少趋势,停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(28)15mg,收率46％；Mp 69-70℃；[α]D 20＝+12.1(c 0.45,CHCl3)；1H NMR(400MHz,CDCl3)δ7.88–7.85(m,2H),7.76–7.73(m,2H),7.43(s,1H),7.27(d,J＝8.6Hz,1H),7.00(d,J＝8.6Hz,1H),6.55(s,2H),5.85(s,1H),5.31(s,1H),5.19(s,1H),4.34(t,J＝6.9Hz,2H),3.91(s,3H),3.80–3.64(m,11H)；13C NMR(150MHz,CDCl3)δ166.9,160.0,153.0,151.2,148.8,137.4,134.2,133.7,132.9,131.2,128.8,125.6,123.4,123.0,113.0,110.7,94.1,62.2,60.3,55.6,55.5,48.4,31.9；ESI-LRMS m/z(％):609.1[M+H]+；ESI-HRMS m/z(％):Calcd for C30H28N2NaO10S[M+Na]+631.1357,found 631.1357.
实施例29 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-吗啉磺酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(29)的合成
Figure PCTCN2017078444-appb-000049
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(14mg,0.162mmol),4-吗啉磺酰氯(15mg,0.081mmol),升至室温反应4d后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.25),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(29)20mg,收率71％；M.p.134～135℃；[α]D 20＝+24.7(c 0.11,CHCl3)；1H NMR(400MHz,CDCl3)δ7.43(d,J＝2.0Hz,1H),7.26(dd,J＝8.5,2.0Hz,1H),6.99(d,J＝8.5Hz,1H),6.57(s,2H),5.86(t,J＝1.6Hz,1H),5.31(s,1H),5.19(s,1H),3.88(s,3H),3.76–3.73(m,13H),3.38(t,J＝4.8Hz,4H)；13C NMR(150MHz,CDCl3)δ160.0,153.0,151.3,148.9,138.5,134.2,132.9,128.6,125.1,122.1, 113.0,110.6,94.2,65.3,62.4,60.3,55.6,55.5,46.3；ESI-LRMS m/z(％):521.2[M+H]+；ESI-HRMS m/z(％):Calcd for C24H29N2O9S[M+H]+521.1588,found 521.1590.
实施例30 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-对乙酰氨基苯磺酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(30)的合成
Figure PCTCN2017078444-appb-000050
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(15mg,0.162mmol),对乙酰氨基苯磺酰氯(21mg,0.081mmol),升至室温反应0.5h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.1),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得淡黄色固体(30)22mg,收率72％；Mp 87-88℃；[α]D 20＝+64.6(c 0.13,CHCl3)；1H NMR(400MHz,CDCl3)δ8.01(s,1H),7.63(d,J＝8.8Hz,2H),7.55(d,J＝8.8Hz,2H),7.23(dd,J＝8.5,1.8Hz,1H),6.98(d,J＝1.8Hz,1H),6.89(d,J＝8.5Hz,1H),6.49(s,2H),5.81(s,1H),5.28(s,1H),5.15(s,1H),3.79(s,3H),3.73(s,6H),3.66(s,3H),2.22(s,3H)；13C NMR(150MHz,CDCl3)δ168.2,159.9,153.1,151.9,148.6,142.8,138.3,133.9,133.0,129.2,129.0,128.1,125.6,121.2,118.2,112.8,110.6,94.1,62.1,60.4,55.5,55.3,24.1；ESI-LRMS m/z(％):569.1[M+H]+；ESI-HRMS m/z(％):Calcd for C28H28N2NaO9S[M+Na]+591.1408,found 591.1409.
实施例31 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-丁磺酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(31)的合成
Figure PCTCN2017078444-appb-000051
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(15mg,0.162mmol),1-丁基磺酰氯(21mg,0.081mmol),升至室温反应0.5h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.25),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(31)24mg,收率82％；M.p.99～100℃；[α]D 20＝+18.6(c 0.35,CHCl3)；1H NMR(400MHz,CDCl3)δ7.39(d,J＝2.1Hz,1H),7.27(dd,J＝8.5,2.1Hz,1H),6.99(d,J＝8.5Hz,1H),6.56(s,2H),5.85(s,1H),5.31(s,1H),5.19(s,1H),3.87(s,3H),3.76(s,3H),3.74(s,6H),3.29(td,J＝6.9,1.6Hz,2H),2.00–1.92(m,2H),1.54–1.45(m,2H),0.97(t,J＝7.4Hz,3H)；13C NMR(100MHz,CDCl3)δ160.7,153.6,151.9,149.5,138.2,134.8,133.6,129.4,126.0,124.0,113.6,111.2,94.8,62.9,61.0,56.1,51.6,29.7,25.5,21.5,13.5； ESI-LRMS m/z(％):492.2[M+H]+；ESI-HRMS m/z(％):Calcd for C24H29NNaO8S[M+Na]+514.1506,found 514.1508.
实施例32 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-苄基磺酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(32)的合成
Figure PCTCN2017078444-appb-000052
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(15mg,0.162mmol),苄基磺酰氯(16mg,0.081mmol),升至室温过夜反应后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.25),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(32)25mg,收率88％；M.p.58～59℃；[α]D 20＝-0.7(c 0.14,CHCl3)；1H NMR(400MHz,CDCl3)δ7.46–7.36(m,5H),7.26(dd,J＝8.4,2.0Hz,1H),7.05(d,J＝2.0Hz,1H),6.99(d,J＝8.4Hz,1H),6.54(s,2H),5.84(s,1H),5.23(s,1H),5.15(s,1H),4.58(q,J＝14.0Hz,2H),3.89(s,3H),3.76(s,3H),3.74(s,6H)；13C NMR(150MHz,CDCl3)δ160.0,153.0,151.4,148.8,137.9,134.2,132.9,130.3,128.7,128.3,126.8,125.5,122.9,112.9,110.6,94.2,62.4,60.3,57.3,55.6,55.5；ESI-LRMS m/z(％):526.1[M+H]+；ESI-HRMS m/z(％):Calcd for C27H31N2O8S[M+NH4]+543.1796,found543.1797.
实施例33 (S)-1-(3,4,5-三甲氧基苯基)-4-[3-(3,4-二甲氧基)苯磺酰氧基-4-甲氧基苯基]-3-亚甲基氮杂环丁烷-2-酮(33)的合成
Figure PCTCN2017078444-appb-000053
化合物1(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(15mg,0.162mmol),3,4-二甲氧基苯磺酰氯(19mg,0.081mmol),升至室温过夜反应后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.15),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥。拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(33)27mg,收率87％；[α]D 20＝+29.3(c 0.14,CHCl3)；M.p.70～71℃；1H NMR(400MHz,CDCl3)δ7.30–7.26(m,2H),7.24(dd,J＝8.5,2.1Hz,1H),7.18(d,J＝2.1Hz,1H),6.87(d,J＝8.5Hz,1H),6.81(d,J＝8.2Hz,1H),6.55(s,2H),5.83(s,1H),5.29(s,1H),5.15(s,1H),3.94(s,3H),3.83(s,3H),3.77(s,3H),3.76(s,6H),3.63(s,3H)；13C NMR(150MHz,CDCl3)δ159.9,153.3,153.0,151.8,148.9,148.4,138.1,134.2,132.9,128.4,126.6,125.5,122.2,122.2,112.7,110.4,110.0,109.5,94.2,62.3,60.3,55.7,55.5,55.3；ESI-LRMS m/z(％):572.2[M+H]+；ESI-HRMS m/z(％):Calcd for C28H29NNaO10S [M+Na]+594.1404,found 594.1405.
实施例34 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-叔丁氧酰氨基乙氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(34)的合成
Figure PCTCN2017078444-appb-000054
化合物1(20mg,0.054mmol),DMAP(3.6mg,0.003mmol),Boc-甘氨酸(38mg,0.216mmol)和2mL DCM置于Schlenk中,最后加入EDC盐酸盐(41mg,0.216mmol),室温搅拌过夜反应,TLC(展开剂：二氯甲烷/甲醇＝70:1)只显示原料1(Rf＝0.3),无新点生成,LC-MS监测原料完全消失,停止反应,直接拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(34)26mg,收率91％；1H NMR(400MHz,CDCl3)δ7.25(dd,J＝8.5,2.0Hz,1H),7.14(s,1H),6.97(d,J＝8.5Hz,1H),6.57(s,2H),5.84(t,J＝1.4Hz,1H),5.30(s,1H),5.18(t,J＝1.4Hz,1H),5.07(s,1H),4.19(d,J＝5.5Hz,2H),3.81(s,3H),3.76(s,3H),3.73(s,6H),1.45(s,9H)；ESI-LRMS m/z(％):551.0[M+Na]+.
实施例35 2-甲氧基-5-[(S)-3-亚甲基-4-氧代-1-(3,4,5-三甲氧基苯基)氮杂环丁烷-2-基]-N-(叔丁氧羰基)-L-丙氨酸苯酯(35)的合成
Figure PCTCN2017078444-appb-000055
化合物1(20mg,0.054mmol),DMAP(3.6mg,0.0324mmol),Boc-L-丙氨酸(80mg,0.432mmol)和2mL无水DCM置于Schlenk中,最后加入EDC盐酸盐(41mg,0.216mmol),室温搅拌过夜反应,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.5)完全消失,有新点生成(Rf＝0.6),停止反应直接拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(35)25mg,收率85％；1H NMR(400MHz,CDCl3)δ7.24(d,J＝8.5Hz,1H),7.14(s,1H),6.95(d,J＝8.5Hz,1H),6.56(s,2H),5.84(s,1H),5.29(s,1H),5.18(s,1H),5.08(d,J＝7.5Hz,1H),4.61–4.49(m,1H),3.80(s,3H),3.75(s,3H),3.72(s,3H),1.55(d,J＝7.2Hz,3H),1.44(s,9H)；ESI-LRMS m/z(％):565.1[M+Na]+.
实施例36 2-甲氧基-5-[(S)-3-亚甲基-4-氧代-1-(3,4,5-三甲氧基苯基)杂氮环丁烷-2-基]-N-(叔丁氧羰基)-L-缬氨酸苯酯(36)的合成
Figure PCTCN2017078444-appb-000056
化合物1(20mg,0.054mmol),DMAP(3.6mg,0.0324mmol),Boc-L-缬氨酸(94mg,0.432mmol)和2mL无水DCM置于Schlenk中,最后加入EDC盐酸盐(41mg,0.216mmol),室温搅拌过夜反应,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.5)完全消失,有新点生成(Rf＝0.6),停止反应直接拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(36)19mg,收率62％；1H NMR(400MHz,CDCl3)δ7.25(d,J＝8.5Hz,1H),7.12(s,1H),6.96(d,J＝8.5Hz,1H),6.57(s,2H),5.85(s,1H),5.30(s,1H),5.18(s,1H),5.06(d,J＝9.0Hz,1H),4.49(dd,J＝9.0,6.4Hz,1H),3.80(s,3H),3.76(s,3H),3.74(s,6H),2.40–2.30(m,1H),1.45(s,9H),1.08(d,J＝6.8Hz,3H),1.02(d,J＝6.8Hz,3H)；ESI-LRMS m/z(％):593.1[M+Na]+.
实施例37 (S)-二苄基[2-甲氧基-5-(3-亚甲基-4-氧代-1-(3,4,5-三甲氧基苯基)氮杂环丁烷-2-基)苯基]磷酸酯(37)的合成
Figure PCTCN2017078444-appb-000057
化合物1(80mg,0.216mmol)2mL无水DCM置于50mL茄型瓶中,冷制0℃,搅拌条件下依次加入60％氢化钠(10.5mg,0.26mmol)(溶液由无色变为黄色),TBPO(173mg,0.32mmol),升至室温,15min后反应液变为白色膏状,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.25),停止反应(1mL水淬灭),加入10mL乙酸乙酯,以水洗涤(3x 10mL),饱和食盐水(10mL)洗,无水硫酸钠干燥。直接拌硅胶柱层析分离纯化,收集对应的洗脱液并蒸干溶剂,得白色固体(37)120mg,收率96％；1H NMR(400MHz,CDCl3)δ7.37–7.23(m,11H),7.15(dd,J＝8.5,1.0Hz,1H),6.91(d,J＝8.5Hz,1H),6.56(s,2H),5.80(t,J＝1.7Hz,1H),5.23(s,1H),5.18–5.07(m,5H),3.78(s,3H),3.73(s,3H),3.70(s,6H)；ESI-LRMS m/z(％):632.2[M+H]+.
实施例38 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氟-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(38)的合成
化合物38的合成方法与化合物1的合成方法类似：
38.1 2-[1-(3-氟-4-甲氧基苯基)-1-羟基甲基]丙烯酸乙酯(38b)的合成
Figure PCTCN2017078444-appb-000058
在50mL茄形瓶中加入化合物3-氟-4-甲氧基苯甲醛38a(175mg,1.14mmol),丙烯酸乙酯(114mg,1.14mmol),DABCO(128mg,1.14mmol),室温搅拌反应5d后,TLC(展开剂：石油醚/乙酸乙酯＝9:1)显示原料(Rf＝0.5)明显减少,有大量新点生成(Rf＝0.2),停止反应,用较粗的柱子,直接将反应液倒到硅胶柱上,用少量甲苯洗涤反应瓶再倒到柱子上,然后用PE及PE/EA 10/1先除去丙烯酸乙酯,继而进行梯度洗脱,收集对应的洗脱液,蒸干溶剂得无色油状物质(38b)207mg,收率71％.1H NMR(400MHz,CDCl3) δ7.13–7.02(m,2H),6.90(t,J＝8.4,1H),6.32(s,1H),5.82(s,1H),5.46(d,J＝5.5Hz,1H),4.16(q,J＝7.1Hz,2H),3.86(s,3H),3.17(d,J＝5.6Hz,1H),1.24(t,J＝7.1Hz,3H)；ESI-LRMS m/z(％):255.1[M+H]+.
38.2 2-[1-(3-氟-4-甲氧基苯基)-1-乙酰氧基甲基]丙烯酸乙酯(38c)的合成
Figure PCTCN2017078444-appb-000059
化合物38b(166mg,0.65mmol),DMAP(8mg,0.065mmol)和TEA(132mg,1.3mmol)溶于5mL无水DCM中,0℃搅拌条件下,缓慢滴加乙酸酐(133mg,1.3mmol),加完后在冰浴中继续搅拌；10min后,TLC(展开剂：石油醚/乙酸乙酯＝7:1)显示原料(Rf＝0.3)完全消失,有新点(Rf＝0.5)生成,缓慢加入饱和碳酸氢钠水溶液3mL淬灭反应；分液,水层用DCM(3x 5mL)萃取,合并有机相,无水硫酸钠干燥。拌硅胶柱层析分离纯化,收集对应的洗脱液,减压蒸干溶剂得无色油状化合物(38c)164mg,收率85％；1H NMR(400MHz,CDCl3)δ7.12–7.07(m,2H),6.91(t,J＝8.6Hz,1H),6.59(s,1H),6.39(s,1H),5.85(s,1H),4.20–4.10(m,2H),3.87(s,3H),2.09(s,2H),1.22(t,J＝7.1Hz,3H)；13C NMR(100MHz,CDCl3)δ169.3,164.8,153.2,150.8,147.7,147.5,139.4,130.7,130.7,125.3,124.0,123.9,115.5,115.3,112.9,72.3,61.0,56.1,21.1,14.0；ESI-LRMS m/z(％):319.1[M+Na]+；ESI-HRMS m/z(％):Calcd for C15H17FNaO5[M+Na]+319.0952,found 319.0954.
38.3 (S)-2-[1-(3-氟-4-甲氧基苯基)-1-(3,4,5-三甲氧基苯基氨基)甲基]丙烯酸乙酯(38d)的合成
Figure PCTCN2017078444-appb-000060
氮气氛围下,三(二亚苄基丙酮)二钯(4.3mg,0.0047mmol)和1e(8.5mg,0.0118mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入化合物38c(140mg,0.47mmol),K2CO3(1.0M水溶液,1.5mL,1.5mmol)和3,4,5-三甲氧基苯胺(134mg,1.41mmol)；室温下搅拌8小时后,TLC(展开剂：石油醚/乙酸乙酯＝2:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.2)生成,用二氯甲烷萃取(3×10mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状物(38d)137mg,收率69％；[α]D 20＝+63.7(c 0.16,CHCl3)；1H NMR(400MHz,CDCl3)δ7.11(t,J＝2.3Hz,1H),7.08(s,1H),6.91(t,J＝8.3Hz,1H),6.38(s,1H),5.92(s,1H),5.81(s,1H),5.31(s,1H),4.21–4.13(m,2H),3.87(s,3H),3.76(s,6H),3.74(s,3H),1.24(t,J＝7.1Hz,3H)；13C NMR(100MHz,CDCl3)δ166.0,153.8,153.5,151.0,147.1,147.0,143.3,140.3,133.6,133.6,130.4,126.1,123.1,123.1,115.1,114.9,113.3,91.1,61.1,60.9,58.5,56.2,55.8,14.0；ESI-LRMS m/z(％):420.2[M+H]+；ESI-HRMS m/z(％):Calcd for C22H27FNO6[M+H]+420.1817,found 420.1817.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
38.4 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氟-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(38)的合成
Figure PCTCN2017078444-appb-000061
化合物38d(137mg,0.33mmol)和Sn[N(TMS)2]2(183mg,0.40mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流6小时后,TLC(展开剂：石油醚/乙酸乙酯＝2:1)显示原料(Rf＝0.6)完全消失,有新点(Rf＝0.5)生成,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状物(38)60mg,收率58％；[α]D 20＝+107.0(c 0.17,CHCl3),97％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝85:15,1.0mL/min,254nm；tR(minor)＝14.68min；tR(major)＝21.68min]；1H NMR(400MHz,CDCl3)δ7.15–7.10(m,2H),6.98–6.94(m,1H),6.57(s,2H),5.84(s,1H),5.30(s,1H),5.16(s,1H),3.88(s,3H),3.76(s,3H),3.74(s,6H)；13C NMR(150MHz,CDCl3)δ160.0,153.0,152.9,151.2,148.9,147.6,147.5,134.2,133.0,128.7,128.7,122.3,122.2,113.9,113.8,113.07,110.4,94.2,62.5,60.3,55.7,55.5；ESI-LRMS m/z(％):374.1[M+H]+；ESI-HRMS m/z(％):Calcd for C20H21eNO5[M+H]+374.1398,found 374.1400.
实施例39 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氯-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(39)的合成
化合物39的合成方法与化合物1的合成方法类似：
39.1 2-[1-(3-氯-4-甲氧基苯基)-1-羟基甲基]丙烯酸乙酯(39b)的合成
Figure PCTCN2017078444-appb-000062
在50mL茄形瓶中加入化合物39a(1.15g,6.8mmol),丙烯酸乙酯(681mg,6.8mmol),DABCO(763mg,6.8mmol),室温搅拌反应4d后,TLC(展开剂：石油醚/乙酸乙酯＝9:1)显示原料(Rf＝0.5)明显减少,有大量新点生成(Rf＝0.2),停止反应,用较粗的柱子,直接将反应液倒到硅胶柱上,用少量甲苯洗涤反应瓶再倒到柱子上,然后用PE及PE/EA 10/1先除去丙烯酸乙酯,继而进行梯度洗脱,收集对应的洗脱液,回收原料440mg,回收率38％,得无色油状产物(39b)700mg,扣除回收原料,收率62％；1H NMR(400MHz,CDCl3)δ7.37(d,J＝2.1Hz,1H),7.23(dd,J＝8.5,2.1Hz,1H),6.89(d,J＝8.5Hz,1H),6.34(s,1H),5.83(s,1H),5.47(d,J＝5.5Hz,1H),4.17(q,J＝7.1Hz,2H),3.88(s,3H),3.10(d,J＝5.6Hz,1H),1.25(t,J＝7.1Hz,3H)；13C NMR(150MHz,CDCl3)δ165.6,153.9,141.2,134.0,127.9,125.5,125.3,121.7,111.2,71.8,60.4,55.5,13.4；ESI-LRMS m/z(％):293.1[M+Na]+；ESI-HRMS m/z(％):Calcd for C13H15ClNaO4[M+Na]+293.0551,found 293.0553.
39.2 2-[1-(3-氯-4-甲氧基苯基)-1-乙酰氧基甲基]丙烯酸乙酯(39c)的合成
Figure PCTCN2017078444-appb-000063
化合物39b(700mg,2.6mmol),DMAP(32mg,0.26mmol)和TEA(523mg,5.2mmol)溶于5mL无水DCM中,0℃搅拌条件下,缓慢滴加乙酸酐(530mg,5.2mmol),加完后在冰浴中继续搅拌；10min后,TLC(展开剂：石油醚/乙酸乙酯＝7:1)显示原料(Rf＝0.3)完全消失,有新点(Rf＝0.5)生成,缓慢加入饱和碳酸氢钠水溶液3mL淬灭反应；分液,水层用DCM(3x 5mL)萃取,合并有机相,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,减压蒸干溶剂得无色油状化合物(39c)640mg,收率79％；1H NMR(400MHz,CDCl3)δ7.36(d,J＝2.1Hz,1H),7.27(dd,J＝8.4,2.0Hz,1H),6.87(d,J＝8.5Hz,1H),6.57(s,1H),6.39(s,1H),5.87(s,1H),4.20–4.08(m,2H),3.87(s,3H),2.09(s,3H),1.22(t,J＝7.1Hz,3H)；13C NMR(150MHz,CDCl3)δ168.8,164.2,154.3,138.8,130.4,128.9,127.1,124.7,121.7,111.1,71.7,60.4,55.5,20.5,13.4；ESI-LRMS m/z(％):335.1[M+Na]+；ESI-HRMS m/z(％):Calcd for C15H17ClNaO5[M+Na]+335.0657,found 335.0659.
39.3 (S)-2-[1-(3-氯-4-甲氧基苯基)-1-(3,4,5-三甲氧基苯基氨基)甲基]丙烯酸乙酯(39d)的合成
Figure PCTCN2017078444-appb-000064
氮气氛围下,三(二亚苄基丙酮)二钯(4.3mg,0.0047mmol)和1e(8.5mg,0.0118mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入底物39c(200mg,0.64mmol),K2CO3(1M水溶液,2mL,2mmol)和3,4,5-三甲氧基苯胺(176mg,0.96mmol)；室温下搅拌4h后,TLC(展开剂：石油醚/乙酸乙酯＝2:1)显示原料(Rf＝0.8)完全消失,有新点(Rf＝0.5)生成,用二氯甲烷萃取(3×10mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状物(39d)182mg,收率65％；[α]D 20＝+70.5(c 0.17,CHCl3)；1H NMR(400MHz,CDCl3)δ7.37(d,J＝2.1Hz,1H),7.24(dd,J＝8.8,2.1Hz,1H),6.89(d,J＝8.8Hz,1H),6.39(s,1H),5.94(s,1H),5.81(s,2H),5.30(s,1H),4.21–4.13(m,2H),3.89(s,3H),3.77(s,6H),3.75(s,3H),1.24(t,J＝7.1Hz,3H)；13C NMR(150MHz,CDCl3)δ165.5,153.9,153.2,142.7,139.6,133.2,129.8,128.5,126.3,125.6,122.0,111.4,90.5,60.5,60.4,57.8,55.6,55.3,13.5；ESI-LRMS m/z(％):436.1[M+H]+；ESI-HRMS m/z(％):Calcd for C22H27ClNO6[M+H]+436.1521,found 436.1523.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
39.4 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氯-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(39)的合成
Figure PCTCN2017078444-appb-000065
化合物39d(182mg,0.42mmol)和Sn[N(TMS)2]2(220mg,0.50mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流3小时后,TLC(展开剂：石油醚/乙酸乙酯 ＝2:1)显示原料(Rf＝0.5)基本消失,有新点(Rf＝0.4)生成,反应液冷却至室温后,浓缩,柱层析纯化,得白色固体(39)110mg，收率68％；Mp 119-120℃；[α]D 20＝+35.1(c 0.29,CHCl3),97％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝80:20,1.0mL/min,254nm；tR(minor)＝10.62min；tR(major)＝13.97min]1H NMR(400MHz,CDCl3)δ7.41(d,J＝2.0Hz,1H),7.25(dd,J＝8.5,2.0Hz,1H),6.92(d,J＝8.5Hz,1H),6.56(s,2H),5.83(s,1H),5.28(s,1H),5.16(s,1H),3.88(s,3H),3.75(s,3H),3.73(s,6H)；13C NMR(150MHz,CDCl3)δ160.1,154.8,153.0,148.8,134.2,133.0,128.9,128.2,125.7,122.6,111.8,110.5,94.2,62.3,60.3,55.6,55.5；ESI-LRMS m/z(％):390.1[M+H]+；ESI-HRMS m/z(％):Calcd for C20H21ClNO5[M+H]+390.1103,found 390.1103.
实施例40 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-甲氧基羰基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(40)的合成
化合物40的合成路线与化合物1类似。
40.1 2-[1-(3-甲氧羰基-4-甲氧基苯基)-1-羟基甲基]丙烯酸乙酯(40b)的合成
Figure PCTCN2017078444-appb-000066
在50mL茄形瓶中加入化合物40a(2.2g,11.3mmol),丙烯酸乙酯(1.13g,11.3mmol),DABCO(1.27g,11.3mmol),室温搅拌反应8d后,TLC(展开剂：石油醚/乙酸乙酯＝5:1)显示原料(Rf＝0.5)明显减少,有新点生成(Rf＝0.3),停止反应,用较粗的柱子,直接将反应液倒到硅胶柱上,用少量甲苯洗涤反应瓶再倒到柱子上,然后用PE及PE/EA 10/1先除去丙烯酸乙酯,继而进行梯度洗脱,收集对应的洗脱液,回收原料800mg,回收率36％,得无色油状产物(40b)1.6g,扣除回收原料,收率75％；1H NMR(400MHz,CDCl3)δ7.74(d,J＝2.0Hz,1H),7.45(dd,J＝8.6,2.0Hz,1H),6.91(d,J＝8.6Hz,1H),6.30(s,1H),5.84(s,1H),5.49(s,1H),4.18–4.07(m,2H),3.85(s,3H),3.83(s,3H),1.21(t,J＝7.1Hz,3H)；13C NMR(150MHz,CDCl3)δ165.8,165.6,158.1,141.3,132.6,131.2,129.4,125.3,119.24,111.4,72.0,60.4,55.5,51.4,13.4；ESI-LRMS m/z(％):317.0[M+Na]+.
40.2 2-[1-(3-甲氧羰基-4-甲氧基苯基)-1-乙酰氧基甲基]丙烯酸乙酯(40c)的合成
Figure PCTCN2017078444-appb-000067
化合物40b(438mg,1.49mmol),DMAP(18mg,0.149mol)和TEA(300mg,2.98mmol)溶于5mL无水DCM中,0℃搅拌条件下,缓慢滴加乙酸酐(304mg,2.98mmol),加完后在冰浴中继续搅拌；10min后,TLC(展开剂：石油醚/乙酸乙酯＝5:1)显示原料(Rf＝0.3)完全消失,有新点(Rf＝0.5)生成,缓慢加入饱和碳酸氢钠水溶液淬灭反应；分液,水层用DCM(3x 5mL)萃取,合并有机相,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,减压蒸干溶剂得无色油状化合物(40c)350mg,收率70％；1H NMR(400MHz,CDCl3)δ7.79(d,J＝2.4Hz,1H),7.50(dd,J＝8.6,2.4Hz,1H),7.26(s,1H),6.94(d,J＝8.6Hz,1H),6.62(s,1H),6.40(s,1H),5.89(s,1H),4.14(dtt,J＝10.8,7.4,3.7Hz,2H),3.89(s,3H),3.88(s,3H),2.10(s,3H),1.21(t,J＝7.1Hz,3H)；13C NMR(150MHz,CDCl3)δ168.8,165.7,164.2,158.5,138.9, 132.7,130.6,129.1,124.6,119.4,111.3,71.8,60.4,55.5,51.4,20.5,13.4；ESI-LRMS m/z(％):359.0[M+Na]+.
40.3 (S)-2-[1-(3-甲氧羰基-4-甲氧基苯基)-1-(3,4,5-三甲氧基苯基氨基)甲基]丙烯酸乙酯(40d)的合成
Figure PCTCN2017078444-appb-000068
氮气氛围下,三(二亚苄基丙酮)二钯(9.2mg,0.01mmol)和1e(18mg,0.025mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入底物40c(350mg,1.04mmol),K2CO3(1.0M水溶液,3mL,3mmol)和3,4,5-三甲氧基苯胺(286mg,1.56mmol)；室温搅拌1h后,TLC(展开剂：石油醚/乙酸乙酯＝2:1)显示原料(Rf＝0.9)完全消失,有新点(Rf＝0.4)生成,用二氯甲烷萃取(3×5mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状化合物(40d)280mg,收率59％；[α]D 20＝+65.2(c 0.34,CHCl3)；1H NMR(400MHz,CDCl3)δ7.77(t,J＝4.9Hz,1H),7.47(dd,J＝8.6,2.1Hz,1H),6.92(d,J＝8.6Hz,1H),6.38(s,1H),5.94(s,1H),5.80(s,2H),5.33(s,1H),4.20–4.03(m,3H),3.87(s,3H),3.85(s,3H),3.74(s,6H),3.72(s,3H),1.21(t,J＝7.1Hz,2H)；13C NMR(150MHz,CDCl3)δ165.8,165.5,158.0,153.2,142.8,139.8,131.9,131.8,130.0,129.9,125.5,119.6,111.7,90.7,60.4,60.3,57.9,55.5,55.3,51.4,13.4；ESI-LRMS m/z(％):460.1[M+H]+.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
40.4 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-甲氧基羰基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(40)的合成
Figure PCTCN2017078444-appb-000069
化合物40d(280mg,0.61mmol)和Sn[N(TMS)2]2(335mg,0.73mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流3.5h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料(Rf＝0.7)少量剩余,有新点(Rf＝0.6生成),反应液冷却至室温后,浓缩,柱层析纯化,得白色固体(40)100mg,收率40％；Mp 112-113℃；[α]D 20＝+5.5(c 0.34,CHCl3)；97％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝75:25,1.0mL/min,254nm；tR(minor)＝10.18min；tR(major)＝15.93min]；1HNMR(400MHz,CDCl3)δ7.89(d,J＝2.1Hz,1H),7.51(dd,J＝8.7,2.1Hz,1H),7.01(d,J＝8.7Hz,1H),6.59(s,2H),5.88(s,1H),5.37(s,1H),5.20(s,1H),3.92(s,3H),3.91(s,3H),3.78(s,3H),3.75(s,6H)；13C NMR(150MHz,CDCl3)δ166.1,160.8,159.6,153.6,149.5,134.8,133.6,131.7,130.8,128.1,120.5,112.9,111.2,94.8,63.1,61.0,56.2,56.1,52.3；ESI-LRMS m/z(％):414.0[M+H]+；ESI-HRMS m/z(％):Calcd for C22H24NO7[M+H]+414.1547,found 414.1550.
实施例41 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氨基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(41)的合成
化合物41的合成路线与化合物1类似：
41.1 2-[1-(3-硝基-4-甲氧基苯基)-1-羟基甲基]丙烯酸乙酯(41b)的合成
Figure PCTCN2017078444-appb-000070
在50mL茄形瓶中加入化合物4-甲氧基-3-硝基苯甲醛(41a)(1.38g,7.62mmol),丙烯酸乙酯(762mg,7.62mmol),DABCO(855mg,7.62mmol),室温搅拌反应3d后,TLC(展开剂：石油醚/乙酸乙酯＝4:1,3次)显示原料(Rf＝0.6)明显减少,有大量新点生成(Rf＝0.5),停止反应,用较粗的柱子,直接将反应液倒到硅胶柱上,用少量甲苯洗涤反应瓶再倒到柱子上,然后用PE及PE/EA 10/1先除去丙烯酸乙酯,继而进行梯度洗脱,收集对应的洗脱液,蒸干溶剂得黄色油状物质(41b)5.9g,收率99％；1H NMR(400MHz,CDCl3)δ7.85(d,J＝2.1Hz,1H),7.57(dd,J＝8.7,2.2Hz,1H),7.07(d,J＝8.7Hz,1H),6.37(s,1H),5.90(s,1H),5.53(d,J＝5.6Hz,1H),4.18(q,J＝7.1Hz,2H),3.95(s,3H),3.43(d,J＝5.6Hz,1H),1.27(t,J＝7.1Hz,3H)；13C NMR(100MHz,DMSO)δ166.1,152.5,141.6,139.4,134.3,132.7,126.5,124.1,113.6,72.1,72.0,61.4,56.8,14.2；ESI-LRMS m/z(％):304.1[M+Na]+；ESI-HRMS m/z(％):Calcd for C13H19N2O6[M+NH4]+299.1238,found 299.1238.
41.2 2-[1-(3-硝基-4-甲氧基苯基)-1-乙酰氧基甲基]丙烯酸乙酯(41c)的合成
Figure PCTCN2017078444-appb-000071
化合物41b(1.94g,6.9mmol),DMAP(100mg,0.69mmol)和TEA(1.4g,13.8mmol)溶于20mL无水DCM中,0℃搅拌条件下,缓慢滴加乙酸酐(1.4g,13.8mmol),加完后在冰浴中继续搅拌；5min后,TLC(展开剂：石油醚/乙酸乙酯＝4:1)显示原料(Rf＝0.1)完全消失,有新点(Rf＝0.2)生成,缓慢加入饱和碳酸氢钠水溶液10mL淬灭反应；分液,水层用DCM(3x10mL)萃取,合并有机相,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,减压蒸干溶剂得无色油状化合物(41c)1.7g,收率76％；1H NMR(400MHz,CDCl3)δ7.84(d,J＝2.2Hz,1H),7.59(dd,J＝8.7,2.2Hz,1H),7.05(d,J＝8.7Hz,1H),6.60(s,1H),6.43(s,1H),5.95(s,1H),4.19–4.10(m,2H),3.94(s,3H),2.10(s,3H),1.23(t,J＝7.1Hz,3H)；ESI-LRMS m/z(％):346.1[M+Na]+；ESI-HRMS m/z(％):Calcd for C15H21N2O7[M+NH4]+341.1343,found341.1344.
41.3 2-[1-(3-叔丁氧基甲酰氨基-4-甲氧基苯基)-1-乙酰氧基甲基]丙烯酸乙酯(41e)的合成
Figure PCTCN2017078444-appb-000072
50mL茄形瓶中加入化合物41c(1.7g,5.2mmol),醋酸(3.2g,52mmol),和20mL 甲醇,搅拌条件下分批加入锌粉(1.03g),加完后回流反应2h,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.6)完全消失,有新点(Rf＝0.5)生成,反应液用硅藻土过滤以除去锌粉,加入饱和碳酸钠水溶液中和多余醋酸,再以乙酸乙酯萃取,无水硫酸钠干燥有机相。加入无水THF 8mL,碳酸钾(45mg,0.323mmol),回流反应16h后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.5)完全消失,有新点生成(Rf＝0.6),停止反应,减压旋干,10mL乙酸乙酯溶解,水洗(5mL x 2),饱和食盐水洗(5mL),无水硫酸钠干燥。过滤浓缩后,柱层析纯化,得无色油状物(41e)858mg,两步收率40％；1H NMR(400MHz,CDCl3)δ8.10(s,1H),7.07(s,1H),7.01(dd,J＝8.3,2.0Hz,1H),6.79(d,J＝8.3Hz,1H),6.62(s,1H),6.39(s,1H),5.85(s,1H),4.20–4.11(m,2H),3.85(s,3H),2.10(s,3H),1.52(s,9H),1.24(t,J＝7.1Hz,3H)；13C NMR(150MHz,CDCl3)δ168.9,164.5,151.9,146.8,139.2,129.8,127.6,124.9,121.4,116.6,108.9,72.7,60.3,55.1,27.7,20.6,13.4；ESI-LRMS m/z(％):416.1[M+Na]+；ESI-HRMS m/z(％):Calcd for C20H31N2O7[M+NH4]+411.2126,found 411.2128.
41.4 (S)-2-[1-(3-叔丁氧基甲酰氨基-4-甲氧基苯基)-1-(3,4,5-三甲氧基苯基氨基)甲基]丙烯酸乙酯(41e)的合成
Figure PCTCN2017078444-appb-000073
氮气氛围下,三(二亚苄基丙酮)二钯(3.8mg,0.0041mmol)和1e(7.2mg,0.01mmol)分别加入一Schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入底物41e(163mg,0.41mmol),K2CO3(1.0M水溶液,1.5mL,1.5mmol)和3,4,5-,三氧基苯胺(114mg,0.62mmol)；室温下搅拌2h后,TLC(展开剂：石油醚/乙酸乙酯＝2:1)显示原料(Rf＝0.7)完全消失,有新点(Rf＝0.4)生成,用二氯甲烷萃取(3×5mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得黄色油状物(41f)173mg,收率81％；[α]D 20＝+56.4(c 0.15,CHCl3)；1H NMR(400MHz,CDCl3)δ8.03(s,1H),7.02(s,1H),6.91(dd,J＝8.4,2.2Hz,1H),6.72(d,J＝8.4Hz,1H),6.30(s,1H),5.89(s,1H),5.75(s,2H),5.27(s,1H),4.16–4.02(m,2H),3.78(s,3H),3.70(s,6H),3.68(s,3H),1.45(s,9H),1.16(t,J＝7.1Hz,3H)；13C NMR(150MHz,CDCl3)δ165.8,153.1,152.0,146.4,143.0,140.1,132.7,129.7,127.8,125.1,120.6,116.5,109.2,90.6,79.8,60.4,60.1,58.4,55.3,55.1,27.7,13.5；ESI-LRMS m/z(％):517.3[M+H]+；ESI-HRMS m/z(％):Calcd for C27H37N2O8[M+H]+517.2544,found 517.2544.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
41.5 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氨基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(41)的合成
Figure PCTCN2017078444-appb-000074
化合物41f(146mg)和Sn[N(TMS)2]2(149mg,0.33mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流6小时后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料(Rf＝0.6)基本消失,有新点(Rf＝0.3)生成,反应液冷却至室温后,浓缩,柱层析纯化,得淡黄色油状物(41)37mg,收率35％；[α]D 20＝+50.0(c 0.14,CHCl3),97％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝80:20,1.0mL/min,254nm；tR(minor)＝19.58min；tR(major)＝17.42min]；1H NMR(400MHz,CDCl3)δ6.79(dd,J＝8.1,1.9Hz,1H),6.75(d,J＝8.1Hz,1H),6.71(d,J＝1.9Hz,1H),6.62(s,2H),5.80(t,J＝1.6Hz,1H),5.24(s,1H),5.15(t,J＝1.6Hz,1H),3.84(s,3H),3.76(s,3H),3.74(s,6H)；13C NMR(150MHz,CDCl3)δ161.1,153.5,150.0,147.8,136.8,134.6,134.0,129.0,117.5,112.5,110.5,110.3,94.9,64.0,60.9,56.1,55.5；ESI-LRMS m/z(％):371.1[M+H]+；ESI-HRMS m/z(％):Calcd for C20H23N2O5[M+H]+371.1601,found 371.1605.
实施例42 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-丙烯酰氨基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(42)的合成
Figure PCTCN2017078444-appb-000075
化合物41(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(11mg,0.108mmol),丙烯酰氯(10mg,0.081mmol),升至室温反应0.5h后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料(Rf＝0.3)完全消失,有新点生成(Rf＝0.25),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥。拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(42)16mg,收率66％；Mp 143-144℃；[α]D 20＝-96.4(c 0.11,CHCl3)；1H NMR(400MHz,CDCl3)δ8.70(s,1H),7.93(s,1H),7.05(dd,J＝8.5,1.9Hz,1H),6.87(d,J＝8.5Hz,1H),6.62(s,2H),6.43(d,J＝16.8Hz,1H),6.29(dd,J＝16.8,10.1Hz,1H),5.83(s,1H),5.79(d,J＝10.1Hz,1H),5.36(s,1H),5.19(s,1H),3.89(s,3H),3.75(s,3H),3.74(s,6H)；13C NMR(150MHz,CDCl3)δ162.8,160.3,152.9,149.1,147.4,133.9,133.2,130.6,128.4,127.3,127.2,121.0,118.7,110.2,110.0,94.2,63.1,60.3,55.5,55.3；ESI-LRMS m/z(％):425.2[M+H]+；ESI-HRMS m/z(％):Calcd for C23H28N3O6[M+NH4]+442.1973,found 442.1975.
实施例43 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-对硝基苯甲酰氨基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(43)的合成
Figure PCTCN2017078444-appb-000076
化合物41(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入 TEA(11mg,0.108mmol),对硝基苯甲酰氯(15mg,0.081mmol),后升至室温反应20h,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示无新点生成,LC-MS显示原料阳离子峰(M+1)消失,大量目标化合物阳离子峰存在(M+1),停止反应(1mL水淬灭),加入10mLDCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得黄色固体(43)25mg,收率89％；Mp234-235℃；[α]D 20＝-81.6(c 0.20,CHCl3)；1H NMR(400MHz,CDCl3)δ8.70(s,1H,NH),8.61(s,1H),8.36(d,J＝8.7Hz,2H),8.05(d,J＝8.7Hz,2H),7.14(dd,J＝8.4,1.8Hz,1H),6.94(d,J＝8.4Hz,1H),6.64(s,2H),5.86(s,1H),5.41(s,1H),5.22(s,1H),3.95(s,3H),3.76(s,9H)；13C NMR(150MHz,CDCl3)δ162.6,160.3,152.9,149.2,149.1,147.7,139.7,134.1,133.1,128.7,127.6,126.9,123.51,121.9,118.6,110.4,110.1,94.3,63.0,60.3,55.5；ESI-LRMS m/z(％):520.2[M+H]+；ESI-HRMS m/z(％):Calcd for C27H25N3NaO8[M+Na]+542.1534,found 542.1534.
实施例44 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氨基磺酰氨基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(44)的合成
Figure PCTCN2017078444-appb-000077
氮气保护条件下,化合物41(20mg,0.054mmol)和氨基磺酰氯(62mg,0.27mmol)置于Schlenk中,加入2.5mL无水二氯甲烷,室温搅拌反应1h,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.2),停止反应(1mL水淬灭),加入10mL乙酸乙酯,以水洗涤(3x 10mL),无水硫酸钠干燥。直接拌硅胶柱层析分离纯化,收集对应的洗脱液并蒸干溶剂,得白色固体(44)11mg,收率42％；Mp 101-102℃；[α]D 20＝-20.3(c 0.35,CHCl3)；1H NMR(400MHz,CDCl3)δ7.60(d,J＝1.9Hz,1H),7.12(dd,J＝8.5,1.9Hz,1H),7.03(s,1H,NH),6.89(d,J＝8.5Hz,1H),6.59(s,2H),5.85(s,1H),5.34(s,1H),5.19(s,1H),4.84(s,2H,NH2),3.86(s,3H),3.75(m,9H)；13C NMR(150MHz,CDCl3)δ160.3,152.9,148.9,148.7,134.2,133.0,128.7,126.4,122.3,118.4,110.7,110.5,94.5,62.8,60.3,55.6,55.4；ESI-LRMS m/z(％):450.1[M+H]+；ESI-HRMS m/z(％):Calcd for C20H24N3O7S[M+H]+450.1329,found 450.1346.
实施例45 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-环丙甲酰氨基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(45)的合成
Figure PCTCN2017078444-appb-000078
化合物41(20mg,0.054mmol)溶于1.5mL无水DCM中,0℃搅拌条件下,依次加入TEA(11mg,0.108mmol),环丙甲酰氯(8.5mg,0.081mmol),升至室温反应0.5h后,TLC(展开 剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.25),1mL水淬灭反应,加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥。拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(45)11mg,收率54％；Mp 181-182℃；[α]D 20＝-77.3(c 0.24,CHCl3)；1H NMR(400MHz,CDCl3)δ8.59(s,1H,NH),8.05(s,1H),7.01(dd,J＝8.5,1.8Hz,1H),6.85(d,J＝8.5Hz,1H),6.61(s,2H),5.81(s,1H),5.32(s,1H),5.18(s,1H),3.89(s,3H),3.75(s,3H),3.73(s,6H),1.63–1.53(m,1H),1.08(s,2H),0.87(dd,J＝7.6,3.1Hz,2H)；13C NMR(150MHz,CDCl3)δ171.4,160.3,152.9,149.1,147.0,133.9,133.2,128.3,127.7,120.4,118.5,110.2,109.9,94.2,63.1,60.3,55.5,55.2,15.5,7.5,7.4；ESI-LRMS m/z(％):439.2[M+H]+；ESI-HRMS m/z(％):Calcd for C24H26N2NaO6[M+Na]+461.1683,found 461.1685.
实施例46 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-乙酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(46)的合成
Figure PCTCN2017078444-appb-000079
在250mL茄形瓶中加入化合物1(0.25g,0.673mmol),4-二甲氨基吡啶(8mg,0.067mmol),三乙胺(0.13mL,0.94mmol)与二氯甲烷10mL,醋酸酐(0.09mL,0.094mmol),室温反应5分钟；TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0.6)；反应液加饱和NaHCO3,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(46)0.23g,收率83％；1H NMR(400MHz,CDCl3)δ7.25(br d,J＝8.4Hz,1H),7.10(br s,1H),6.97(d,J＝8.4Hz,1H),6.57(s,1H),5.84(s,1H),5.30(s,1H),5.18(s,1H),3.83(s,3H),3.76(s,3H),3.73(s,6H),2.29(s,3H).13C NMR(151MHz,CDCl3)δ168.0,160.2,153.0,151.0,149.0,139.6,134.1,133.1,128.3,124.6,121.1,112.2,110.4,94.2,62.7,60.3,55.5,55.4,20.0；ESI-LRMS m/z(％):414.1[M+H]+.
实施例47 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-苄氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(47)的合成
Figure PCTCN2017078444-appb-000080
在100mL茄形瓶中加入化合物1(0.15g,0.4mmol),溴化苄(58μL,0.5mmol),碳酸钾(72mg,0.5mmol)与DMF 5mL,100℃反应8小时；TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.5)消失,生成新点(Rf＝0.65)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝3:1),得到白色固体(47)0.146g,收率78％；1H NMR(400MHz,CDCl3)δ7.40–7.23(m,5H),6.98(dd,J＝8.2,1.6Hz,1H),6.92–6.82(m,2H),6.54(s,2H),5.78(s,1H),5.24(s,1H),5.11(s,2H),5.07(s,1H),3.89(s,3H),3.77(s,3H),3.70(s,6H).13C NMR(151MHz,CDCl3)δ160.3,152.9,149.7,149.1,147.9,135.9,134.0, 133.2,128.1,127.9,127.3,126.7,119.6,111.5,111.2,110.1,94.1,70.4,63.2,60.3,55.4；ESI-LRMS m/z(％):462.2[M+H]+.
实施例48 (S)-1-(3,4-二甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮的(48)合成
化合物48的合成路线与化合物1类似：
48.1 (S)-2-[1-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-1-(3,4-二甲氧基苯基氨基)甲基]丙烯酸乙酯(48a)的合成
Figure PCTCN2017078444-appb-000081
氮气氛围下,三(二亚苄基丙酮)二钯(5.7mg,0.0062mmol)和1e(11.1mg,0.016mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入1d(250mg,0.62mmol),K2CO3(1.0M水溶液,3mL,3mmol)和3,5-二氧基苯胺(142mg,0.93mmol)；室温下搅拌1.5小时后,TLC(展开剂：石油醚/乙酸乙酯＝5:1)显示原料(Rf＝0.6)完全消失,有新点(Rf＝0.2)生成,用二氯甲烷萃取(3×5mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状物(48a)220mg,收率72％；[α]D 20＝-5.1(c 0.26,CHCl3)；1H NMR(400MHz,CDCl3)δ6.91(dd,J＝8.3,1.9Hz,1H),6.84(d,J＝2.0Hz,1H),6.79(d,J＝8.3Hz,1H),6.70(d,J＝8.6Hz,1H),6.33(s,1H),6.22(d,J＝2.5Hz,1H),6.08(dd,J＝8.6,2.5Hz,1H),5.88(s,1H),5.24(s,1H),4.21–4.08(m,2H),3.88–3.71(m,9H),1.28–1.16(m,3H),0.97(s,9H),0.11(d,J＝9.2Hz,6H)；13C NMR(150MHz,CDCl3)δ165.8,149.8,149.3,144.4,141.1,140.3,132.8,124.7,120.1,119.6,112.5,111.5,103.8,99.0,60.1,58.5,56.1,55.1,54.9,25.1,17.8,13.5,-5.2；ESI-LRMS m/z(％):502.2[M+H]+.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
48.2 (S)-1-(3,4-二甲氧基苯基)-4-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(48b)的合成
Figure PCTCN2017078444-appb-000082
化合物48a(220mg)和Sn[N(TMS)2]2(231mg,0.53mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流4小时后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.7)基本消失,有新点(Rf＝0.5)生成,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状化合物(48b)147mg,收率74％；[α]D 20＝+29.9(c 0.32,CHCl3)；1H NMR(400MHz,CDCl3)δ7.23(s,1H),6.94(dd,J＝8.2,1.7Hz,1H),6.82(d,J＝8.4Hz,2H),6.68(d,J＝8.6Hz,1H),6.57(dd,J＝8.6,2.0Hz,1H),5.78(s,1H),5.25(s,1H),5.11(s,1H),3.86–3.72(m,9H),0.94(s,6H),0.08(s,3H),0.07(s,3H)；13C NMR(150MHz,CDCl3)δ160.1,150.8,149.5,148.7,145.2,144.9,130.9,128.2,119.6,118.8,111.7,110.8,109.4,108.0,101.5,62.9,55.5,55.3,54.9,25.0,17.8,-5.2,-5.3；ESI-LRMS m/z(％):456.2[M+H]+.
48.3 (S)-1-(3,4-二甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮的(48)合成
Figure PCTCN2017078444-appb-000083
化合物48b(1117mg,0.26mmol)溶于5mL THF,冷至冰浴下,TBAF(136mg,0.52mmol)溶于1mL THF,由分液漏斗缓慢滴加,滴完后继续搅拌15min后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.1)生成,减压旋干,10mL乙酸乙酯溶解,水洗(5mL x 2),饱和食盐水洗(5mL),无水硫酸钠干燥3h,过滤浓缩后,柱层析纯化,得白色固体(48)72mg,收率82％；Mp 74-75℃；[α]D 20＝+1.9(c 0.32,CHCl3)；99％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝75:25,1.0mL/min,254nm；tR(minor)＝11.78min；tR(major)＝16.22min]；1H NMR(400MHz,CDCl3)δ7.28(d,J＝2.1Hz,1H),6.94(d,J＝1.7Hz,1H),6.87(dd,J＝8.2,1.7Hz,1H),6.81(d,J＝8.2Hz,1H),6.67(d,J＝8.6Hz,1H),6.54(dd,J＝8.6,2.1Hz,1H),5.91(s,1H),5.76(s,1H),5.26(s,1H),5.11(s,1H),3.85(s,3H),3.81(s,3H),3.78(s,3H)；13C NMR(150MHz,CDCl3)δ160.1,149.4,148.8,146.4,145.6,145.2,130.9,129.0,118.0,112.3,110.8,110.3,109.5,107.8,101.6,62.9,55.5,55.4,55.3；ESI-LRMS m/z(％):342.1[M+H]+；ESI-HRMS m/z(％):Calcd for C20H26NO4[M+H]+342.1336,found 342.1339.
实施例49 (S)-1-(3,5-二甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(49)的合成
化合物49和合成路线与化合物1类似：
49.1 (S)-2-[1-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-1-(3,5-二甲氧基苯基氨基)甲基]丙烯酸乙酯(49a)的合成
Figure PCTCN2017078444-appb-000084
氮气氛围下,三(二亚苄基丙酮)二钯(5.7mg,0.0062mmol)和1e(11.1mg,0.016mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入1d(250mg,0.62mmol),K2CO3(1.0M水溶液,3mL,3mmol)和3,5-二氧基苯胺(142mg,0.93mmol)；室温下搅拌3.5小时后,TLC(展开剂：石油醚/乙酸乙酯＝8:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.4)生成,用二氯甲烷萃取(3×5mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状物(49a)270mg,收率88％；[α]D 20＝+27.7(c 0.27,CHCl3)；1H NMR(400MHz,CDCl3)δ6.89(dd,J＝8.3,2.1Hz,1H),6.82–6.78(m,2H),6.35(s,1H),5.88(d,J＝2.1Hz,2H),5.77(d,J＝2.1Hz,2H),5.28(s,1H),4.21–4.05(m,2H),3.78(s,3H),3.72(s,6H),1.20(t,J＝7.1Hz,3H),0.97(s,9H),0.12(s,6H)；13C NMR(150MHz,CDCl3)δ166.2,161.6,150.5,148.7,145.1,140.5,133.2,125.4,120.7,120.3,112.2,92.4,90.1,60.7,58.5,55.5,55.1,25.7,18.5, 14.,-4.6；ESI-LRMS m/z(％):502.2[M+H]+；ESI-HRMS m/z(％):Calcd for C27H40NO6Si[M+H]+502.2619,found 502.2619.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
49.2 (S)-1-(3,5-二甲氧基苯基)-4-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(49b)的合成
Figure PCTCN2017078444-appb-000085
化合物49a(270mg)和Sn[N(TMS)2]2(284mg,0.65mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流6小时后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.7)基本消失,有新点(Rf＝0.5)生成,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状化合物(49b)213mg,收率76％；[α]D 20＝+39.5(c 0.20,CHCl3)；1H NMR(400MHz,CDCl3)δ6.94(dd,J＝8.3,2.1Hz,1H),6.83–6.81(m,2H),6.53(d,J＝2.2Hz,2H),6.16(t,J＝2.2Hz,1H),5.81(t,J＝1.7Hz,1H),5.25(s,1H),5.18–5.10(m,1H),3.79(s,3H),3.70(s,6H),0.94(s,9H),0.09(s,3H),0.08(s,3H)；13C NMR(150MHz,CDCl3)δ161.2,161.1,151.4,150.1,145.6,139.2,128.8,120.2,119.3,112.3,110.7,96.7,95.8,63.6,55.5,55.3,25.7,18.4,-4.7；ESI-LRMS m/z(％):456.2[M+H]+；ESI-HRMS m/z(％):Calcd for C25H34NO75Si[M+H]+456.2201,found456.2202.
49.3 (S)-1-(3,5-二甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(49)的合成
Figure PCTCN2017078444-appb-000086
化合物49b(172mg,0.38mmol)溶于5mL THF,冷至冰浴下,TBAF(198mg,0.76mmol)溶于2mL THF,由分液漏斗缓慢滴加,滴完后继续搅拌15min后,TLC(展开剂：石油醚/乙酸乙酯＝2:1)显示原料(Rf＝0.7)完全消失,有新点(Rf＝0.3)生成,减压旋干,10mL乙酸乙酯溶解,水洗(5mL x 2),饱和食盐水洗(5mL),无水硫酸钠干燥。过滤浓缩后,柱层析纯化,得无色油状物(49)116mg,收率90％；[α]D 20＝+49.6(c 0.12,CHCl3),99％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝80:20,1.0mL/min,254nm；tR(minor)＝13.41min；tR(major)＝14.97min]；1H NMR(400MHz,CDCl3)δ6.93(d,J＝2.0Hz,1H),6.87(dd,J＝8.2,2.0Hz,1H),6.81(d,J＝8.2Hz,1H),6.53(d,J＝2.2Hz,2H),6.15(t,J＝2.2Hz,1H),5.94(brd,1H),5.80(t,J＝1.2Hz,1H),5.25(s,1H),5.13(s,1H),3.84(s,3H),3.70(s,6H)；13C NMR(150MHz,CDCl3)δ160.6,160.5,149.3,146.5,145.6,138.6,128.9,118.0,112.2,110.4,110.3,95.9,95.2,63.0,55.3,54.7；ESI-LRMS m/z(％):342.1[M+H]+；ESI-HRMS m/z(％):Calcd for C19H20NO5[M+H]+342.1336,found 342.1337.
实施例50 (S)-1-(2,4-二甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(50)的合成
化合物50的合成路线与化合物1类似：
50.1 (S)-2-[1-(3-叔丁基二甲基硅氧基-4-甲氧基)苯基-1-(2,4-二甲氧基苯基氨基)甲基]丙烯酸乙酯(50a)的合成
Figure PCTCN2017078444-appb-000087
氮气氛围下,三(二亚苄基丙酮)二钯(3.8mg,0.0042mmol)和1e(7.56mg,0.0105mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入化合物1d(170mg,0.42mmol),K2CO3(1.0M水溶液,1.5mL,1.5mmol)和2,4-二甲氧基苯胺(104mg,0.68mmol)；室温下搅拌4小时后,TLC(展开剂：石油醚/乙酸乙酯＝10:1)展开5次显示原料(Rf＝0.8)完全消失,有新点(Rf＝0.7)生成,用二氯甲烷萃取(3×5mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状物(50a)180mg,收率86％；1H NMR(400MHz,CDCl3)δ6.92(dd,J＝8.3,2.0Hz,1H),6.84(d,J＝2.0Hz,1H),6.79(d,J＝8.3Hz,1H),6.44(d,J＝2.4Hz,1H),6.41(d,J＝8.5Hz,1H),6.37–6.30(m,2H),5.85(s,1H),5.24(s,1H),4.23–4.05(m,2H),3.79(s,3H),3.78(s,3H),3.74(s,3H),1.23(q,J＝7.5Hz,3H),0.97(s,9H),0.12(s,6H)；13C NMR(151MHz,CDCl3)δ166.5,152.1,150.4,147.9,145.0,141.1,133.6,131.2,129.0,128.4,125.1,120.9,120.4,112.1,111.3,103.7,99.1,60.7,58.7,55.8,55.5,55.5,25.7,18.5,14.1,-4.6；ESI-LRMS m/z(％):502.2[M+H]+.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
50.2 (S)-1-(2,4-二甲氧基苯基)-4-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(50b)的合成
Figure PCTCN2017078444-appb-000088
化合物50a(180mg)和Sn[N(TMS)2]2(189mg,0.43mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流4.5小时后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.7)基本消失,有新点(Rf＝0.5)生成,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状化合物(50b)77mg,收率47％；1H NMR(400MHz,CDCl3)δ7.61(d,J＝8.7Hz,1H),6.84(dd,J＝8.2,2.0Hz,1H),6.76–6.73(m,2H),6.47–6.38(m,1H),6.36(d,J＝2.4Hz,1H),5.76(s,1H),5.60(s,1H),5.08(s,1H),6.76–6.73(m,6H),3.67(s,3H),0.94(s,9H),0.09(t,J＝24.3Hz,6H)；ESI-LRMS m/z(％):456.2[M+H]+.
50.3 (S)-1-(2,4-二甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(50)的合成
Figure PCTCN2017078444-appb-000089
化合物50b(55mg,0.12mmol)溶于5mL THF,冷至冰浴下,TBAF(63mg,0.24mmol)溶于1mL THF,由分液漏斗缓慢滴加,滴完后继续搅拌15min后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料(Rf＝0.8)完全消失,有新点(Rf＝0.5)生成,减压旋干,10mL乙酸乙酯溶解,水洗(5mL x 2),饱和食盐水洗(5mL),无水硫酸钠干燥。过滤浓缩后,柱层析纯化,得棕色油状物(50)37mg,收率90％；1H NMR(400MHz,CDCl3)δ7.64(d,J＝8.7Hz,1H),6.88(d,J＝2.0Hz,1H),6.79(dd,J＝8.3,2.0Hz,1H),6.75(d,J＝8.3Hz,1H),6.43(dd,J＝8.7,2.5Hz,1H),6.36(d,J＝2.5Hz,1H),5.75(t,J＝1.5Hz,1H),5.66(d,J＝8.3Hz,1H),5.62(s,1H),5.08(s,1H),3.83(s,3H),3.74(s,3H),3.69(s,3H)；13C NMR(150MHz,CDCl3)δ162.3,158.5,152.4,151.4,146.6,145.7,131.0,124.7,118.7,118.4,113.2,110.6,109.4,104.6,99.7,66.3,55.9,55.5；ESI-LRMS m/z(％):342.0[M+H]+.
实施例51 (S)-1-(3-甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(51)的合成
化合物51的合成路线与化合物1类似：
51.1 (S)-2-[1-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-1-(3-甲氧基苯基氨基)甲基]丙烯酸乙酯(51a)的合成
Figure PCTCN2017078444-appb-000090
氮气氛围下,三(二亚苄基丙酮)二钯(3.8mg,0.0042mmol)和1e(7.56mg,0.0105mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入1d(170mg,0.42mmol),K2CO3(1.0M水溶液,1.5mL,1.5mmol)和3-甲氧基苯胺(84mg,0.68mmol)；室温下搅拌4小时后,TLC(展开剂：石油醚/乙酸乙酯＝5:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.45)生成,用二氯甲烷萃取(3×5mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状物(51a)170mg,87％；[α]D 20＝+118.9(c 0.14,CHCl3)；1H NMR(400MHz,CDCl3)δ7.07(t,J＝8.1Hz,1H),6.92(dd,J＝8.3,2.1Hz,1H),6.84–6.80(m,2H),6.36(s,1H),6.30(dd,J＝8.1,1.8Hz,1H),6.21(d,J＝8.0Hz,1H),6.14(t,J＝2.2Hz,1H),5.90(s,1H),5.30(s,1H),4.18–4.12(m,2H),3.80(s,3H),3.76(s,3H),1.23(t,J＝7.1Hz,3H),0.99(s,9H),0.14(s,6H)；13C NMR(150MHz,CDCl3)δ165.7,160.1,149.9,147.6,144.4,139.9,132.6,129.2,124.7,120.1,119.7,111.5,105.9,102.2,98.9,60.1,57.8,54.9,54.4,25.1,17.8,13.5,-5.2；ESI-LRMS m/z(％):472.1[M+H]+；ESI-HRMS m/z(％):Calcd for C26H38NO5Si[M+H]+472.2514,found 472.2514.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
51.2 (S)-1-(3-甲氧基苯基)-4-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-3-亚甲基氮杂 环丁烷-2-酮(51b)的合成
Figure PCTCN2017078444-appb-000091
化合物51a(170mg)和Sn[N(TMS)2]2(190mg,0.43mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流4.5小时后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.7)基本消失,有新点(Rf＝0.5)生成,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状化合物(51b)106mg,收率69％；[α]D 20＝+36.5(c 0.12,CHCl3)；1H NMR(400MHz,CDCl3)δ7.15(t,J＝8.2Hz,1H),7.04(s,1H),6.97(dd,J＝8.3,1.9Hz,1H),6.86–6.84(m,3H),6.61(dd,J＝8.3,1.7Hz,1H),5.84(s,1H),5.30(s,1H),5.17(s,1H),3.81(s,3H),3.76(s,3H),0.96(s,9H),0.11(s,3H),0.10(s,3H)；13C NMR(150MHz,CDCl3)δ160.5,159.5,150.8,149.4,144.8,138.1,129.2,128.1,119.6,118.7,111.6,110.1,109.5,108.9,102.4,62.8,54.9,54.6,25.1,17.8,-5.3；ESI-LRMS m/z(％):426.2[M+H]+；ESI-HRMS m/z(％):Calcd for C24H32NO4Si[M+H]+426.2095,found 426.2096.
51.3 (S)-1-(3-甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(51)的合成
Figure PCTCN2017078444-appb-000092
化合物51b(106mg,0.25mmol)溶于5mL THF,冷至冰浴下,TBAF(130mg,0.50mmol)溶于1mL THF,由分液漏斗缓慢滴加,滴完后继续搅拌15min后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料(Rf＝0.8)完全消失,有新点(Rf＝0.5)生成,减压旋干,10mL乙酸乙酯溶解,水洗(5mL x 2),饱和食盐水洗(5mL),无水硫酸钠干燥。过滤浓缩后,柱层析纯化,得白色固体(51)63mg,收率77％；Mp 104-105℃；[α]D 20＝+61.0(c0.21,CHCl3)；99％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝75:25,1.0mL/min,254nm；tR(minor)＝10.45min；tR(major)＝12.04min]；1H NMR(400MHz,CDCl3)δ7.14(t,J＝8.2Hz,1H),7.04(s,1H),6.95(d,J＝1.9Hz,1H),6.89(dd,J＝8.2,1.9Hz,1H),6.84(s,1H),6.82(s,1H),6.60(dd,J＝8.2,1.6Hz,1H),5.82(s,1H),5.73(brd,1H),5.29(s,1H),5.15(s,1H),3.88(s,3H),3.75(s,3H)；13C NMR(150MHz,CDCl3)δ161.1,160.1,145.0,147.0,146.2,138.7,129.9,129.6,118.6,112.8,110.9,110.8,110.1,109.5,103.1,63.5,56.0,55.3；ESI-LRMS m/z(％):334.1[M+Na]+；ESI-HRMS m/z(％):Calcd for C18H18NO4[M+H]+312.1230,found 312.1232.
实施例52 (S)-1-(4-甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(52)的合成
化合物52的合成路线与化合物1类似：
52.1 (S)-2-[1-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-1-(4-甲氧基苯基氨基)甲基]丙 烯酸乙酯(52a)的合成
Figure PCTCN2017078444-appb-000093
氮气氛围下,三(二亚苄基丙酮)二钯(5mg,0.0054mmol)和1e(10mg,0.0108mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入1d(220mg,0.54mmol),K2CO3(1.0M水溶液,1.5mL,1.5mmol)和4-甲氧基苯胺(100mg,0.81mmol)；室温下搅拌1h后,TLC(展开剂：石油醚/乙酸乙酯＝5:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.45)生成,用二氯甲烷萃取(3×5mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,淡黄色油状物(52a)186mg,收率73％；[α]D 20＝-2.3(c 0.30,CHCl3)；1H NMR(400MHz,CDCl3)δ6.91(dd,J＝8.2,1.8Hz,1H),6.86–6.69(m,4H),6.53(d,J＝8.8Hz,2H),6.33(s,1H),5.88(s,1H),5.22(s,1H),4.23–4.07(m,2H),3.89(s,1H),3.78(s,3H),3.73(s,3H),1.22(t,J＝7.1Hz,3H),0.97(s,9H),0.12(s,6H)；13C NMR(150MHz,CDCl3)δ167.4,151.8,151.5,144.4,141.9,141.4,127.1,126.8,123.6,121.8,114.2,111.2,60.3,55.2,54.8,41.4,25.1,25.0,17.7,13.7,-5.4；ESI-LRMS m/z(％):472.2[M+H]+；ESI-HRMS m/z(％):Calcd for C26H38NO5Si[M+H]+472.2514,found 472.2515.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
52.2 (S)-1-(4-甲氧基苯基)-4-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(52b)的合成
Figure PCTCN2017078444-appb-000094
化合物52a(186mg)和Sn[N(TMS)2]2(208mg,0.47mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流4.5小时后,TLC(展开剂：石油醚/乙酸乙酯＝5:1)显示原料(Rf＝0.5)基本消失,有新点(Rf＝0.1)生成,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状化合物(52b)130mg,收率77％；[α]D 20＝+68.0(c 0.10,CHCl3)；1H NMR(400MHz,CDCl3)δ7.28(d,J＝9.1Hz,2H),6.93(dd,J＝8.3,1.8Hz,1H),6.83–6.77(m,4H),5.78(s,1H),5.25(s,1H),5.11(s,1H),3.79(s,3H),3.74(s,3H),0.94(s,9H),0.09(s,3H),0.07(s,3H)；13C NMR(150MHz,CDCl3)δ160.1,155.6,150.8,149.5,144.8,130.5,128.2,119.7,118.8,117.9,113.7,111.6,109.3,62.7,54.8,25.1,17.8,-5.2,-5.3；ESI-LRMS m/z(％):426.2[M+H]+；ESI-HRMS m/z(％):Calcd for C24H32NO4Si[M+H]+426.2095,found 426.2096.
52.3 (S)-1-(4-甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(52)的合成
Figure PCTCN2017078444-appb-000095
化合物52b(100mg,0.25mmol)溶于5mL THF,冷至冰浴下,TBAF(130mg,0.5mmol)溶于1mL THF,由分液漏斗缓慢滴加,滴完后继续搅拌15min后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.4)完全消失,有新点(Rf＝0.1)生成,减压旋干,10mL乙酸乙酯溶解,水洗(5mL x 2),饱和食盐水洗(5mL),无水硫酸钠干燥。过滤浓缩后,柱层析纯化,得白色固体(52)63mg,收率86％；Mp 180-181℃；[α]D 20＝+113.4(c 0.10,CHCl3)；97％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝75:25,1.0mL/min,254nm；tR(minor)＝15.28min；tR(major)＝19.43min]；1H NMR(400MHz,CDCl3)δ7.28(d,J＝8.9Hz,2H),6.98–6.74(m,5H),5.81–5.73(m,2H),5.26(s,1H),5.10(s,1H),3.87(s,3H),3.73(s,3H)；13C NMR(150MHz,CDCl3)δ160.0,155.6,149.5,146.3,145.5,130.5,129.0,117.9,117.9,113.8,112.3,110.3,109.4,62.7,55.3,54.8；ESI-LRMS m/z(％):334.1[M+Na]+；ESI-HRMS m/z(％):Calcd for C18H18NO4[M+H]+312.1230,found 312.1231.
实施例53 (S,Z)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-[2-(三甲基硅基)亚乙基]氮杂环丁烷-2-酮(53)和(S,E)-1-(3,4,5-三甲氧基苯基-4-(3-羟基-4-甲氧基苯基-3-[2-(三甲基硅基)亚乙基]氮杂环丁烷-2-酮(54)的合成
Figure PCTCN2017078444-appb-000096
向50mL Schlenk管中加入化合物1(40mg,0.1mmol),无水甲苯1mL,氮气保护下,加入Grubb’s二代催化剂(12mg,0.01mmol),烯丙基三甲基硅烷(86μL,0.5mmol),80℃反应8h；TLC(展开剂：PE/EA＝2:1)显示生成新点(Rf＝0.8)；加水,EA萃取,饱和食盐水洗,硫酸钠干燥,柱层析分离(展开剂：PE/EA＝5:1),得到白色固体顺式产物(53)24mg,收率49％；1H NMR(400MHz,CDCl3)δ6.96(d,J＝1.7Hz,1H),6.90(dd,J＝8.2Hz,1.7Hz,1H),6.83(d,J＝8.2Hz,1H),6.59(s,2H),5.72(br s,1H),5.64(t,J＝9.2Hz,1H),5.17(s,1H),3.88(s,3H),3.75(s,3H),3.73(s,6H),2.17–2.00(m,2H),0.07(d,J＝18.0Hz,9H)；13C NMR(150MHz,CDCl3)δ161.81,152.82,146.13,145.45,137.81,133.83,133.42,130.22,129.89,118.01,112.33,110.14,93.65,62.29,60.32,55.34,21.24,-2.38；ESI-LRMS m/z(％):458.2[M+H]+；白色固体反式产物(54)17mg,收率35％；1HNMR(400MHz,CDCl3)δ7.00(d,J＝1.8Hz,1H),6.94(d,J＝8.2Hz,1.8Hz,1H),6.84(d,J＝8.2Hz,1H),6.57(s,2H),6.35(t,J＝8.7Hz,1H),5.74(s,1H),5.23(s,1H),3.89(s,3H),3.75(s,3H),3.73(s,6H),1.53–1.34(m,2H),-0.05(s,9H)；13C NMR(150MHz,CDCl3)δ161.2,152.8,146.2,145.6,138.3,133.7,133.5,129.4,126.6,118.3,112.6,110.3,93.8,62.4,60.3,55.4,19.5,-2.3；ESI-LRMS m/z(％):458.2[M+H]+.
实施例54 (S,Z)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚乙基氮杂环 丁烷-2-酮(55)的合成
Figure PCTCN2017078444-appb-000097
在100mL茄形瓶中加入化合物53(8mg,0.018mmol),TBAF(7mg,0.026mmol),THF 1mL,冰浴反应30分钟；TLC(展开剂：PE/EA＝4:1)展开两次,显示原料点(Rf＝0.2)消失,生成新点(Rf＝0.1)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,柱层析分离(展开剂：PE/EA＝3:1),得到白色固体(55)6mg,收率90％；1H NMR(400MHz,CDCl3)δ6.98(d,J＝1.8Hz,1H),6.91(dd,J＝8.2Hz,1.8Hz,1H),6.80(d,J＝8.2Hz,1H),6.71(s,2H),5.62(br s,1H),5.64(q,J＝9.2Hz,1H),5.27(s,1H),3.89(s,3H),3.76(s,3H),3.70(s,6H),2.07(d,J＝9.2Hz,2H)；ESI-LRMS m/z(％):386.2[M+H]+.
实施例55 (S,E)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚乙基氮杂环丁烷-2-酮(56)的合成
Figure PCTCN2017078444-appb-000098
在100mL茄形瓶中加入化合物54(10mg,0.022mmol),TBAF(9mg,0.034mmol),THF 1mL,冰浴反应30分钟；TLC(展开剂：PE/EA＝4:1)展开两次,显示原料点(Rf＝0.2)消失,生成新点(Rf＝0.1)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,柱层析分离(展开剂：PE/EA＝3:1),得到白色固体(56)8mg,收率95％；1H NMR(400MHz,CDCl3)δ7.01(d,J＝1.8Hz,1H),6.96(d,J＝8.2Hz,1.8Hz,1H),6.82(d,J＝8.2Hz,1H),6.57(s,2H),6.45(q,J＝8.7Hz,1H),5.64(br s,1H),5.11(s,1H),3.81(s,3H),3.74(s,3H),3.70(s,6H),2.09(d,J＝8.7Hz,,2H).13C NMR；ESI-LRMS m/z(％):386.2[M+H]+.
实施例56 (S,Z)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-(苯亚甲基)氮杂环丁烷-2-酮(57)的合成
Figure PCTCN2017078444-appb-000099
向50mL Schlenk管中加入化合物1(16mg,0.043mmol),二氯乙烷1mL,通氮气除氧30分钟,加入Grubb’s二代催化剂(5mg,0.006mmol),苯乙烯(13μL,0.11mmol),氮气保护下,60℃反应12h；TLC(展开剂：PE/EA＝2:1)显示生成新点(Rf＝0.7)；加水,EA 萃取,饱和食盐水洗,硫酸钠干燥,柱层析分离(展开剂：PE/EA＝4:1),原料回收5mg,回收率31％；同时得到黄色固体(57)10mg,收率51％；1H NMR(400MHz,CDCl3)δ7.99(d,J＝7.2Hz,2H),7.41-7.31(m,3H),7.02(d,J＝2.0Hz,1H),6.96(dd,J＝8.2,2.0Hz,1H),6.86(d,J＝8.2Hz,1H),6.68(s,2H),6.29(s,1H),5.70(s,1H),5.28(s,1H),3.89(s,3H),3.78(s,3H),3.76(s,6H)；ESI-LRMS m/z(％):448.2[M+H]+.
实施例57 (S,Z)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-[(4-叔丁基苯基)亚甲基]氮杂环丁烷-2-酮(58)的合成
Figure PCTCN2017078444-appb-000100
向50mL Schlenk管中加入化合物1(10mg,0.027mmol),无水二氯乙烷1mL,通氮气除氧30分钟,加入Grubb’s二代催化剂(2mg,0.003mmol),对叔丁基苯乙烯(13μL,0.081mmol),氮气保护下,60℃反应12h；TLC(展开剂：PE/EA＝2:1)显示生成新点(Rf＝0.7)；加水,EA萃取,饱和食盐水洗,硫酸钠干燥,柱层析分离(展开剂：PE/EA＝4:1),得到黄色固(58)4mg,收率30％；原料回收5mg,回收率50％；Mp 77–78℃；[α]D 20＝+131.8(c 1.0,CHCl3).1H NMR(400MHz,CDCl3):δ7.92(d,J＝8.4Hz,2H),7.42(d,J＝8.4Hz,2H),7.01(d,J＝1.9Hz,1H),6.95(dd,J＝8.2,1.9Hz,1H),6.85(d,J＝8.2Hz,1H),6.68(s,2H),6.28(s,1H),5.68(s,1H),5.27(s,1H),3.89(s,3H),3.77(s,3H),3.76(s,6H),1.31(s,9H).13C NMR(150MHz,CDCl3):δ159.9,152.9,152.4,146.4,145.6,139.1,133.8,133.6,130.8,130.0,129.6,129.1,125.0,118.2,112.5,110.3,94.0,76.6,76.4,76.2,61.8,60.4,55.4,55.4,34.2,30.5.ESI-MS(m/z):504.2(M+H+).ESI-HRMS(m/z):calcd forC30H33NO6+H+[M+H]+,504.2381；found,504.2376.
实施例58 (3R,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-(2-三甲基硅基乙基)氮杂环丁烷-2-酮(59)的合成
Figure PCTCN2017078444-appb-000101
在100mL茄形瓶中加入化合物53和54混合物(38mg,0.083mmol),10％Pd/C 4mg,甲醇1mL,常压氢气下,室温反应12h,TLC(展开剂：PE/EA＝4:1)展开两次显示原料点(Rf＝0.2)消失,生成新点(Rf＝0.3)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝4:1),得到白色固体(59)33mg,收率87％；1H NMR(400MHz,CDCl3)δ6.86-6.82(m,2H),6.77(d,J＝8.2Hz,1H),6.55(s,2H),5.70(s,1H),5.07(d,J＝5.6Hz,1H),3.89(s,3H),3.81–3.68(m,10H),3.46(dt,J＝9.5,6.0Hz,1H),0.91–0.82(m,1H),0.48(td,J＝14.0,4.8Hz,1H),0.21(td,J＝14.0,3.7Hz,1H),-0.21(s,9H)；ESI-LRMS m/z(％):482.2[M+Na]+.
实施例59 (3R,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-(4-羟基苄基) 氮杂环丁烷-2-酮(60)的合成
Figure PCTCN2017078444-appb-000102
向50mL Schlenk管中加入化合物1(20mg,0.054mmol),无水甲苯1mL,氮气保护下,加入Grubb’s二代催化剂(5mg,0.006mmol),对羟基苯乙烯(41mg,0.34mmol),80℃反应12h；TLC(展开剂：PE/EA＝2:1)显示生成新点(Rf＝0.15)；加水,EA萃取,饱和食盐水洗,硫酸钠干燥,蒸干快速柱层析粗分后(展开剂：PE/EA＝4:1),加入10％Pd/C 3mg,甲醇1mL,常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.15)消失,生成新点(Rf＝0.19)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(60)17mg,收率83％；1H NMR(400MHz,CDCl3)δ6.85–6.69(m,5H),6.65(d,J＝8.4Hz,2H),6.56(s,2H),5.72(s,1H),5.26(br s,1H),5.07(d,J＝5.6Hz,1H),3.91(s,3H),3.83–3.74(m,4H),3.71(s,6H),2.84(dd,J＝14.8,7.2Hz,1H),2.44(dd,J＝14.8,8.5Hz,1H)；ESI-LRMS m/z(％):466.2[M+H]+.
实施例60 (3S,4S)-1-(3,4,5-三甲氧基苯基)-3-(4-羟甲基-1H-1,2,3-三氮唑-1-基)-4-(3-羟基-4-甲氧基苯基)氮杂环丁烷-2-酮(61)的合成
Figure PCTCN2017078444-appb-000103
向100mL茄形瓶中加入化合物75、炔丙醇(5μL,0.086mmol)、Vc-Na(2mg,0.009mmol)、水合硫酸铜(1mg,0.004mmol)、乙醇1mL和水0.2mL,室温反应12h；TLC(展开剂：PE/EA＝1：3)显示生成新点(Rf＝0.15)；加水,EA萃取,饱和食盐水洗,硫酸钠干燥,快速柱层析粗分后,加入10％Pd/C 3mg,甲醇1mL,常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝1:3)显示原料点(Rf＝0.15)消失,生成新点(Rf＝0.1)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝1:4),得到白色固体(61)11mg,收率84％；1H NMR(400MHz,CDCl3)δ；1H NMR(400MHz,CDCl3)δ7.75(s,1H),6.98(d,J＝1.6Hz,1H),6.94(dd,J＝8.0,1.6Hz,1H),6.89(d,J＝8.0Hz,1H),6.59(s,2H),5.76(s,1H),5.55(d,J＝1.6Hz,1H),5.31(d,J＝1.6Hz,1H),4.85(s,2H),3.92(s,3H),3.79(s,3H),3.74(s,6H)；13C NMR(150MHz,CDCl3)δ159.0,153.7,147.6,146.7,135.5,132.5,127.6,121.5,118.4,112.0,111.2,95.65,72.0,71.5,67.92,66.2,65.5,64.1,63.5,61.0,59.9,58.8,58.5,56.7,56.2,56.1,55.9,54.7,50.7.
实施例61 (3R,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-(二甲氨基甲基)氮杂环丁烷-2-酮(62)的合成
Figure PCTCN2017078444-appb-000104
在100mL Schlenk管中加入化合物1(16mg,0.043mmol),盐酸二甲胺(11mg,0.13mmol),DBU(19μL,0.13mmol)与甲醇1mL,橡胶塞密封,回流反应8小时；TLC(展开剂：PE/EA＝1:5)显示原料点(Rf＝0.99)消失,生成新点(Rf＝0.1)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到淡黄色固体(62)16mg,收率89％；1H NMR(400MHz,CDCl3)δ6.85-6.83(m,2H),6.77(d,J＝7.1Hz,1H),6.55(s,2H),5.12(d,J＝5.8Hz,1H),3.90(s,3H),3.82–3.74(m,4H),3.71(s,6H),2.44(dd,J＝13.3,6.8Hz,1H),2.25(dd,J＝13.3,5.8Hz,1H),2.17(s,6H)；13C NMR(150MHz,CDCl3)δ166.0,152.8,146.1,145.3,133.8,133.1,126.9,118.4,112.9,110.1,94.3,60.3,57.6,55.4,55.3,53.8,53.0,44.9.
实施例62 (3R,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-(N-苄基氨基甲基)氮杂环丁烷-2-酮(63)的合成
Figure PCTCN2017078444-appb-000105
在100mL Schlenk管中加入化合物1(0.5g,1.35mmol),苄胺(0.18mL,1.63mmol)与甲醇12mL,回流反应12小时；TLC(展开剂：PE/EA＝1:1)显示生成新点(Rf＝0.1)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,柱层析分离(展开剂：PE/EA＝2:1),回收原料0.22g；同时得到淡黄色固体(63)0.15g,收率23％；1H NMR(400MHz,CDCl3)δ7.29–7.16(m,3H),7.05(d,J＝6.6Hz,2H),6.85(d,J＝1.4Hz,1H),6.78(q,J＝8.3Hz,2H),6.53(s,2H),5.10(d,J＝5.6Hz,1H),3.90(s,3H),3.84–3.74(m,4H),3.71(s,6H),3.63(d,J＝13.2Hz,1H),3.48(d,J＝13.2Hz,1H),2.79(dd,J＝12.2,6.3Hz,1H),2.60(dd,J＝12.2,9.2Hz,1H)；；13C NMR(150MHz,CDCl3)δ165.7,152.9,146.1,145.4,138.8,133.8,133.0,127.7,127.3,126.6,126.3,118.0,112.4,110.2,94.3,60.3,56.9,55.5,55.30,54.1,53.2,44.0.
实施例63 (3R,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-(3-羧基丙酰氧基)氮杂环丁烷-2-酮(64)的合成
Figure PCTCN2017078444-appb-000106
冰浴下,在100mL茄形瓶中加入化合物72(23mg,0.049mmol),丁二酸酐(6mg,0.006mmol),DIPEA(15μL,mmol),DMAP 1mg与无水DCM 1mL,室温反应6小时； TLC(展开剂：PE/EA＝1:2)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0.03)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,加入10％Pd/C 3mg,甲醇1mL,常压氢气下,室温反应12小时,TLC(展开剂：DCM/MeOH＝10:1)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0.3)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：DCM/MeOH＝10:1),得到白色固体(64)18mg,收率77％；1H NMR(400MHz,CDCl3)δ6.86(d,J＝1.7Hz,1H),6.84–6.78(m,2H),6.57(s,2H),5.95(d,J＝4.9Hz,1H),5.25(d,J＝4.9Hz,1H),3.88(s,3H),3.77(s,3H),3.73(s,6H),2.54–2.43(m,1H),2.42–2.35(m,2H),2.29–2.18(m,1H).
实施例64 (3R,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-(乙氧羰基甲氧基)氮杂环丁烷-2-酮(65)的合成
Figure PCTCN2017078444-appb-000107
冰浴下,在100mL茄形瓶中加入化合物72(15mg,0.032mmol),溴乙酸乙酯(6μL,0.05mmol),NaH(2mg,0.05mmol)与无水THF 1mL,室温反应1小时；TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0.6)消失,生成新点(Rf＝0.8)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,加入10％Pd/C 3mg,甲醇1mL,常压氢气下,室温反应24小时,TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0.6)消失,生成新点(Rf＝0.5)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(65)7mg,收率47％；1H NMR(400MHz,CDCl3)δ7.01(d,J＝1.8Hz,1H),6.93(dd,J＝8.2,1.8Hz,1H),6.85(d,J＝8.2Hz,1H),6.57(s,2H),5.67(s,1H),5.11(dd,J＝11.4,5.0Hz,2H),4.14(d,J＝16.5Hz,1H),4.03(d,J＝16.5Hz,1H),3.89(s,3H),3.76(s,3H),3.72(s,6H),1.25(t,J＝7.1Hz,6H)；13C NMR(150MHz,CDCl3)δ168.6,163.1,152.9,146.3,145.1,134.2,132.5,125.4,119.4,113.7,109.9,94.7,82.0,66.5,60.9,60.4,60.3,55.5,55.3,29.1,13.5.
实施例65 (3S,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-溴氮杂环丁烷-2-酮(66)和(R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)氮杂环丁烷-2-酮(67)的合成
Figure PCTCN2017078444-appb-000108
在5mL微波反应管中加入化合物73(50mg,0.092mmol),TBAB(59mg,0.18mmol)和DMF 1mL,微波加热170℃反应3h.TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.15)消失,生成新点(Rf＝0.7)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,快速柱层析分离粗分后直接加入加入10％Pd/C 3mg,甲醇1mL,常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.7)消失,生成两个新点(Rf＝0.4,0.5)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝3:1),得到Rf＝0.5的脱苄基产物为白色固体(66)10mg,收率53％；1H NMR(400MHz,CDCl3)δ6.93(d,J＝1.9Hz,1H),6.90(dd,J＝8.2,1.9Hz,1H),6.86(d,J＝8.2Hz,1H),6.54(s,2H),5.73(s,1H),4.87(d,J＝1.9Hz,1H),4.59(d,J＝1.9Hz,1H),3.91(s,3H),3.76(s,3H),3.72(s,6H)；Rf＝0.4脱苄基同时得到溴被还原产物为白色固体(67)5mg,收率32％；1H NMR(400MHz,CDCl3)δ6.96(d,J＝2.1Hz,1H),6.89(dd,J＝8.3,2.1Hz,1H),6.84(d,J＝8.3Hz,1H),6.55(s,2H),5.67(s,1H),4.87(dd,J＝5.6,2.6Hz,1H),3.89(s,3H),3.76(s,3H),3.72(s,6H),3.51(dd,J＝15.2,5.6Hz,1H),2.93(dd,J＝15.2,2.6Hz,1H).
实施例66 (3S,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-羟甲基氮杂环丁烷-2-酮(68)和(3R,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-羟甲基氮杂环丁烷-2-酮(69)的合成
Figure PCTCN2017078444-appb-000109
氮气保护下下,在50mL Schlenk中加入化合物1(11mg,0.030mmol),联硼酸频哪醇酯(10mg,0.039mmol),CuCl(0.3mg,0.03mmol),三苯基膦(1mg,0.038mmol),甲醇(1.5μL,0.037mmol),叔丁醇锂(0.3mg,0.037mmol)与无水THF 1mL,室温反应12小时；TLC(展开剂：PE/EA＝1:2)显示仅原料点(Rf＝0.4)等高处有点；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,加入NaBO3·4H2O(23mg,0.15mmol),THF 1mL和水1mL,室温反应12h,TLC(展开剂：PE/EA＝1:3)显示原料点(Rf＝0.9)消失,生成两新点(Rf＝0.17,0.22)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,柱层析分离(展开剂：PE/EA＝1:1),得到反式产物为白色固体(69)4.5mg,收率39％；1H NMR(400MHz,CDCl3)δ6.97(d,J＝1.5Hz,1H),6.91(dd,J＝8.2Hz,1.5Hz,1H),6.84(d,J＝8.2Hz,1H),6.56(s,2H),5.70(s,1H),4.91(d,J＝1.8Hz,1H),4.15(dd,J＝12.1,4.5Hz,1H),4.00(dd,J＝12.1,3.7Hz,1H),3.90(s,3H),3.79–3.75(m,4H),3.73(s,6H),3.28(br d,J＝2.3Hz,1H)；13C NMR(151MHz,CDCl3)δ165.60,153.50,146.78,146.32,134.57,133.72,130.77,117.94,112.16, 110.99,94.91,62.14,60.93,58.89,57.49,56.06；得到顺式产物(68)为白色固体7mg,收率61％；1H NMR(400MHz,CDCl3)δ6.90(d,J＝1.2Hz,1H),6.87–6.81(m,2H),6.55(s,2H),5.75(s,1H),5.16(d,J＝5.3Hz,1H),3.90(s,3H),3.86–3.75(m,5H),3.73(s,6H),3.64(dd,J＝11.3,7.8Hz,1H)；13C NMR(151MHz,CDCl3)δ165.18,153.56,146.73,146.13,134.68,133.60,127.13,118.47,112.90,110.93,95.06,60.96,58.15,57.27,56.72,56.15,55.98.
实施例67 (3R,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-甲基氮杂环丁烷-2-酮(70)的合成
Figure PCTCN2017078444-appb-000110
在100mL茄形瓶中加入化合物1(15mg,0.04mmol),10％Pd/C 3mg,甲醇1mL常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0.33)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(70)15mg,收率99％；1H NMR(400MHz,CDCl3)δ6.86–6.77(m,2H),6.72(d,J＝8.3Hz,1H),6.56(s,2H),5.71(s,1H),5.06(d,J＝5.8Hz,1H),3.89(s,3H),3.77(s,3H),3.72(s,6H),3.66-3.59(m,1H),0.91(d,J＝7.6Hz,3H).
实施例68 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-苄氧基-4-甲氧基苯基)氮杂环丁烷-2,3-二酮(71)的合成
Figure PCTCN2017078444-appb-000111
在100mL茄形瓶中加入化合物84(0.2g,0.41mmol),高碘酸钠(0.13g,0.62mmol),甲醇4mL与水1mL,室温反应3小时；TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0)消失,生成新点(Rf＝0.2)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝2:1),得到黄色固体(71)0.15g,收率79％；1H NMR(400MHz,CDCl3)δ7.32–7.18(m,3H),6.90(s,2H),6.56(d,J＝6.8Hz,1H),6.05(s,1H),5.26(s,1H),4.95(d,J＝6.8Hz,1H),4.26(s,3H),4.13(s,3H),4.07(s,6H),2.44(s,3H).
实施例69 (3R,4S)-1-(3,4,5-三甲氧基苯基)-3-羟基-4-(3-苄氧基-4-甲氧基苯基)氮杂环丁烷-2-酮(72)的合成
Figure PCTCN2017078444-appb-000112
在100mL茄形瓶中加入化合物71(0.18g,0.39mmol),甲醇4mL,冰浴下加入硼氢化钠(18mg,0.47mmol),保持冰浴反应1小时；TLC(展开剂：PE/EA＝2:1,2次)显示原料点(Rf＝0.2)消失,生成新点(Rf＝0.1)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝1:1),得到白色固体(72)0.35g,收率90％；1H NMR(400MHz,CDCl3)δ7.40–7.28(m,5H),6.94(d,J＝8.3Hz,1H),6.91(dd,J＝8.3,1.6Hz,1H),6.83(br d,J＝1.6Hz,1H),6.54(s,2H),5.16(s,2H),5.14(d,J＝5.2Hz,1H),5.10((br d,J＝5.2Hz,1H),3.91(s,3H),3.80(s,3H),3.70(s,6H).
实施例70 (3R,4S)-1-(3,4,5-三甲氧基苯基)-3-甲磺酰氧基-4-(3-苄氧基-4-甲氧基苯基)氮杂环丁烷-2-酮(73)的合成
Figure PCTCN2017078444-appb-000113
在100mL茄形瓶中加入化合物72(0.2g,0.43mmol),三乙胺(0.09mL,0.64mmol),二氯甲烷4mL,冰浴下加入甲烷磺酰氯(0.05mL,0.64mmol),逐渐升至室温反应1小时；TLC(展开剂：PE/EA＝2:1,两次)显示原料点(Rf＝0.1)消失,生成新点(Rf＝0.2)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝5:1),得到白色固体(73)0.2g,收率86％；1H NMR(400MHz,CDCl3)δ7.36(d,J＝7.7Hz,2H),7.33–7.20(m,3H),6.99–6.84(m,3H),6.47(s,2H),5.76(d,J＝5.0Hz,1H),5.21(d,J＝5.0Hz,1H),5.13(s,2H),3.88(s,3H),3.77(s,3H),3.67(s,6H),2.66(s,3H)；13C NMR(101MHz,CDCl3)δ160.3,153.7,150.8,148.3,136.8,135.5,132.6,128.8,128.2,127.5,123.8,121.5,113.7,111.9,95.4,79.4,71.2,61.7,61.2,56.28,56.2,38.9.
实施例71 (3R,4S)-1-(3,4,5-三甲氧基苯基)-3-甲烷磺酰氧基-4-(3-羟基-4-甲氧基苯基)氮杂环丁烷-2-酮(74)的合成
Figure PCTCN2017078444-appb-000114
在100mL茄形瓶中加入化合物73(11mg,0.02mmol),10％Pd/C 3mg,甲醇1mL常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0.4)消失,生 成新点(Rf＝0.1)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝1:1),得到白色固(74)9mg,收率98％；1H NMR(400MHz,CDCl3)δ6.96–6.85(m,3H),6.56(s,2H),5.80(d,J＝5.1Hz,1H),5.70(s,1H),5.28(d,J＝5.1Hz,1H),3.90(s,3H),3.77(s,3H),3.73(s,6H),2.91(s,3H)；13C NMR(151MHz,CDCl3)δ159.5,153.0,146.8,145.4,131.8,123.8,119.5,113.3,110.1,94.8,78.6,60.7,60.3,55.5,55.3,38.4.
实施例72 (3S,4S)-1-(3,4,5-三甲氧基苯基)-3-叠氮基-4-(3-苄氧基-4-甲氧基苯基)氮杂环丁烷-2-酮(75)的合成
Figure PCTCN2017078444-appb-000115
在100mL茄形瓶中加入化合物73(0.5g，0.92mmol),叠氮化钠(90mg，1.38mmol),DMF 10mL,100℃反应48小时；TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.2)消失,生成新点(Rf＝0.4)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝5:1),得到黄色液体(75)0.24g,收率53％；[α]D 20＝-50.4(c 1.0,CHCl3).1H NMR(400MHz,CDCl3):δ7.38–7.23(m,5H),6.94–6.88(m,2H),6.80(br s,1H),6.43(s,2H),5.12(s,2H),4.67(d,J＝1.4Hz,1H),4.39(d,J＝1.4Hz,1H),3.89(s,3H),3.76(s,3H),3.65(s,6H).13C NMR(150MHz,CDCl3):δ160.7,152.9,150.0,148.1,135.8,134.5,132.2,127.9,127.4,126.8,126.6,118.8,111.5,111.0,94.5,71.7,70.5,62.5,60.3,55.5,55.4.ESI-MS(m/z):491.1(M+H+).ESI-HRMS(m/z):calcd for C26H26N4O6+H+[M+H]+,491.1925；found,491.1924.
实施例73 (3S,4S)-1-(3,4,5-三甲氧基苯基)-3-氨基-4-(3-苄氧基-4-甲氧基苯基)氮杂环丁烷-2-酮(76)的合成
Figure PCTCN2017078444-appb-000116
在100mL茄形瓶中加入化合物75(0.23g，0.47mmol),氯化亚锡(0.28g，1.4mmol),甲醇8mL,9％盐酸2mL,回流反应1小时；TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0)；反应液加碳酸氢钠,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝5:1),得到白色固体(76)0.14g,收率64％；Mp 64–65℃；[α]D 20＝+7.9(c 1.0,CHCl3).1H NMR(400MHz,CDCl3):δ7.39–7.21(m,5H),6.97–6.82(m,3H),6.47(s,2H),5.12(s,2H),4.50(s,1H),3.98(s,1H),3.89(s,3H),3.77(s,3H),3.66(s,6H).13C NMR(150MHz,CDCl3):δ167.9,153.5,150.1,148.6,136.6,134.6,133.6,129.2,128.5,128.0,127.3,119.2,112.1,111.6,95.0,71.1,69.7,66.8,60.9,56.1,56.0.ESI-MS(m/z):465.1(M+H+).ESI-HRMS(m/z):calcd for C26H28N2O6+H+[M+H]+,465.2020；found,465.2035.
实施例74 (3S,4S)-1-(3,4,5-三甲氧基苯基)-3-乙酰氨基-4-(3-羟基-4-甲氧基苯基)氮杂环丁烷-2-酮(77)的合成
Figure PCTCN2017078444-appb-000117
在100mL茄形瓶中加入化合物76(29mg，0.062mmol),DIPEA(16μL，0.093mmol),醋酐(8μL，0.08mmol)与二氯甲烷2mL,室温反应1h；TLC(展开剂：PE/EA＝1:5)显示原料点(Rf＝0.1)消失,生成新点(Rf＝0.3)；反应液加饱和NaHCO3,用DCM萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；整个加入10％Pd/C 3mg,甲醇1mL常压氢气下室温反应12小时,TLC(展开剂：PE/EA＝1:5)显示原料点(Rf＝0.3)消失,生成新点(Rf＝0.25)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(77)15mg,收率58％；1H NMR(400MHz,CDCl3)δ7.32–7.18(m,3H),6.90(s,2H),6.56(d,J＝6.8Hz,1H),6.05(s,1H),5.26(s,1H),4.95(d,J＝6.8Hz,1H),4.26(s,3H),4.13(s,3H),4.07(s,6H),2.44(s,3H).
实施例75 (3S,4S)-1-(3,4,5-三甲氧基苯基)-3-苯甲酰氨基-4-(3-羟基-4-甲氧基苯基)氮杂环丁烷-2-酮(78)的合成
Figure PCTCN2017078444-appb-000118
在100mL茄形瓶中加入化合物76(32mg，0.069mmol),DIPEA(19μL，0.11mmol),苯甲酸酐(23mg，0.1mmol)与二氯甲烷5mL,室温反应4h；TLC(展开剂：PE/EA＝1:5)显示原料点(Rf＝0.1)消失,生成新点(Rf＝0.4)；反应液加饱和NaHCO3,用DCM萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,加入10％Pd/C3mg,甲醇1mL，常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝1:5)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0.33)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(78)26mg,收率80％；1H NMR(400MHz,CDCl3)δ7.82(d,J＝7.2Hz,2H),7.53(t,J＝7.3Hz,1H),7.44(t,J＝7.6Hz,2H),7.00(d,J＝6.7Hz,1H),6.97(d,J＝2.0Hz,1H),6.93(dd,J＝8.2,2.0Hz,1H),6.85(d,J＝8.2Hz,1H),6.55(s,2H),5.69(s,1H),5.00(d,J＝2.1Hz,1H),4.77(dd,J＝6.7,2.1Hz,1H),3.89(s,3H),3.75(s,3H),3.70(s,6H).
实施例76 (3S,4S)-1-(3,4,5-三甲氧基苯基)-3-甲磺酰氨基-4-(3-羟基-4-甲氧基苯基)氮杂环丁烷-2-酮(79)的合成
Figure PCTCN2017078444-appb-000119
在100mL茄形瓶中加入化合物76(12mg，0.026mmol),DIPEA(5μL，0.031mmol),甲磺酰氯(2μL，0.028mmol)与二氯甲烷1mL,室温反应2h；TLC(展开剂：PE/EA＝1:5)显示原料点(Rf＝0.1)消失,生成新点(Rf＝0.3)；反应液加饱和NaHCO3,用DCM萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,加入10％Pd/C3mg,甲醇1mL，常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝1:5)显示原料点(Rf＝0.3)消失,生成新点(Rf＝0.26)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(79)15mg,收率99％；1H NMR(400MHz,CDCl3)δ6.75–6.65(m,2H),6.62(d,J＝1.5Hz,1H),6.14(s,2H),5.62(t,J＝8.0Hz,1H),4.92(d,J＝2.8Hz,2H),4.53(d,J＝2.0Hz,1H),4.17(dd,J＝8.8,2.0Hz,1H),3.70(s,3H),3.59(s,3H),3.45(s,6H),2.92(s,3H).
实施例77 (3R,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-苄氧基-4-甲氧基苯基)-3-羟基-3-羟甲基氮杂环丁烷-2-酮(84)的合成的合成
Figure PCTCN2017078444-appb-000120
在100mL茄形瓶中加入化合物47(0.93g,2mmol),锇酸钾(10mg,0.03mmol),NMO(0.62mL,3mmol)与丙酮5mL,水0.2mL,室温反应8小时；TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.6)消失,生成新点(Rf＝0)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝1:4),得到白色固体84 0.91g,收率92％；Mp 123–124℃；[α]D 20＝+61.8(c 1.0,CHCl3).1H NMR(400MHz,CDCl3):δ7.34–7.21(m,5H),6.90–6.77(m,2H),6.72(s,1H),6.43(s,2H),5.10(s,2H),4.94(s,1H),4.53(br d,J＝30.9Hz,1H),3.88(s,3H),3.77(s,3H),3.63(s,6H),3.53(dd,J＝12.3,4.3Hz,1H),3.27(dd,J＝12.3,5.5Hz,1H).13C NMR(150MHz,CDCl3):δ166.5,152.8,149.5,147.4,135.9,134.3,132.3,127.9,127.4,126.6,124.3,119.1,112.0,111.3,95.0,85.7,70.4,66.9,61.6,60.3,55.4,55.4.ESI-MS(m/z):496.1(M+H+).ESI-HRMS(m/z):calcd for C27H29NO8+H+[M+H]+,496.1966；found,496.1978..
实施例78 (3R,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-羟基-3-羟甲基氮杂环丁烷-2-酮(80)的合成的合成
Figure PCTCN2017078444-appb-000121
在100mL茄形瓶中加入化合物84(11mg,0.022mmol),10％Pd/C 3mg,甲醇1mL常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝1:2)显示原料点(Rf＝0.6)消失,生成新点(Rf＝0.3)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝1:2),得到白色固体(80)8mg,收率89％；1H NMR(400MHz,CDCl3)δ6.84-6.82(m,2H),6.76(br d,J＝8.1Hz,1H),6.52(s,2H),5.80(s,1H),5.01(s,1H),4.53(br s,1H),3.89(s,3H),3.82–3.62(m,10H),3.45(d,J＝12.3Hz,1H),2.49(br s,1H).13C NMR(151MHz,CDCl3)δ166.6,152.9,146.2,145.4,134.3,132.3,125.2,117.8,112.1,110.3,95.1,85.7,76.6,76.4,76.2,66.8,61.6,60.3,55.5,55.3.
实施例79 (3R,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-甲氧基-3-甲氧甲基氮杂环丁烷-2-酮(81)的合成
Figure PCTCN2017078444-appb-000122
在冰浴下,在100mL茄形瓶中加入化合物84(15mg,0.03mmol),硫酸二甲酯(15μL,0.15mmol),NaH(3mg,0.075mmol)与无水THF 1mL,室温反应1小时；TLC(展开剂：PE/EA＝1:2)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0.9)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,加入10％Pd/C 3mg,甲醇1mL,常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0.6)消失,生成新点(Rf＝0.5)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(81)11mg,收率84％；1H NMR(400MHz,CDCl3)δ6.85(d,J＝1.4Hz,1H),6.82(d,J＝8.2Hz,1H),6.77(dd,J＝8.2,1.4Hz,1H),6.58(s,2H),5.06(s,1H),3.89(s,3H),3.78(s,3H),3.72(s,6H),3.63(s,3H),3.51(d,J＝11.0Hz,1H),3.37(d,J＝11.0Hz,1H),3.03(s,3H)；13C NMR(150MHz,CDCl3)δ163.7,152.9,145.9,145.0,134.3,132.3,125.7,118.4,112.6,109.8,95.1,91.8,68.8,63.1,60.3,58.7,55.5,55.3,53.1.
实施例80 (3R,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-乙酰氧基-4-甲氧基苯基)-3-羟基-3-羟甲基氮杂环丁烷-2-酮(82)的合成
Figure PCTCN2017078444-appb-000123
在100mL茄形瓶中加入化合物46(96mg,0.22mmol),锇酸钾(3mg,0.01mmol), NMO(0.1mL,0.48mmol)与丙酮5mL,水0.2mL,室温反应8小时；TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.6)消失,生成新点(Rf＝0)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝1:3),得到白色固体(82)93mg,收率95％；1H NMR(400MHz,CDCl3)δ7.10(d,J＝8.2Hz,1H),6.98(s,1H),6.95(d,J＝8.2Hz,1H),6.52(s,2H),5.04(s,1H),3.82(s,3H),3.78(s,3H),3.71-3.69(m,7H),3.46(d,J＝12.2Hz,1H),2.28(s,3H).ESI-LRMS m/z(％):470.1[M+Na]+.
实施例81 (3R,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-乙酰氧基-4-甲氧基苯基)-3-乙酰氧基-3-乙酰氧甲基氮杂环丁烷-2-酮(83)的合成
Figure PCTCN2017078444-appb-000124
在100mL茄形瓶中加入化合物82(6.5mg,0.015mmol),4-二甲氨基吡啶0.5mg,三乙胺(5μL,0.03mmol)与二氯甲烷5mL,醋酸酐(3.5μL,0.03mmol),室温反应30分钟；TLC(展开剂：PE/EA＝1:4)显示原料点(Rf＝0.2)消失,生成新点(Rf＝0.8)；反应液加饱和NaHCO3,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝1:1),得到白色固体(83)6mg,收率85％；1H NMR(400MHz,CDCl3)δ7.24(d,J＝8.6Hz,1H),7.13(s,1H),6.95(d,J＝8.6Hz,1H),6.54(s,2H),5.29(s,1H),4.31(d,J＝12.7Hz,1H),4.24(d,J＝12.7Hz,1H),3.83(s,3H),3.78(s,3H),3.71(s,6H),2.28(s,3H),2.21(s,3H),1.99(s,3H)；ESI-LRMS m/z(％):532.1[M+H]+.
实施例82 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基苯基)-3-亚甲基氮杂环丁烷-2-酮(85)的合成
化合物85的合成路线与化合物1类似：
82.1 3-叔丁基二甲基硅氧基苯甲醛(85b)的合成
Figure PCTCN2017078444-appb-000125
500mL三口瓶中加入3-羟基苯甲醛(85a)16g(0.13mol),DMAP(320mg,2.6mmol),无水DCM(150mL),无水三乙胺(23mL,0.182mol),冷至冰浴下,TBSCl(20g,0.156mol)溶于无水DCM(50mL)中,由滴液漏斗缓慢加入；滴完后撤去冰浴,室温搅拌两小时；加入饱和碳酸氢钠水溶液淬灭反应,分出有机相,水相用DCM萃取,无水硫酸钠干燥3h,湿法上柱,分离纯化,收集对应的洗脱液,蒸干溶剂得无色油状化合物(85b)30g,收率97％；1H NMR(400MHz,CDCl3)δ9.95(s,1H),7.47(d,J＝7.6Hz,1H),7.40(t,J＝7.6Hz,1H),7.32(s,1H),7.12–7.08(m,1H),0.99(s,9H),0.22(s,6H)；ESI-LRMS m/z(％):237.1[M+H]+.
82.2 2-[1-(3-叔丁基二甲基硅氧基苯基)-1-羟基甲基]丙烯酸乙酯(85c)的合成
Figure PCTCN2017078444-appb-000126
在50mL茄形瓶中加入化合物85b(30g,0.114mol),丙烯酸乙酯(13.7g,0.136mol),DABCO(15.3g,0.136mol),室温搅拌反应20d后,TLC(展开剂：石油醚/乙酸乙酯＝10:1)显示原料(Rf＝0.6)明显减少,有大量新点生成(Rf＝0.2),停止反应,用较粗的柱子,直接将反应液倒到硅胶柱上,用少量甲苯洗涤反应瓶再倒到柱子上,然后用PE及PE/EA 10/1先除去丙烯酸乙酯,继而进行梯度洗脱,收集对应的洗脱液,回收原料8.3g,回收率28％,得无色油状产物(85c)17g,扣除回收原料,收率55％；1H NMR(400MHz,CDCl3)δ7.11(t,J＝7.8Hz,1H),6.89(d,J＝7.6Hz,1H),6.81(s,1H),6.69(dd,J＝7.8,2.0Hz,1H),6.24(s,1H),6.24(s,1H),5.79(s,1H),5.42(d,J＝5.2Hz,1H),4.05(q,J＝7.1Hz,2H),3.76(d,J＝5.2Hz,1H),1.14(t,J＝7.1Hz,3H),0.95(s,9H),0.15(s,6H)；ESI-LRMS m/z(％):337.2[M+H]+.
82.3 2-[1-(3-叔丁基二甲基硅氧基苯基)-1-乙酰氧基甲基]丙烯酸乙酯(85d)的合成
Figure PCTCN2017078444-appb-000127
化合物85c(16.6g,0.049mol),DMAP(600mg,0.0049mol)和TEA(9.9g,0.098mmol)溶于50mL无水DCM中,0℃搅拌条件下,缓慢滴加乙酸酐(10g,0.098mmol),加完后在冰浴中继续搅拌；10min后,TLC(展开剂：石油醚/乙酸乙酯＝5:1)显示原料(Rf＝0.3)完全消失,有新点(Rf＝0.5)生成,缓慢加入饱和碳酸氢钠水溶液淬灭反应；分液,水层用DCM(3x 50mL)萃取,合并有机相,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,减压蒸干溶剂得无色油状化合物(85d)8.9g,收率47％；1H NMR(400MHz,CDCl3)δ7.17(t,J＝7.9Hz,1H),6.95(d,J＝7.6Hz,1H),6.83(s,1H),6.78–6.73(m,1H),6.62(s,1H),6.37(s,1H),5.77(s,1H),4.14(q,J＝7.1Hz,2H),2.07(s,3H),1.20(t,J＝7.1Hz,3H),0.96(s,9H),0.17(s,6H)；13C NMR(150MHz,CDCl3)δ182.1,177.8,168.5,152.9,152.2,142.3,142.2,138.4,133.4,132.8,132.2,85.7,73.7,38.5,33.9,33.8,31.1,26.9,8.4；ESI-LRMS m/z(％):401.2[M+Na]+；ESI-HRMS m/z(％):Calcd for C20H34NO5Si[M+H]+396.2201,found 396.2201.
82.4 (S)-2-[1-(3-叔丁基二甲基硅氧基苯基)-1-(3,4,5-三甲氧基苯基氨基)甲基]丙烯酸乙酯(85e)的合成
Figure PCTCN2017078444-appb-000128
氮气氛围下,三(二亚苄基丙酮)二钯(22mg,0.023mmol)和1e(44mg,0.060mmol)分别加入一Schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入底物85d(900mg,2.38mmol),K2CO3(1.0M水溶液,7.14mL,7.14mmol)和3,4,5-三甲氧基苯胺(653mg,3.57mmol)；室温搅拌,过夜反应后,TLC(展开剂：石油醚/乙酸乙酯＝ 5:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.2)生成,用二氯甲烷萃取(3×50mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状物(85e)1.12g,收率94％；[α]D 20＝+27.0(c 0.16,CHCl3)；1H NMR(400MHz,CDCl3)δ7.19(t,J＝7.9Hz,1H),6.96(d,J＝7.7Hz,1H),6.83(t,J＝2.0Hz,1H),6.75(dd,J＝7.7,2.0Hz,1H),6.37(s,1H),5.91(s,1H),5.82(s,2H),5.33(s,1H),4.21–4.12(m,2H),3.76(s,6H),3.75(s,3H),1.23(t,J＝7.1Hz,3H),0.95(d,J＝4.9Hz,9H),0.15(s,6H)；13C NMR(150MHz,CDCl3)δ166.2,156.0,153.8,143.5,142.1,140.8,130.5,129.7,126.0,120.4,119.4,119.2,91.4,61.1,60.9,59.2,55.9,25.7,18.2,14.1,-4.4；ESI-LRMS m/z(％):502.2[M+H]+；ESI-HRMS m/z(％):Calcd for C27H40NO6Si[M+H]+502.2620,found 502.2619.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
82.5 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-叔丁基二甲基硅氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(85f)的合成
Figure PCTCN2017078444-appb-000129
化合物85e(1.094g)和Sn[N(TMS)2]2(183mg,0.40mmol)加入一Schlenk管中,加入无水甲苯(10mL),加热回流6小时后,TLC(展开剂：石油醚/乙酸乙酯＝5:1)显示产物与原料极性相近,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状化合物(85f)753mg,收率76％；[α]D 20＝+44.3(c 0.24,CHCl3)；1H NMR(400MHz,CDCl3)δ7.27–7.20(m,1H),6.99(d,J＝7.7Hz,1H),6.83–6.78(m,2H),6.57(s,2H),5.81(t,J＝1.7Hz,1H),5.30(s,1H),5.16–5.15(m,1H),3.75(s,3H),3.71(s,6H),0.91(s,9H),0.11(s,3H),0.10(s,3H)；13C NMR(150MHz,CDCl3)δ160.7,156.4,153.6,149.6,138.0,134.8,133.7,130.1,120.7,119.8,118.2,110.7,95.0,63.7,60.9,56.0,25.6,18.2,-4.4,-4.5；ESI-LRMS m/z(％):456.1[M+H]+；ESI-HRMS m/z(％):Calcd for C25H34NO5Si[M+H]+456.2201,found456.2201.
82.6 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基苯基)-3-亚甲基氮杂环丁烷-2-酮(85)的合成
Figure PCTCN2017078444-appb-000130
化合物85f(753mg)溶于10mL THF,冷至冰浴下,TBAF(863mg,3.3mmol)溶于3mL THF,由分液漏斗缓慢滴加,滴完后继续搅拌15min后,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料(Rf＝0.8)完全消失,有新点(Rf＝0.5)生成,减压旋干,10mL乙酸乙酯溶解,水洗(5mL x 2),饱和食盐水洗(5mL),无水硫酸钠干燥3h,过滤浓缩后,柱层析纯化,得淡黄色固体(85)489mg,收率86％；Mp 150-151℃；[α]D 20＝+101.5(c 0.13,CHCl3),98％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/ i-PrOH＝80:20,1.0mL/min,254nm；tR(minor)＝12.09min；tR(major)＝8.43min]；1H NMR(400MHz,CDCl3)δ7.59(s,1H),7.22–7.21(m,1H),6.94(d,J＝7.6Hz,1H),6.88–6.86(m,2H),6.56(s,2H),5.73((t,J＝1.7Hz,1H),5.31(s,1H),5.14(dd,J＝1.7,1.2Hz,1H),3.74(s,3H),3.66(s,6H)；13C NMR(100MHz,CDCl3)δ161.4,157.4,153.5,148.7,137.7,133.5,130.3,119.1,116.6,113.0,111.5,95.1,64.1,60.9,56.0；ESI-LRMS m/z(％):342.1[M+H]+；ESI-HRMS m/z(％):Calcd for C19H20NO5[M+H]+342.1336,found342.1337.
实施例83 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(86)的合成
Figure PCTCN2017078444-appb-000131
20mL茄型瓶中加入化合物85(20mg,0.059mmol),碳酸钾(9.7mg,0.071mmol)和丙酮(3mL),搅拌条件下慢慢加入硫酸二甲酯(9.6mg,0.077mmol),回流反应0.5h后,TLC(展开剂：石油醚/乙酸乙酯＝2:1)显示原料(Rf＝0.2)完全消失,有新点生成(Rf＝0.4),停止反应,过滤出去反应液中固体成分,直接拌硅胶加压蒸干,柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得白色固体(86)20mg,收率96％；Mp 122-123℃；[α]D 20＝+64.3(c 0.29,CHCl3)；1H NMR(400MHz,CDCl3)δ7.30(t,J＝7.7Hz,1H),6.99(d,J＝7.7Hz,1H),6.89(m,2H),6.59(s,2H),5.83(s,1H),5.33(s,1H),5.17(s,1H),3.78(s,3H),3.76(s,3H),3.73(s,6H)；13C NMR(150MHz,CDCl3)δ160.2,159.6,152.9,148.8,137.5,134.1,133.2,129.6,118.6,113.6,111.6,110.3,94.2,63.3,60.3,55.4,54.7；ESI-LRMS m/z(％):356.0[M+H]+；ESI-HRMS m/z(％):Calcd for C20H22NO5[M+H]+356.1492,found 356.1494.
实施例84 (S)-1-(3,4,5-三甲氧基苯基)-4-(4-甲基苯基)-3-亚甲基氮杂环丁烷-2-酮(87)的合成
化合物84的合成与化合物1的合成类似：
84.1 2-[1-(4-甲基苯基)-1-羟基甲基]丙烯酸乙酯(84b)的合成
Figure PCTCN2017078444-appb-000132
在50mL茄形瓶中加入化合物对甲苯甲醛84a(5g,0.042mol),丙烯酸乙酯(4.17g,0.042mol),DABCO(4.67g,0.042mol),室温搅拌反应12d后,TLC(展开剂：石油醚/乙酸乙酯＝10:1)显示原料(Rf＝0.5)明显减少,有大量新点生成(Rf＝0.2),停止反应,用较粗的柱子,直接将反应液倒到硅胶柱上,用少量甲苯洗涤反应瓶再倒到柱子上,然后用PE及PE/EA 10/1先除去丙烯酸乙酯,继而进行梯度洗脱,收集对应的洗脱液,蒸干溶剂得无色油状物质(84b)6.7g,收率73％；1H NMR(400MHz,CDCl3)δ7.25(d,J＝8.0Hz,2H),7.15(d,J＝8.0Hz,2H),6.32(s,1H),5.83(s,1H),5.52(d,J＝5.4Hz,1H),4.16(q,J＝7.1Hz,2H),3.07(d,J＝5.4Hz,1H),2.33(s,3H),1.24(t,J＝7.1Hz,3H)；ESI-LRMS m/z(％):221.1[M+H]+.
84.2 2-[1-(4-甲基苯基)-1-乙酰氧基甲基]丙烯酸乙酯(84c)的合成
Figure PCTCN2017078444-appb-000133
化合物84b(2g,8.93mmol),DMAP(108mg,0.893mmol)和TEA(1.8g,17.86mmol)溶于15mL无水DCM中,0℃搅拌条件下,缓慢滴加乙酸酐(1.8g,17.86mmol),加完后在冰浴中继续搅拌；10min后,TLC(展开剂：石油醚/乙酸乙酯＝10:1)显示原料(Rf＝0.2)完全消失,有新点(Rf＝0.4)生成,缓慢加入饱和碳酸氢钠水溶液淬灭反应；分液,水层用DCM(3x 15mL)萃取,合并有机相,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,减压蒸干溶剂得无色油状化合物(84c)1.95g,收率83％；1HNMR(400MHz,CDCl3)δ7.28(d,J＝7.9Hz,2H),7.15(d,J＝7.9Hz,2H),6.67(s,1H),6.39(s,1H),5.84(s,1H),4.15(q,J＝6.6Hz,2H),2.33(s,3H),2.09(s,3H),1.22(t,J＝7.1Hz,3H)；ESI-LRMS m/z(％):263.1[M+H]+.
84.3 (S)-2-[1-(4-甲基苯基)-1-(3,4,5-三甲氧基苯基氨基)甲基]丙烯酸乙酯(84d)的合成
Figure PCTCN2017078444-appb-000134
氮气氛围下,三(二亚苄基丙酮)二钯(6mg,0.0064mmol)和1e(12mg,0.016mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入底物84c(167mg,0.64mmol),K2CO3(1.0M水溶液,2mL,2mmol)和3,4,5-三甲氧基苯胺(175mg,0.96mmol)；室温下搅拌4小时后,TLC(展开剂：石油醚/乙酸乙酯＝2:1)显示原料(Rf＝0.7)完全消失,有新点(Rf＝0.5)生成,用二氯甲烷萃取(3×5mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状液体(84d)127mg,收率52％；[α]D 20＝+88.9(c 0.10,CHCl3)；1H NMR(400MHz,CDCl3)δ7.26(d,J＝7.9Hz,2H),7.14(d,J＝7.9Hz,2H),6.38(s,1H),5.95(s,1H),5.82(s,2H),5.35(s,1H),4.21–4.05(m,3H),3.76(s,6H),3.75(s,3H),2.33(s,3H),1.22(t,J＝7.1Hz,3H)；13C NMR(150MHz,CDCl3)δ166.3,153.8,143.6,140.8,137.7,137.5,130.5,129.4,127.3,125.7,91.2,61.1,60.8,59.1,55.9,21.1,14.1；ESI-LRMS m/z(％):386.2[M+H]+；ESI-HRMS m/z(％):Calcd forC22H28NO5[M+H]+386.1962,found 386.1964.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
84.4 (S)-1-(3,4,5-三甲氧基苯基)-4-(4-甲基苯基)-3-亚甲基氮杂环丁烷-2-酮(84)的合成
Figure PCTCN2017078444-appb-000135
化合物84d(127mg,0.33mmol)和Sn[N(TMS)2]2(218mg,0.50mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流6小时后,TLC(展开剂：石油醚/乙酸乙 酯＝2:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.45)生成,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状化合物(84)101mg,收率89％；[α]D 20＝+87.5(c 0.12,CHCl3),97％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝85:15,1.0mL/min,254nm；tR(minor)＝9.04min；tR(major)＝11.43min]；1H NMR(400MHz,CDCl3)δ7.29(d,J＝8.0Hz,2H),7.19(d,J＝8.0Hz,2H),6.58(s,2H),5.83(t,J＝1.7Hz,1H),5.33(s,1H),5.16–5.12(m,1H),3.76(s,3H),3.72(s,6H),2.35(s,3H)；13C NMR(150MHz,CDCl3)δ160.9,153.5,149.9,138.9,134.7,133.8,133.5,129.8,126.8,110.7,94.9,63.9,60.9,56.1,21.2；ESI-LRMS m/z(％):340.1[M+H]+；ESI-HRMS m/z(％):Calcd for C20H22NO4[M+H]+340.1543,found 340.1550.
实施例85 (S)-1-(3,4,5-三甲氧基苯基)-4-(4-异丙基苯基)-3-亚甲基氮杂环丁烷-2-酮(88)的合成
化合物88的合成与化合物1的合成类似：
85.1 2-[1-(4-异丙基苯基)-1-羟基甲基]丙烯酸乙酯(88b)的合成
Figure PCTCN2017078444-appb-000136
在50mL茄形瓶中加入化合物4-异丙基苯甲醛88a(2g,13.5mmol),丙烯酸乙酯(1.35g,13.5mmol),DABCO(1.5g,13.5mmol),室温搅拌反应13d后,TLC(展开剂：石油醚/乙酸乙酯＝10:1)显示原料(Rf＝0.7)明显减少,有大量新点生成(Rf＝0.2),停止反应,用较粗的柱子,直接将反应液倒到硅胶柱上,用少量甲苯洗涤反应瓶再倒到柱子上,然后用PE及PE/EA 10/1先除去丙烯酸乙酯,继而进行梯度洗脱,收集对应的洗脱液,回收原料0.6g,回收率30％,得无色油状产物(88b)1.1g,扣除回收原料,收率47％；1H NMR(400MHz,CDCl3)δ7.29(d,J＝8.2Hz,2H),7.20(d,J＝8.2Hz,2H),6.33(s,1H),5.83(t,J＝1.1Hz,1H),5.54(s,1H),4.17(q,J＝7.1Hz,2H),2.99(s,1H),2.89(dt,J＝13.8,6.9Hz,1H),1.26–1.23(m,9H)；ESI-LRMS m/z(％):249.1[M+H]+.
85.2 2-[1-(4-异丙基苯基)-1-乙酰氧基甲基]丙烯酸乙酯(88c)的合成
Figure PCTCN2017078444-appb-000137
化合物88b(1.1g,4.44mmol),DMAP(54mg,0.444mmol)和TEA(0.9g,8.88mmol)溶于15mL无水DCM中,0℃搅拌条件下,缓慢滴加乙酸酐(0.9g,8.88mmol),加完后在冰浴中继续搅拌；10min后,TLC(展开剂：石油醚/乙酸乙酯＝10:1)显示原料(Rf＝0.2)完全消失,有新点(Rf＝0.4)生成,缓慢加入饱和碳酸氢钠水溶液淬灭反应；分液,水层用DCM(3x 15mL)萃取,合并有机相,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,减压蒸干溶剂得无色油状化合物(88c)946mg,收率74％；1H NMR(400MHz,CDCl3)δ7.28(d,J＝7.9Hz,2H),7.15(d,J＝7.9Hz,2H),6.67(s,1H),6.39(s,1H),5.84(s,1H),4.25–4.00(m,2H),2.33(s,3H),2.09(s,3H),1.22(t,J＝7.1Hz,3H)；ESI-LRMS m/z(％):291.1[M+H]+.
85.3 (S)-2-[1-(4-异丙基基苯基)-1-(3,4,5-三甲氧基苯基氨基)甲基]丙烯酸乙酯(88d)的合成
Figure PCTCN2017078444-appb-000138
氮气氛围下,三(二亚苄基丙酮)二钯(4.7mg,0.0052mmol)和1e(9.3mg,0.0129mmol)分别加入一schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入底物88c(150mg,0.52mmol),K2CO3(1.0M水溶液,1.5mL,1.5mmol)和3,4,5-三甲氧基苯胺(143mg,0.78mmol)；室温下搅拌24小时后,TLC(展开剂：石油醚/乙酸乙酯＝5:1)显示原料(Rf＝0.7)完全消失,有新点(Rf＝0.3)生成,用二氯甲烷萃取(3×5mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得淡黄色油状物(88d)210mg,收率98％；[α]D 20＝+67.5(c 0.17,CHCl3)；1H NMR(400MHz,CDCl3)δ7.28(d,J＝8.2Hz,2H),7.19(d,J＝8.2Hz,2H),6.38(s,1H),5.97(s,1H),5.82(s,2H),5.35(s,1H),4.19–4.13(m,2H),3.76(s,6H),3.75(s,3H),2.95–2.82(m,1H),1.26–1.19(m,9H)；13C NMR(150MHz,CDCl3)δ166.4,153.8,148.5,143.6,140.8,138.0,130.4,127.4,126.8,125.6,91.2,61.1,60.8,59.1,55.9,33.8,24.0,14.1；ESI-LRMS m/z(％):414.1[M+H]+.
以0.005当量的三苯基膦替换1e做配体，其他操作完全相同,可得相应外消旋体。
85.4 (S)-1-(3,4,5-三甲氧基苯基)-4-(4-异丙基苯基)-3-亚甲基氮杂环丁烷-2-酮(88)的合成
Figure PCTCN2017078444-appb-000139
化合物88d(110mg)和Sn[N(TMS)2]2(140mg,0.32mmol)加入一Schlenk管中,加入无水甲苯(5mL),加热回流3.5小时后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.45)生成,反应液冷却至室温后,浓缩,柱层析纯化,得白色固体(88)21mg,收率21％；Mp 99-100℃；[α]D 20＝+47.7(c 0.11,CHCl3)；92％ee[determined by HPLC analysis using a Chiralcel AD-H column；n-Hex/i-PrOH＝75:25,1.0mL/min,254nm；tR(minor)＝5.30min；tR(major)＝6.59min]；1H NMR(400MHz,CDCl3)δ7.32(d,J＝8.2Hz,2H),7.23(d,J＝8.2Hz,2H),6.58(s,2H),5.83(t,J＝1.7Hz,1H),5.34(s,1H),5.19–5.15(m,1H),3.76(s,3H),3.71(s,6H),2.89(dq,J＝13.7,6.9Hz,1H),1.24(s,3H),1.22(s,3H)；13C NMR(150MHz,CDCl3)δ161.0,153.5,149.9,149.8,134.7,133.9,133.8,127.2,126.9,110.7,95.0,64.0,60.9,56.0,33.9,23.9；ESI-LRMS m/z(％):368.2[M+H]+；ESI-HRMS m/z(％):Calcd for C20H26NO4[M+H]+368.1856,found 368.1859.
实施例86 (3R,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-丙烯酰氧基-4-甲氧基苯基)-3-丙烯酰氧基-3-丙烯酰氧甲基氮杂环丁烷-2-酮(89)的合成
Figure PCTCN2017078444-appb-000140
在100mL茄形瓶中加入化合物80(21mg,0.052mmol),丙烯酰氯(42μL,0.52mmol),三乙胺(70μL,0.52mmol),DMAP 1mg与无水DCM 1mL,室温反应0.5小时；TLC(展开剂：PE/EA＝1:2)显示原料点(Rf＝0.3)消失,生成新点(Rf＝0.95)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(89)14mg,收率48％；1H NMR(400MHz,CDCl3)δ7.28(d,J＝10.4Hz,1H),7.20(s,1H),6.98(d,J＝8.5Hz,1H),6.65–6.51(m,4H),6.43–6.17(m,3H),6.13–5.96(m,3H),5.84(d,J＝10.5Hz,1H),5.38(s,1H),4.42(s,2H),3.83(s,3H),3.79(s,3H),3.73(s,6H)；；13C NMR(150MHz,CDCl3)δ164.5,164.3,163.0,160.7,153.0,151.1,139.2,134.5,133.0,132.3,132.2,131.2,126.9,126.8,126.5,125.2,123.6,121.8,112.0,94.9,88.0,64.1,60.3,55.5,55.4.
实施例87 (3S,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-烯丙酰氧基-4-甲氧基苯基)-3-烯丙酰氨基氮杂环丁烷-2-酮(90)的合成
Figure PCTCN2017078444-appb-000141
在100mL茄形瓶中加入化合物76(17mg,0.045mmol),丙烯酰氯(11μL,0.13mmol),三乙胺(19μL,0.13mmol),DMAP 1mg与无水DCM 1mL,室温反应3小时；TLC(展开剂：PE/EA＝1:3)显示原料点(Rf＝0.2)消失,生成新点(Rf＝0.95)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(90)15mg,收率69％；1H NMR(400MHz,CDCl3)δ7.24(dd,J＝8.5Hz,2.1Hz,1H),7.12(d,J＝2.1Hz,1H),6.99(d,J＝8.5Hz,1H),6.71(d,J＝6.8Hz,1H),6.59(d,J＝17.3Hz,1H),6.51(s,2H),6.40–6.25(m,2H),6.15(dd,J＝17.0,10.3Hz,1H),6.02(d,J＝10.4Hz,1H),5.72(d,J＝10.4Hz,1H),5.02(d,J＝2.2Hz,1H),4.57(dd,J＝6.8,2.2Hz,1H),3.82(s,3H),3.76(s,3H),3.70(s,6H)；13C NMR(150MHz,CDCl3)δ165.8,163.9,163.7,153.5,151.6,140.1,134.8,133.4,132.9,129.4,128.8,128.3,127.4,124.7,121.1,113.1,95.2,66.2,62.5,60.9,56.1.
实施例88 (S)-1-(3,4,5-三甲氧基苯基)-2-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷(91)的合成
Figure PCTCN2017078444-appb-000142
向50mL Schlenk管中加入无水THF 3mL,冰浴,氮气保护,缓缓加入三氯化铝(36mg,0.27mmol),四氢锂铝(10mg,0.27mmol),氮气保护下逐渐升至室温反应1小时；加入化合物1(10mg,0.027mmol),室温反应3h后,TLC(展开剂：PE/EA＝2:1)显示生成新点(Rf＝0.46)；反应液加饱和氯化铵,用乙酸乙酯萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝5:1),得到白色固体(91)5mg,收率51％；1H NMR(400MHz,CDCl3)δ6.79(d,J＝1.1Hz 1H),6.73(dd,J＝8.1,1.1Hz,1H),6.63(d,J＝8.1Hz,1H),5.94(s,2H),5.19(s,1H),4.92(s,1H),4.73(s,1H),3.89–3.74(m,14H).
实施例89(3S,4R)-2-(3,4,5-三甲氧基苯基)-3-(3-乙酰氧基-4-甲氧基苯基)-2-氮杂-5,7-二氧杂螺[3.4]辛烷-1,6-二酮(92)的合成
Figure PCTCN2017078444-appb-000143
在100mL茄形瓶中加入化合物82(18mg,0.04mmol),4-二甲氨基吡啶0.5mg,三乙胺(14μL,0.1mmol)与二氯甲烷5mL,草酰氯(5μL,0.058mmol),室温反应3小时；TLC(展开剂：PE/EA＝1:4)显示原料点(Rf＝0.2)消失,生成新点(Rf＝0.9)；反应液加饱和NaHCO3,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝1:1),得到白色固体(92)12mg,收率63％；1H NMR(400MHz,CDCl3)δ7.26(d,J＝1.0Hz,1H),7.15–7.00(m,2H),6.53(s,2H),5.33(s,1H),4.55(d,J＝9.9Hz,1H),4.05(d,J＝9.9Hz,1H),3.86(s,3H),3.78(s,3H),3.72(s,6H),2.30(s,3H)；13C NMR(101MHz,CDCl3)δ168.8,160.4,153.9,152.8,152.7,141.0,135.9,132.1,123.4,113.7,96.0,90.3,67.1,64.5,61.2,56.4,56.3.
实施例90 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-乙酰氧基-4-甲氧基苯基)氮杂环丁烷-2,3-二酮(93)的合成
Figure PCTCN2017078444-appb-000144
在100mL茄形瓶中加入化合物82(0.25g,0.56mmol),高碘酸钠(0.18g,0.84mmol),甲醇4mL与水1mL,室温反应3小时；TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0)消失,生成新点(Rf＝0.2)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水 洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝1:1),得到黄色固体(93)0.18g,收率80％；1H NMR(400MHz,CDCl3)δ7.17(dd,J＝8.5,2.2Hz,1H),7.04(d,J＝2.2Hz,1H),6.99(d,J＝8.5Hz,1H),6.74(s,2H),5.49(s,1H),3.83(s,3H),3.81(s,3H),3.76(s,6H),2.30(s,3H)；13C NMR(101MHz,DMSO)δ209.3,191.0,168.8,160.5,153.9,152.4,140.6,136.6,132.6,125.13,124.2,121.7,113.3,96.1,74.7,61.2,56.4,56.3,20.9.
实施例91 (3R,4S)-1-(3,4,5-三甲氧基苯基)-3-甲基-3-羟基-4-(3-乙酰氧基-4-甲氧基苯基)氮杂环丁烷-2-酮(94)的合成
Figure PCTCN2017078444-appb-000145
向50mL Schlenk管中加入化合物93(10mg,0.024mmol),抽换三次氮气,加入无水THF 1mL,冰浴下加入MgCl(3M in THF)(1.2μL,0.036mmol),氮气保护下,室温反应1h；TLC(展开剂：PE/EA＝1:2)显示生成新点(Rf＝0.2)；加水,EA萃取,饱和食盐水洗,硫酸钠干燥,柱层析分离(展开剂：PE/EA＝1:1),得到白色固体(94)6mg,收率58％；1H NMR(400MHz,CDCl3)δ7.17(dd,J＝8.5,2.1Hz,1H),7.02-7.00(m,2H),6.59(s,2H),4.90(s,1H),3.84(s,3H),3.78(s,3H),3.73(s,6H),2.30(s,3H),1.70(s,3H).
实施例92 (3R,4S)-1-(3,4,5-三甲氧基苯基)-3-甲基-3-羟基-4-(3-羟基-4-甲氧基苯基)氮杂环丁烷-2-酮(95)的合成
Figure PCTCN2017078444-appb-000146
在100mL茄形瓶中加入化合物94(6mg,0.014mmol),70％水合肼(1.5μL,0.03mmol),MeOH 1mL,室温反应1小时；TLC(展开剂：PE/EA＝1:3)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0.3)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；柱层析分离(展开剂：PE/EA＝1:2),得到白色固体(95)5mg,收率96％；1H NMR(400MHz,CDCl3)δ6.84(d,J＝1.5Hz,1H),6.83(d,J＝8.3Hz,1H),6.76(dd,J＝8.3,1.5Hz,1H),6.57(s,2H),5.61(br s,1H),4.78(s,1H),3.89(s,3H),3.78(s,3H),3.72(s,6H),1.69(s,3H).；ESI-LRMS m/z(％):390.1[M+H]+.
实施例93 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-对甲氧基苯甲酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(96)的合成
Figure PCTCN2017078444-appb-000147
化合物1(20mg,0.054mmol)溶于2mL DCM中,加入TEA(15μL,0.108mmol),对甲氧基苯甲酸(17mg,0.108mmol),EDCI(21mg,0.108mmol),DMAP(1mg,0.008mmol),室温反应0.5h,TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料1(Rf＝0.3)完全消失,有新点生成(Rf＝0.4),停止反应(1mL水淬灭),加入10mL DCM,以水(2x 10mL)洗涤,饱和食盐水(10mL)洗,无水硫酸钠干燥3h,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得无色油状液体(96)25mg,收率93％；[α]D 20＝+31.7(c 1.0,CHCl3)；1H NMR(CDCl3,400MHz)δ:7.95(d,J＝8.7Hz,1H),7.11～7.05(m,3H),6.84～6.78(m,3H),6.43(s,2H),5.68(s,1H),5.15(s,1H),5.04(s,1H),3.71(s,3H),3.63(s,3H),3.59～3.58(m,9H)；13C NMR(CDCl3,150MHz)δ:163.5,163.3,160.2,153.0,151.3,149.0,139.9,134.1,133.1,131.8,128.3,124.5,121.4,120.8,113.2,112.3,110.4,94.2,62.8,60.3,55.5,55.4,54.9.MS(ESI)m/z(％):506.1[M+H]+；HRMS(ESI)calcd for C28H27NNaO8[M+Na]+528.1629,found 528.1631.
实施例94 (3S,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-甲基氮杂环丁烷-2-酮(97)的合成
Figure PCTCN2017078444-appb-000148
化合物69(210mg,0.54mmol)溶于5mL MeCN中,加入碳酸钾(112mg,0.81mmol),氯化苄(75μL,0.65mmol),回流反应8h,TLC(展开剂：PE/EA＝1:3)显示原料点(Rf＝0.22)消失,生成新点(Rf＝0.5)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,溶于5mL无水THF中，加入四溴化碳(537mg,1.62mmol),三苯基膦(425mg,1.62mmol),室温反应4h,TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0.1)消失,生成新点(Rf＝0.5)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,溶于5mL乙醇中，加入无水醋酸钠(132mg,1.62mmol),10％钯碳(20mg),常压通氢气室温反应8h,TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0.5)消失,生成新点(Rf＝0.4)；反应液过滤除去钯碳，拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干得到白色固体64mg，收率32％；熔点:54–55℃；[α]D 20＝+12.8(c 0.5,CHCl3).1H NMR(400MHz,CDCl3)δ6.93(d,J＝1.5Hz,1H),6.87(dd,J＝8.3,1.5Hz,1H),6.83(d,J＝8.3Hz,1H),6.54(s,2H),5.69(s,1H),4.44(d,J＝2.2Hz,1H),3.89(s,3H),3.76(s,3H),3.72(s,6H),3.11(qd,J＝7.3,2.2Hz,1H),1.45(d,J＝7.3Hz,3H).13C NMR(150MHz,CDCl3)δ167.7,152.9,146.1,145.6,133.7,133.5,130.5,117.1,111.4,110.3,94.1,62.2,60.3,55.4,54.5,12.4.ESI-MS(m/z):374.0(M+H+).
实施例95 化合物(R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(98)的合成
Figure PCTCN2017078444-appb-000149
95.1 (R)-2-[1-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-1-(3,4,5-三甲氧基苯基氨基)甲基]丙烯酸乙酯(98b)的合成
氮气氛围下,三(二亚苄基丙酮)二钯(11mg,0.012mmol)和98a(21mg,0.031mmol)分别加入一Schlenk管中,加入无水CH2Cl2(5mL),室温下搅拌10分钟后,先后加入底物1d(0.5g,1.23mmol),K2CO3(1.0M水溶液,2.1mL,2.1mmol)和3,4,5-三氧基苯胺(0.34g,0.26mmol)；室温下搅拌2h后,TLC(展开剂：石油醚/乙酸乙酯＝9:1)显示原料(Rf＝0.5)完全消失,有新点(Rf＝0.2)生成,用二氯甲烷萃取(3×10mL),无水硫酸钠干燥,过滤浓缩后,柱层析纯化,得黄色油状物(98b)0.62g,收率95％.
95.2 (R)-1-(3,4,5-三甲氧基苯基)-4-(3-叔丁基二甲基硅氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(98c)的合成
化合物98b(0.62g)和Sn[N(TMS)2]2(0.76g,1.77mmol)加入一Schlenk管中,加入无水甲苯(20mL),加热回流8小时后,TLC(展开剂：石油醚/乙酸乙酯＝3:1)显示原料(Rf＝0.8)消失,有新点(Rf＝0.5)生成,反应液冷却至室温后,浓缩,柱层析纯化,得无色油状化合物(98c)0.52g,收率93％.
95.3 (R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(98)的合成
化合物98c(0.52g,1.1mmol)溶于15mL THF，冷至冰浴下，TBAF(0.42g,1.6mmol)溶于5mL THF，缓慢滴加，滴完后继续搅拌15min后，TLC(展开剂：石油醚/乙酸乙酯＝1:1)显示原料(Rf＝0.7)完全消失，有新点(Rf＝0.5)生成，减压旋干，20mL乙酸乙酯溶解，水洗(20mL x 2)，饱和食盐水洗(20mL)，无水硫酸钠干燥。过滤浓缩后，柱层析纯化，得无色液体(98)0.29g,收率73％。[α]D 20＝-37.9(c 0.77,CHCl3)；1H NMR(400MHz,CDCl3):δ6.93(d,J＝1.7Hz,1H),6.87(dd,J＝8.2,1.7Hz,1H),6.82(d,J＝8.2Hz,1H),6.58(s,2H),5.79(s,1H),5.26(s,1H),5.13(s,1H),3.84(s,3H),3.73(s,3H),3.71(s,6H).13C NMR(150MHz,CDCl3):δ160.3,152.9,149.2,146.5,145.6,134.1,133.2,128.9,118.1,112.3,110.3,110.0,94.31,63.0,60.3,55.5,55.4.ESI-MS(m/z):372.1(M+H+).ESI-HRMS(m/z):calcd for C20H21NO6+H+[M+H]+,372.1442；found,372.1441.
实施例96 (3S,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-羟甲基氮杂环丁烷-2-酮(99)和(3S,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-羟甲基氮杂环丁烷-2-酮(100)的合成
Figure PCTCN2017078444-appb-000150
氮气保护下下,在50mL Schlenk中加入化合物98(220mg,0.6mmol),联硼酸频哪醇酯(0.2g,0.78mmol),CuCl(6mg,0.06mmol),三苯基膦(20mg,0.076mmol),甲醇(30μL,0.74mmol),叔丁醇锂(6mg,0.074mmol)与无水THF 3mL,室温反应12小时；TLC(展开剂：PE/EA＝1:2)显示仅原料点(Rf＝0.4)等高处有点；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,加入NaBO3·4H2O(0.46g,3mmol),THF 2mL和水1mL,室温反应2h,TLC(展开剂：PE/EA＝1:3)显示原料点(Rf＝0.9)消失,生成两新点(Rf＝0.17,0.22)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,柱层析分离(展开剂：PE/EA＝1:1),得到反式产物为无色液体(99)48mg,收率21％；[α]D 20＝-80.1(c 1.68,CHCl3)；1H NMR(400MHz,CDCl3)δ6.97(d,J＝1.5Hz,1H),6.91(dd,J＝8.2Hz,1.5Hz,1H),6.84(d,J＝8.2Hz,1H),6.56(s,2H),5.70(s,1H),4.91(d,J＝1.8Hz,1H),4.15(dd,J＝12.1,4.5Hz,1H),4.00(dd,J＝12.1,3.7Hz,1H),3.90(s,3H),3.79–3.75(m,4H),3.73(s,6H),3.28(br d,J＝2.3Hz,1H)；13C NMR(151MHz,CDCl3)δ165.60,153.50,146.78,146.32,134.57,133.72,130.77,117.94,112.16,110.99,94.91,62.14,60.93,58.89,57.49,56.06；得到顺式产物(100)为白色固体95mg,收率41％；[α]D 20＝-116.9(c 1.0,CHCl3)；1H NMR(400MHz,CDCl3)δ6.90(d,J＝1.2Hz,1H),6.87–6.81(m,2H),6.55(s,2H),5.75(s,1H),5.16(d,J＝5.3Hz,1H),3.90(s,3H),3.86–3.75(m,5H),3.73(s,6H),3.64(dd,J＝11.3,7.8Hz,1H)；13C NMR(151MHz,CDCl3)δ165.18,153.56,146.73,146.13,134.68,133.60,127.13,118.47,112.90,110.93,95.06,60.96,58.15,57.27,56.72,56.15,55.98.
实施例97 (3S,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-甲基氮杂环丁烷-2-酮(101)的合成
Figure PCTCN2017078444-appb-000151
在100mL茄形瓶中加入化合物98(15mg,0.04mmol),10％Pd/C 3mg,甲醇1mL常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0.33)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白 色固体(101)13mg,收率95％；[α]D 20＝‐102.8(c 1.64,CHCl3)；1H NMR(400MHz,CDCl3)δ6.86–6.77(m,2H),6.72(d,J＝8.3Hz,1H),6.56(s,2H),5.71(s,1H),5.06(d,J＝5.8Hz,1H),3.89(s,3H),3.77(s,3H),3.72(s,6H),3.66-3.59(m,1H),0.91(d,J＝7.6Hz,3H).
实施例98 (3R,4S)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-甲基氮杂环丁烷-2-酮(102)的合成
Figure PCTCN2017078444-appb-000152
化合物99(30mg,0.078mmol)溶于2mL MeCN中,加入碳酸钾(16mg,0.12mmol),氯化苄(10μL,0.09mmol),回流反应8h,TLC(展开剂：PE/EA＝1:3)显示原料点(Rf＝0.22)消失,生成新点(Rf＝0.5)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,溶于2mL无水THF中，加入四溴化碳(77mg,0.23mmol),三苯基膦(61mg,0.23mmol),室温反应4h,TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0.1)消失,生成新点(Rf＝0.5)；反应液加水,用EA萃取三次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥；蒸干,溶于5mL乙醇中，加入无水醋酸钠(19mg,0.23mmol),10％钯碳(3mg),常压通氢气室温反应8h,TLC(展开剂：PE/EA＝1:1)显示原料点(Rf＝0.5)消失,生成新点(Rf＝0.4)；反应液过滤除去钯碳，拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干得到白色固体9mg，收率32％；熔点:54–55℃；[α]D 20＝+14.5(c0.65,CHCl3).1H NMR(400MHz,CDCl3)δ6.93(d,J＝1.5Hz,1H),6.87(dd,J＝8.3,1.5Hz,1H),6.83(d,J＝8.3Hz,1H),6.54(s,2H),5.69(s,1H),4.44(d,J＝2.2Hz,1H),3.89(s,3H),3.76(s,3H),3.72(s,6H),3.11(qd,J＝7.3,2.2Hz,1H),1.45(d,J＝7.3Hz,3H).13C NMR(150MHz,CDCl3)δ167.7,152.9,146.1,145.6,133.7,133.5,130.5,117.1,111.4,110.3,94.1,62.2,60.3,55.4,54.5,12.4.ESI-MS(m/z):374.0(M+H+).
实施例99 (3R,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-氨基-4-甲氧基苯基)-3-甲基氮杂环丁烷-2-酮(103)的合成
Figure PCTCN2017078444-appb-000153
在100mL茄形瓶中加入化合物41(20mg,0.05mmol),10％Pd/C 3mg,甲醇1mL常压氢气下,室温反应12小时,TLC(展开剂：PE/EA＝2:1)显示原料点(Rf＝0.4)消失,生成新点(Rf＝0.3)；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(103)17mg,收率85％；熔点：75–76℃；[α]D 20＝+120.7(c 1.0,CHCl3).1H NMR(400MHz,CDCl3)δ6.74(d,J＝8.1Hz,1H),6.61–6.52(m,4H),5.01(d,J＝5.8Hz,1H),3.84(s,3H),3.76(s,3H),3.71(s,6H),3.59(dq,J＝7.6,5.8Hz,1H),0.91(d,J＝7.6Hz,3H).13C NMR(100MHz,CDCl3):δ168.8,153.9,146.5,146.2,134.0,132.2,128.2,118.6,113.5, 110.0,95.1,61.3,58.8,56.9,56.4,49.5,29.8,9.7.ESI-MS(m/z):373.1(M+H+).
实施例100 (3S,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-氨基-4-甲氧基苯基)-3-甲基氮杂环丁烷-2-酮(104)的合成
Figure PCTCN2017078444-appb-000154
在100mL茄形瓶中加入化合物41f(100mg,0.19mmol),(S)-Ir-PHOX 10mg,二氯甲烷5mL常压氢气下,室温反应12小时；反应液加水,用二氯甲烷萃取3次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥,蒸干后和Sn[N(TMS)2]2(100mg,0.33mmol)加入Schlenk管中,加入无水甲苯(5mL),加热回流3小时,待反应液冷却至室温后,柱层析纯化,得无色固体(104)46mg,收率65％；熔点:62–64℃；[α]D 20＝-19.1(c 1.0,CHCl3).1H NMR(400MHz,CDCl3)δ6.82(d,J＝8.0Hz,1H),6.65–6.55(m,4H),4.50(d,J＝2.1Hz,1H),3.88(s,3H),3.79(s,3H),3.70(s,6H),3.32(qd,J＝7.1,2.1Hz,1H),1.41(d,J＝7.1Hz,3H).13C NMR(150MHz,CDCl3)δ167.8,152.8,146.0,145.6,133.0,133.8,130.3,117.0,111.5,110.4,94.2,62.1,60.5,55.5,54.1,12.3.ESI-MS(m/z):373.1(M+H+).
实施例101 (S)-5-(3,4,5-三甲氧基苯基)-6-(3-羟基-4-甲氧基苯基)-5-氮杂螺[2.3]己烷-4-酮(105)的合成
Figure PCTCN2017078444-appb-000155
在100mL茄形瓶中加入化合物1g(390mg,0.81mmol),TMSCH2N2(1.2mL,2M in hexane,2.4mmol),二氯甲烷5mL,室温反应12小时；反应液加水,用二氯甲烷萃取3次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥,蒸干后加入THF 5mL,TBAF(210mg,0.81mmol),室温反应3小时；反应液加水,用乙酸乙酯萃取3次,有机相合并后用饱和食盐水洗一次,Na2SO4干燥,柱层析纯化,得黄色固体(105)160mg,收率53％；熔点:120–122℃；[α]D 20＝+63.2(c 1.0,CHCl3).1H NMR(400MHz,CDCl3)δ6.92(dd,J＝8.1,2.0Hz,1H),6.88(d,J＝8.1Hz,1H),6.77(d,J＝2.0Hz,1H),4.72(s,1H),3.85(s,3H),3.76(s,3H),3.68(s,6H),2.15–2.08(m,2H),1.45–1.38(m,2H).13C NMR(150MHz,CDCl3)δ167.6,152.6,146.2,145.8,133.1,133.8,130.4,117.5,111.0,110.5,94.1,62.2,60.4,55.5,54.0,22.9,18.1,16.3.ESI-MS(m/z):386.1(M+H+).
实施例102 (3R,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-羟基-4-甲氧基苯基)-3-乙基氮杂环丁烷-2-酮(106)的合成
Figure PCTCN2017078444-appb-000156
在100mL茄形瓶中加入化合物105(20mg,0.05mmol),10％Pd/C 3mg,甲醇1mL,常压氢气下,室温反应12小时；过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到白色固体(106)15mg,收率75％；熔点:85–87℃；[α]D 20＝+130.3(c 1.0,CHCl3).1H NMR(400MHz,CDCl3)δ6.85–6.78(m,2H),6.70(d,J＝8.1Hz,1H),6.60(s,2H),5.01(d,J＝5.7Hz,1H),3.88(s,3H),3.76(s,3H),3.70–3.63(m,7H),1.24–1.20(m,1H),0.87(d,J＝7.5Hz,3H)..13C NMR(150MHz,CDCl3)δ167.6,152.8,147.5,146.4,133.4,133.6,130.1,117.5,111.2,110.4,94.0,62.3,61.4,55.5,54.2,22.7,19.0,18.2,16.6.ESI-MS(m/z):388.1(M+H+).
实施例103 (S)-1-(3,4,5-三甲氧基苯基)-4-(3-氟磺酰氧基-4-甲氧基苯基)-3-亚甲基氮杂环丁烷-2-酮(107)的合成
Figure PCTCN2017078444-appb-000157
化合物1(光学纯度99％,20mg,0.054mmol)溶于1.5mL无水二氯甲烷中,0℃搅拌条件下,依次加入三乙胺(11mg,0.108mmol)和氟磺酰氟(过量),后升至室温反应1h.停止反应,用1mL水淬灭,加入10mL二氯甲烷,分别以水(2×10mL)、饱和食盐水(10mL)洗涤,无水硫酸钠干燥.过滤,减压蒸除溶剂,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得纯品107为淡黄色油状液体,收率86％；[α]D 20＝+30.0(c 0.34,CHCl3)；1H NMR(CDCl3,400MHz)δ:7.39(br s,2H),7.13～7.00(m,1H),6.53(s,2H),5.87(s,1H),5.33(s,1H),5.19(s,1H),3.91(s,3H),3.76(s,3H),3.73(s,6H)；13C NMR(CDCl3,150MHz)δ:159.9,153.1,151.0,148.6,138.3,134.4,132.7,129.0,127.2,120.8,113.6,110.8,94.2,62.0,60.3,55.8,55.4；MS(ESI)m/z(％):454.1(M+H)+；HRMS(ESI)calcd for C20H21FNO8S[M+H]+454.0966,found 454.0968.
实施例104 (3R,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-氟磺酰氧基-4-甲氧基苯基)-3-甲基氮杂环丁烷-2-酮(108)的合成
Figure PCTCN2017078444-appb-000158
在100mL茄形瓶中加入化合物107(18mg,0.04mmol),10％Pd/C(3mg),甲醇1 mL常压氢气下,室温反应12小时,过滤除去Pd/C后蒸干,柱层析分离(展开剂：PE/EA＝2:1),得到黄色固体(108)17mg,收率95％；[α]D 20＝-98.8(c 1.64,CHCl3)；1H NMR(400MHz,CDCl3)δ6.86–6.77(m,2H),6.72(d,J＝8.3Hz,1H),6.56(s,2H),5.71(s,1H),5.06(d,J＝5.8Hz,1H),3.89(s,3H),3.77(s,3H),3.72(s,6H),3.66-3.59(m,1H),0.91(d,J＝7.6Hz,3H).
实施例105 (3S,4R)-1-(3,4,5-三甲氧基苯基)-4-(3-氟磺酰氧基-4-甲氧基苯基)-3-甲基氮杂环丁烷-2-酮(109)的合成
Figure PCTCN2017078444-appb-000159
在100mL茄形瓶中加入化合物97(20mg,0.054mmol)和1.5mL无水二氯甲烷,0℃搅拌条件下,依次加入三乙胺(11mg,0.108mmol)和过量氟磺酰氟,后升至室温反应1h.停止反应,用1mL水淬灭,加入10mL二氯甲烷,分别以水(2×10mL)、饱和食盐水(10mL)洗涤,无水硫酸钠干燥.过滤,减压蒸除溶剂,拌硅胶柱层析分离纯化,收集对应的洗脱液,蒸干溶剂得无色油状液体(109)19mg，收率79％；[α]D 20＝+10.5(c 0.5,CHCl3)；1H NMR(CDCl3,400MHz)δ7.15(d,J＝1.8Hz,1H),7.03(dd,J＝8.1,1.8Hz,1H),6.93(d,J＝8.1Hz,1H),6.55(s,2H),4.48(d,J＝2.1Hz,1H),3.88(s,3H),3.79(s,3H),3.70(s,6H),3.12-3.04(m,1H),1.47(d,J＝7.2Hz,3H).13C NMR(150MHz,CDCl3)δ168.7,152.7,146.5,145.4,133.8,133.7,130.5,117.0,111.5,110.5,94.6,62.4,60.3,55.5,54.4,12.8.MS(ESI)m/z(％):456.1(M+H)+；HRMS(ESI)calcd for C20H23FNO8S[M+H]+456.1128,found 456.1125.
实施例106 目标化合物体外抑制人肿瘤细胞增殖活性检测实验
肿瘤细胞接种于96孔板中于37℃，5％CO2的环境下培养24小时，然后加入6个不同浓度的样品，并以阳性样品紫杉醇、CA-4为阳性对照。继续培养48小时后，加入MTT，4小时后弃去上清液，加入DMSO溶解紫色结晶，在酶标仪上540nm处测得OD值，并计算抑制率。化合物半数抑制浓度IC50数值，根据6个浓度的抑制率计算所得。
表1.二芳基-β-内酰胺类目标化合物抑制肿瘤细胞增殖活性(IC50,μM)
Figure PCTCN2017078444-appb-000160
Figure PCTCN2017078444-appb-000161
Figure PCTCN2017078444-appb-000162
Figure PCTCN2017078444-appb-000163
说明：抗肿瘤活性由MTT法测定，数据均是三次测量的平均值。A2780和SKOV-3是人卵巢癌细胞株；MDA-MB-231是人乳腺癌细胞株；Hela是人宫颈癌细胞株。
实施例107 抑制微管蛋白聚集实验：体外微管蛋白自组装实验
采用浊度法检测受试化合物69、70和97对体外微管聚集的抑制作用，检测试剂盒购自美国Cytoskeleton,Inc.。具体步骤如下，微管聚集体系内含0.1M PIPES，pH＝6.6，10mM MgCl2，1mM GTP，1mM EGTA，3.4M甘油。反应液先在冰上预孵育，加入不同浓度的受试化合物，并设置DMSO(4％，v/v)组作为阴性对照组，Colchicine处理组作为阳性对照。加入10mM微管蛋白后，立刻置于37℃进行聚集反应，保持37℃，并用分光光度计(Synergy H4Hybrid)在340nm下每隔1min测定吸光度，共测30min，根据吸光度绘制曲线图。结果显示化合物69、70和97能明显抑制微管聚集，IC50分别为3.5、1.7和1.6μM。
表2.化合物69、70和97的微管聚集抑制活性(IC50，μM)
Figure PCTCN2017078444-appb-000164
实施例108 抑制微管蛋白聚集实验：免疫印迹分析
Hela细胞用不同浓度的受试化合物处理6小时，并设置DMSO组作为阴性对照组，收集细胞并用PBS洗涤2次后用含有微管稳定剂的细胞裂解液消化(含有100mM PIPES,pH 6.8,1mM MgCl2,2mM EGTA,0.5％NP-40,2M glycerol,5μM紫杉醇和蛋白激酶抑制剂)。消化液用15000转/分离心15分钟。吸出上清液，并将沉淀溶解于SDS消化缓冲液。分别取等量上清和沉淀的蛋白样品经SDS-PAGE电泳后转移至PVDF膜。用含5％BSA的TBST缓冲液固定后，加入α-微管蛋白一抗，随后加入HRP标记的二抗进行孵育，最后通过化学发光试剂进行显色。结果显示化合物1、69、70和97能明显抑制微管聚集，使微管蛋白维持解聚状态。
实施例109 抑制微管蛋白聚集实验：免疫荧光检测微管蛋白形态实验
培养肿瘤细胞，将细胞接种到预先处理(2M NaOH浸泡2h，75％乙醇浸泡30min)过的盖玻片上，待细胞贴壁后，将盖玻片置于24孔板中，加入培养基，常规培养24h后，加入不同浓度的受试化合物处理24h，同时设置DMSO处理组作为阴性对照。弃去培养基，细胞用PBS洗涤2次，加入甲醇固定15min，PBS洗涤3次，用0.1％triton通透15min，PBS洗涤3次。用5％BSA室温封闭1h，加入一抗4℃孵育过夜，PBST漂洗3次，每次5min。加入荧光二抗，室温避光孵育1h，PBST漂洗3次，每次5min。滴加封片剂一滴(含DAPI)封片，用共聚焦显微镜观察微管蛋白的形态，检测受试化合物对微管结构的影响，并拍摄相关照片。结果显示化合物1、69、70和97能明显抑制微管的聚集。
实施例110 抑制血管生成实验
将细胞外基质胶MatriGel(BD Biosciences，美国)用PBS按1:1稀释混匀后，加入24孔板。置于37℃、5％CO2培养箱中孵育1小时，待凝胶形成后，将HUVEC细胞按浓度3×104个/孔接种到凝胶上，加入含10％PBS的DMEM培养基，并加入测试样品，同时设置CA-4为阳性对照，DMSO为阴性对照，于37℃、5％CO2培养箱中培养12h，用倒置相差显微镜下观察毛细血管形成情况，并拍摄相关照片。结果显示化合物69、70和97能明显抑制HUVEC细胞生成毛细血管样结构(图4)。
实施例111 基质胶塞实验
在含有100ng/mL人源重组VEGF-A165的基质胶中，在4℃加入测试化合物，同时设置DMSO处理组作为阴性对照。然后将此混合物按0.5mL/只，在6周大的Balb/C裸鼠(n＝4)的背部皮下注射以形成基质胶塞。2周后处死老鼠，回收基质胶塞，拍摄照片。结果显示化合物69、70和97能明显抑制由VEGF介导的新生血管生成(图5)。
实施例112 菌落抑制实验
6孔板中，每孔种1000个细胞，待细胞贴壁后，加入不同浓度的受试样品处理细胞48h，并设阳性对照组(CA-4)、阴性对照组(空白溶剂)，更换新鲜培养基继续培养7-10天后，弃去培养基，细胞用甲醇固定，并用Giemsa或结晶紫染料染色，显微镜下计数大于50个细胞形成的集落数。结果显示化合物69、70和97能明显抑制肿瘤细胞菌落的形成(图6)。
实施例113 体外细胞周期实验
6孔板中，按2×105个/孔的数量接种细胞，待细胞贴壁后，用不同浓度的受试样品处理细胞24h，同时设阳性对照组(CA-4)、阴性对照组(空白溶剂)。收集细胞，用PBS冲洗两次，再用75％乙醇-20℃固定过夜后，用PI染色后采用流式细胞仪进行测试。结果如图7所示，化合物69、70和97能明显将细胞阻滞于G2/M期。
实施例114 体外细胞周期相关蛋白检测实验
A.Western blot实验：选择不同浓度化合物处理肿瘤细胞后(同时设计DMSO为阴性对照)，收集并用裂解液裂解细胞。蛋白样品经加热变性后，电泳分离，转膜，封闭， 依次经一抗反应、二抗反应后，曝光显色。
B.荧光定量PCR实验：用不同浓度受试样品处理细胞后，使用定量PCR试剂盒(Takara)以及荧光定量PCR仪，检测细胞内相关蛋白mRNA水平。
结果显示化合物69、70和97能明显促进磷酸化组蛋白H3、细胞周期蛋白B1、有丝分裂检验点蛋白BuBR1表达(图8)。
实施例115 体外细胞凋亡实验
6孔板中，按2×105个/孔的数量接种细胞，待细胞贴壁后，用不同浓度的受试样品处理细胞24h，同时设阳性对照组(CA-4)、阴性对照组(空白溶剂)。收集细胞，用PBS冲洗两次，再用75％乙醇-20℃固定过夜后，用PI和Annexin V双染色，细胞染色后采用流式细胞仪进行测试。结果显示化合物69、70和97能明显促进细胞凋亡(图9)。
实施例116 体外凋亡相关蛋白检测实验
A.Western blot实验：选择不同浓度化合物处理肿瘤细胞后(同时设计DMSO为阴性对照)，收集并用裂解液裂解细胞。蛋白样品经加热变性后，电泳分离，转膜，封闭，依次经一抗反应、二抗反应后，曝光显色。
B.荧光定量PCR实验：用不同浓度受试样品处理细胞后，使用定量PCR试剂盒(Takara)以及荧光定量PCR仪，检测细胞内相关蛋白mRNA水平。
结果显示化合物69、70和97能明显促进促凋亡蛋白Bax、抑癌基因p53、剪切的DNA修复酶的表达(图10)。
实施例117 急性毒性测试实验
ICR小鼠单独饲养于无病原笼子内。每组小鼠(10只，5雌5雄)腹腔注射待测药物69(95,70,50,35和25mg/kg)、70(500,425,350,275和200mg/kg)或97(275,200,150,125和100mg/kg)，或安慰剂(8.3％蓖麻油与8.3％乙醇的PBS溶液)。70、69溶解于蓖麻油乙醇(1:1,v/v)中，然后用PBS(1:5,v/v)稀释。每天记录小鼠死亡情况，持续14天。结果显示化合物69的LD50为61.5mg/kg，化合物70的LD50大于500mg/kg,化合物97的LD50为136.5mg/kg。
表3.化合物69、70和97对小鼠的急性毒性
Figure PCTCN2017078444-appb-000165
Figure PCTCN2017078444-appb-000166
69:LD50＝61.5mg/kg；70:LD50>500mg/kg；97:LD50＝136.5mg/kg.
实施例118 动物水平和组织水平的肿瘤治疗作用和机理研究
培养卵巢癌细胞系A2780，当细胞处于生长旺盛期进行接种，按每只2×106个细胞腹腔注射接种于6周龄Balb/C裸鼠，建立裸鼠肿瘤转移模型，SPF条件下生长。待裸鼠皮下移植瘤长至体积约100mm3，将已经建成的荷瘤小鼠随机分成4组，每组10只，经腹腔注射，分别给予不同浓度的7.5mg/kg的69、12.5mg/kg的70、4mg/kg和8mg/kg的97、5mg/kg或10mg/kg紫杉醇和空白对照(与急性毒性测试相同)，记录瘤生长大小。肿瘤体积按以下公式计算：肿瘤体积(mm3)＝a×b2×0.52(这里a代表最长的直径，b代表最短的直径，0.52是经验系数)。当空白对照组肿瘤体积平均体积达到2000mm3后，处死裸鼠，剥离瘤组织，称取瘤质量，计算抑瘤率：
抑瘤率＝(1-实验组平均瘤质量/对照组平均瘤质量)×100％
裸鼠内脏组织用H&E染色进行进一步观察。
结果显示化合物70、69和97在体内能明显抑制肿瘤生长，并且对小鼠体重无明显影响。组织染色也显示出化合物70、69和97能够使肿瘤组织产生坏死区(图中箭头标示)，并且对肝、肾、脾的组织染色并未观察到非正常区域。
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 (10)

  1. 一种具有通式Ⅰ结构的二芳基-β-内酰胺类化合物，及其药学上可接受的盐，水合物，溶剂合物或前药：
    Figure PCTCN2017078444-appb-100001
    其特征在于，
    R1为一个或多个位于环上的选自下组的基团：取代或未取代的C1-C4烷氧基、C1-C4烷基、卤素、氨基、羟基、羧基、取代或未取代的C2-C10酰氧基、C2-C10酯基、甲氧甲酰基、烯丙氧基、炔丙氧基、磺酰氧基、烷氨基、酰氨基、磺酰氨基或者2-3个上述相同或不同基团的组合；
    R2为一个或多个位于环上的选自下组的基团：取代或未取代的C1-C6烷氧基、取代或未取代的C1-C6烷基、卤素、氨基、羟基、羧基、氟磺酰氧基、烯丙氧基、炔丙氧基、C1-C4烷胺基、C2-C10酯基、取代或未取代的C1-C6烷基-羟基、取代或未取代的C6-C10的芳基、取代或未取代的5-12元杂芳基、-OTBS、-CH2-R、-OR、-O(C＝O)R、-O-(SO2)-R、-O(PO)-R2、-NH(C＝O)R、-NH-(SO2)-R；
    R3和R4各自独立地选自下组：取代或未取代的C1-C6烷基、氢原子、酰氧基、羟基、羧基、环丙基、氨基、取代或未取代的C1-C4烷胺基、磺酰氧基、取代或未取代的C1-C4烷氧基、取代或未取代的C1-C6烷基-羟基、取代或未取代的芳基、取代或未取代的吗啉基、-CH2-R、-OR、-O(C＝O)R、-O-(SO2)-R、-O(PO)-R2、-NH(C＝O)R、-NH-(SO2)-R；
    或者R3和R4共同构成＝CHR、-OC(＝O)OCH2-、＝O、C3-C6环烷基、C3-C6杂环基、或取代或未取代的-(CH2)n-，其中n选自下组：1、2、3、4、5或6；
    R5和R6各自独立地为H；或者R5和R6共同构成＝CHR、-OC(＝O)OCH2-、＝O、＝S；
    其中，所述的R选自下组：乙烯基、卤素、氨基、羟基、羧基、氟磺酰氧基、甲基磺酰基(Ms)、取代或未取代的C1-C4烷氧基、取代或未取代的C1-C6烷基、取代或未取代的C3-C6环烷基、取代或未取代的C1-C4烷胺基、取代或未取代的C2-C10酯基、取代或未取代的C1-C6烷基-羟基、取代或未取代的C6-C10的芳基、取代或未取代的5-12元杂芳基；
    所述的取代指基团上的一个或多个氢原子被选自下组的取代基取代：C1-C4烷氧基、C1-C4烷基、卤素、C2-C10酰氧基、C2-C10酯基、羟基、环丙基、乙烯基、氨基、氧基(＝O)、吗啉基、磺酰氧基、C1-C6酰胺基、-NO2、-NHBoc、-NHCbz、-NHC(＝O)Me、-OBn、-NHBn、-SiMe3、未取代或被1-3个选自下组的取代基取代的苯基或吡啶基：C1-C4烷氧基、C1-C4烷基、卤素、羟基。
    在另一优选例中，所述的取代苯基是指苯环被1-5个选自下组的取代基取代：硝基、氟原子或甲氧基。
  2. 如权利要求1所述的化合物，其特征在于，所述的化合物具有如下式I-1所示的结构：
    Figure PCTCN2017078444-appb-100002
    其中，各基团的定义如权利要求1中所述。
  3. 如权利要求1所述的化合物，其特征在于，所述的化合物具有如下式I-2所示的结构：
    Figure PCTCN2017078444-appb-100003
    其中，各基团的定义如权利要求1中所述。
    在另一优选例中，所述的化合物具有如下式I-3、I-4、I-5、I-6或I-7所示的结构：
    Figure PCTCN2017078444-appb-100004
    其中，各基团的定义如权利要求1中所述。
  4. 如下式所示的化合物：
    Figure PCTCN2017078444-appb-100005
  5. 如权利要求1所述的化合物，其特征在于，所述的化合物选自下组：
    Figure PCTCN2017078444-appb-100006
    Figure PCTCN2017078444-appb-100007
    Figure PCTCN2017078444-appb-100008
    Figure PCTCN2017078444-appb-100009
    Figure PCTCN2017078444-appb-100010
  6. 如权利要求1所述的药物组合物的用途，其特征在于，用于制备治疗或预防选自下组的疾病的药物组合物：与微管相关蛋白聚集有关的哺乳动物疾病、与血管生成相关的哺乳动物疾病。
  7. 如权利要求6所述的用途，其特征在于，所述的与微管相关蛋白聚集有关的哺乳动物疾病为肿瘤。
  8. 如权利要求6所述的用途，其特征在于，所述的肿瘤选自下组：甲状腺癌、头颈部鳞状细胞癌、宫颈癌、卵巢癌、乳腺癌、结直肠癌、胰腺癌、食管癌、骨肉瘤、肾癌、胃癌、肺癌、肝癌、黑色素瘤、淋巴瘤、前列腺癌、肾癌、膀胱癌、脑胶质瘤、鼻咽癌，神经内分泌癌、未分化癌、间质肉瘤、绒癌、恶性葡萄胎、恶性畸胎瘤、良性肿瘤。
  9. 一种药物组合物，其特征在于，所述的药物组合物包括：(i)有效量的式I化合物，及其药学上可接受的盐，水合物，溶剂合物或前药；和(ii)药学上可接受的载体。
  10. 一种式I化合物的制备方法，其特征在于，所述方法包括步骤：
    Figure PCTCN2017078444-appb-100011
    在惰性溶剂中，用式If化合物进行反应，得到式Ig化合物；
    和任选的步骤：
    Figure PCTCN2017078444-appb-100012
    用所述的式Ig化合物制备得到式I化合物。
PCT/CN2017/078444 2016-03-29 2017-03-28 二芳基-β-内酰胺类化合物及其制备方法和在制药中的用途 WO2017167183A1 (zh)

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