CN107903210B - Small molecule inhibitor SLD4650 and application thereof in pharmacy - Google Patents
Small molecule inhibitor SLD4650 and application thereof in pharmacy Download PDFInfo
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- CN107903210B CN107903210B CN201711420932.1A CN201711420932A CN107903210B CN 107903210 B CN107903210 B CN 107903210B CN 201711420932 A CN201711420932 A CN 201711420932A CN 107903210 B CN107903210 B CN 107903210B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
- C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
- C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D215/48—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
Abstract
The invention provides a small molecule inhibitor SLD4650, which has a structural formula as follows:
the invention relates to application of the small molecule inhibitor SLD4650 in preparing a medicament for inhibiting spermine oxidase. The result shows that SLD4650 changes the growth cycle of human small cell lung cancer A549 cells, changes the expression content of A549 cell cyclins, and can induce A549 cells to undergo apoptosis.
Description
Technical Field
The invention provides a small molecule inhibitor for inhibiting spermine oxidase, and simultaneously, the small molecule inhibitor is applied to the preparation of drugs for treating tumor diseases.
Background
Polyamines (putrescine, spermidine and spermine) are small molecular organic compounds widely existing in eukaryotic cells and participate in important physiological functions such as cell differentiation, proliferation and gene regulation. Research shows that high polyamine content in cells is necessary for the rapid growth of tumor cells, so that a polyamine metabolic pathway is gradually a new target point for antitumor treatment and drug design. Spermine oxidase (SMO) is a key enzyme involved in polyamine catabolism, an FAD (flavin adenine dinucleotide) -dependent oxidase that uses oxidation of spermine (Spm) as its first substrate degradation pathway. The products of this enzyme are spermidine, aminopropionaldehyde and H2O2. Recent studies have found that SMO expression is continuously up-regulated in cells of affected tissues in various chronic inflammatory environments, thereby causing intracellular active oxygen content increase and DNA damage, and this process is closely related to the occurrence of various tumors, so SMO is a potential new molecular target for antitumor therapy.
With the continuous and deep understanding of the anti-tumor molecular mechanism of polyamine analogs, more and more spermine analogs are synthesized, and some have entered clinical trials. Among all spermine analogs, BENSpm is the most intensive study, which has cytotoxic effects mainly on lung cancer, breast cancer and the like. However, BENSpm was found to be not significantly effective as a single agent in the secondary clinical study for the treatment of advanced breast cancer, and extensive experiments in this study showed that BENSpm was synergistic with other conventional chemotherapeutic agents when used in combination. The second generation of the spermine analog CPENSpm has lower toxic effect and more remarkable effect compared with the first generation, but CPENSpm and BENSpm have the same problems and need to be combined with other chemotherapeutic drugs. Although BENSpm and CPENSpm have good clinical application prospects in preclinical experimental results, the results are slightly effective in first-stage and second-stage clinical trials. Therefore, a need exists for new small molecule inhibitors of spermine oxidase to meet the need of anti-tumor drug development.
Disclosure of Invention
Based on the research background, a pharmacophore model with the effect of inhibiting the SMO activity is constructed based on an Spm and SMO compound by using a computer-aided drug design technology, the obtained pharmacophore model is taken as a questioning structure, virtual screening is carried out in a chemical database, so that candidate molecules which can inhibit the SMO activity theoretically are obtained, and a screening result is analyzed and evaluated by using a molecular docking technology. Obtaining the small molecule compound, and evaluating the activity of the small molecule compound at the protein level in vitro, thereby obtaining the small molecule inhibitor for inhibiting the SMO activity. Next, further validation is carried out at the cellular level, and the mechanism of the antitumor action of these small molecule inhibitors is explored.
Based on the complex structure of Spd and SMO, key interactions formed between small molecule inhibitors and proteins were analyzed. In the software construction process, a pharmacophore module is used for establishing a pharmacophore model of the interaction between SMO and Spd. And carrying out molecular docking on the compound obtained by searching the pharmacophore model by using docking software. On the basis of butt joint scoring, the structural diversity of the compound is combined to obtain the inhibitor.
The specific structural formula of the small molecule inhibitor SLD4650 is as follows:
the small molecule inhibitor SLD4650 is applied to preparing a medicament for inhibiting spermine oxidase.
The small molecule inhibitor SLD4650 is applied to preparing a medicament for inhibiting human small cell lung cancer.
The application of the preparation of the medicament for inhibiting the human small cell lung cancer, in particular to the application of the preparation of the medicament for inhibiting the growth and the reproduction of the human small cell lung cancer A549 cells.
The application of the preparation of the medicament for inhibiting the human small cell lung cancer, in particular to the application of the preparation of the medicament for inhibiting the migration of the human small cell lung cancer A549 cells.
Drawings
Figure 1 screens the pharmacophore model for SMO small molecule inhibitors.
FIG. 2 is a graph comparing binding patterns of SLD4650 and Spd.
Fig. 3 chemiluminescence assay for inhibition of spermine oxidase activity by small molecule SLD 4650: p <0.01 compared to 0 mM.
Figure 4 HPLC assay of polyamine content in a549 cells: p <0.01, x: p < 0.05.
FIG. 5 MTT method for detecting the growth inhibition effect of small molecule drug SLD4650 on A549 cells.
FIG. 6 the Transwell method measures the ability of A549 cells to migrate in vitro,
a is a normally cultured A549 cell;
and B, adding SLD4650 micromolecule medicine with the final concentration of 80 mu M into the cell culture solution, and treating the A549 cells after 48 hours of action.
FIG. 7 flow cytometry is adopted to detect the influence of small molecule drug SLD4650 on the growth cycle of A549 cells,
a is a normally cultured A549 cell;
b, adding SLD4650 micromolecule medicine with the final concentration of 40 mu M into the cell culture solution, and treating the A549 cells after 48 hours of action;
c, adding SLD4650 micromolecule medicine with the final concentration of 40 mu M into the cell culture solution, and acting for 72h to obtain the A549 cells.
FIG. 8 flow cytometry is used to detect the effect of small molecule drug SLD4650 on the growth cycle of A549 cells,
a is a normally cultured A549 cell;
b, adding SLD4650 micromolecule medicine with the final concentration of 80 mu M into the cell culture solution, and treating the A549 cells after 48 hours of action;
c, adding SLD4650 micromolecule medicine with the final concentration of 80 mu M into the cell culture solution, and acting for 72h to obtain the A549 cells.
FIG. 9 immunoblotting to detect the change in the content of the cycle protein in A549 cells,
1. a549 cells in normal culture;
2. and adding SLD4650 micromolecule medicine with the final concentration of 80 mu M into the cell culture solution, and treating the A549 cells after 48 hours of action.
FIG. 10AnnexinV-FITC/PI double-staining flow cytometry for detecting A549 cell apoptosis induced by small molecule drug SLD4650
A is the A549 cells cultured normally,
and B, adding SLD4650 micromolecule medicine with the final concentration of 80 mu M into the cell culture solution, and treating the A549 cells after 48 hours of action.
Detailed Description
A small molecule inhibitor SLD4650 has a specific structural formula:
the drug effect group results are as follows:
in the complex structure, much hydrogen bonding interaction is formed between Spd and SMO. To ensure the rationality of the pharmacophore model, we retained only two key pairs of hydrogen bonding signature elements. In addition, in order to ensure the structural diversity, a group of acceptor amino acid-based hydrogen bond characteristic elements P1-Ser O is added after the characteristics of the active pocket are analyzed. The details are as follows (fig. 1): two donor centers-P1-Ser O and P3-Glu O.
Molecular docking results
And (3) carrying out molecular docking on the compound obtained by the pharmacophore model, and grading according to docking energy to obtain the compound which is combined in an active pocket defined by docking. FIG. 2 is a diagram of the binding pattern defined therein as SLD 4650.
Effect of Small molecule inhibitors on spermine oxidase Activity
In vitro experiment detection of activity of SLD4650 in effectively inhibiting spermine oxidase
Constructing a prokaryotic expression vector, inducing bacteria to express spermine oxidase protein, obtaining the protein and purifying. After SLD4650 micromolecule medicines with different concentrations and spermine are mixed, spermine oxidase is added, and the activity of the enzyme is detected by a chemiluminescence method. The results showed that the enzyme activity was gradually decreased with the increase of the drug concentration of SLD4650, and thus SLD4650 was effective in inhibiting the activity of spermine oxidase, as shown in fig. 3.
After treating A549 cells with SLD4650 with the final concentration of 80 mu M for 48h, collecting the cells and cracking, extracting polyamine components in the cells, and detecting the content of polyamine in the cells through high performance liquid phase analysis, the result shows that compared with control cells, after SLD4650 treatment, the content of spermine in the cells is increased, and the content of spermidine is reduced, so that SLD4650 inhibits the activity of spermine oxidase in the cells, and the spermine cannot be effectively converted into spermidine, thereby interfering the metabolism of polyamine in the cells, as shown in figure 4.
Research on antitumor activity of small molecule inhibitor
SLD4650 is effective in inhibiting growth and reproduction of human small cell lung cancer A549 cells
The MTT method detects the inhibition effect of SLD4650 on A549 cells, and the result shows that SLD4650 effectively inhibits cell proliferation and has time and dose dependence, wherein the inhibition rate of the cell is up to 63.79% after 72h of 160 mu M treatment, as shown in figure 5.
SLD4650 is effective in inhibiting migration of A549 cells
After 48h of treatment of A549 cells with SLD4650 at a final concentration of 80. mu.M, the ability of A549 cells to migrate was examined in vitro using the Transwell method, and it was shown that SLD4650 was effective in inhibiting cell migration, as shown in FIG. 6.
Research on anti-tumor action mechanism of small molecule inhibitor
SLD4650 altering the growth cycle of human small cell lung carcinoma A549 cells
After cells were treated with SLD4650 at final concentrations of 40. mu.M and 80. mu.M for 48h and 72h, respectively, the cells were harvested, the genome was stained with PI, and the percentage of cells in different growth cycles was determined by flow cytometry, showing that SLD4650 decreased the number of cells in G0/G1 phase and increased the number of cells in S phase, changing the growth cycle of the cells, as shown in FIGS. 7 and 8.
SLD4650 altering A549 cyclin expression levels
After treating A549 cells with SLD4650 with the final concentration of 80 mu M for 48h, collecting the cells and cracking the cells to obtain cell proteins, detecting the expression amount of the Cyclin in the cell proteins by using an immunoblotting method, and showing that the Cyclin B1 and the p21 protein are remarkably increased compared with control cells and are consistent with the change of the cell growth cycle, as shown in figure 9.
SLD4650 induces apoptosis in A549 cells
After 48h of treatment of A549 cells with SLD4650 at a final concentration of 80. mu.M, the cells were harvested, stained with the annexin V-FITC/PI double stain kit, and the number of annexin V-FITC/PI double stained cells increased from 1.61% to 12.45% compared to control cells as measured by an up-flow cytometer. The resulting SLD4650 was effective in inducing apoptosis in a549 cells, as shown in fig. 10.
Claims (4)
1. The application of a small molecule inhibitor SLD4650 in preparing a medicament for inhibiting spermine oxidase is disclosed, wherein the structural formula of the small molecule inhibitor is as follows:
2. the application of a small molecule inhibitor SLD4650 in preparing a medicament for inhibiting human non-small cell lung cancer is disclosed, wherein the structural formula of the small molecule inhibitor is as follows:
3. the use of claim 2, wherein the small molecule inhibitor is used for preparing a medicament for inhibiting the growth and reproduction of human non-small cell lung cancer A549 cells.
4. The use of claim 2, wherein the small molecule inhibitor is used for preparing a medicament for inhibiting the migration of human non-small cell lung cancer A549 cells.
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| WO2023141866A1 (en) * | 2022-01-27 | 2023-08-03 | Janssen Pharmaceutica Nv | Pyrazolopyrimidines as modulators of spermine oxidase |
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| CN109942467B (en) * | 2019-04-04 | 2020-09-25 | 泉州师范学院 | Small molecule inhibitor AZIN32 and application thereof in pharmacy |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1237963A (en) * | 1996-09-25 | 1999-12-08 | 曾尼卡有限公司 | Qinoline derivatives inhibiting effect of growth factors such as VEGF |
| WO2006094237A2 (en) * | 2005-03-03 | 2006-09-08 | Sirtris Pharmaceuticals, Inc. | Acridine and quinoline dervatives as sirtuin modulators |
| WO2008012782A2 (en) * | 2006-07-27 | 2008-01-31 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Labelled analogues of halobenzamides as radiopharmaceuticals |
| WO2012158435A1 (en) * | 2011-05-17 | 2012-11-22 | Han-Jie Zhou | Compositions and methods for jamm protein inhibition |
| CN102827142A (en) * | 2011-06-17 | 2012-12-19 | 中国科学院上海药物研究所 | Quinoline-8-formamide compound as well as preparation method and applications thereof |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1237963A (en) * | 1996-09-25 | 1999-12-08 | 曾尼卡有限公司 | Qinoline derivatives inhibiting effect of growth factors such as VEGF |
| WO2006094237A2 (en) * | 2005-03-03 | 2006-09-08 | Sirtris Pharmaceuticals, Inc. | Acridine and quinoline dervatives as sirtuin modulators |
| WO2008012782A2 (en) * | 2006-07-27 | 2008-01-31 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Labelled analogues of halobenzamides as radiopharmaceuticals |
| WO2012158435A1 (en) * | 2011-05-17 | 2012-11-22 | Han-Jie Zhou | Compositions and methods for jamm protein inhibition |
| CN102827142A (en) * | 2011-06-17 | 2012-12-19 | 中国科学院上海药物研究所 | Quinoline-8-formamide compound as well as preparation method and applications thereof |
Non-Patent Citations (1)
| Title |
|---|
| RN：1120230-78-5;美国化学会;《STN ON THE WEB》;20090313;第2页 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023141866A1 (en) * | 2022-01-27 | 2023-08-03 | Janssen Pharmaceutica Nv | Pyrazolopyrimidines as modulators of spermine oxidase |
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