CN110128477B - Nucleolar stress-based platinum compounds - Google Patents
Nucleolar stress-based platinum compounds Download PDFInfo
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- CN110128477B CN110128477B CN201810214963.XA CN201810214963A CN110128477B CN 110128477 B CN110128477 B CN 110128477B CN 201810214963 A CN201810214963 A CN 201810214963A CN 110128477 B CN110128477 B CN 110128477B
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- PNQYAMWGTGWJDW-UHFFFAOYSA-N tert-butyl n-(3-aminopropyl)-n-methylcarbamate Chemical compound NCCCN(C)C(=O)OC(C)(C)C PNQYAMWGTGWJDW-UHFFFAOYSA-N 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 229940045145 uridine Drugs 0.000 description 1
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Abstract
The invention discloses a platinum compound based on nucleolar stress, which is a platinum anti-tumor compound prepared by performing nitrile-secondary amine Click reaction on cis-chloro-mononitrile diaminoplatinum and an RNA polymerase I inhibitor containing primary amine. The platinum anti-tumor compound has good RNA polymerase I inhibition activity and tumor selectivity, thereby showing the anti-tumor activity superior to that of the classical platinum drugs, reducing the toxic and side effects and overcoming the clinical platinum drug resistance.
Description
Technical Field
The invention belongs to the technical field of platinum compounds, and particularly relates to a nucleolar stress-based platinum anti-tumor compound, an anti-tumor effect, toxic and side effects alleviation and clinical platinum drug resistance overcoming research.
Background
Cancer (Cancer) is a big killer threatening mankind at present world time, and has a very profound practical significance for the research of Cancer treatment. Platinum antineoplastic drugs are an important class of metal chemotherapeutic drugs and are applied to more than half of chemotherapy methods. The representative medicine cisplatin has wide anti-tumor spectrum and strong effect, has synergistic effect with various anti-tumor medicines, has no cross drug resistance, has obvious curative effect on ovarian cancer and testicular cancer, and has over 90 percent of cure rate in the treatment of the testicular cancer. However, the development of the traditional platinum drugs is severely limited by serious adverse reactions (such as bone marrow suppression, leukopenia, ototoxicity, neurotoxicity, hepatotoxicity and the like) and tumor drug resistance (including primary drug resistance and acquired drug resistance) of the traditional platinum drugs, so that the development of novel platinum anti-tumor drugs is necessary.
Monofunctional platinum compounds represent a very advantageous class of anticancer agents. Such complexes contain only one labile ligand and may form only one covalent bond with DNA. Unlike complexes which can form monofunctional adducts and bifunctional adducts, such compounds are designed to form at most one bond with DNA. The monofunctional platinum compound can change the action mechanism of the classical platinum drugs so as to radically improve the drug resistance of the classical platinum drugs.
Nucleolus is a kind of tumor treating target with great potential. Almost all types of tumors are accompanied by an increase in the volume and number of nucleoli. Tumor cells can grow unregulated due to cell cycle disorders, often accompanied by a significant increase in the nucleoli and a large amount of intracellular rRNA and protein synthesis. The biological function of the nucleolus is closely related to the synthetic release of proteins. Nucleoli is the central control of ribosome synthesis and assembly, and its transcription and disassembly are the result of changes in the level of RNA polymerase I (RNA polymerase I, Pol I) transcription. The abnormal regulation of nucleolar morphology reflects the overactivity of rDNA transcription, and in brief, the acceleration of ribosome synthesis in tumor cells aims to enhance the proliferation and growth capacity of the tumor cells. In addition, nucleolus can regulate the activity of various vital proto-oncogenes and cancer suppressor genes, which plays an important role in tumor transformation. If the ribosome biosynthesis is rapidly inhibited by an inhibitor molecule, tumor cells can be specifically killed with less influence on normal cells, and the method is non-genotoxic and can be used for treating tumors. Ribosome biosynthesis is dependent on continuous ribosomal DNA (rDNA) transcription, and accelerated transcription of rDNA is a necessary condition for survival of tumor cells and can also cause nucleolar abnormality of tumor cells. If the transcription of rRNA of the tumor cells is interfered, nucleolus stress reaction of the tumor cells can be caused, nucleolus disintegration and ribosomal protein accumulation are caused, a p53 channel is further activated, and finally the apoptosis of the tumor cells is caused; however, in normal cells, because no variation or deletion of cancer suppressor genes exists, the regulation of cell cycle check points can be started, and normal cell apoptosis cannot be caused. Therefore, the selective inhibition of rDNA transcription and the rapid inhibition of ribosome biosynthesis are promising in the aspect of tumor treatment, and the core member RNA polymerase I in the process is considered to be a brand new target with great development potential. With the continuous improvement of nucleolus understanding, in the development of novel anti-tumor drugs, small molecule inhibitors which target Pol I mediated rDNA transcription so as to inhibit ribosome organisms have achieved great success.
The research of combining the molecular targeted drug and the platinum drug to form the twin drug is a new generation of metal anti-tumor drug which is expected to improve the targeting property, selectivity and safety of the platinum drug and overcome the tumor drug resistance of the platinum drug. Based on the research result of the existing RNA polymerase I inhibitor, the selective RNA polymerase I inhibitor and the platinum drug are combined to form a twin drug by combining the drug resistance mechanism of the platinum drug, so that the following advantages are achieved: (1) from the chemical structure, the introduction of the Pol I inhibitor can change the physical and chemical properties of the platinum compound, such as electrical property, polarity, logP value and the like, and further change the uptake mode of the platinum drug so as to increase the platinum content in cells; meanwhile, the combination rate of the platinum drugs and sulfur-containing proteins such as GSH, Trx, GPx and the like in cells can be changed, and the pre-target drug resistance of the platinum drugs can be overcome. (2) From the point of action targets, 45S rDNA exists in a large number of tandem repeat sequences in nucleolus, the tandem repeat sequences are G-rich sequences, and the structural characteristics cause the rDNA to have higher affinity for platinum compounds, so a large number of rDNA adducts which obstruct replication and transcription can be formed; meanwhile, when the platinum compound forms an adduct with rDNA in the nucleolar region, the DNA damage of the region is difficult to be eliminated by a Nucleotide Excision Repair (NER) mechanism of transcription coupling mediated by RNA polymerase II (pol II), so that the platinum drug can better play a role. The NER repair mechanism is one of the main repair mechanisms of platinum compound DNA damage and one of the important reasons of platinum drug resistance. The Pol I inhibitor is introduced into the platinum drug, so that the platinum drug can be directly introduced into DNA of nucleolus region of cells but not on chromosome DNA existing around nucleolus, and further a large amount of platinized rDNA injury which is difficult to be repaired by NER protein is formed, and the platinum drug escape from NER related repair mechanism is facilitated, therefore, the target resistance of the platinum drug can be fundamentally overcome. (3) From the characteristic point of view of tumor cells, the tumor cells are more dependent on the biosynthesis of ribosome than normal cells, and the tiny rRNA synthesis is blocked, so that the tumor cells can generate nucleolar stress, the intracellular ribosomal protein is accumulated, and then the p53 pathway is activated to cause apoptosis. Meanwhile, due to the inactivation of cancer suppressor genes in tumor cells, the regulation of cell cycle check points is lacked, and once the rRNA synthesis is blocked, the programmed death of cells is directly started; the cancer suppressor gene in normal cells can play a normal role, and normal cell apoptosis can not be caused because the regulation of cell cycle check points can be started. The selectivity of the platinum drug to tumor tissues can be further improved by utilizing the characteristics of the Pol I selective inhibitor, the toxicity is reduced, and the continuous tumor deterioration caused by the genotoxicity of the platinum drug can be greatly reduced.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides a nucleolar stress-based platinum compound and a preparation method and application thereof, and compared with the existing platinum compound in the literature, the platinum anti-tumor compound (namely, a monofunctional platinum compound) has targeting property, selectivity and safety and overcomes the advantage of tumor resistance of the classical platinum drugs.
The technical purpose of the invention is realized by the following technical scheme:
the nucleolar stress-based platinum compound has a structure shown in the following chemical formula:
centered on the divalent metal platinum (Pt), with two NH groups3One Cl-And the side chain NH to form coordination, so that the whole is positively charged, and the platinum compound is formed with the nitrate radical which is negatively charged.
Wherein R is1Is methyl or ethyl, R2Is methyl or ethyl, L is a chain group with 2-4 carbons, namely- (CH)2) 2-4 CH2And (4) forming.
Preferably R1Is methyl, R2Is methyl, L is a chain radical having 3 carbons, i.e., - (CH)2)3 CH2And (4) forming.
The method for producing a platinum compound is characterized by reacting a substance B with a substance P, wherein the substance B is in excess relative to the substance P, and the molar ratio of the substance B to the substance P is (1-1.2): the reaction temperature is minus 20 to minus 4 ℃, preferably minus 10 to 0 ℃, the reaction time is 20 to 100 hours, preferably 60 to 80 hours, a closed environment is adopted in the reaction process, magnetic stirring is selected, 100 to 300 revolutions per minute is selected, an organic solvent is selected to provide a reaction environment for the substance B and the substance P, and the solubility and the dispersibility of the substance B and the substance P in the organic solvent need to be considered, such as N, N-dimethylformamide, N-dimethylacetamide and tetrahydrofuran.
In the technical scheme, a substance P is prepared by reacting cisplatin and a nitrile substance in hydrochloric acid to generate a substance P'; uniformly dispersing the substance P' in an organic solvent, and adding silver nitrate for reaction to obtain a substance P; filtering the precipitate to obtain an organic solvent containing the substance P; the amount of nitrile is in excess relative to cisplatin, so that cisplatin is all involved in the reaction; substance P' and silver nitrate are in equimolar ratio.
In the technical scheme, the substance B is prepared by reacting the substance B2 with trifluoroacetic acid, and NaHCO is added after the reaction is finished3Until no bubbles are generated in the reaction solution, transferring the reaction solution to a separating funnel,with dichloromethane as solvent and saturated NaHCO3Washed 2 times and the dichloromethane layer was collected. Drying with anhydrous sodium sulfate, rotary steaming, and passing through column to obtain; the substance B2 is prepared from a substance B0 and a substance B1, O-benzotriazole-N, N, N ', N' -tetramethyluronium tetrafluoroborate and N, N-diisopropylethylamine are used as catalysts, and organic solvents are selected to provide a reaction environment for the substance B0 and the substance B1, such as N, N-dimethylformamide, N, N-dimethylacetamide and tetrahydrofuran;
catalyst O-benzotriazole-N, N, N ', N' -tetramethylurea tetrafluoroboric acid and N, N-diisopropylethylamine
And (3) reacting the substance B2 with trifluoroacetic acid, wherein the trifluoroacetic acid is excessive to ensure the deprotection effect of the terminal Boc, reacting at room temperature of 20-25 ℃ for 1-40 hours, and selecting inert protective gas to protect the reaction system.
When the substance B0 and the substance B1 react, the molar ratio of the substance B0 to the substance B1 is 1: (1-2) in excess of substance B1 to ensure that substance B0 is fully involved in the reaction; the reaction is carried out at room temperature of 20-25 ℃ for 1-40 hours, and inert protective gas is selected to protect the reaction system.
The substance B0 is obtained by commercial method or by reflux reaction of 2-chloronicotinic acid and 3-amino-2-naphthoic acid, and reflux is carried out under acidic condition, for example, hydrochloric acid is added after dissolving by ethanol, reflux is carried out for 20-72 h at room temperature of 20-25 ℃, and the molar ratio of 2-chloronicotinic acid to 3-amino-2-naphthoic acid is equal.
The application of the platinum compound in preparing antitumor drugs.
Compared with the prior art, the platinum compound has obviously better antitumor activity than classical platinum drugs such as cisplatin in cell lines such as HeLa, MCF-7, A549, NCI-H460, MDA-MB-231, HepG2 and the like. The toxicity of the platinum compound in normal cell systems such as MRC-5 and the like is obviously lower than that of classical platinum drugs such as cisplatin and the like. The platinum compound still keeps excellent anti-tumor activity in cisplatin-resistant cell lines such as A549/cis, NCI-H460/cis and the like. The invention can enrich in nucleolar region, inhibit DNA replication and global RNA transcription, inhibit the transcription of 47S pre-rRNA, target nucleolar or RNA polymerase I, and reduce the expression of corresponding proteins NCL and RPA 194.
Drawings
FIG. 1 is a NMR spectrum of B1 prepared in example of the present invention.
FIG. 2 is a NMR carbon spectrum of B1 prepared in example of the present invention.
FIG. 3 is a NMR spectrum of P1' prepared in example of the present invention.
FIG. 4 is a liquid phase purity detection profile of P1-B1 prepared in the examples of the present invention.
FIG. 5 is a NMR spectrum of P1-B1 prepared in example of the present invention.
FIG. 6 is a NMR carbon spectrum of P1-B1 prepared in an example of the invention.
FIG. 7 is a high resolution mass spectrum of P1-B1 prepared in an example of the invention.
FIG. 8 is a graph showing the results of tests on the effect of P1-B1 on DNA replication and global RNA transcription prepared in the examples of the present invention.
FIG. 9 is a graph showing the results of a test for the effect of P1-B1 on 47S pre-rRNA transcription, prepared in the example of the present invention.
FIG. 10 is a graphical representation of the results of a test of the effect of P1-B1 on RPA194 and NCL expression, prepared in an example of the present invention.
FIG. 11 is a graph showing the results of the γ -H2AX test on the effect of P1-B1 on RPA194, NCL and expression prepared in the examples of the present invention.
Detailed Description
The technical scheme of the invention is further explained by combining specific examples.
Instrument list
| Name (R) | Model number | Manufacturer of the product |
| Nuclear magnetic resonance tester | AVANCE III 400MHz | Bruker, Switzerland |
| High resolution mass spectrometry | Agilent 6520Q-TOF LC/MS | Agilent, USA |
| Electronic analytical balance | BP211D | Sartorius germany |
| High performance liquid chromatograph | SPD-20A | Shimadzu Japan Ltd |
| Laser confocal microscope | FV-1000 | Olympus et al, Japan |
List of raw materials and reagents
Example 1
Synthesis and characterization of monofunctional platinum antitumor compound P1B1 based on nucleolar stress
As shown in the above chemical formula, 1.68g (10.68mmol) of 2-chloronicotinic acid (substance 1 in the chemical formula) and 2g (10.68mmol) of 3-amino-2-naphthoic acid (substance 2 in the chemical formula) are weighed into a round-bottomed flask, 70mL of ethanol is added to dissolve, 0.9mL (29.13mmol) of hydrochloric acid (aqueous hydrogen chloride solution with concentration of 2mol/L) is added, the mixture is refluxed at room temperature of 20-25 ℃ for 72 hours, the progress of the reaction is monitored by thin layer chromatography, and after the reaction is almost completed, the mixture is left to stand and cool. And filtering the cooled reaction solution, taking the solid, washing the solid with absolute ethyl alcohol, and drying to obtain an orange solid. And (3) uniformly dispersing the obtained orange solid in ethanol, refining, refluxing at 80 ℃ for 2h, filtering, and drying to obtain a substance 3 in the chemical formula, wherein the substance is an orange solid in a macroscopic view.
100mg (0.344mmol) of substance 3 and 166mg (0.517mmol) of TBTU (O-benzotriazole-N, N, N ', N' -tetramethyluronium tetrafluoroborate) were weighed into a round-bottomed flask and 5mL of DMF was added, as shown in the above formula. mu.L DIPEA (N, N-diisopropylethylamine) was added with stirring, and after stirring for fifteen minutes at 20-25 ℃ 128mg (0.679mmol) of 3- (N-Boc-N-methylamino) propylamine were added, the system changed from an orange-red suspension to a deep-red clear solution. Stirring under argon gas at room temperature for 24 h. After the reaction is finished, pouring the reaction liquid into cold water to obtain orange-red suspension, filtering, washing with cold water, and drying filter residues to obtain an orange solid substance 4.
The solid 4 was dissolved in dichloromethane and 20% (volume percent) aqueous trifluoroacetic acid was added and stirred at room temperature for 2h and monitored by thin layer chromatography. After the reaction is finished, saturated NaHCO is added dropwise3Until no bubbles are generated, the reaction solution is transferred to a separating funnel and then dichloromethane is addedAlkane as solvent, saturated NaHCO3Washed 2 times and the dichloromethane layer was collected. Dried over anhydrous sodium sulfate, rotary evaporated, and passed through a column to give B1 as a yellow solid, 90.5mg, 75.2% yield.1HNMR(400MHz，CDCl3)δppm 11.31(s，1H),9.03(s，1H)，8.90(d，J＝7.2Hz，1H)，8.69(d，J＝6.8Hz，1H)，8.15(s，1H)，8.06(d,J＝8.4Hz,1H),7.98(d,J＝8.5Hz,1H),7.63(t,J＝7.5Hz,1H),7.52(t,J＝7.4Hz,1H),6.85(t,J＝7.0Hz,1H),3.69(q,J＝6.1Hz,2H),2.85(t,J＝6.5Hz,2H),2.54(s,3H),2.04-1.98(m,2H).13C NMR(101MHz,CDCl3)δppm163.01,159.58,145.70,141.31，140.07，137.61,131.03,129.79,129.62,129.55,129.27,127.69,126.22,125.97,123.53,116.48,115.60,110.86,49.93,37.98,36.79,30.01。
As shown in the above chemical formula, 60mg of cisplatin (0.2mmol) was weighed, added to 6mL of dilute hydrochloric acid having pH 4, and 2mL of acetonitrile was added to the solution, followed by thorough stirring. The solution was heated to reflux until the yellow suspension turned to a colorless clear solution (2 h in use). The solution was spin dried to give a yellow solid residue. The solid residue solution was taken up in 3mL of dry absolute methanol to give a yellow suspension. Filtering the solution through a microporous filter membrane, and slowly dripping the filtrate into 60mL of dry anhydrous ether to generate a large amount of white precipitate. The suspension was stirred well for 2h, filtered, the filter cake washed with a small amount of ether and dried in vacuo to give P1', 52mg as a white solid in 76.2% yield.1HNMR(400MHz,D2O,ppm)，δ2.55(s,3H)。
As shown in the above chemical formula, the above compound P1' (50mg,0.15mmol) was weighed, added to 2mL of dry DMF, and AgNO was added3(25mg,0.15mmol) and stirred for 30min in the dark, resulting in a white precipitate (silver chloride). The reaction was filtered and the filtrate was cooled to-10 ℃ (s.ding, x.qiao, g.l.kucera and u.bibach, C)humicommunications, 2013,49,2415-2417), yielding P1. Compound B1(60mg,0.17mmol) was weighed into 2mL of dry DMF solution and the solution was slowly added dropwise to the above DMF filtrate containing substance P1 and stirred under sealed conditions at-10 ℃ for 4 days. After the reaction, the reaction solution was slowly dropped into 40mL of dry anhydrous ether to produce an orange-yellow precipitate. And fully stirring the suspension for 24 hours, filtering, washing a filter cake by using a small amount of anhydrous ether, and drying to obtain a yellow solid. The resulting solid was recrystallized from a small amount of methanol to give P1-B1 as a yellow powder 70mg, 64.1% yield, 97% purity.1H NMR(400MHz,DMSO-d6)δppm 10.93(s,1H),9.05(s,1H),8.88(d,J＝7.0Hz,1H),8.48(d,J＝6.4Hz,1H),8.39(s,1H),8.26(d,J＝8.0Hz,1H),8.13(d,J＝8.1Hz,1H),7.70(t,J＝7.1Hz,1H),7.58(t,J＝7.1Hz,1H),7.03(t,J＝6.7Hz,1H),5.80(s,1H),4.87-4.51(m,1H),4.19(s,2H),3.91(s,2H),3.50(s,4H),3.01(s,3H),2.01-1.87(m,2H).13C NMR(101MHz,DMSO-d6)δppm 164.67,162.54,158.97,145.35,141.04,139.49,136.82,130.22,129.91,129.41,129.07,128.65,127.37,125.92,125.64,123.22,115.54,110.97,50.80,46.99,36.77,36.59.MS(ESI,positive-ion mode):m/z:calcd for C23H29ClN7O2Pt[M]+:665.1719；found:[M+1]+:666.1693。
Example 2 evaluation of antitumor Activity of P1-B1 and cisplatin
The MTT colorimetric method is one of the methods for detecting the growth and survival of cells in experimental research. In mitochondria of living cells, exogenous MTT can be reduced to formazan, a crystal substance which is purple blue and difficult to dissolve in water, by succinate dehydrogenase in cells, and formazan is deposited in cytoplasm of cells, but dead cells do not have the phenomenon. When formazan encounters DMSO, formazan deposited in cells will be dissolved by DMSO. Detecting formazan optical density value (OD value) at 570nm wavelength by enzyme labeling instrument to indirectly reflect the number of living cells, forming MTT crystal with positive correlation with the number of living cells, and processing data by Graphpad Prism 5 software to calculate IC50Value and plot, IC50Values refer to the half inhibitory concentration of the drug being measured on cell growth.
Experimental results show that the monofunctional platinum compound disclosed by the invention has obviously better antitumor activity than classical platinum drugs such as cisplatin in cell lines such as A549, HeLa, MCF-7, NCI-H460, MDA-MB-231, HepG2 and the like, as shown in the following table. The toxicity of the monofunctional platinum compound in normal cell systems such as MRC-5 and the like is obviously lower than that of classical platinum drugs such as cisplatin and the like. The monofunctional platinum compound still maintains excellent antitumor activity in cisplatin-resistant cell lines such as A549cisR, NCI-H460cisR and the like.
IC of P1-B1 and cisplatin in various cell lines at 72h50Value ofa
| Cell lines | CDDP | P1-B1(FIb) |
| A549 | 7.49±0.70 | 5.42±0.48(1.38) |
| A549cisR | 29.52±1.33 | 1.66±0.20(17.8) |
| RFc | 3.94 | 0.31 |
| NCI-H460 | 4.75±1.10 | 1.84±0.10(2.58) |
| NCI-H460cisR | 14.02±0.21 | 3.22±1.37(4.35) |
| RF | 2.95 | 1.75 |
| HeLa | 7.07±0.98 | 3.21±1.07(2.20) |
| MCF-7 | 8.63±1.39 | 0.93±0.30(9.27) |
| MRC-5 | 2.39±0.30 | 16.46±1.70(0.15) |
| SIe(MRC-5/A549) | 0.32 | 3.04 |
| SI(MRC-5/MCF-7) | 0.28 | 17.70 |
aIC50Values ± standard deviation (μ M) were obtained from a one-half dilution method with triplicate wells per group, repeated 1 time, and administered for 72 hours incubation to fit the dose-effect curve.bFI (multiple of growth) is IC50(CDDP) and IC50(P1B 1).cRF (drug resistance coefficient) is IC50(A549cisR) and IC50(A549) Or IC50(NCI-H460cisR) and IC50(NCI-H460).dND, not determined。eSI (selection index) is IC50(MRC-5) and IC50(A549) Or IC50(MRC-5) and IC50(MCF-7).
Example 3
Based on copper-catalyzed azido-alkynyl addition reaction (CuAAC reaction), thymidine analogue 5-ethynyl-2' -deoxyuridine is introduced into a system so as to solve the investigation of DNA replication level and further research the intracellular action mechanism of the monofunctional platinum compound. 5-ethynyl-2' -deoxyuridine (EdU) can be substituted for deoxythymine to be incorporated into a newly synthesized DNA double strand in the cell proliferation process, and because the EdU has an alkynyl group which can be labeled by using a CuAAC reaction, the EdU can be selectively labeled by using a fluorescent molecule modified by azido, and the intracellular DNA synthesis level and the S-phase cell proportion can be effectively detected. The transcription inhibition effect is an important mechanism of the anti-tumor effect of platinum drugs, similar to EdU, 5-ethynyl-2' -uridine (EU) is introduced, and based on CuAAC reaction, Alexa Fluor 647-azide dye is used for labeling by a fluorescence post-labeling method to monitor the global transcription level in cells.
Experimental results show that the monofunctional platinum compound of the present invention can inhibit DNA replication and global RNA transcription, as shown in FIG. 8. The red fluorescence generated in the cells due to the binding of EdU involved in DNA replication and Alexa Fluor 647-azide shows large difference in different cells, which is closely related to the cell cycle state of the cells: in S phase cells with active DNA replication, due to the existence of a large amount of newly synthesized DNA, a large amount of EdU is incorporated into a newly synthesized DNA chain, so that the cells can be marked by Alexa Fluor 647-azide to show remarkable red fluorescence; in the G0/Gl phase cells at the preparation stage of replication or the G2/M phase cells after completion of replication, red fluorescence cannot be observed because EdU cannot participate in the DNA double strand due to the absence of DNA replication. In the treated cells, the percentage of cells stained with red fluorescence was significantly reduced, indicating that cisplatin, BMH-21, B1, and P1-B1 all inhibited DNA replication (EdU data in the figure). During RNA transcription, EU is incorporated into the newly synthesized RNA strand, marked by Alexa Fluor 647-azide and exhibits significant red fluorescence. The cells in the control group showed high brightness in the nucleolus region and dark red fluorescence in other regions of the nucleus, indicating that the rRNA transcription is more active. In the cells of the treated group, the red fluorescence was reduced to different degrees, and it is noted that the nucleolar region of the cells of BMH-21, B1 and P1-B1 treated group has almost no red fluorescence, and other regions of the cell nucleus have no obvious change compared with the control group, which indicates that BMH-21, B1 and P1-B1 selectively inhibit the transcription of rRNA, but cisplatin does not have the effect (shown in the EdU data). P1-B1 overcomes the defect of cisplatin resistance and is related to selective inhibition of rRNA transcription.
Example 4
FISH (fluorescence in hybridization) technology is an important nonradioactive in situ hybridization technique. The basic principle is as follows: if the target DNA on the chromosome or DNA fiber section to be detected is homologously complementary to the nucleic acid probe used, the target DNA and the nucleic acid probe are denatured, annealed and renatured to form a hybrid of the target DNA and the nucleic acid probe. We labeled 47S pre-rRNA 5-external transformed spacer (5 ' -ETS) and internal transformed spacer 2(ITS2) with Cy5 and Cy3, respectively, and then performed FISH experiments (C.Carron, M.F.O ' Donohue, V.Choesmel, M.Faubladier and P.E.Gleizes, Nucleic acids research,2011,39, 280-; D.Drygin, A.Siddiqui-Jain, S.O ' Brien, M.Schwaebe, A.Lin, J.Bliesage, C.B.Ho, C.Proffit, K.Trent, J.P.P.initten, J.K.Lim, D.Von Hoff, K.K.and R.69, Rirese, G1, 765, 3).
Experimental results show that the monofunctional platinum compound can inhibit the transcription of 47S pre-rRNA, and as shown in figure 9, 47S pre-rRNA is marked as red and green fluorescence in cells of a control group; among the cells of the treated group, the 47S pre-rRNA of the cells treated by the cisplatin exhibited red and green fluorescence with almost the same intensity as that of the cells of the control group, indicating that the cisplatin had no significant effect on the generation of the 47S pre-rRNA; the red and green fluorescence of 47S pre-rRNA of BMH-21, B1 and P1-B1 treated cells was significantly darkened or even disappeared, indicating that the transcription of 47Spre-rRNA was significantly inhibited by BMH-21, B1 and P1-B1.
Example 5
We examined the effect of monofunctional platinum compounds of the invention on the expression of nucleolin RPA194, NCL and the DNA damage-associated protein gamma-H2 AX using Western blotting (Western Blot) and Immunofluorescence (IF) experiments. RPA194 is the catalytic subunit of RNA Pol I, which plays an important role in rDNA transcription, NCL is nucleolin, which is widely distributed in nucleoli and involved in the biosynthesis of ribosomes, and gamma-H2 AX is a marker protein for DNA damage.
The monofunctional platinum compounds of the present invention can target nucleolus or RNA polymerase I, reduce the expression of the corresponding proteins NCL and RPA194, and up-regulate γ -H2AX, as shown in FIGS. 10-11. Western blot experiment was performed to examine the effect of different concentrations of cisplatin, BMH-21 and P1-B1 on the expression of RPA194 and NCL in cells. BMH-21 and P1-B1 can effectively reduce the expression of RPA194 and NCL, and cisplatin has little influence on the expression of RPA194 and NCL.
The cells were stained with a green secondary antibody labeled with Alexa Fluor 488 by immunofluorescence, and the expression level and positional change of RPA194 and NCL in the cells were detected to determine the presence or absence of nucleolar stress. The experimental results show that the expression levels and positions of RPA194 and NCL of cells treated by cisplatin are not obviously changed compared with those of cells of a control group, and the expression levels and positions of RPA194 and NCL of cells treated by BMH-21 and P1-B1 are obviously changed. Wherein RPA194 is condensed into a highlight dot-shaped area from the dispersion of the control group; the position of NCL varies significantly, spreading from the nucleolar region throughout the nucleus, indicating that P1-B1, unlike cisplatin, produces a typical nucleolar stress response. Staining with a secondary antibody labeled with red Alexa Fluor 594, detecting the expression level of gamma-H2 AX in cells, and further detecting whether DNA double strand break damage exists. The results showed that there was almost no expression of γ -H2AX in the control and BMH-21 treated cells; in cells treated by cisplatin and P1-B1, the cell nucleus is stained by Alexa Fluor 594 and shows red fluorescence, and gamma-H2 AX is expressed, which indicates that P1-B1 can generate DNA damage, double-strand breaks of cell DNA and has the effect characteristic of platinum drugs.
The preparation of the platinum compound can be realized by adjusting the process parameters according to the content of the invention, and the platinum compound has anticancer performance basically consistent with the embodiment. The invention has been described in an illustrative manner, and it is to be understood that any simple variations, modifications or other equivalent changes which can be made by one skilled in the art without departing from the spirit of the invention fall within the scope of the invention.
Claims (8)
1. The nucleolar stress-based platinum compound is characterized by having a structure shown in the following chemical formula: wherein a divalent metal platinum (Pt) is used as a center, and two NH groups3One Cl-And side chain NH to form coordination, so that the whole is positively charged and forms platinum compounds with nitrate radical with negative charge, R1Is methyl or ethyl, R2Is methyl or ethyl, L is a chain group with 2-4 carbons
2. A nucleolar stress-based platinum-based compound according to claim 1, characterised in that R1Is methyl, R2Is methyl, L is a chain radical having 3 carbons, i.e., - (CH)2)3—。
3. The preparation method of the nucleolar stress-based platinum compound is characterized in that the nucleolar stress-based platinum compound is obtained by reacting a substance B with a substance P, and the molar ratio of the substance B to the substance P is (1-1.2): 1, the reaction temperature is minus 20 to minus 4 ℃;
4. The method for preparing a nucleolar stress-based platinum compound according to claim 3, wherein the reaction temperature is-10 to 0 ℃, the reaction time is 20 to 100 hours, a closed environment is adopted in the reaction process, magnetic stirring is selected, 100 to 300 revolutions per minute is performed, and an organic solvent is selected to provide a reaction environment for the substance B and the substance P.
5. The method for preparing a nucleolar stress-based platinum compound as claimed in claim 4, wherein the reaction time is 60 to 80 hours, and the organic solvent is N, N-dimethylformamide, N-dimethylacetamide, or tetrahydrofuran.
6. A method of preparing a nucleolar stress-based platinum-based compound according to claim 3, wherein substance P is formed by reacting cisplatin with a nitrile in hydrochloric acid; uniformly dispersing the substance P' in an organic solvent, and adding silver nitrate for reaction to obtain a substance P;
7. Use of a nucleolar stress-based platinum compound according to claim 1 for the preparation of an anti-tumour medicament.
8. Use of nucleolar stress-based platinum compounds for the preparation of antitumoral drugs according to claim 7, characterized by enrichment of the nucleolar region and inhibition of DNA replication and global RNA transcription, inhibition of the transcription of 47S pre-rRNA, targeting nucleolar or RNA polymerase I, reducing the expression of the corresponding proteins NCL and RPA 194.
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| Alkaline Phosphatase Inhibition by a Series of Pyrido[2,l-b]quinazolines: A Possible Relationship with Cromolyn-like Antiallergy Activity;Charles F. Schwender等;《J. Med. Chem.》;19821231;第25卷(第6期);第742-745页 * |
| Design, Synthesis, and Structure−Activity Relationships of Pyridoquinazolinecarboxamides as RNA Polymerase I Inhibitors;Laureen Colis等;《J. Med. Chem.》;20140521;第57卷;第4950-4961页 * |
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