CN104910196A - Chiral tetranuclear copper-based organic metal framework, and preparation method and applications thereof - Google Patents

Chiral tetranuclear copper-based organic metal framework, and preparation method and applications thereof Download PDF

Info

Publication number
CN104910196A
CN104910196A CN201510272482.0A CN201510272482A CN104910196A CN 104910196 A CN104910196 A CN 104910196A CN 201510272482 A CN201510272482 A CN 201510272482A CN 104910196 A CN104910196 A CN 104910196A
Authority
CN
China
Prior art keywords
tnsbcu
metal framework
organic metal
chirality
copper base
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.)
Granted
Application number
CN201510272482.0A
Other languages
Chinese (zh)
Other versions
CN104910196B (en
Inventor
刘杰
秦秀英
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jinan University
Original Assignee
Jinan University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Jinan University filed Critical Jinan University
Priority to CN201510272482.0A priority Critical patent/CN104910196B/en
Publication of CN104910196A publication Critical patent/CN104910196A/en
Application granted granted Critical
Publication of CN104910196B publication Critical patent/CN104910196B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F1/00—Compounds containing elements of Groups 1 or 11 of the Periodic Table
    • C07F1/005—Compounds containing elements of Groups 1 or 11 of the Periodic Table without C-Metal linkages
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Landscapes

  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Plant Pathology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention belongs to the technical field of metal organic material preparation, and specifically discloses a chiral tetranuclear copper-based organic metal framework, and a preparation method and applications thereof. The structural formula of the chiral tetranuclear copper-based organic metal framework is [Cu4(C17H16NO3Br)2(C17H17NO3Br)2(H2O)2]2(NO3)4.3(H2O), wherein C17H16NO3Br and C17H17NO3Br are anion ligands. The chiral tetranuclear copper-based organic metal framework is a candidate of antitumor drugs, an excellent non-viral gene vector, and is a novel multifunctional antitumor drug candidate.

Description

A kind of chirality four core copper base organic metal framework and its preparation method and application
Technical field
The present invention relates to metallo organic material preparing technical field, be specifically related to a kind of chirality four core copper base organic metal framework and its preparation method and application.
Background technology
For a long time, tumour is still one of death main in world wide.Although existing more ripe methods for the treatment of, but very eurypalynous tumour lacks early symptom, shows as different clinical indication, simultaneously insensitive to radiation and chemotherapy, especially many chemotherapeutics can damage the immunity system of body, can produce toxic side effect and resistance to patient.Particularly tumour occurs to middle and advanced stage, and often occur that the speed of antineoplaston is unable to catch up with the speed of metastases diffusion far away, therefore, antineoplaston is faced with stern challenge always.For reaching good oncotherapy effect, novel effective antitumour medicine and combination therapy strategy extremely urgent.
The growth of tumour, deterioration, invasion and m etastasis and new vascular generation are closely related.Malignant tumour can derive the vascular system of self and obtain nutrition, and tumour cell promotes again tumor vascular generation, this means that tumor vessel and tumour cell play an effect of mutually promoting in the growth of tumour after obtaining nutrition.In addition, when tumour occurs, vascular endothelial growth factor (VEGF) is overexpression, suppresses or lowers the expression of VEGF, just can close " switch " of tumor-blood-vessel growth, the growth of Tumor suppression.So, VEGF, tumor vessel and tumour cell become antineoplastic important target spot, and these important target spots all interact, connect each other, this makes the antitumor drug of single target spot be difficult to reach best result for the treatment of, therefore grind and obtain the medicine with Mutiple Targets antitumor action and become grand strategy target, also will administration and personalised drug development provide new thinking as required for following antitumor drug.
Metallic organic framework (Metal-Organic Frameworks MOFs) utilizes multiple tooth organic ligand and metal ion containing oxygen, nitrogen to pass through freely to assemble the coordination chemistry thing with periodic network structure formed.In recent years, because MOFs can show regulatable porousness and abnormal high-ratio surface, multiple fields of biological medicine can be widely used in, as the encapsulation of medicine, transmission, transport and release, even utilize MOFs to achieve gene therapy disease.Research shows, RNA disturbs the expression of reticent VEGF to be proved and inhibits tumor-blood-vessel growth and hinder the growth of tumour, in addition, utilize MOFs jointly to deliver and discharge anti-tumor predrug and the siRNA relevant to tumour achieves combination therapy, enhancing antitumous effect.Combined action is antitumor is a new and effective antitumor strategy, is an important breakthrough in oncotherapy research.
Copper is a vital trace element in all organisms, and copper complex is effective cytotoxic drug.Copper activity compound is different from the platinum medicine of current use in mechanism of action, bio distribution and cytotoxicity, and they resist those chemosensitivities difference or traditional platinum medicine to create drug-fast cancer be effective, are effective at least substantially.But finding no so far and closing copper basigamy compound self, as antitumor drug, and can be not only excellent siRNA carrier, target vascular therapy generates, vascular endothelial cell and tumour cell, the report of powerful antitumous effect.
Summary of the invention
Technical problem to be solved by this invention is, provides a kind of and self not only can be used as antitumor drug but also can be used as the chirality four core copper base organic metal framework of carrier.
Technical problem to be solved by this invention, is achieved by the following technical programs:
A kind of chirality four core copper base organic metal framework, its structural formula is [Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o), wherein C 17h 16nO 3br and C 17h 17nO 3br is anion ligand; Described chirality four core copper base organic metal framework exists with crystalline form, belongs to oblique system, C2 spacer; Shown in its single crystal structure figure structure as arbitrary in Fig. 2.
Preferably,
Described structural formula is L-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o), L-TNSBCu is abbreviated as;
Or D-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o), D-TNSBCu is abbreviated as;
Wherein L-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o) crystalline structure belongs to oblique system, C2 spacer, β=108.740 °; z=2; D-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o) crystalline structure belongs to oblique system, C2 spacer, β=108.747 °; z=2.
The preparation method of described chirality four core copper base organic metal framework, comprises following steps:
S1. by phenylalaninol and highly basic mixing, add organic solvent stirring and dissolving, then add 5-bromo-2-hydroxy 3-methoxybenzene formaldehyde, constant temperature stirs 1 ~ 4h, and stopped reaction, obtains part SB, i.e. Na 2(C 17h 16nO 3and Na (C Br) 17h 17nO 3br);
S2. by part SB and soluble copper salt mixing, be dissolved in ethanol, then add many pyridines, isothermal reaction 8 ~ 24h, after reaction terminates, obtain chirality four core copper base organic metal framework.
Further, by the chirality four core copper base organic metal framework obtained in water-soluble or aqueous ethanolic solution, nano level chirality four core copper base organic metal framework can be obtained.
Preferably, the mol ratio of the phenylalaninol described in S1., highly basic and 5-bromo-2-hydroxy 3-methoxybenzene formaldehyde is 1 ~ 2:1 ~ 3:1 ~ 2.
Most preferably, the mol ratio of the phenylalaninol described in S1., highly basic and 5-bromo-2-hydroxy 3-methoxybenzene formaldehyde is 1:2:1.
Preferably, the phenylalaninol described in S1. is the phenylalaninol of L-phenylalaninol, D-phenylalaninol or non-resolving chiral.
Preferably, the organic solvent described in S1. is absolute alcohol, and thermostat temperature is 45 ~ 60 DEG C.
Most preferably, the organic solvent described in S1. is anhydrous methanol or dehydrated alcohol, and thermostat temperature is 50 DEG C, and constant temperature time is 2.5h.
Preferably, the mol ratio of the part SB described in S2., soluble copper salt and many pyridines is 1 ~ 3:2 ~ 6:1 ~ 2.
Most preferably, the mol ratio of the part SB described in S2., soluble copper salt and many pyridines is 2:5:2.
Preferably, the mantoquita described in S2. is cupric nitrate; Described many pyridines are preferably phenanthroline, dipyridyl, or do not add pyridine.
Preferably, the ethanol described in S2. to be volume fraction be 95% ethanol.
Preferably, the thermostat temperature described in S2. is 55 DEG C, and the reaction times is 12h.
Described chirality four core copper base organic metal framework is preparing the application in antitumor drug.
Described chirality four core copper base organic metal framework is as the application of angiogenesis inhibitor.
Described chirality four core copper base organic metal framework is as the application of antitumor drug or genophore.
Preferably, described chirality four core copper base organic metal framework is as the application of the carrier of VEGF-siRNA.
Further preferably, described chirality four core copper base organic metal framework and VEGF-siRNA mass ratio are 1 ~ 100:1.
Most preferably, L – TNSBCu and VEGF-siRNA mass ratio are 5 ~ 10:1; D – TNSBCu and VEGF-siRNA mass ratio are 20 ~ 40:1.
Beneficial effect: (1) chirality four of the present invention core copper base organic metal framework has the ability of inhibition tumor cell propagation significantly, and suppresses vascular endothelial cell proliferation effect; (2) L-TNSBCu and the D – TNSBCu of chirality of the present invention has the cytotoxicity of broad-spectrum to tumour cell, and has chiral selectivity to the toxicity of some tumour cell, large to cytotoxicity than D-type of L-type; (3) L-TNSBCu and D-TNSBCu of the chirality described in has the ability of supported V EGF-siRNA, and the VEGF-siRNA carrier loaded by L-TNSBCu and D-TNSBCu can from RNAse enzymolysis.Therefore, L-TNSBCu and D – TNSBCu of the present invention is the candidate of antitumor drug, is again good non-viral gene vector, is a kind of novel multi-functional antitumor drug candidate.
Accompanying drawing explanation
Fig. 1 is chirality four core copper base organic metal framework scanning electron microscope (SEM) photograph of the present invention; Wherein (a) scanning electron microscope (SEM) photograph that is L-TNSBCu; B scanning electron microscope (SEM) photograph that () is D-TNSBCu.
Fig. 2 is chirality four core copper base organic metal framework single crystal structure figure of the present invention; Wherein (a) single crystal structure figure that is L-TNSBCu; B single crystal structure figure that () is D-TNSBCu.
Fig. 3 is chirality four core copper base organic metal framework transmission electron microscope picture of the present invention; Wherein (a) and (b) transmission electron microscope picture that is L-TNSBCu; Wherein (c) and (d) transmission electron microscope picture that is D-TNSBCu.
Fig. 4 is the sepharose gel electrophoresis experiment of L-TNSBCu or D-TNSBCu; Wherein (a) is the electrophoresis experiment figure detecting L-TNSBCu or D-TNSBCu supported V EGF-siRNA; B electrophoresis experiment figure that () is degraded from RNAse for detecting L-TNSBCu or D-TNSBCu supported V EGF-siRNA; C () discharges the electrophoresis experiment figure of VEGF-siRNA for detecting L-TNSBCu or D-TNSBCu.
To be chirality four core copper base organo-metallic frame of the present invention to sprout restraining effect result figure to mouse arterial ring Fig. 5.
Embodiment
Explain the present invention further below in conjunction with specific embodiment, but embodiment does not limit in any form to the present invention.
The preparation of embodiment 1 L-TNSBCu
(1) L-phenylalaninol (5.0mmol) and potassium hydroxide (10.0mmol) are placed in the ground flask of 100mL, add the anhydrous methanol of 25mL, reflux condensation mode, and 50 DEG C of constant temperature, magnetic agitation are dissolved.Then, add 5-bromo-2-hydroxy 3-methoxybenzene formaldehyde (5.0mmol), continue to stir at 50 DEG C of constant temperature, reflux condensation mode, reaction 2.5h, solution is in yellow.Stopped reaction, steams most anhydrous methanol with Rotary Evaporators, obtains yellow solid, filters.The a small amount of anhydrous methanol of solid is washed, anhydrous diethyl ether washing, then vacuum-drying 2 days, obtains yellow solid, i.e. the schiff base ligand (L-SB) of L-type.
(2) in the round-bottomed flask of 100mL, by ligand L-SB (0.2mmol) and Cu (NO 3) 23H 2o (0.3mmol) mixes, with the dissolve with ethanol of 17mL 95%, reflux condensation mode, 50 DEG C of constant temperature, magnetic agitation are dissolved, after reaction 15min, by the dissolve with ethanol of phenanthroline (0.2mmol) with 2mL 95%, join in above-mentioned solution, continue 55 DEG C of constant temperature, magnetic agitation, back flow reaction 12h.After reaction terminates, filter.Bottle-green filtrate is left standstill, naturally volatilizees.Behind some skies, obtain the green monocrystalline of hexagonal plate pattern, i.e. L-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o) (L-TNSBCu is abbreviated as).Its scanning electron microscope (SEM) photograph is as shown in Fig. 1 (a).
L-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o) crystalline structure belongs to oblique system, C2 spacer, β=108.740 °; its single crystal structure figure is as shown in Fig. 2 (a).
The preparation of embodiment 2 D-TNSBCu
(1) D-phenylalaninol (5.0mmol) and potassium hydroxide (10.0mmol) are placed in the ground flask of 100mL, add the anhydrous methanol of 10mL, reflux condensation mode, and 50 DEG C of constant temperature, magnetic agitation are dissolved.Then, add 5-bromo-2-hydroxy 3-methoxybenzene formaldehyde (5.0mmol), have yellow mercury oxide at once, continuing to stir at 50 DEG C of constant temperature, reflux condensation mode, there is a large amount of yellow mercury oxide in reaction 2.5h.Stopped reaction, steams a small amount of anhydrous methanol with Rotary Evaporators, obtains yellow solid, filters.The a small amount of anhydrous methanol of solid is washed, anhydrous diethyl ether washing, then vacuum-drying 2 days, obtains yellow solid, i.e. the schiff base ligand (D-SB) of D-type,
(2) in the round-bottomed flask of 100mL, by part D-SB (0.2mmol) and Cu (NO 3) 23H 2o (0.3mmol) mixes, with the dissolve with ethanol of 15mL 95%, and reflux condensation mode, 50 DEG C of constant temperature, magnetic agitation are dissolved, after reaction 30min, phenanthroline (0.2mmol) is joined in above-mentioned solution, continues 55 DEG C of constant temperature, magnetic agitation, back flow reaction 12h.After reaction terminates, filter.Bottle-green filtrate is left standstill, naturally volatilizees.Behind some skies, obtain the green monocrystalline of hexagonal plate pattern, i.e. D-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o) (D-TNSBCu is abbreviated as).Its scanning electron microscope (SEM) photograph is as shown in Fig. 1 (b).
D-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o) crystalline structure belongs to oblique system, C2 spacer, β=108.747 °; its single crystal structure figure is as shown in Fig. 2 (b).
The preparation of embodiment 3 nano level L-TNSBCu and D-TNSBCu monocrystalline
Monocrystalline second alcohol and water (3:2) mixed solvent of L-TNSBCu and D-TNSBCu is fully dissolved, is prepared into saturated solution, is placed on copper mesh lightly, naturally dries, obtain nano level L-TNSBCu and D-TNSBCu.With the pattern of L-TNSBCu and D-TNSBCu nanoparticle on transmission electron microscope observing copper mesh.See Fig. 3 (b) respectively, shown in 3 (d).
The preparation of embodiment 4 nano level L-TNSBCu and D-TNSBCu monocrystalline
Solvent is made with water, respectively the monocrystalline distilled water of L-TNSBCu and D-TNSBCu is dissolved, be prepared into supersaturated solution, after standing sedimentation, get supernatant liquid 10 μ L, then add with 5 μ L distilled water diluting mixings, hold over night, supernatant liquid 15 μ L then got by D-TNSBCu sample, and hold over night obtains nano level L-TNSBCu and D-TNSBCu monocrystalline.See Fig. 3 (a) respectively, shown in 3 (c).
The preparation of embodiment 5 nano-complex L-TNSBCu/VEGF-SiRNA
(1) the DEPC water getting the VEGF-SiRNA 20 μ L of 4 μ L 20 μMs dilutes.
(2) title complex L-TNSBCu ethanol/water (volume ratio is 3:2) makes saturated solution.
(3) get the VEGF-SiRNA solution that the saturated L-TNSBCu solution of 5 μ L and 10 μ L dilute, mix, 37 ° of constant-temperature incubations 3h, drips on copper mesh and dries, obtain L-TNSBCu/VEGF-SiRNA.
The preparation of embodiment 6 nano-complex D-TNSBCu/VEGF-SiRNA
(1) the DEPC water getting the VEGF-SiRNA 20 μ L of 4 μ L 20 μMs dilutes.
(2) title complex D-TNSBCu ethanol/water (volume ratio is 3:2) makes saturated solution.
(3) get the VEGF-SiRNA solution that the saturated D-TNSBCu solution of 5 μ L and 10 μ L dilute, mix, 37 ° of constant-temperature incubations 3h, drips on copper mesh and dries, obtain D-TNSBCu/VEGF-SiRNA.
Embodiment 7 agarose gel electrophoresis is tested
Prepare L-TNSBCu/VEGF-siRNA and D-TNSBCu/VEGF-siRNA mixture respectively: first respectively L-TNSBCu and D-TNSBCu is dissolved with distilled water, form saturated solution 1ug/ μ L, the stock solution of the preparation 0.5 μ g/ μ L got; To specifications by the solution of VEGF-siRNA with DEPC water dilution about 0.264 μ g/ μ L (20 μMs); L – TNSBCu/VEGF-siRNA mixture and D – TNSBCu/VEGF-siRNA mixture is prepared respectively according to mass ratio (L/D – TNSBCu:VEGF-siRNA) 80:1,40:1,20:1,10:1,5:1,2.5:1, with siRNA and Naked-siRNA for reference, the cumulative volume 20 μ L of each sample, then at 37 DEG C of constant-temperature incubation 2.5h.Then carry out agarose gel electrophoresis, adjust voltage 100V, electrophoresis is about 20min.The results are shown in Figure 4.
If Fig. 4 (a) is the electrophoresis experiment figure detecting L-TNSBCu or D-TNSBCu supported V EGF-siRNA.By finding out in Fig. 4 (a) that two reference siRNA and Naked-siRNA are after electrophoresis, band is clear, bright, the VEGF-siRNA of hatching without L-TNSBCu or D-TNSBCu is described, migration amount in electrophoresis does not change, and after VEGF-siRNA is hatched by L-TNSBCu or D-TNSBCu respectively, the band of VEGF-siRNA, the mass ratio different according to VEGF-siRNA and L-TNSBCu or D-TNSBCu, there is band brightness deterioration, even without phenomenon, which illustrate the ability that L-TNSBCu and D-TNSBCu has supported V EGF-siRNA to some extent, enable in electrophoresis the VEGF-siRNA of free migration reduce to some extent.
The electrophoresis experiment figure that Fig. 4 (b) degrades from RNAse for detecting L-TNSBCu or D-TNSBCu supported V EGF-siRNA.Concrete grammar is with reference to above-mentioned electrophoresis method, and it is that the RNAse solution A 2 μ L of 0.25mg/mL carries out electrophoresis experiment that each sample adds concentration again.As can be seen from Fig. 4 (b), only have reference siRNA after electrophoresis, have clear, bright VEGF-siRNA band to occur, all there is not any VEGF-siRNA band in remaining loading hole, illustrate those not by the carrier loaded VEGF-siRNA of L-TNSBCu or D-TNSBCu all by RNAse enzymolysis so there is not VEGF-siRNA band.In addition, DNAse A also can make not carrier loaded by L-TNSBCu or D-TNSBCu VEGF-siRNA enzymolysis.
Fig. 4 (c) discharges the electrophoresis experiment figure of VEGF-siRNA for detecting L-TNSBCu or D-TNSBCu, concrete grammar is with reference to above-mentioned electrophoresis method, DNAse A with after excessive EDTA inactivation, each sample add again 1.0% SDS 2.0 μ L and 5 × PBS solution 5uL carry out electrophoresis experiment.The result of Fig. 4 (c) shows, for L – TNSBCu, when the mass ratio of L – TNSBCu and VEGF-siRNA is between 5 ~ 10, best carrying capacity is had to VEGF-siRNA because this scope is in L – TNSBCu, the amount of the VEGF-siRNA of load is maximum, and the amount of the VEGF-siRNA be therefore finally released is also maximum; For D – TNSBCu, when the mass ratio of D – TNSBCu and VEGF-siRNA is between 20 ~ 40, best carrying capacity is had to VEGF-siRNA because this scope is in D – TNSBCu, the amount of the VEGF-siRNA of load is maximum, and the amount of the VEGF-siRNA be therefore finally released is also maximum.
L-TNSBCu and D-TNSBCu that the present embodiment indicates chirality has the ability of supported V EGF-siRNA, and the best combination of L – TNSBCu supported V EGF-siRNA is than being 5:1 (mass ratio); The best combination of D – TNSBCu supported V EGF-siRNA is than being 20:1 (mass ratio); The VEGF-siRNA carrier loaded by L-TNSBCu and D-TNSBCu can from RNAse enzymolysis.
Embodiment 8 L – TNSBCu and the experiment of D – TNSBCu vitro cytotoxicity
Mtt assay: get the tumour cell being in logarithmic phase, adjustment viable cell concentrations is 2 × 10 4/ ml is added on 96 well culture plates, and every hole 100 μ l, cultivates 24h after adherent in incubator, then adds different concns given the test agent 100 μ l respectively, and application of sample group establishes 4 multiple holes, puts 37 DEG C, 5%CO 2cultivate 48h, then add MTT (5mg/ml) 20 μ l/ hole, centrifugally after 4h abandon supernatant liquor, add dimethyl sulfoxide (DMSO) (DMSO) 100 μ l/ hole, vibration about 10min, measures OD value by microplate reader under 570nm wavelength.Calculate cell survival rate, by its half-inhibition concentration of computed in software IC 50.
The OD value of the actual OD value/negative control hole of cell survival rate (%)=medicine feeding hole;
Cell inhibitory rate (%)=100%-cell survival rate;
The ability of different cell proliferation is suppressed with MTT technique study L – TNSBCu and D – TNSBCu.Selected cell has gastric carcinoma cells (MGC803), human liver cancer cell (HepG2), human cervical carcinoma cell (HeLa), human pancreatic cancer cell (Canpan-2), human lung carcinoma cell (A549), Human umbilical vein endothelial cells (HUVECs).Analyze from table 1, L – TNSBCu and D – TNSBCu is to test cell MGC803, the toxicity of Canpan-2 and A549R has significant chiral selectivity, namely L – TNSBCu is larger than the toxicity of D – TNSBCu, HeLa cell is had to the chiral selectivity of certain degree, and significant chiral selectivity is not had to HepG2 cell and vascular endothelial cell.It should be noted that especially, L – TNSBCu and D – TNSBCu has the effect of significant Inhibit proliferaton to HUVECs cell, and HUVECs cell plays important role in tumor vascular generation, this illustrates that L – TNSBCu and D – TNSBCu can have very large impact to the generation of blood vessel to their restraining effect.Therefore, L – TNSBCu of the present invention and D – TNSBCu all have suppress significantly survey the ability of tumor cell proliferation, and to vascular endothelial cell, also there is very strong Inhibit proliferaton effect.
Table 1.L – TNSBCu and D – TNSBCu is to the IC of tumour cell 50value
The generation Inhibition test of embodiment 9 mouse artery microtubule
Experimental technique is as follows:
(1) the matrigel ECGM substratum (another supplement is containing the VEGF of 20ng/mL) of high density is diluted one times, and then the micropore middle berth of 96 orifice plates of precooling enters the matrigel of 60 μ L dilutions, (CO was avoided in attention in 25 minutes to hatch solidification at 37 DEG C 2ventilating pit).
(2) aseptically, the thorax artery of SD rat in the ECGM substratum of serum-free, by periphery fat and reticular tissue totally disallowable, the section of cutting into 1.0 ~ 2.0mm, puts down gently in matrigel.
(3) cover thorax artery ring with the matrigel that 60 μ L dilute again, hatch solidification 25 minutes at 37 DEG C;
(4) ECGM substratum 100 μ L is added, every sample adds VEGF makes concentration be 40ng/mL, in medicine group, add L-TNSBCu2.0 μM and L-TNSBCu/VEGF-siRNA2.0 μM simultaneously respectively, cultivate in cultivation, within every 2 days, change the nutrient solution of an identical component.4d, takes pictures under inverted microscope 4 × object lens, record analysis.
The result of experiment as shown in Figure 5, 4d observes, there is a large amount of microtubules significantly in VEGF (40ng/mL) group, present a quick growth tendency, relative to VEGF group, the arterial ring that medicine group [VEGF (40ng/mL)+L-TNSBCu (2.0 μMs)] processed, few a lot of obviously than VEGF group of the growing amount of microtubule, and the arterial ring that medicine group [VEGFF (40ng/mL)+2.0 μMs of L-TNSBCu/VEGF-siRNA] processed, the generation of microtubule is inhibit more significantly than the arterial ring of L-TNSBCu process, this demonstrate the effect that carrier L-TNSBCu self has angiogenesis inhibiting significantly, and when its load after VEGF-siRNA, because VEGF-siRNA is to the reticent effect of VEGF gene, inhibit the expression of VEGF, make it L-TNSBCu and VEGF-siRNA and create synergy, enhance the effect of L-TNSBCu angiogenesis inhibiting.

Claims (10)

1. a chirality four core copper base organic metal framework, is characterized in that, its structural formula is [Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o), molecular weight is 3784.25; Wherein C 17h 16nO 3br and C 17h 17nO 3br is anion ligand; Described chirality four core copper base organic metal framework exists with crystalline form, belongs to oblique system, C2 spacer.
2. chirality four core copper base organic metal framework according to claim 1, it is characterized in that, described structural formula is L-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2or D-[Cu O) 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o); Wherein L-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o) crystalline structure belongs to oblique system, C2 spacer, β=108.740 °; z=2; D-[Cu 4(C 17h 16nO 3br) 2(C 17h 17nO 3br) 2(H 2o) 2] 2(NO 3) 43 (H 2o) crystalline structure belongs to oblique system, C2 spacer, β=108.747 °; z=2.
3. the preparation method of the chirality four core copper base organic metal framework described in claim 1 or 2, is characterized in that, comprise following steps:
S1. by phenylalaninol and highly basic mixing, add organic solvent stirring and dissolving, then add 5-bromo-2-hydroxy 3-methoxybenzene formaldehyde, constant temperature stirs 1 ~ 4h, and stopped reaction, obtains part SB, i.e. Na 2(C 17h 16nO 3and Na (C Br) 17h 17nO 3br);
S2. by part SB and soluble copper salt mixing, be dissolved in ethanol, then add many pyridines, isothermal reaction 8 ~ 24h, after reaction terminates, obtain chirality four core copper base organic metal framework.
4. preparation method according to claim 3, is characterized in that, by the chirality four core copper base organic metal framework that obtains in water-soluble or aqueous ethanolic solution, obtains nano level chirality four core copper base organic metal framework.
5. preparation method according to claim 3, is characterized in that, the mol ratio of the phenylalaninol described in S1., highly basic and 5-bromo-2-hydroxy 3-methoxybenzene formaldehyde is 1 ~ 2:1 ~ 3:1 ~ 2; S1. the phenylalaninol described in is the phenylalaninol of L-phenylalaninol, D-phenylalaninol or non-resolving chiral.
6. preparation method according to claim 3, is characterized in that, the mol ratio of the part SB described in S2., soluble copper salt and many pyridines is 1 ~ 3:2 ~ 6:1 ~ 2.
7. preparation method according to claim 3, is characterized in that, the mantoquita described in S2. is cupric nitrate; Described many pyridines are preferably phenanthroline, dipyridyl, or do not add pyridine.
8. the chirality four core copper base organic metal framework described in claim 1 or 2 is preparing the application in antitumor drug.
9. the chirality four core copper base organic metal framework described in claim 1 or 2 is as the application of angiogenesis inhibitor.
10. the chirality four core copper base organic metal framework described in claim 1 or 2 is as the application of antitumor drug or genophore.
CN201510272482.0A 2015-05-25 2015-05-25 A kind of chiral copper-based organic metal framework of four core and its preparation method and application Expired - Fee Related CN104910196B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510272482.0A CN104910196B (en) 2015-05-25 2015-05-25 A kind of chiral copper-based organic metal framework of four core and its preparation method and application

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510272482.0A CN104910196B (en) 2015-05-25 2015-05-25 A kind of chiral copper-based organic metal framework of four core and its preparation method and application

Publications (2)

Publication Number Publication Date
CN104910196A true CN104910196A (en) 2015-09-16
CN104910196B CN104910196B (en) 2017-06-16

Family

ID=54079686

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510272482.0A Expired - Fee Related CN104910196B (en) 2015-05-25 2015-05-25 A kind of chiral copper-based organic metal framework of four core and its preparation method and application

Country Status (1)

Country Link
CN (1) CN104910196B (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106215197A (en) * 2016-07-25 2016-12-14 南开大学 A kind of zeolite type metal-organic framework material based on gadolinium ion and preparation method thereof
CN110483790A (en) * 2019-07-25 2019-11-22 深圳大学 A kind of copper base metal-organic frame nanosphere and the preparation method and application thereof
CN111018893A (en) * 2019-11-25 2020-04-17 桂林医学院 Sulfur-containing amino alcohol Schiff base copper complex and preparation and application thereof
CN114394964A (en) * 2022-02-09 2022-04-26 沈阳化工大学 Preparation method of high-yield berberberrubine

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1950217A1 (en) * 2007-01-26 2008-07-30 Université de Neuchâtel Organometallic compounds for the treatment of cancer
CN103242349A (en) * 2013-04-12 2013-08-14 聊城大学 5-chlorosalicylaldehyde Schiff base tetra-nucleus copper complex as well as preparation method and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1950217A1 (en) * 2007-01-26 2008-07-30 Université de Neuchâtel Organometallic compounds for the treatment of cancer
CN103242349A (en) * 2013-04-12 2013-08-14 聊城大学 5-chlorosalicylaldehyde Schiff base tetra-nucleus copper complex as well as preparation method and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
XIN LI. ET AL: ""Syntheses and structures of tetracopper(II) complexes with an N-benzoate-N"-[3-(2-hydroxylethylammino)propyl]oxamide ligand:reactivity towards DNA, cytotoxic and antimicrobial activities"", 《NEW J. CHEM》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106215197A (en) * 2016-07-25 2016-12-14 南开大学 A kind of zeolite type metal-organic framework material based on gadolinium ion and preparation method thereof
CN110483790A (en) * 2019-07-25 2019-11-22 深圳大学 A kind of copper base metal-organic frame nanosphere and the preparation method and application thereof
CN111018893A (en) * 2019-11-25 2020-04-17 桂林医学院 Sulfur-containing amino alcohol Schiff base copper complex and preparation and application thereof
CN114394964A (en) * 2022-02-09 2022-04-26 沈阳化工大学 Preparation method of high-yield berberberrubine
CN114394964B (en) * 2022-02-09 2023-09-15 沈阳化工大学 Preparation method of high-yield berberine

Also Published As

Publication number Publication date
CN104910196B (en) 2017-06-16

Similar Documents

Publication Publication Date Title
CN104910196A (en) Chiral tetranuclear copper-based organic metal framework, and preparation method and applications thereof
CN106831375B (en) Chalcone compound with anti-tumor activity and preparation method and application thereof
CN104610372A (en) Synthesis method and application of platinum (II) complex using 2-benzoylpyridine as ligand
Zhang et al. Two novel chiral tetranucleate copper-based complexes: Crystal structures, nanoparticles, and inhibiting angiogenesis and the growth of human breast cancer by regulating the VEGF/VEGFR2 signal pathway in vitro
CN102688493B (en) Pharmaceutical composition containing resveratrol, resveratrol derivative and Bc1-2 inhibitor, and application thereof
Hou et al. Two novel chiral tetranucleate copper-based complexes: syntheses, crystal structures, inhibition of angiogenesis and the growth of human breast cancer in vitro and in vivo
CN104610373A (en) Platinum (II) complex employing terpyridyl derivative as ligand and synthesis method and application of complex
CN102631683A (en) Application of functionalized nano-selenium in tumor angiogenesis inhibiting and antitumor drugs
CN102688489B (en) Pharmaceutical composition containing triptolide, triptolide derivative and Bc1-2 inhibitor and application thereof
CN111848690A (en) Anticancer tetravalent platinum complex capable of inhibiting inflammation and immune escape and preparation method thereof
CN103463097A (en) Preparation and application of human serum albumin-ruthenium inorganic medicine compound
CN107698609B (en) Method for synthesizing anti-tumor metal prodrug based on human serum albumin IB subdomain and application
CN114605475B (en) Oral Pt (IV) anticancer prodrugs containing 3-bromopyruvate ligand axially
CN107488198B (en) A kind of couroupitine A platinum complex and its synthetic method and application
CN112870189B (en) Application of 2-acyl-1-dimethylaminomethyl ferrocene derivative in preparation of drug for targeted therapy of hepatocellular carcinoma
CN104650121A (en) Synthesis method and application of complex [Zn(H2L2)2](H2O)5 with anticancer activity
CN103463643B (en) The preparation of human serum albumin-ruthenium inorganic medicine compound and application thereof
CN104649930B (en) Coordination compound with anticancer activity [Zn (H2L4)2].(H2O) synthesis and application
CN102688490B (en) Pharmaceutical composition containing evodiamine, evodiamine derivative and Bc1-2 inhibitor, and the application
CN108440577B (en) Mixed copper-based complex and preparation method and application thereof
CN103463641B (en) The preparation of human serum albumin-ruthenium inorganic medicine compound and application thereof
CN104447397B (en) The synthesis of 3,5-Dibromosalicylaldehyde contracting AMPD schiff bases with anticancer activity and application
CN104650157A (en) Synthesis method and application of a complex [Ni(H2L2)2](H2O)2 with anticancer activity
CN104961752A (en) Binuclear copper (II) and zoledronic acid coordination complex and application thereof
CN116549476B (en) Compound, composition and application thereof in preparation of drugs with tumor angiogenesis inhibiting effect

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20170616

Termination date: 20210525

CF01 Termination of patent right due to non-payment of annual fee