CN115850277B - 骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物及其制备方法及应用 - Google Patents
骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物及其制备方法及应用 Download PDFInfo
- Publication number
- CN115850277B CN115850277B CN202211701584.6A CN202211701584A CN115850277B CN 115850277 B CN115850277 B CN 115850277B CN 202211701584 A CN202211701584 A CN 202211701584A CN 115850277 B CN115850277 B CN 115850277B
- Authority
- CN
- China
- Prior art keywords
- skeleton
- compound
- cells
- nmr
- spiro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 235000005126 Peganum harmala Nutrition 0.000 title claims abstract description 25
- 240000005523 Peganum harmala Species 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims description 5
- 150000001875 compounds Chemical class 0.000 claims abstract description 78
- 239000003814 drug Substances 0.000 claims abstract description 14
- 238000006845 Michael addition reaction Methods 0.000 claims abstract description 4
- 238000007363 ring formation reaction Methods 0.000 claims abstract description 4
- 206010028980 Neoplasm Diseases 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 5
- 229940125904 compound 1 Drugs 0.000 claims description 2
- 229940125782 compound 2 Drugs 0.000 claims description 2
- 241000159204 Peganum Species 0.000 claims 2
- 239000012467 final product Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 13
- 229940079593 drug Drugs 0.000 abstract description 10
- 238000012216 screening Methods 0.000 abstract description 7
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 abstract description 6
- 206010060862 Prostate cancer Diseases 0.000 abstract description 6
- 208000032839 leukemia Diseases 0.000 abstract description 6
- 201000005249 lung adenocarcinoma Diseases 0.000 abstract description 6
- 208000000236 Prostatic Neoplasms Diseases 0.000 abstract description 5
- 230000005764 inhibitory process Effects 0.000 abstract description 4
- 239000003960 organic solvent Substances 0.000 abstract description 4
- 230000004071 biological effect Effects 0.000 abstract description 3
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 229930014626 natural product Natural products 0.000 abstract description 3
- 125000001424 substituent group Chemical group 0.000 abstract description 3
- 230000004614 tumor growth Effects 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 2
- 238000003786 synthesis reaction Methods 0.000 abstract description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 93
- 210000004027 cell Anatomy 0.000 description 64
- 238000004896 high resolution mass spectrometry Methods 0.000 description 32
- 238000002844 melting Methods 0.000 description 32
- 230000008018 melting Effects 0.000 description 32
- 239000007787 solid Substances 0.000 description 32
- 230000006907 apoptotic process Effects 0.000 description 9
- 230000000144 pharmacologic effect Effects 0.000 description 8
- 102000003952 Caspase 3 Human genes 0.000 description 7
- 108090000397 Caspase 3 Proteins 0.000 description 7
- 101710179684 Poly [ADP-ribose] polymerase Proteins 0.000 description 6
- 102100023712 Poly [ADP-ribose] polymerase 1 Human genes 0.000 description 6
- 229920000776 Poly(Adenosine diphosphate-ribose) polymerase Polymers 0.000 description 6
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- RERZNCLIYCABFS-UHFFFAOYSA-N harmaline Chemical compound C1CN=C(C)C2=C1C1=CC=C(OC)C=C1N2 RERZNCLIYCABFS-UHFFFAOYSA-N 0.000 description 6
- 230000005012 migration Effects 0.000 description 6
- 238000013508 migration Methods 0.000 description 6
- 238000011282 treatment Methods 0.000 description 6
- 210000004881 tumor cell Anatomy 0.000 description 6
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 210000000170 cell membrane Anatomy 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 229910052801 chlorine Inorganic materials 0.000 description 4
- 239000000460 chlorine Substances 0.000 description 4
- 230000005757 colony formation Effects 0.000 description 4
- 210000001700 mitochondrial membrane Anatomy 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 3
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 3
- 101000721661 Homo sapiens Cellular tumor antigen p53 Proteins 0.000 description 3
- 101000911513 Homo sapiens Uncharacterized protein FAM215A Proteins 0.000 description 3
- 206010027476 Metastases Diseases 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 102100026728 Uncharacterized protein FAM215A Human genes 0.000 description 3
- 230000001640 apoptogenic effect Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000013078 crystal Substances 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 231100000263 cytotoxicity test Toxicity 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000009401 metastasis Effects 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- INAAIJLSXJJHOZ-UHFFFAOYSA-N pibenzimol Chemical compound C1CN(C)CCN1C1=CC=C(N=C(N2)C=3C=C4NC(=NC4=CC=3)C=3C=CC(O)=CC=3)C2=C1 INAAIJLSXJJHOZ-UHFFFAOYSA-N 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 102000010565 Apoptosis Regulatory Proteins Human genes 0.000 description 2
- 108010063104 Apoptosis Regulatory Proteins Proteins 0.000 description 2
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 2
- BXNJHAXVSOCGBA-UHFFFAOYSA-N Harmine Chemical group N1=CC=C2C3=CC=C(OC)C=C3NC2=C1C BXNJHAXVSOCGBA-UHFFFAOYSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 2
- 229910052794 bromium Inorganic materials 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 230000012292 cell migration Effects 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 2
- 229960004316 cisplatin Drugs 0.000 description 2
- 125000001559 cyclopropyl group Chemical group [H]C1([H])C([H])([H])C1([H])* 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000009510 drug design Methods 0.000 description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 2
- 229910052731 fluorine Inorganic materials 0.000 description 2
- 239000011737 fluorine Substances 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 2
- 230000003389 potentiating effect Effects 0.000 description 2
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000012192 staining solution Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229910052717 sulfur Inorganic materials 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 101100329869 Nitrosomonas europaea (strain ATCC 19718 / CIP 103999 / KCTC 2705 / NBRC 14298) cyt gene Proteins 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- DPKHZNPWBDQZCN-UHFFFAOYSA-N acridine orange free base Chemical compound C1=CC(N(C)C)=CC2=NC3=CC(N(C)C)=CC=C3C=C21 DPKHZNPWBDQZCN-UHFFFAOYSA-N 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 238000011394 anticancer treatment Methods 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000009925 apoptotic mechanism Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 102000055102 bcl-2-Associated X Human genes 0.000 description 1
- 108700000707 bcl-2-Associated X Proteins 0.000 description 1
- DZBUGLKDJFMEHC-UHFFFAOYSA-N benzoquinolinylidene Natural products C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004611 cancer cell death Effects 0.000 description 1
- 238000010370 cell cloning Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000003436 cytoskeletal effect Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000012137 double-staining Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 229960005542 ethidium bromide Drugs 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000000861 pro-apoptotic effect Effects 0.000 description 1
- 201000001514 prostate carcinoma Diseases 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000003642 reactive oxygen metabolite Substances 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000025915 regulation of apoptotic process Effects 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 210000004926 tubular epithelial cell Anatomy 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Landscapes
- Nitrogen Condensed Heterocyclic Rings (AREA)
Abstract
本发明公开了一种骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物，具体以各种取代的骆驼蓬灵骨架化合物1和与α‑异硫氰酸酯羰基化合物2发生Michael加成环化反应，获得骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物3，该骨架化合物是第一例天然产物骆驼蓬碱骨架拼接的多螺环化合物,可以为生物活性筛选提供化合物源，对药物的筛选和制药行业具有重要的应用价值。本发明操作简单易行，原料合成便宜易得，可以在各种有机溶剂中进行，也具有较好的空气稳定性，适用性广，对于各种取代基都有很好的兼容性。且该类骨架化合物对人白血病细胞(K562)、人肺腺癌细胞(A549)、人前列腺癌细胞(PC‑3)具有肿瘤生长抑制活性作用。
Description
技术领域
本发明涉及化学技术和药学技术领域，尤其是一种骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物及其制备方法及应用。
背景技术
根据药物设计的活性骨架拼接和迁越原理，把两个或多个具有生物活性骨架拼接成一个潜在生物活性的多骨架分子在有机化学和医药化学中是极其重要的研究领域。骆驼蓬灵，英文名称：Harmaline，是从天然植物骆驼蓬中分离出来的一种天然植物生物碱。主要用于抑制革兰氏阳性菌和真菌活性。这些化合物在解除病痛、经济发展中起着重大作用。因此，利用药物设计中优势骨架重组策略，合成一系列新的潜在活性的骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物具有重要的研究意义。特别说明的是:所合成的骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物3，该骨架化合物是第一例天然产物骆驼蓬碱骨架拼接的多螺环化合物,可以为生物活性筛选提供化合物源，对药物的筛选和制药行业具有重要的应用价值(如图6所示)。
发明内容
本发明的目的是：提供一种骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物及其制备方法与应用，它是一类重要的医药中间体类似物和药物分子类似物，对药物筛选和制药行业具有重要的应用价值，且其合成方法非常经济简便。
本发明还发现该类化合物在制备防治肿瘤疾病药物中的应用。
本发明是这样实现的：一种骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物，该化合物具有如下通式(Ⅰ)的结构：
式中，R1为甲氧基或氯或氢；R2为氟、氯、溴、乙基、甲基或环丙基；X为C、O或S；
具体为如下结构式之一:
骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物的制备方法，在有机溶剂中，在碱性催化剂三乙胺的作用下，将各种取代的骆驼蓬灵骨架化合物1和与α-异硫氰酸酯羰基化合物2发生Michael加成环化反应，获得骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物3。
合成路线举例如下：
其中合成路线中的化合物，其取代基满足式中，R1为甲氧基或氯或氢；R2为氟、氯、溴、乙基、甲基或环丙基；X为C、O或S。
反应机理如下：
骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物在制备防治肿瘤疾病药物中的应用。
通过采用上述技术方案，在有机溶剂中，在碱性催化剂三乙胺的作用下，以各种取代的骆驼蓬灵骨架化合物1和与α-异硫氰酸酯羰基化合物2发生Michael加成环化反应，获得骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物3，该骨架化合物是第一例天然产物骆驼蓬碱骨架拼接的多螺环化合物,可以为生物活性筛选提供化合物源，对药物的筛选和制药行业具有重要的应用价值。且该骨架化合物对人白血病细胞(K562)、人肺腺癌细胞(A549)、人前列腺癌细胞(PC-3)具有抑制活性的作用。本发明操作简单易行，原料合成便宜易得，可以在各种有机溶剂中进行，也具有较好的空气稳定性，适用性广，对于各种取代基都有很好的兼容性。
附图说明
图1及图2为本发明的实施例的化合物3aa谱图数据；
图3及图4为本发明的实施例的化合物3fa谱图数据；
图5为本发明的实施例的化合物3ba和3de单晶图；
图6为本发明所合成的化合物的设计思路；
图7为本发明3cb诱导A549细胞凋亡的变化图；
图7中，3cb诱导A549细胞凋亡的变化。将A549细胞以每孔1×105个细胞的密度接种在六孔板中，并用3cb处理48小时以观察凋亡的变化。(A)A549细胞形态图。(B)A549细胞的AO/EB双染色谱。(C)A549细胞的Hoechst 33258染色谱；
图8为本发明3cb对A549细胞ROS和MMP变化的影响图；
图8中，A549细胞以每孔1×105个细胞的密度接种在六孔板中，并用3cb处理48小时(A)表明3cb诱导A549细胞中活性氧产生的变化。(B)3cb诱导线粒体膜电位的变化；
图9为本发明3cb对A549细胞集落形成的影响图；
图9中，A549细胞以每孔200个细胞的密度接种，并响应3cb培养14天。(A)结晶紫染色的A549细胞集落的代表图。(B)A549细胞的单克隆形成率(％)。(与对照组相比，*p<0.05，*p<0.01)。
图10为本发明3cb对A549细胞迁移的影响图；
图10中，(A)A549细胞以每孔1×104个细胞的密度接种，直至细胞融合生长，并在用移液管刮擦后在3cb培养24小时；0小时和24小时的细胞划痕表示图。(B)A549细胞的迁移率(％)。(C)以每孔3×104个细胞的密度接种的A549细胞的Traswell迁移曲线，Traswell插入物培养24小时以响应3cb。(与对照组相比，*p<0.05，*p<0.01)。
图11为本发明化合物3cb对A549细胞中凋亡相关蛋白表达的影响图；
图11中，(A)在用指定浓度的化合物3cb处理A549细胞48小时后，p53、Bax、Bcl-2、Cyt c、前半胱天冬酶3、切割的半胱天冬蛋白酶3、PARP、切割的PARP的代表性Western印迹。(B-H)受化合物3cb影响的A549细胞中这些促凋亡和抗凋亡蛋白的定量。进行了三个独立的实验。所有数据均以平均值±SEM表示(与对照组相比，*p<0.05，*p<0.01)。
具体实施方式
本发明的实施例：在反应管中依次加入骆驼蓬灵骨架化合物1(0.2mmol)，α-异硫氰酸酯羰基化合物2(0.3mmol)，Et3N(10mol％)和3.0mL甲苯，室温中搅拌反应1h,TLC检测基本反应完全,上样经柱层析[洗脱剂：V(石油醚)：V(乙酸乙酯)＝3:1]纯化得化合物3aa:黄色固体,熔点：159.9-160.4℃；产率87％,15:1dr。
核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.36(s,3H),2.35-2.43(m,1H),2.47-2.50(m,1H),2.53(s,3H),2.93-3.00(m,1H),3.46(s,3H),4.47-4.51(m,1H),6.37-3.40(m,1H),6.46(d,J＝2.0Hz,1H),6.82(d,J＝7.6Hz,1H),6.90-6.94(m,1H),7.04(d,J＝8.8Hz,1H),7.21-7.25(m,1H),7.40(d,J＝7.2Hz,1H),8.45(br s,1H),9.95(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.3,23.4,26.3,55.6,66.4,72.1,95.3,109.0,109.2,109.8,119.2,120.8,122.4,124.3,127.3,130.8,131.5,137.9,145.3,156.0,174.2,181.5；HRMS(ESI-TOF)m/z:Calcd.for C23H22N4NaO2S[M+Na]+:441.1356；Found:441.1353.
表1为一种骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物的化学结构
表2为一种骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物的化学结构
化合物3ab至3ka的制备方法同化合物3aa，投料比与化合物3aa相同，可得到化合物3ab至3ka，反应产率见表1和表2，但需强调的是本发明的化合物不限于表1和表2所表示的内容。
本实施例制备化合物3ab:黄色固体,熔点：144.5-145.4℃；产率79％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.76(s,3H),2.75-2.87(m,2H),3.26-3.33(m,1H),3.68(s,3H),4.72-4.77(m,2H),5.22(d,J＝16.0Hz,1H),6.53-6.64(m,4H),6.70(d,J＝8.0Hz,1H),6.96-7.01(m,1H),7.25-7.30(m,2H),7.32-7.36(m,2H),7.43(d,J＝7.2Hz,2H),9.30(br s,1H),10.27(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.1,21.9,44.1,55.5,68.5,70.5,94.8,108.6,108.8,109.7,119.3,120.5,124.2,127.8,127.9,128.1,128.9,129.9,131.9,136.3,137.4,142.0,156.0,173.0,180.8；HRMS(ESI-TOF)m/z:Calcd.for C29H26N4NaO2S[M+Na]+:517.1669；Found:517.1673.
本实施例制备化合物3ba:黄色固体,熔点：156.5-157.3℃；产率81％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.67(s,3H),2.69-2.76(m,1H),2.80(s,3H),2.83-2.85(m,1H),3.25-3.32(m,1H),4.79-4.83(m,1H),6.99-7.12(m,3H),7.19-7.26(m,2H),7.46(d,J＝7.6Hz,1H),7.50-7.54(m,1H),7.70(d,J＝7.2Hz,1H),8.77(br s,1H),10.44(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.3,23.4,26.3,66.4,72.1,109.3,109.8,111.9,118.6,122.5,124.4,126.4,127.4,130.8,133.0,137.1,145.4,174.2,181.5；HRMS(ESI-TOF)m/z:Calcd.for C22H20N4NaOS[M+Na]+:411.1250；Found:411.1253.
本实施例制备化合物3bb:黄色固体,熔点：147.2-148.1℃；产率79％,2:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(CDCl3,400MHz)δ:1.69(s,3H),2.72-2.77(m,1H),2.87-2.95(m,1H),3.20-3.28(m,1H),4.55(d,J＝14.8Hz,1H),4.82-4.87(m,1H),5.23(d,J＝14.8Hz,1H),6.39(s,1H),6.50(s,1H),.53-6.57(m,1H),6.64-6.66(m,1H),6.72-6.77(m,2H),6.91-6.97(m,3H),7.27-7.29(m,1H),7.35-7.44(m,5H)；13C NMR(CDCl3,100MHz)δ:20.8,22.2,40.0,44.3,69.5,70.9,108.9,110.4,110.7,118.6,119.8,122.5,123.2,124.7,125.9,126.5,128.1,128.6,129.5,130.5,131.4,135.9,136.3,141.0,172.9,181.8；HRMS(ESI-TOF)m/z:Calcd.for C28H24N4NaOS[M+Na]+:487.1563；Found:487.1563.
本实施例制备化合物3bc:黄色固体,熔点：185.3-186.2℃；产率76％,4:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.71(s,3H),2.67-2.75(m,1H),2.78(s,3H),2.81-2.86(m,1H),3.25-3.32(m,1H),4.77-4.82(m,1H),6.99-7.14(m,3H),7.24(d,J＝8.0Hz,1H),7.36-7.41(m,1H),7.46(d,J＝7.6Hz,1H),7.68-7.71(m,1H),8.77(br s,1H),10.48(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.3,23.0,26.5,66.5,72.3,109.4,110.6(d,JCF＝8.2Hz),111.8,115.3(d,JCF＝26.1Hz),117.1(d,JCF＝23.1Hz),118.7,119.2,121.8,125.9(d,JCF＝8.1Hz),126.3,132.7,137.0,141.6,158.8(d,JCF＝235.4Hz),174.1,181.6；HRMS(ESI-TOF)m/z:Calcd.for C22H19FN4NaOS[M+Na]+:429.1156；Found:429.1155.
本实施例制备化合物3bd:黄色固体,熔点：152.6-152.9℃；产率80％,2:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.73(s,3H),2.81-2.83(m,2H),3.28-3.30(m,1H),4.73(d,J＝14.4Hz,1H),6.41(d,J＝7.2Hz,1H),6.54-6.59(m,1H),6.89-6.98(m,3H),7.07(d,J＝8.0Hz,1H),7.37(d,J＝7.6Hz,1H),8.95(br s,1H),10.52(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.0,22.1,29.7,29.8,68.5,70.3,108.7,111.5,117.7(d,JCF＝20.2Hz),118.7,119.0,120.1,121.9,123.4,126.0,129.5,131.3,133.0,136.6,146.9(d,JCF＝240.3Hz),172.4,180.3；HRMS(ESI-TOF)m/z:Calcd.forC22H19FN4NaOS[M+Na]+:429.1156；Found:429.1158.
本实施例制备化合物3be:黄色固体,熔点：198.7-199.5℃；产率81％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.63(s,3H),2.38(s,3H),2.62-2.70(m,1H),2.72(s,3H),2.76-2.80(m,1H),3.20-3.27(m,1H),4.72-4.76(m,1H),6.94-7.03(m,3H),7.18(d,J＝8.0Hz,1H),7.28(d,J＝8.0Hz,1H),7.41(d,J＝7.6Hz,1H),7.51(s,1H),8.73(br s,1H),10.39(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:14.6,21.2,23.3,26.3,60.2,66.3,72.1,109.3,109.5,111.9,118.6,119.1,121.7,124.3,126.3,128.0,131.0,131.4,133.0,137.0,143.0,174.1,181.5；HRMS(ESI-TOF)m/z:Calcd.forC23H22N4NaOS[M+Na]+:425.1407；Found:425.1412.
本实施例制备化合物3bf:黄色固体,熔点：146.9-147.5℃；产率82％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(CDCl3,400MHz)δ:1.28-1.32(m,3H),1.71(s,3H),2.73-2.78(m,1H),2.94-3.02(m,1H),3.22-3.29(m,1H),3.89-4.00(m,2H),4.81-4.85(m,1H),6.51-6.59(m,3H),6.79-6.84(m,1H),6.97-7.02(m,3H),7.07(s,1H),7.35(d,J＝7.2Hz,1H)；13C NMR(CDCl3,100MHz)δ:14.1,20.7,22.3,37.8,40.3,69.6,70.7,110.9,118.6(d,JCF＝19.2Hz),120.0,121.1,122.8,123.8(d,JCF＝6.4Hz),126.1,131.5,136.2,146.9(d,JCF＝244.3Hz),172.7,181.8；HRMS(ESI-TOF)m/z:Calcd.for C23H21FN4NaOS[M+Na]+:443.1312；Found:443.1315.
本实施例制备化合物3bg:黄色固体,熔点：178.7-179.3℃；产率78％,4:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:0.61-0.65(m,3H),1.64(s,3H),2.38(s,3H),2.61-2.69(m,1H),2.76-2.80(m,1H),3.10-3.15(m,1H),3.19-3.26(m,1H),3.41-3.43(m,1H),4.72-4.77(m,1H),6.93-7.03(m,3H),7.14(d,J＝7.6Hz,1H),7.27(d,J＝8.0Hz,1H),7.40(d,J＝8.0Hz,1H),7.51(s,1H),8.71(br s,1H),10.38(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:12.4,21.2,21.4,23.0,33.9,66.2,72.4,109.3,109.4,111.9,118.6,119.0,121.7,124.1,126.2,128.2,131.1,131.2,132.7,136.8,142.0,173.5,181.7；HRMS(ESI-TOF)m/z:Calcd.for C24H24N4NaOS[M+Na]+:439.1563；Found:439.1560.
本实施例制备化合物3bh:黄色固体,熔点：157.5-157.9℃；产率81％,5:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.77(s,3H),1.81(s,3H),2.79-2.95(m,2H),3.28-3.33(m,1H),4.72-4.77(m,2H),5.20(d,J＝12.4Hz,1H),6.36(s,1H),6.58(d,J＝8.0Hz,1H),6.78(d,J＝8.0Hz,1H),6.90-7.00(m,2H),7.06(d,J＝8.0Hz,1H),7.25-7.35(m,3H),7.40-7.43(m,3H),9.31(br s,1H),10.39(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:20.8,21.0,21.9,44.0,68.7,70.7,108.9,109.4,111.4,118.6,119.0,121.8,125.1,126.1,127.7,127.8,128.9,130.0,131.1,133.2,136.3,136.5,139.9,173.0,181.3；HRMS(ESI-TOF)m/z:Calcd.for C29H26N4NaOS[M+Na]+:501.1720；Found:501.1723.
本实施例制备化合物3ca:黄色固体,熔点：158.9-159.5℃；产率85％,5:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.68(s,3H),2.43(s,3H),2.66-2.73(m,1H),2.77(s,3H),2.81-2.86(m,1H),3.24-3.31(m,1H),4.77-4.82(m,1H),7.00-7.08(m,2H),7.25(d,J＝8.8Hz,1H),7.35(d,J＝8.0Hz,1H),7.52-7.56(m,2H),8.79(br s,1H),10.65(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.2,23.2,26.3,55.4,66.2,72.1,109.4,109.6,113.4,118.0,121.6,123.8,124.1,127.5,128.0,131.1,131.5,134.8,135.5,143.0,174.1,181.7；HRMS(ESI-TOF)m/z:Calcd.for C23H21ClN4NaOS[M+Na]+:459.1017；Found:459.1014.
本实施例制备化合物3cb:黄色固体,熔点：147.6-148.5℃；产率75％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.78(s,3H),2.83-2.85(m,2H),3.29-3.33(m,1H),4.72-4.76(m,2H),5.23(d,J＝16.0Hz,1H),6.54-6.59(m,2H),6.70(d,J＝8.0Hz,1H),6.94-6.99(m,2H),7.05(d,J＝8.8Hz,1H),7.27-7.35(m,3H),7.42-7.45(m,3H),9.36(br s,1H),10.70(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.0,21.8,44.1,68.4,70.4,108.8,109.8,113.0,118.1,121.7,122.4,123.7,124.1,127.2,127.8,127.9,128.9,130.0,135.0,135.1,136.2,142.0,172.8,180.9；HRMS(ESI-TOF)m/z:Calcd.forC28H23ClN4NaOS[M+Na]+:521.1173；Found:521.1172.
本实施例制备化合物3cc:黄色固体,熔点：162.1-162.7℃；产率77％,4:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.76(s,3H),1.82(s,3H),2.80-2.87(m,2H),3.28-3.34(m,1H),4.70-4.75(m,2H),5.18(d,J＝16.4Hz,1H),6.33(s,1H),6.59(d,J＝8.0Hz,1H),6.79(d,J＝7.6Hz,1H),6.95-6.98(m,1H),7.06(d,J＝8.8Hz,1H),7.27-7.36(m,3H),7.38-7.44(m,2H),7.46(d,J＝2.0Hz,1H),9.34(br s,1H),10.62(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:20.7,21.8,44.0,68.6,70.7,109.0,109.5,112.9,118.0,121.8,123.6,125.0,127.2,127.6,127.8,128.9,130.1,131.2,134.9,135.1,136.3,139.8,172.9,181.3；HRMS(ESI-TOF)m/z:Calcd.for C29H25ClN4NaOS[M+Na]+:535.1330；Found:535.1326.
本实施例制备化合物3cd:黄色固体,熔点：165.5-166.4℃；产率84％,2:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:0.66-0.70(m,3H),1.69(s,3H),2.43(s,3H),2.64-2.72(m,1H),2.81-2.85(m,1H),3.16-3.30(m,2H),3.42-3.47(m,1H),4.77-4.82(m,1H),7.01-7.08(m,2H),7.20(d,J＝8.8Hz,1H),7.31-7.33(m,1H),7.52-7.56(m,2H),8.78(br s,1H),10.65(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:12.4,21.2,21.3,22.9,34.0,66.0,72.3,109.5,113.4,118.0,121.6,123.8,124.0,127.4,128.2,131.2,131.3,134.6,135.3,142.0,173.5,181.8；HRMS(ESI-TOF)m/z:Calcd.forC24H23ClN4NaOS[M+Na]+:473.1173；Found:473.1176.
本实施例制备化合物3ce:黄色固体,熔点：155.4-155.7℃；产率74％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.23-1.26(m,3H),1.73(s,3H),2.77-2.87(m,2H),3.26-3.34(m,1H),3.70-3.79(m,1H),3.97-4.06(m,1H),4.69-4.73(m,1H),6.34-6.37(m,1H),6.58-6.63(m,1H),6.95-7.09(m,3H),7.45(s,1H),9.11(br s,1H),10.60(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:14.0,20.8,22.0,37.5,55.4,68.5,70.4,109.0,113.0,117.9(d,JCF＝20.2Hz),118.1,120.5,121.9,123.4(d,JCF＝8.4Hz),123.7,127.2,128.7(d,JCF＝9.0Hz),131.1,131.2,134.8,135.0,146.6(d,JCF＝241.3Hz),172.4,180.8；HRMS(ESI-TOF)m/z:Calcd.for C23H20ClFN4NaOS[M+Na]+:477.0923；Found:477.0927.
本实施例制备化合物3cf:黄色固体,熔点：164.7-165.6℃；产率75％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.86(s,3H),2.85-2.97(m,2H),3.34-3.40(m,1H),4.77-4.82(m,1H),6.63-6.71(m,3H),7.00-7.03(m,1H),7.08-7.15(m,2H),7.52-7.57(m,2H),7.61-7.63(m,2H),7.68-7.72(m,2H),9.18(br s,1H),10.77(brs,1H)；13CNMR(DMSO-d6,100MHz)δ:21.0,22.0,55.4,68.6,70.3,109.2,109.5,113.1,118.2,121.8,123.0,123.8,124.6,127.3,127.5,127.8,128.7,128.7,130.0,130.1,134.7,135.1,135.2,143.0,172.3,180.7；HRMS(ESI-TOF)m/z:Calcd.for C27H21ClN4NaOS[M+Na]+:507.1017；Found:507.1014.
本实施例制备化合物3da:黄色固体,熔点：160.3-161.5℃；产率85％,15:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:0.65-0.69(m,3H),1.89-1.95(m,1H),2.43-2.48(m,1H),2.66(s,3H),2.68-2.73(m,1H),2.80-2.84(m,1H),3.25-3.32(m,1H),4.90-4.94(m,1H),6.95-7.04(m,3H),7.15-7.23(m,2H),7.43-7.49(m,2H),7.65(d,J＝7.2Hz,1H),8.78(br s,1H),10.44(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:9.9,21.0,26.2,30.4,43.3,69.4,73.8,109.7,111.5,111.9,118.6,119.0,122.4,126.2,127.5,130.2,130.9,137.2,145.3,174.5,183.9；HRMS(ESI-TOF)m/z:Calcd.forC23H22N4NaOS[M+Na]+:425.1407；Found:425.1413.
本实施例制备化合物3db:黄色固体,熔点：172.5-173.3℃；产率80％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:0.85-0.89(m,3H),2.06-2.12(m,1H),2.58-2.67(m,1H),2.80-2.93(m,2H),3.42-3.43(m,1H),4.70(d,J＝16.4Hz,1H),4.93-4.97(m,1H),5.26(d,J＝16.0Hz,1H),6.50-6.54(m,1H),6.61(d,J＝8.0Hz,1H),6.67(d,J＝6.8Hz,1H),6.86-6.97(m,3H),7.07(d,J＝8.0Hz,1H),7.27-7.43(m,6H),9.33(br s,1H),10.52(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:10.3,21.0,29.8,42.1,44.1,71.7,71.8,109.6,111.5,118.6,118.9,121.7,122.3,123.6,126.0,127.8,127.9,128.8,128.9,129.7,131.7,136.2,136.7,141.5,172.8,181.8；HRMS(ESI-TOF)m/z:Calcd.forC29H26N4NaOS[M+Na]+:501.1720；Found:501.1723.
本实施例制备化合物3dc:黄色固体,熔点：172.5-173.3℃；产率80％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:0.83-0.87(m,3H),1.99-2.05(m,1H),2.52-2.56(m,1H),2.76-2.82(m,1H),2.86-2.91(m,1H),3.34-3.39(m,1H),4.89-4.94(m,1H),6.46(d,J＝6.8Hz,1H),6.51-6.56(m,1H),6.87-6.97(m,3H),7.09(d,J＝8.0Hz,1H),7.37(d,J＝7.6Hz,1H),8.95(br s,1H),10.54(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:10.2,21.0,29.7,29.9,42.1,71.5,71.8,109.7,111.5,117.5(d,JCF＝19.6Hz),118.6,118.9,119.7,121.8,123.2(d,JCF＝6.3Hz),125.8,128.9(d,JCF＝8.1Hz),131.3,131.9,136.7,146.7(d,JCF＝241.1Hz),172.3,181.4；HRMS(ESI-TOF)m/z:Calcd.forC23H21FN4NaOS[M+Na]+:443.1312；Found:443.1317.
本实施例制备化合物3dd:黄色固体,熔点：153.7-154.4℃；产率80％,2:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(CDCl3,400MHz)δ:0.73-0.76(m,3H),1.75-1.81(m,1H),2.35(s,3H),2.52-2.57(m,1H),2.59(s,3H),2.77-2.82(m,1H),2.85-2.93(m,1H),3.29-3.37(m,1H),4.96-5.01(m,1H),6.16(s,1H),6.65(d,J＝8.0Hz,1H),7.00-7.10(m,3H),7.16-7.19(m,1H),7.29(s,1H),7.37(s,1H),7.43(d,J＝7.2Hz,1H)；13C NMR(CDCl3,100MHz)δ:8.3,19.7,20.2,25.1,29.0,42.3,68.6,73.0,107.6,109.7,112.3,118.1,118.8,121.5,125.3,127.2,127.3,130.4,131.3,135.4,141.4,173.2,183.6；HRMS(ESI-TOF)m/z:Calcd.for C24H24N4NaOS[M+Na]+:439.1563；Found:439.1557.
本实施例制备化合物3de:黄色固体,熔点：158.4-158.9℃；产率81％,7:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:0.45-0.48(m,3H),0.67-0.71(m,3H),1.90-1.95(m,1H),2.38(s,3H),2.44-2.48(m,1H),2.65-2.73(m,1H),2.79-2.83(m,1H),3.02-3.07(m,1H),3.26-3.36(m,2H),4.89-4.94(m,1H),6.91-7.03(m,3H),7.16(d,J＝8.0Hz,1H),7.25(d,J＝7.6Hz,1H),7.43(d,J＝7.6Hz,1H),7.48(s,1H),8.75(br s,1H),10.39(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:9.8,12.3,21.0,21.2,30.0,33.8,43.5,69.2,74.2,109.3,111.6,111.9,118.5,118.9,121.7,123.8,126.1,128.3,129.9,131.0,131.1,137.0,142.0,173.8,184.3；HRMS(ESI-TOF)m/z:Calcd.forC25H26N4NaOS[M+Na]+:453.1720；Found:453.1725.
本实施例制备化合物3df:黄色固体,熔点：157.9-158.9℃；产率79％,4:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:0.85-0.88(m,3H),1.25-1.29(m,3H),2.04-2.10(m,1H),2.54-2.60(m,1H),2.83-2.93(m,2H),3.36-3.38(m,1H),3.72-3.81(m,1H),4.03-4.08(m,1H),4.92-4.97(m,1H),6.48(d,J＝7.2Hz,1H),6.57-6.62(m,1H),6.92-7.01(m,3H),7.12(d,J＝8.0Hz,1H),7.41(d,J＝7.6Hz,1H),9.11(br s,1H),10.48(br s,1H)；13CNMR(DMSO-d6,100MHz)δ:10.2,13.9,20.9,29.8,37.6,42.3,71.5,71.9,109.9,111.5,117.7(d,JCF＝20.2Hz),118.6,119.0,120.1,121.9,123.2(d,JCF＝6.3Hz),125.9,128.1(d,JCF＝9.1Hz),131.2,132.0,136.7,146.7(d,JCF＝240.4Hz),172.3,181.6；HRMS(ESI-TOF)m/z:Calcd.for C24H23FN4NaOS[M+Na]+:457.1469；Found:457.1473.
本实施例制备化合物3ea:黄色固体,熔点：155.6-155.9℃；产率79％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(CDCl3,400MHz)δ:0.06-0.09(m,3H),0.40-0.47(m,1H),0.55-0.62(m,1H),1.26-1.30(m,1H),2.00-2.05(m,1H),2.81-2.86(m,1H),3.02-3.11(m,1H),3.64-3.71(m,1H),4.70(d,J＝15.2Hz,1H),4.89-4.94(m,1H),5.32(d,J＝15.2Hz,1H),6.36(s,1H),6.57-6.61(m,1H),6.66(s,1H),6.70-6.74(m,2H),6.88(d,J＝7.6Hz,1H),7.01-7.08(m,3H),7.39-7.51(m,4H),7.59(d,J＝7.2Hz,2H)；13C NMR(CDCl3,100MHz)δ:1.42,3.28,16.4,20.5,39.8,44.5,71.4,74.3,108.9,110.6,112.7,118.6,119.7,122.7,122.9,125.1,125.6,126.5,126.7,128.4,128.5,129.3,130.4,135.7,136.2,141.5,173.2,181.2；HRMS(ESI-TOF)m/z:Calcd.for C30H26N4NaOS[M+Na]+:513.1720；Found:513.1720.
本实施例制备化合物3eb:黄色固体,熔点：144.7-145.9℃；产率80％,2:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(CDCl3,400MHz)δ:0.16-0.20(m,1H),0.38-0.43(m,1H),0.47-0.51(m,1H),0.70-0.77(m,1H),1.55-1.61(m,1H),2.43(s,3H),2.77(s,3H),2.84-2.89(m,1H),2.95-3.03(m,1H),3.44-3.51(m,1H),4.94-4.99(m,1H),6.26(s,1H),6.79(d,J＝8.0Hz,1H),7.09-7.19(m,3H),7.28(d,J＝6.0Hz,1H),7.44(s,1H),7.51(d,J＝7.6Hz,1H),7.64(s,1H)；13C NMR(CDCl3,100MHz)δ:2.51,3.16,17.1,20.7,21.2,26.2,41.4,70.1,73.2,108.5,110.8,112.7,119.2,119.9,122.7,122.8,126.2,127.7,128.7,131.4,132.4,136.2,142.1,173.9,183.6；HRMS(ESI-TOF)m/z:Calcd.for C25H24N4NaOS[M+Na]+:451.1563；Found:451.1567.
本实施例制备化合物3ec:黄色固体,熔点：142.3-143.2℃；产率78％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(CDCl3,400MHz)δ:0.19-0.24(m,1H),0.40-0.44(m,1H),0.50-0.57(m,1H),0.70-0.77(m,4H),1.56-1.60(m,1H),2.43(s,3H),2.84-2.89(m,1H),2.95-3.04(m,1H),3.06-3.12(m,1H),3.42-3.48(m,1H),3.52-3.57(m,1H),4.95-4.99(m,1H),6.23(s,1H),6.79(d,J＝8.0Hz,1H),7.08-7.16(m,3H),7.26-7.27(m,1H),7.43(s,1H),7.51(d,J＝7.6Hz,1H),7.65(s,1H)；13C NMR(CDCl3,100MHz)δ:2.53,3.15,12.3,17.0,20.7,21.2,34.5,41.6,69.8,73.4,108.5,110.8,112.6,119.2,119.9,122.7,122.8,126.2,127.8,128.9,131.4,132.2,136.0,141.3,173.3,183.8；HRMS(ESI-TOF)m/z:Calcd.for C26H26N4NaOS[M+Na]+:465.1720；Found:465.1723.
本实施例制备化合物3fa:黄色固体,熔点：164.7-165.5℃；产率82％,3:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.62(s,3H),2.77-2.92(m,3H),3.12(d,J＝14.2Hz,1H),3.22-3.29(m,1H),4.69-4.73(m,1H),7.01-7.11(m,3H),7.25(d,J＝7.6Hz,1H),7.51-7.58(m,2H),7.69-7.73(m,1H),7.87(d,J＝7.6Hz,1H),8.81(brs,1H),10.28(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.2,21.6,38.5,68.5,74.3,109.6,111.8,118.8,119.4,122.1,124.9,126.3,126.6,128.3,133.1,135.6,136.5,136.6,151.1,180.3,201.5；HRMS(ESI-TOF)m/z:Calcd.for C22H19N3NaOS[M+Na]+:396.1141；Found:396.1145.
本实施例制备化合物3ga:黄色固体,熔点：175.4-175.7℃；产率83％,2:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.61(s,3H),2.73-2.93(m,3H),3.07(d,J＝17.2Hz,1H),3.22-3.29(m,1H),4.68-4.72(m,1H),7.06-7.12(m,2H),7.25(d,J＝7.6Hz,1H),7.54-7.58(m,2H),7.69-7.73(m,1H),7.87(d,J＝7.6Hz,1H),8.84(brs,1H),10.51(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.2,21.5,38.4,68.4,74.2,109.7,113.3,118.2,122.1,124.1,124.9,126.5,127.4,128.3,134.9,135.1,135.5,136.6,151.0,180.3,201.4；HRMS(ESI-TOF)m/z:Calcd.for C22H18ClN3NaOS[M+Na]+:430.0751；Found:430.0746.
本实施例制备化合物3ha:黄色固体,熔点：172.9-173.5℃；产率75％,4:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(CDCl3,400MHz)δ:0.72-0.76(m,3H),1.58-1.64(m,1H),2.36-2.41(m,1H),2.75-3.01(m,4H),3.27-3.34(m,1H),4.92-4.97(m,1H),6.29(s,1H),6.61(s,1H),6.83-6.85(m,1H),6.91(d,J＝7.6Hz,1H),7.05-7.07(m,2H),7.43-7.47(m,2H),7.52-7.55(m,1H),7.82(d,J＝7.6Hz,1H)；13C NMR(CDCl3,100MHz)δ:8.7,20.3,29.5,38.0,41.5,71.5,109.9,111.8,117.7,119.1,122.0,123.3,124.9,125.4,127.5,128.2,133.8,134.7,135.6,149.0,180.2,199.7；HRMS(ESI-TOF)m/z:Calcd.forC23H21N3NaOS[M+Na]+:410.1298；Found:410.1295.
本实施例制备化合物3ia:黄色固体,熔点：155.6-156.3℃；产率90％,20:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.61(s,3H),2.10-2.16(m,1H),2.23-2.28(m,1H),2.70-2.74(m,2H),3.13-3.20(m,1H),3.77(s,3H),4.14-4.21(m,1H),4.29-4.34(m,1H),4.54-4.58(m,1H),6.68-6.70(m,1H),6.86(d,J＝2.0Hz,1H),7.35(d,J＝8.4Hz,1H),8.95(br s,1H),11.00(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.2,21.7,32.2,55.7,65.3,68.0,69.4,95.2,109.7,110.7,119.5,120.9,130.6,137.8,156.6,174.0,180.7；HRMS(ESI-TOF)m/z:Calcd.for C18H19N3NaO3S[M+Na]+:380.1039；Found:380.1042.
本实施例制备化合物3ib:黄色固体,熔点：142.2-143.1℃；产率88％,10:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.57(s,3H),2.05-2.13(m,1H),2.35-2.39(m,1H),2.67-2.79(m,2H),3.04-3.11(m,2H),3.20-3.24(m,1H),3.77(s,3H),4.49-4.53(m,1H),6.68-6.70(m,1H),6.89(d,J＝2.0Hz,1H),7.35(d,J＝8.4Hz,1H),8.97(br s,1H),10.80(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:20.9,21.6,26.4,33.5,55.7,67.8,77.7,95.3,109.7,111.1,119.5,121.0,130.7,137.9,156.6,181.6,204.9；HRMS(ESI-TOF)m/z:Calcd.for C18H19N3NaO2S2[M+Na]+:396.0811；Found:396.0815.
本实施例制备化合物3ja:黄色固体,熔点：147.7-148.8℃；产率88％,8:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:1.59(s,3H),2.05-2.13(m,1H),2.34-2.39(m,1H),2.72-2.85(m,2H),3.00-3.06(m,1H),3.09-3.24(m,2H),4.52-4.57(m,1H),7.02-7.06(m,1H),7.12-7.16(m,1H),7.40(d,J＝8.4Hz,1H),7.48(d,J＝7.6Hz,1H),9.01(br s,1H),11.0(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:21.0,21.5,26.5,33.6,67.8,77.6,111.1,112.2,118.9,119.6,122.5,126.6,132.2,137.1,181.6,204.8；HRMS(ESI-TOF)m/z:Calcd.for C17H17N3NaOS2[M+Na]+:366.0705；Found:366.0708.
本实施例制备化合物3ka:黄色固体,熔点：160.4-161.3℃；产率89％,20:1dr；核磁共振和高分辨质谱测试等结果如下：1H NMR(DMSO-d6,400MHz)δ:0.64-0.67(m,3H),1.90-2.04(m,2H),2.16-2.37(m,3H),2.62-2.70(m,1H),2.90-3.00(m,2H),3.11-3.17(m,1H),4.77-4.81(m,1H),7.03-7.07(m,1H),7.13-7.16(m,1H),7.41(d,J＝8.0Hz,1H),7.52(d,J＝8.0Hz,1H),8.91(br s,1H),11.21(br s,1H)；13C NMR(DMSO-d6,100MHz)δ:9.52,21.6,26.1,28.3,34.2,43.7,70.3,79.1,112.2,112.9,118.9,119.6,122.5,126.4,129.5,136.9,182.1,204.1；HRMS(ESI-TOF)m/z:Calcd.for C18H19N3NaOS2[M+Na]+:380.0862；Found:380.0863.
本发明的式(1)化合物具有重要的生物活性，体外对人白血病细胞(K562)、人肺腺癌细胞(A549)、人前列腺癌细胞(PC-3)的细胞毒性试验表明：此类式(1)所示结构的骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物对肿瘤细胞生长具有抑制作用，有可能发展成为新的防治肿瘤药物。但需强调的是本发明的化合物不限于人白血病细胞(K562)、人肺腺癌细胞(A549)、人前列腺癌细胞(PC-3)表示的细胞毒性。
药理实施例1：化合物3bb、3bh、3ca、3cb、3cc、3cd、3ce、3cf、3dc和3de对人白血病细胞(K562)、人肺腺癌细胞(A549)、人前列腺癌细胞(PC-3)的细胞毒性
人白血病细胞(K562)、人肺腺癌细胞(A549)、人前列腺癌细胞(PC-3)分别用RPMI-1640培养基培养，培养基中含10％的胎牛血清，100U/mL的青霉素和100U/mL链霉素。细胞以每孔5000个细胞的浓度加入到96孔中，在37℃含5％CO2潮湿空气的培养箱中培养24小时。
细胞存活率的测定用改良MTT法。细胞经过24小时的孵育后，分别将新配的化合物3bb、3bh、3ca、3cb、3cc、3cd、3ce、3cf、3dc和3de的二甲基亚砜溶液以浓度梯度加入到各孔中，使孔中化合物最终浓度分别为5μmol/L,10μmol/L,20μmol/L,40μmol/L和80μmol/L。48小时后，每孔加入10μL MTT(5mg/mL)的磷酸盐缓冲液，再继续在37℃培养4小时后，离心5分钟除去未转化的MTT，每孔中加入150μL二甲基亚砜。以溶解还原的MTT晶体甲臜(formazan)，用酶标仪在490nm波长测定OD值。结果如下表3所示。
表3
a IC50:The concentration which results in 50％of tumour cellproliferation inhibition after 48h of compounds treatment.Data wererepresented as means±SEM obtained in at least three independentexperiments.b Cisplatin used as positive control.
c Harmaline used as positive control.
药理实施例2：化合物3cb对HK-2和MRC-5进行细胞毒性试验
肾小管上皮细胞HK-2和胚胎肺成纤维细胞MRC-5属于正常人体细胞，选化合物3cb对HK-2和MRC-5进行细胞毒性试验。细胞存活率的测定用改良MTT法，具体方法如药理实施例1。结果如表4所示，化合物3cb对正常细胞3cb显示出几乎无毒的细胞毒性作用。
表4
a IC50:The concentration which results in 50％of tumour cellproliferation inhibition after 48h of compounds treatment.Data wererepresented as means±SEM obtained in at least three independent experiments.
b Cisplatin used as positive control.
c Harmaline used as positive control.
药理实施例3：化合物3cb诱导A549细胞凋亡的变化
用化合物3cb处理的A549细胞在对照细胞中显示出长纺锤形。如图7中(A)所示，在低浓度下，相邻细胞之间的接触消失，粘附能力降低，细胞膜起泡，细胞骨架解体细胞变圆。高浓度时，随着细胞膜向内折叠，细胞器向内折叠，凋亡小体逐渐形成。在荧光显微镜下观察，如图7中(B)所示，在对照组中，吖啶橙可以穿透正常细胞膜并使细胞核显示绿色荧光。在用3cb处理的凋亡细胞中，由于染色质凝结或碎裂成不同大小的碎片而形成凋亡小体。溴化乙锭染色密集且强烈的橙红色荧光或橙红色碎片颗粒。当细胞凋亡发生时，DNA也随之改变。抗肿瘤药物被证明会导致DNA损伤，而细胞毒性药物主要通过破坏肿瘤细胞中的DNA来发挥作用，而影响DNA化学结构的药物在临床实践中已经可用。在倒置荧光显微镜下观察，如图7中(C)所示，正常细胞能够穿透少量Hoechst 33258染色溶液，而对照组中的细胞被观察到染色低。细胞膜渗透性增强，Hoechst 33258染色溶液能够大量穿透细胞并与DNA结合，染色致密且染色明亮的蓝色荧光。
药理实施例4：化合物3cb对A549细胞ROS和MMP变化的影响
研究了化合物3cb在A549细胞中诱导的活性氧产生和线粒体膜电位变化，结果如图8中(A)所示。绿色荧光逐渐增加，表明3cb诱导的ROS释放随着药物浓度的增加而增加。在图8中(B)中，JC-1在对照线粒体的基质中积累，并形成产生红色荧光的聚合物，从而导致更高的线粒体膜电位。与对照组相比，3cb处理后，红色荧光逐渐减少，绿色荧光逐渐增加，表明3cb显著下调了线粒体膜电位。
药理实施例5：化合物3cb对A549细胞集落形成的影响
集落形成是测定细胞增殖能力的有效方法之一。单个细胞的增殖潜力可以通过计数菌落形成率来定量评估，以便独立地确定其增殖能力和生存能力。研究了3cb的菌落形成能力，结果如图9所示。与对照组中形成的菌落数量相比，药物组中的菌落数量逐渐减少。对照组的单细胞克隆效率为100％，药物组的克隆效率分别为82.67％、41.53％和12.63％。结果表明，化合物3cb能显著影响A549细胞的增殖。
药理实施例6：化合物3cb对A549细胞迁移的影响
肿瘤转移是癌症患者高死亡率的主要原因。识别调节肿瘤细胞的迁移、分化和生长，并在预防和治疗恶性肿瘤中发挥关键作用。癌细胞迁移的增强可导致肿瘤的侵袭和转移。在本研究中，通过伤口划痕试验、transwell插入物研究了3cb对肿瘤细胞转移的影响，结果如图10所示。化合物3cb可以在低浓度下影响A549细胞的迁移能力，并在10μM时显著抑制A549细胞迁移能力。
药理实施例7：化合物3cb对A549细胞中凋亡相关蛋白表达的影响
细胞凋亡是癌细胞死亡的主要机制。许多癌症治疗依赖于诱导有效的细胞凋亡，而细胞凋亡的复杂调节为药物开发人员在有效的抗癌治疗中利用细胞凋亡诱导创造了相当大的障碍。因此，必须阐明3cb的细胞凋亡机制。Western blot检测3cb细胞凋亡相关蛋白的表达。
如图11所示，p53、Bax、Cyt c、切割的Caspase 3、切割的PARP表达显著上调，Bcl-2、Pro-Caspase 3、PARP表达明显下调。结果表明，3cb通过激活p53蛋白表达来调节Bax/Bcl-2比值和Cyt-c表达。Caspase 3酶原在近端被切割成活性Caspase3，并且被激活的切割Caspase3进一步切割PARP，从而激活凋亡标记蛋白切割PARP。
Claims (3)
1.一种骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物，其特征在于：该化合物具有如通式(Ⅰ)所示的结构：
其选自如下化合物：
2.一种如权利要求1所述的骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物的制备方法，其特征在于：由相应的骆驼蓬灵骨架化合物1与α-异硫氰酸酯羰基化合物2发生Michael加成环化反应，生成最终产物骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物(I)；
合成路线如下：
3.一种如权利要求1所述的骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物在制备防治肿瘤疾病药物中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211701584.6A CN115850277B (zh) | 2022-12-28 | 2022-12-28 | 骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物及其制备方法及应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211701584.6A CN115850277B (zh) | 2022-12-28 | 2022-12-28 | 骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物及其制备方法及应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115850277A CN115850277A (zh) | 2023-03-28 |
| CN115850277B true CN115850277B (zh) | 2024-03-08 |
Family
ID=85655683
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211701584.6A Active CN115850277B (zh) | 2022-12-28 | 2022-12-28 | 骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物及其制备方法及应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115850277B (zh) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20180067965A (ko) * | 2016-12-13 | 2018-06-21 | 성균관대학교산학협력단 | 신규 스파이로아이소인돌리논 유도체, 이의 제조방법 및 이를 유효성분으로 포함하는 암 예방 및 치료용 조성물 |
| CN109705130A (zh) * | 2018-11-23 | 2019-05-03 | 贵州大学 | 二氢色原酮骨架拼接多环吡咯螺环氧化吲哚化合物及其制备方法及应用 |
| CN110755428A (zh) * | 2019-11-04 | 2020-02-07 | 五邑大学 | 骆驼蓬灵在制备降脂药物中的应用 |
-
2022
- 2022-12-28 CN CN202211701584.6A patent/CN115850277B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20180067965A (ko) * | 2016-12-13 | 2018-06-21 | 성균관대학교산학협력단 | 신규 스파이로아이소인돌리논 유도체, 이의 제조방법 및 이를 유효성분으로 포함하는 암 예방 및 치료용 조성물 |
| CN109705130A (zh) * | 2018-11-23 | 2019-05-03 | 贵州大学 | 二氢色原酮骨架拼接多环吡咯螺环氧化吲哚化合物及其制备方法及应用 |
| CN110755428A (zh) * | 2019-11-04 | 2020-02-07 | 五邑大学 | 骆驼蓬灵在制备降脂药物中的应用 |
Non-Patent Citations (2)
| Title |
|---|
| Neelakandan Vidhya Lakshmi 等.An expedient approach for the synthesis of dispiropyrrolidine bisoxindoles, spiropyrrolidine oxindoles and spiroindane-1,3-diones through 1,3-dipolar cycloaddition reactions.《Tetrahedron Letters》.第51卷1064-1068. * |
| 通过1，3 － 偶极环加成反应 合成3 － 吡咯螺环氧化吲哚的研究进展;周英等;《山地农业生物学报》;第34卷(第2期);009-013 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115850277A (zh) | 2023-03-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR20230127294A (ko) | 코로나 바이러스 감염을 치료하기 위한 방법 및 변형된뉴클레오사이드 | |
| JP7410087B2 (ja) | ５員環および６員環が縮合したアザ環式芳香族系化合物、その製造方法、医薬組成物およびその応用 | |
| EA032362B1 (ru) | Производные индола в качестве ингибиторов репликации вирусов денге | |
| CN101171250A (zh) | 具有抗菌活性的氮杂环丁烷和吡咯的三环衍生物 | |
| CN115554303B (zh) | 一种三环类化合物、其制备方法及其应用 | |
| EP3611170B1 (en) | Deuterated compounds and medical use thereof as antianxiety agents | |
| EP3424913B1 (en) | 2,3-epoxy succinyl derivative, preparation method therefor, and uses thereof | |
| Wei et al. | Synthesis and biological evaluation of indazole derivatives as anti-cancer agents | |
| CN115850277B (zh) | 骆驼蓬灵骨架拼接的多螺环吡咯啉酮类衍生物及其制备方法及应用 | |
| TW201609683A (zh) | 阿可拉定化合物的晶型、含有該晶型的藥物及用途 | |
| CN113773316B (zh) | 一种tnik抑制剂及其制备方法和用途 | |
| CN108727437B (zh) | 一类姜黄素芳基金属配合物及其合成方法和应用 | |
| Cao et al. | Microwave-assisted synthesis of polypyridyl ruthenium (ii) complexes as potential tumor-targeting inhibitors against the migration and invasion of Hela cells through G2/M phase arrest | |
| EP4043467A1 (en) | Class of functional molecules targeting proteolysis pathways, preparation and application thereof | |
| Kim et al. | Synthesis of 6-chloroisoquinoline-5, 8-diones and pyrido [3, 4-b] phenazine-5, 12-diones and evaluation of their cytotoxicity and DNA topoisomerase II inhibitory activity | |
| CN114685504B (zh) | 一种Fascaplysin衍生物及其制备方法与应用 | |
| AU2789999A (en) | Farnesyl-protein transferase inhibitors | |
| JP7353500B2 (ja) | アクリル酸トリプトリド、その調製方法および用途 | |
| CN111808105B (zh) | 含有并环基团的嘧啶酮并吡唑类化合物、其制备方法及用途 | |
| CN114105856B (zh) | 邻羟基苯乙酰拼接双吲哚甲基类化合物及其制备方法及应用 | |
| CN106279058B (zh) | 3,4-二芳基-1,2,5-噁二唑氧化物的制备及用途 | |
| JP2022542449A (ja) | Ｓｔａｔ３二機能性リン酸化部位を標的とするトリアロマティック化合物のクラスおよびその応用 | |
| TW593306B (en) | Klainetins and their derivatives, method for their preparation and use thereof | |
| CN108586426A (zh) | 一种烷氧基联苯/查尔酮杂合类化合物、其制备方法及医药用途 | |
| CN114933628B (zh) | 一种靶向p53突变体的甾体类化合物及其制备方法、及在治疗肿瘤中的应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |