N-heterocyclic dithiocarbamate Fe (III) compound as well as preparation method and application thereof
By developing N heterocyclic dithiocarbamate Fe(III) compounds, the toxicity and resistance of existing metal coordination compounds in cancer treatment are solved, and efficient tumor elimination is achieved in photothermal therapy , demonstrated anti-cancer activity and high photothermal conversion efficiency better than cisplatin.
Patent Information
- Application Number
- CN202411355671.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-27
- Publication Date
- 2025-01-03
AI Technical Summary
The existing metal coordination compounds have toxicity and drug resistance problems in cancer treatment, and the targeting ability of photothermal therapy is poor, resulting in limited therapeutic effects.
Developed an N heterocyclic dithiocarbamate Fe(III) compound that not only exhibits hemolytic properties comparable to cisplatin, but also exhibits anti-cancer activity better than cisplatin and is at 650 nm in near infrared It has high photothermal conversion efficiency under laser light.
The compound significantly outperforms the anti-cancer activity of cisplatin at the micromolar level and exhibits efficient tumor elimination potential in photothermal therapy, with good biocompatibility and selectivity.
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Figure CN119241428A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to an Fe(III) compound, in particular to an N-heterocyclic dithiocarbamate Fe(III) compound and a preparation method and application thereof. Background Art
[0002] Metal coordination complexes offer biological and chemical diversity distinct from organic drugs. For decades, cisplatin and its derivatives have been used in more than 50% of cancer patient treatment regimens, however, the attendant toxicity as well as acquired and intrinsic drug resistance limit their widespread clinical application. These unresolved shortcomings have stimulated the search for metal-based anticancer compounds.
[0003] As one of the redox elements essential for human health and proper functioning, iron (Fe) plays a key role in regulating several important biological processes such as DNA synthesis and electron transport. Iron exists in two common oxidation states: Fe(II) and Fe(III). Iron also has the ability to change from one oxidation state to another by accepting or donating electrons, which allows it to play a role in a variety of biological pathways. Emerging studies have shown that tumor cells have a higher demand for iron than normal cells and that proteins involved in iron storage, export, and import may be altered in tumor cells. Therefore, the binding of iron to important cellular biochemical processes makes its complexes very useful in the design and development of effective anticancer drugs.
[0004] Cancer remains a major public health problem worldwide. Photothermal therapy is an innovative tumor treatment modality that ablates local solid tumors by converting light into heat and exhibits good anti-tumor effects. Despite advances in clinical treatments, limited therapeutic efficacy remains to be overcome due to tumor metastasis, poor targeting ability, strong side effects, and susceptibility to drug resistance. Therefore, various emerging treatment strategies, especially multimodal treatments, such as compounds that have both chemotherapy and photothermal therapy, show great potential in improving the therapeutic efficiency of tumor elimination. Summary of the invention
[0005] The present invention provides an N-heterocyclic dithiocarbamate Fe(III) compound, which not only shows hemolytic activity comparable to that of cisplatin, but also shows anticancer activity superior to that of cisplatin. In addition, the compound achieves a high photothermal conversion efficiency in aqueous solution under near-infrared 650nm laser irradiation, showing the potential for photothermal therapy. This study expands the research field of iron compounds.
[0006] The technical solution for achieving the purpose of the present invention is as follows:
[0007] An N-heterocyclic dithiocarbamate Fe(III) compound having the chemical formula [FeIII (L)]·ClO4·H2O, where HL is (E)-methyl 2-(quinoline-8-methylene)hydrazineaminodithiocarboxylate ligand; the Fe(III) metal center is coordinated with the ligand HL, the imine hydrogen on HL will be removed, and the ligand will become an anion, so it is represented by L in the chemical formula.
[0008] The structural formula of the compound is as follows:
[0009]
[0010] X-ray single crystal diffraction shows that the Fe(III) compound belongs to the tetragonal system, space group P - 421c; unit cell parameters are: a 14.5612(15),b 14.5612(15),c 13.2614(6),α(°)90,β(°)90,γ(°)90; The Fe(III) in this compound forms a hexacoordinated octahedral structure with the thiol mode of two HLs.
[0011] The preparation method of the N-heterocyclic dithiocarbamate Fe (III) compound of the present invention is as follows:
[0012] Quinoline-8-carboxaldehyde, methylhydrazine dithiocarbamate and Fe(ClO4)2·6H2O were dissolved in the organic solution at a molar ratio of 1:1:0.5 and stirred at room temperature for 40-60 minutes;
[0013] Then, the Fe(III) compound is obtained by crystallization at room temperature.
[0014] During preparation, raw materials can be added in multiples of the above ratio.
[0015] In the above preparation method, the organic solvent is one or a combination of two or more of methanol, ethanol and chloroform.
[0016] When the organic solvent is methanol, its amount is preferably such that it can dissolve the raw materials participating in the reaction. Usually, based on 1 mmol of Fe(ClO4)2·6H2O compound, all the raw materials participating in the reaction are usually dissolved in 10 to 50 mL of methanol.
[0017] Another object of the present invention is to provide the use of the Fe(III) compound in the preparation of anti-tumor chemotherapy drugs.
[0018] Another object of the present invention is to provide the use of the Fe(III) compound in the preparation of photothermal drugs.
[0019] Advantages of the present invention:
[0020] (1) The N-heterocyclic dithiocarbamate Fe(III) compound of the present invention has simple preparation conditions, is easy to separate, and has high yield and purity.
[0021] (2) The Fe(III) compound of the present invention exhibits hemolytic activity comparable to that of cisplatin and exhibits good biocompatibility.
[0022] (3) The Fe(III) compound of the present invention exhibits better anticancer activity than cisplatin and has better selectivity.
[0023] (4) The Fe(III) compound of the present invention also exhibits photothermal conversion performance under 650nm laser irradiation and can be used for photothermal therapy and for preparing photothermal drugs. BRIEF DESCRIPTION OF THE DRAWINGS
[0024] Figure 1 It is the crystal structure of the Fe(III) compound of the present invention.
[0025] Figure 2 UV-Vis stability graphs of the example Fe(III) compounds in methanol (left) and 1640 medium (right).
[0026] Figure 3 The hemolysis rates observed in the examples are those of Fe(III) compounds at different concentrations.
[0027] Figure 4 The aqueous solutions of Fe(III) compounds with different concentrations in the embodiment were illuminated by a 650 nm laser with a power density of 0.55 W / cm 2 The photothermal curve below.
[0028] Figure 5 The light-to-heat conversion efficiency is calculated based on the relationship between the cooling time and the negative natural logarithm of the driving force temperature in this embodiment. DETAILED DESCRIPTION
[0029] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments and drawings, but the present invention is not limited thereto.
[0030] Example
[0031] Methanol, ethanol, quinoline-8-carboxaldehyde, methylhydrazine dithiocarbamate, and Fe(ClO4)2·6H2O were purchased from Sigma-Aldrich. All other reagents were commercially available and used without further purification.
[0032] (1) Synthesis and structural characterization of Fe(III) compounds:
[0033] 1 mmol of quinoline-8-carboxaldehyde and 1 mmol of methylhydrazine dithiocarbamate were added to 20 mL of methanol, and the mixed liquid was stirred at room temperature for 10 min. Then, 10 mL of a methanol solution containing 0.5 mmol of Fe(ClO4)2·6H2O was added and stirred at room temperature for 50 min to obtain a dark brown mixed solution.
[0034] The dark brown mixed solution was placed at room temperature, and the dark black flaky Fe(III) compound was obtained by slowly evaporating the solvent, with a yield of 63%. ESI-MS (m / z): 575.99 = [M-ClO4-H2O] + , fit:575.99.
[0035] Single crystal diffraction data for Fe(III) compounds were collected using a Bruker APEX-IICCD diffractometer equipped with a graphite monochromator. Radiation source. The experimental structure of this compound was elucidated by direct method and the heavy atom anisotropy was refined using Olex-2 software. We generated all hydrogen atoms using riding model geometry.
[0036] The single crystal data of Fe(III) compounds have been deposited in the Cambridge Crystallographic Data Centre (CCDC) database, and the corresponding CCDC number is 2385395.
[0037] Selected crystallization data are listed in Tables 1 and 2.
[0038] Table 1. Crystallographic data of Fe(III) compounds.
[0039]
[0040]
[0041] Table 2. Bond lengths of Fe(III) compounds and bond angles (°).
[0042]
[0043] X-ray single crystal diffraction shows that the Fe(III) compound belongs to the tetragonal system, space group P - 421c.
[0044] like Figure 1 As shown, the Fe(III) in the compound forms a six-coordinated octahedral structure with the thiol mode of the two HLs.
[0045] (2) UV-Vis study of Fe(III) compounds:
[0046] Ultraviolet-visible spectroscopy (UV-Vis) is often used for qualitative analysis, structural analysis, purity testing, and stability determination. Here, a certain concentration of Fe(III) compounds is taken and added to methanol and 1640 medium. Then, their absorption spectra are measured using UV-Vis spectroscopy.
[0047] The experiment was divided into two groups: 7.5 μM of Fe(III) compound was added to methanol, mixed thoroughly, and then 3 ml of the solution was taken for measurement in a cuvette, with the methanol removed as the blank baseline.
[0048] For the next group, 7.5 μM of Fe(III) compound was added to complete 1640 medium, mixed thoroughly, and then 3 ml of the obtained solution was taken out for measurement in a cuvette, with complete 1640 medium as blank baseline.
[0049] The scanning wavelength range was set from 280 to 850 nm, fast scanning was selected, and measurements were taken at 0 min, 5 min, 10 min, 30 min, 60 min, 120 min, 12 h, and 24 h.
[0050] like Figure 2 As shown, the main absorption peak of Fe(III) compounds is in the 300-500nm band, which is mainly the spin-allowed ligand center n→π* and / or π→π* transition. Importantly, at room temperature, the time-dependent absorption spectra of Fe(III) compounds in methanol and 1640 medium did not detect significant changes within 24 hours, which indicates that Fe(III) compounds can maintain a relatively stable structure in these two media.
[0051] (3) Study on the anticancer activity of Fe(III) compounds:
[0052] MTT is mainly used for cytotoxicity, cell proliferation, and activity determination of cells cultured in vitro after drugs or other interventions. First, cells in the logarithmic growth phase are collected, the cell suspension concentration is adjusted to 5000–8000 cells per well, and evenly distributed in a 96-well plate, 180 μL per well. Then, the plate is placed in a 5% CO2, 37°C incubator overnight until the cells form a monolayer covering 50%–70% of the bottom of the well. Next, different concentrations of drugs are added (20 μL per well) and cultured for another 48 hours. Cisplatin is used as a positive control. After the incubation period, 10 μL of MTT solution (5 mg / mL) is added to each well and cultured for another 4 hours. Then, the culture medium in each well is carefully removed, 100 μL of dimethyl sulfoxide (DMSO) is added, and the wells are shaken at low speed on a shaker for 10 minutes to completely dissolve the crystals. Finally, the absorbance value of each well is measured at OD 570nm using an enzyme-linked immunosorbent assay.
[0053] Iron is found in high levels in several types of human cancer, suggesting that iron compounds may have potential as anticancer agents. Two human lung cancer cell lines, A549 and HCC827, and human normal lung fibroblasts WI-38 / VA13 were selected for in vitro anticancer activity testing. The half maximal inhibitory concentration (IC 50 ) The test results are shown in Table 3. The results show that the Fe(III) compound of the present invention exhibits significant anticancer activity at the micromolar level and is significantly better than the anticancer activity of cisplatin. Importantly, the IC of the Fe(III) compound in normal lung fibroblasts WI-38 / VA13 is 50 The value (2.62 μM) was greater than that of lung cancer cells A549 (0.41 μM) and HCC827 (0.54 μM), which indicated that the Fe(III) compound had a certain selectivity.
[0054] Table 3. IC values of Fe(III) compounds against two human lung cancer cell lines 50 (μΜ) value.
[0055]
[0056] (4) Research on the detection of hemolytic reaction of Fe(III) compounds:
[0057] Hemolysis analysis was performed according to existing methods (Bioorganic Chemistry, 2024, 144, 107122). In short, fresh whole human blood is commercially available (Innovative Research, Novi, MI). After taking out the human blood sample, it was centrifuged and carefully aspirated to remove the supernatant containing plasma and platelets, and then washed three times with sterile PBS, and the red blood cells were diluted 10 times with PBS. Then 900 μL of the dilution was taken and placed in a 1.5 ml centrifuge tube for later use.
[0058] The drug concentration was set into six groups (blank control, 5μM, 10μM, 20μM, 40μM and positive control) and added to the diluent in sequence. PBS was then added to make the total volume reach 1mL. After thorough mixing, the solution was placed in a 37°C incubator for 4 hours. The sample was taken out and centrifuged, photographed, and then the supernatant was taken into a 96-well plate and placed in an ELISA reader set to 570nm for absorbance detection.
[0059] Calculation of hemolysis rate: Hemolysis rate % = (As-An) / (Ap-An) × 100%; As, sample; An, negative control; Ap, positive control.
[0060] Hemolysis experiments can determine the toxic effects of drugs and the safety and effectiveness of drug use. According to the requirements of ISO / Technical Report 7406, materials with good biocompatibility require hemoglobin dissolution levels below 5%. The positive control (PC) completely dissolves all red blood cells, and the solution appears as a red transparent liquid with no red blood cells remaining, and the dissolution rate reaches 100%. The dissolution rate of Fe(III) compounds is within 5%, such as Figure 3 As shown, the compound has excellent biocompatibility. In addition, the dissolution rate of the Fe(III) compound is comparable to that of the clinical anticancer drug cisplatin, which also shows that the Fe(III) compound has the potential to be used in anticancer.
[0061] (6) Study on the photothermal effect of Fe(III) compounds:
[0062] To measure the photothermal effect of Fe(III) compounds, a 650 nm laser was used at 0.55 W / cm 2 The power density was irradiated with aqueous dispersions of Fe(III) compounds of different concentrations (0, 0.025, 0.05, 0.10, 0.25 mM) until the temperature stopped rising; the sample was then allowed to cool naturally. The working distance was 10 cm and the spot area was 2 cm2. The temperature change was measured by a thermal recorder.
[0063] The sample was irradiated with a 650 nm laser with a power density of 0.55 W / cm 2 , until the temperature reaches a stable state. Figure 4 As shown, the temperature of the aqueous solution of Fe(III) compound of 0.025, 0.05, 0.10 and 0.25 mM samples increased by 15.0, 19.9, 24.1 and 27.3 °C, respectively. In contrast, the temperature of the pure water control sample did not increase significantly during this period. The photothermal transduction of 0.10 mM Fe(III) compound was observed using Roper's method.
[0064] The heat transfer rate of the system is calculated by recording the change in temperature over time. Figure 5 As shown, the photothermal conversion efficiency of the Fe(III) compound was 55.9%. These results indicate that it can be used for effective thermal-based tumor cell elimination.
Claims
1. An N-heterocyclic dithiocarbamate Fe(III) compound, characterized in that: The chemical formula of this compound is [Fe III (L)]·ClO4·H2O, where HL is ( E )-methyl 2-(quinoline-8-methylene)hydrazinecarbamodithioate ligand; The structural formula of the compound is as follows:
2. X-ray single crystal diffraction shows that the Fe(III) compound belongs to the tetragonal system, space group P- 421 c ; The unit cell parameters are: a (Å) 14.5612(15), b (Å) 14.5612(15), c (Å) 13.2614(6), α ( o )90, β ( o ) 90, γ ( o ) 90; the Fe(III) in the compound forms a six-coordinated octahedral structure with the thiol mode of two HLs.
3. The method for preparing the N-heterocyclic dithiocarbamate Fe(III) compound according to claim 1, characterized in that: The preparation method is: Quinoline-8-carboxaldehyde, methylhydrazine dithiocarbamate and Fe(ClO4)2·6H2O were dissolved in the organic solution at a molar ratio of 1:1:0.5 and stirred at room temperature for 40-60 minutes; Then, crystallization is performed at room temperature to obtain an N-heterocyclic dithiocarbamate Fe (III) compound.
4. The method for preparing the N-heterocyclic dithiocarbamate Fe(III) compound according to claim 2, characterized in that: The organic solvent is one or a combination of two or more of methanol, ethanol and chloroform.
5. The method for preparing the N-heterocyclic dithiocarbamate Fe(III) compound according to claim 2, characterized in that: The dosage of the organic solvent is based on Fe(ClO4)2·6H2O, and the ratio of the organic solvent to Fe(ClO4)2·6H2O is 10~50 mL:1 mmol.
6. The use of the N-heterocyclic dithiocarbamate Fe(III) compound as claimed in claim 1, characterized in that: The application of the Fe(III) compound in the preparation of anti-tumor chemotherapy drugs.
7. The use of the N-heterocyclic dithiocarbamate Fe(III) compound as claimed in claim 1, characterized in that: Application of the Fe(III) compound in the preparation of photothermal drugs.