A binuclear metal ruthenium photocatalyst with full spectrum absorption and its preparation method and application
A dual ruthenium complex with broad-spectrum light absorption, synthesized using specific ligands, addresses the limitations of short-wavelength light therapies by providing effective anti-cancer and anti-bacterial treatments with enhanced tissue penetration and oxidative capabilities.
Patent Information
- Application Number
- CN202211729817.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2022-12-30
- Publication Date
- 2025-03-21
- Estimated Expiration
- 2042-12-30
AI Technical Summary
Current anti-cancer and anti-bacterial treatments face challenges due to limited tissue penetration of short-wavelength light-based photodynamic therapy, which can also damage biological systems, necessitating the development of metal complexes that absorb longer wavelengths like red light for deeper tissue penetration and effective treatment.
A dual ruthenium complex with a specific structure, synthesized through a multi-step process involving 4’-bromotripyridine, 2,6-diquinoline, and 2,5-dithien-3-ylpyrazine-1,4-diketone, exhibiting broad-spectrum light absorption and catalytic activity.
The dual ruthenium complex demonstrates potent photodynamic therapy effects against drug-resistant non-small cell lung cancer cells and Staphylococcus aureus, with enhanced light penetration and oxidative capabilities, offering a new approach for developing anti-cancer and anti-bacterial metal-based photodynamic agents.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the field of pharmaceutical chemistry technology, relates to the field of metal complex photosensitizer anti-tumor and antibacterial technology, and specifically relates to a binuclear metal ruthenium photocatalyst with full spectrum absorption, and a preparation method and application thereof. Background Art
[0002] Antimicrobial drugs are drugs that can inhibit or kill bacteria and are used to prevent and treat bacterial infections. Since the discovery and use of penicillin in 1928, antimicrobial drugs represented by penicillin have saved the lives of countless patients. Currently, commonly used antimicrobial drugs in clinical practice include antibiotics extracted from microorganisms and synthetic antimicrobial drugs, which are used to prevent and treat bacterial infections. The emergence of bacterial resistance has reduced the efficacy of antimicrobial drugs, so the development of new antimicrobial drugs is an effective means to deal with the increasingly serious resistance. Bacterial resistance has become a public health crisis that needs to be addressed urgently around the world.
[0003] Photosensitizer-based photodynamic therapy is a very promising cancer treatment strategy, and it is also being gradually applied to antibacterial research. Photosensitizers in photodynamic therapy are usually non-toxic or low-toxic in the dark, so the side effects on normal tissues are minimal. However, most of the currently used organometallic photosensitizers have a maximum wavelength of short-wavelength excitation, and ultraviolet light or short-wavelength visible light cannot penetrate deeply into tissues for biomedical applications, and ultraviolet light can also damage biological systems. In contrast, red light or near-infrared light (NIR) is more suitable for biomedical applications.
[0004] Therefore, the development of metal photosensitizers with long-wavelength light absorption can significantly overcome the problem of insufficient penetration depth in photodynamic therapy, and can be used for photodynamic therapy of tumors in deep tissues. At the same time, by inducing NADH / NADPH oxidation, it can be applied to photodynamic antibacterial research. The development of new red light therapy Ru(II) complexes for anti-tumor and antibacterial purposes is of great significance. Summary of the invention
[0005] In order to overcome the above-mentioned deficiencies of the prior art, the primary purpose of the present invention is to provide a binuclear metal ruthenium complex with full spectrum absorption.
[0006] The second object of the present invention is to provide the preparation of the above binuclear metal ruthenium complex.
[0007] The third object of the present invention is to provide the application of the binuclear ruthenium complex. The complex has a strong photocatalytic therapeutic effect on cisplatin-resistant strains of human non-small cell lung cancer (A549 / DDP) and Staphylococcus aureus (S. aureus).
[0008] The above first object of the present invention is achieved by the following technical solutions:
[0009] The present invention provides a binuclear ruthenium metal complex, wherein the binuclear ruthenium metal complex has a structure as shown in formula (I):
[0010]
[0011] The present invention also provides a method for preparing the binuclear ruthenium complex, wherein the preparation of the binuclear ruthenium complex comprises the following steps:
[0012] S1, generating a precursor compound Ru(tpy-Br)Cl3 by reacting 4'-bromo-2,2':6',2"-terpyridine with ruthenium(III) chloride hydrate;
[0013] S2, reacting the precursor compound Ru(tpy-Br)Cl3 of step S1 with 2,6-di(quinolin-8-yl)pyridine to generate a ruthenium complex [Ru(dqp)(tpy-Br)](PF6)2 after ion replacement with ammonium hexafluorophosphate;
[0014] S3. The novel binuclear ruthenium complex [Ru(dqp)(tpy-Br)](PF6)2 of step S2 is reacted with 2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophene-2-yl)pyrrolo[3,4-c]pyrrole-1,4-(2H,5H)-dione to obtain the novel binuclear ruthenium complex.
[0015] Preferably, step S1 is specifically as follows: 4'-bromo-2,2':6',2"-terpyridine and ruthenium (III) chloride hydrate are dissolved in an ethanol solution, and then refluxed at 75-95°C for 8-16h, and cooled to room temperature. The reaction formula of this step is as follows:
[0016]
[0017] Further, the molar ratio of 2,2':6',2"-terpyridine to ruthenium (III) chloride hydrate in step S1 is 1:1.2;
[0018] Furthermore, the heating reflux temperature in step S1 is 85° C. and the reaction time is 12 h.
[0019] Preferably, step S2 is specifically as follows: the precursor Ru(tpy-Br)Cl3 and 2,6-di(quinolin-8-yl)pyridine are dissolved in ethylene glycol solution, refluxed at 180-220°C for 10-30 minutes, cooled to room temperature, and then ammonium hexafluorophosphate is added for ion replacement, and purified by filtering, washing, drying and purification. The reaction formula of this step is as follows:
[0020]
[0021] Furthermore, in step S2, the molar ratio of the precursor Ru(tpy-Br)Cl3 to 2,6-di(quinolin-8-yl)pyridine is 1:1.
[0022] Furthermore, the heating reflux temperature in step 24 is 200° C. and the time is 20 min.
[0023] Preferably, step S3 is specifically as follows: under argon protection, [Ru(dqp)(tpy-Br)](PF6)2,2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophene-2-yl)pyrrolo[3,4-c]pyrrole-1,4-(2H,5H)-dione and tetrakis(triphenylphosphine)palladium are dissolved in toluene / N,N-dimethylformamide, and stirred at 100-135°C for 18-24h. After the reaction is completed, cool to room temperature, add saturated brine, filter and collect the precipitate, wash with water and toluene, and dry. The reaction formula of this step is as follows:
[0024]
[0025] Furthermore, in step S3, the molar ratio of 2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophene-2-yl)pyrrolo[3,4-c]pyrrole-1,4-(2H,5H)-dione, [Ru(dqp)(tpy-Br)](PF6)2, and tetrakis(triphenylphosphine)palladium is 1:2:0.1.
[0026] Furthermore, the heating reflux temperature in step S3 is 115° C. and the time is 20 h.
[0027] The third object of the present invention is achieved by the following technical solutions:
[0028] The present invention also provides the use of the binuclear metal ruthenium complex in the preparation of anti-tumor photocatalytic drugs.
[0029] Preferably, the tumor is lung cancer,
[0030] Furthermore, the human lung cancer cell line is a cisplatin-resistant strain of human non-small cell lung cancer (A549 / DDP).
[0031] The present invention also provides a binuclear metal ruthenium complex anti-tumor photocatalytic drug, wherein the drug uses the above binuclear metal ruthenium complex as a main active ingredient.
[0032] The present invention also provides a binuclear ruthenium complex anti-tumor metal photosensitizer, wherein the photosensitizer uses the above binuclear ruthenium complex as a main active ingredient.
[0033] Preferably, the drug or photosensitizer further comprises a pharmaceutically acceptable carrier and / or excipient. That is, the drug or photosensitizer uses a binuclear metal ruthenium complex as a main active ingredient, mixed with a pharmaceutically acceptable carrier and / or excipient to prepare a composition, and prepared into a clinically acceptable dosage form.
[0034] Furthermore, the excipient refers to diluents, adhesives, lubricants, disintegrants, solubilizers, stabilizers and other pharmaceutical matrices that can be used in the pharmaceutical field.
[0035] Furthermore, the carrier is an acceptable functional pharmaceutical excipient in the pharmaceutical field, including surfactants, suspending agents, emulsifiers and some new pharmaceutical polymer materials, such as cyclodextrin, chitosan, polylactic acid (PLA), polyglycolic acid-polylactic acid copolymer (PLGA), hyaluronic acid, etc.
[0036] Furthermore, the present invention has no particular limitation on the dosage form of the above-mentioned drugs or photosensitizers, and they can be prepared into tablets, capsules, suppositories, powder injections, etc., which are well known to those skilled in the art. The prepared preparations can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally or topically). If some drugs are unstable under gastric conditions, they can be prepared into enteric-coated tablets.
[0037] The present invention also provides the use of the binuclear metal ruthenium complex in the preparation of antibacterial photocatalytic drugs.
[0038] Preferably, the bacterium is Staphylococcus aureus (S. aureus).
[0039] The present invention also provides a binuclear metal ruthenium complex antibacterial photocatalytic drug, wherein the drug uses the above binuclear metal ruthenium complex as a main active ingredient.
[0040] The present invention also provides a binuclear ruthenium complex antibacterial metal photosensitizer, wherein the photosensitizer uses the above binuclear ruthenium complex as a main active ingredient.
[0041] Preferably, the drug or photosensitizer further comprises a pharmaceutically acceptable carrier and / or excipient. That is, the drug or photosensitizer uses a binuclear metal ruthenium complex as a main active ingredient, mixed with a pharmaceutically acceptable carrier and / or excipient to prepare a composition, and prepared into a clinically acceptable dosage form.
[0042] Furthermore, the excipient refers to diluents, adhesives, lubricants, disintegrants, solubilizers, stabilizers and other pharmaceutical matrices that can be used in the pharmaceutical field.
[0043] Furthermore, the carrier is an acceptable functional pharmaceutical excipient in the pharmaceutical field, including surfactants, suspending agents, emulsifiers and some new pharmaceutical polymer materials, such as cyclodextrin, chitosan, polylactic acid (PLA), polyglycolic acid-polylactic acid copolymer (PLGA), hyaluronic acid, etc.
[0044] Furthermore, the present invention has no particular limitation on the dosage form of the above-mentioned drugs or photosensitizers, and they can be prepared into tablets, capsules, suppositories, powder injections, etc., which are well known to those skilled in the art. The prepared preparations can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally or topically). If some drugs are unstable under gastric conditions, they can be prepared into enteric-coated tablets.
[0045] Compared with the prior art, the present invention has the following beneficial effects:
[0046] The invention discloses a binuclear ruthenium complex with full spectrum absorption, which has good photocatalytic proliferation inhibition ability (IC 50 0.08μM), with a high phototherapy index (PI>1250), and also has a photocatalytic therapeutic effect on Staphylococcus aureus (S.aureus), and has the ability to inhibit the proliferation of Staphylococcus aureus (MIC 90 6.25μM), and can produce superoxide anions and singlet oxygen under light conditions, and has photocatalytic oxidation ability for NADH / NADPH in tumor cells and bacteria, which is of great significance for the study of binuclear ruthenium complexes for anti-tumor and antibacterial drugs, and provides a new idea for the clinical development of binuclear ruthenium complexes for anti-tumor and antibacterial metal photosensitizers. BRIEF DESCRIPTION OF THE DRAWINGS
[0047] Figure 1 This is the UV absorption spectrum of the new binuclear ruthenium complex;
[0048] Figure 2 The ability of novel binuclear ruthenium complexes to photocatalytically generate superoxide anions;
[0049] Figure 3 The ability of novel binuclear ruthenium complexes to photocatalytically generate singlet oxygen;
[0050] Figure 4 The photocatalytic oxidation ability of the novel binuclear ruthenium complex for NADH;
[0051] Figure 5 The photocatalytic oxidation of NADPH by a novel binuclear ruthenium complex;
[0052] Figure 6A novel binuclear ruthenium complex was developed to combat the dark toxicity and phototoxicity of cisplatin-resistant human non-small cell lung cancer cell line (A549 / DDP);
[0053] Figure 7 A new binuclear ruthenium complex is used to combat the dark toxicity and phototoxicity of Staphylococcus aureus (S. aureus). DETAILED DESCRIPTION
[0054] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in each embodiment of the present invention described below can be combined with each other as long as they do not conflict with each other.
[0055] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.
[0056] Example 1 Preparation of a novel binuclear ruthenium complex
[0057] The structural formula of the binuclear ruthenium complex is as follows:
[0058]
[0059] The synthesis of the binuclear ruthenium complex comprises the following steps:
[0060] (1) Ru(tpy-Br)Cl3 is generated by the reaction of 4'-bromo-2,2':6',2"-terpyridine with ruthenium(III) chloride hydrate.
[0061] 4'-Bromo-2,2':6',2"-terpyridine (312 mg, 1 mmol) and ruthenium (III) chloride hydrate (249 mg, 1.2 mmol) were dissolved in 15 mL of ethanol solution, refluxed at 85 °C for 12 h and then cooled to room temperature. The reaction solution was filtered, and the filter cake was washed with ethanol and n-hexane and then vacuum dried to obtain a brown solid powder Ru(tpy-Br)Cl3 (449 mg, 86%).
[0062] The chemical reaction equation above is as follows:
[0063]
[0064] (2) Ru(tpy-Br)Cl3 reacts with 2,6-di(quinolin-8-yl)pyridine to generate [Ru(dqp)(tpy-Br)](PF6)2
[0065] The precursor Ru(tpy-Br)Cl3 (260 mg, 0.5 mmol) and 2,6-di(quinolin-8-yl)pyridine (167 mg, 0.5 mmol) were dissolved in 20 mL of ethylene glycol solution, stirred at 200 °C for 20 min and then cooled to room temperature. 50 mL of saturated NH4PF6 aqueous solution was added and stirred at room temperature for 1 h. The filter cake was washed with water and ethanol and dried in vacuo to obtain a ruthenium complex [Ru(dqp)(tpy-Br)](PF6)2 as a brown powder (264 mg, 51%).
[0066] The chemical reaction equation above is as follows:
[0067]
[0068] The H NMR spectrum of the product is: 1 NMR(600MHz, DMSO-d6)δ9.04(s,2H),8.77(dd,J=7.6,1.4Hz,2H),8.57–8.45(m,5H),8.40(ddd,J=5.5,1.5,0.7Hz,2H),8.32(dd,J=8.3,1.4Hz,2H),8.1 1(dd,J=8.2,1.3Hz,2H),7.91(td,J=7.8,1.5Hz,2H),7.86(t,J=7.8Hz,2H), 7.33(dd,J=5.3,1.4Hz,2H),7.30–7.25(m,2H),7.14(dd,J=8.2,5.2Hz,2H).
[0069] (3) [Ru(dqp)(tpy-Br)](PF6)2 and 2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophen-2-yl)pyrrolo[3,4-c]pyrrole-1,4-(2H,5H)-dione (CAS No.: 1392422-47-7) were reacted to form a novel binuclear ruthenium complex
[0070] [Ru(dqp)(tpy-Br)](PF6)2 (207 mg, 0.2 mmol), 2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophene-2-yl)pyrrolo[3,4-c]pyrrole-1,4-(2H,5H)-dione (85 mg, 0.1 mmol) and tetrakis(triphenylphosphine)palladium (12 mg, 0.01 mmol) were dissolved in 15 mL of toluene / N,N-dimethylformamide (V / V=2:1) and stirred at 115°C for 20 h under argon protection. After the reaction was completed, the mixture was cooled to room temperature, saturated brine was added, the precipitate was collected by filtration, washed with water and toluene, and vacuum dried to obtain 207 mg (85%) of dark green solid powder.
[0071] The chemical reaction equation above is as follows:
[0072]
[0073] The mass spectrum of the product is: ESI-MS [CH3OH, m / z]: 464 [M-4PF6 - ] 4+ ;
[0074] The H NMR spectrum of the product is: 1 H NMR(500MHz,DMSO-d6)δ9.02(s,4H),8.82(dd,J=14.6,5.8Hz,6H),8.71(d,J=8.1Hz,4H),8.5 4(dt,J=8.4,4.5Hz,8H),8.43(d,J=5.5Hz,4H),8.37–8.32(m,4H),8.13(d,J=8.2Hz,4H),7.98 (t,J=7.9Hz,4H),7.89(t,J=7.8Hz,4H),7.40(d,J=4.8Hz,4H),7.32(t,J=6.7Hz,4H),7.15(d d,J=8.2,5.4Hz,4H),1.41–1.15(m,22H),0.89(dt,J=7.7,3.9Hz,6H),0.80(t,J=6.9Hz,6H).
[0075] Experimental Example 1 Performance Test of Binuclear Ruthenium Complex
[0076] 1. Absorption spectra of binuclear ruthenium complexes in different solvents
[0077] The binuclear ruthenium complex of Example 1 was prepared into a 10 μM sample solution using phosphate buffered saline (PBS), dichloromethane (DCM), and acetonitrile (MeCN) as solvents, and then the ultraviolet absorption spectrum of the binuclear ruthenium complex was recorded using a double-beam ultraviolet-visible spectrophotometer. Figure 1 The absorbance in different solvents is shown, indicating that it has good light absorption ability in organic solvents and the absorption wavelength can be significantly red-shifted in PBS.
[0078] 2. Determination of the ability of binuclear ruthenium complexes to generate superoxide anions
[0079] In order to detect the ability of the binuclear ruthenium complex synthesized in Example 1 to photocatalytically generate superoxide anions, dihydrorhodamine 123 (DHR 123 for short) was used to measure the ability of the novel binuclear ruthenium complex to generate superoxide anions. When superoxide anions are generated in the solution, DHR 123 will immediately capture the superoxide anions in the solution, and DHR 123 is oxidized to generate the fluorescent derivative rhodamine 123, so that bright green fluorescence (Ex / Em=500 / 536nm) is emitted, wherein the increase in fluorescence intensity can indicate that superoxide anions are generated in the solution. The change in the fluorescence spectrum of the mixed solution of the sample to be tested and DHR 123 under different illumination times can reflect the ability to generate superoxide anions.
[0080] An aqueous solution containing a binuclear ruthenium complex (5 μM) and a DHR 123 reagent (5 μM) was placed in a cuvette and its superoxide anion generation capacity was measured under dark and light conditions. Figure 2 As shown, the binuclear ruthenium complex has the ability to generate superoxide anions after light irradiation.
[0081] 3. Determination of the ability of binuclear ruthenium complexes to generate singlet oxygen
[0082] In order to detect the ability of the binuclear ruthenium complex synthesized in Example 1 to photocatalytically generate singlet oxygen, the singlet oxygen probe 9,10-anthryl-bis(methylene)dimalonic acid (ABDA for short) was used to measure the ability of the new binuclear ruthenium complex to generate singlet oxygen. When singlet oxygen is generated in the solution, ABDA will immediately capture the singlet oxygen in the solution and react to generate an endogenous oxidation product, causing the characteristic absorption peak of ABDA to decrease, wherein the rate of decrease of the ABDA absorption peak is the rate of generation of singlet oxygen. The change of the ultraviolet-visible absorption spectrum of the sample to be tested and the ABDA mixed solution under different illumination times can reflect the ability to generate singlet oxygen.
[0083] Two aqueous solutions containing the same binuclear ruthenium complex (5 μM) and ABDA reagent (200 μM) were placed in a cuvette and their singlet oxygen generation abilities were measured under light and dark conditions, respectively. Figure 3 As shown, the binuclear ruthenium complex has the ability to generate singlet oxygen after light irradiation.
[0084] 4. Determination of the ability of binuclear ruthenium complexes to photocatalytically oxidize NADH / NADPH
[0085] Under light irradiation, the metal complex can oxidize reduced coenzyme I (NADH) and reduced coenzyme II (NADPH) into their oxidized states NADPH and NADPH. + and NADP +Therefore, the ruthenium complex (5 μM) and NADH or NADPH (A 339nm =1.0) in a cuvette, and its ability to oxidize NADH / NADPH under light and dark conditions can be measured respectively. Figure 4 , 5 As shown, the ruthenium complex has obvious photocatalytic oxidation ability for NADH and NADPH.
[0086] 5. Photodynamic therapy effect of a novel binuclear ruthenium complex on cisplatin-resistant human non-small cell lung cancer cell lines
[0087] Resazurin solution is blue and is usually used as an acid-base indicator (orange at pH 3.8 to dark purple at pH 6.5) and a redox indicator. When performing cell activity tests, resazurin can penetrate into cells and be irreversibly reduced to pink by living cells, while red fluorescent resorufin appears. The absorbance or fluorescence intensity of resorufin is positively correlated with the number of cells and the reducing ability, so cell proliferation can be analyzed using an enzyme-linked immunofluorescence detector.
[0088] The steps of the Resazurin experiment are as follows:
[0089] (1) First, revive one tube of A549 / DDP tumor cells and culture them with fresh complete culture medium (DMEM medium + 10% fetal bovine serum + 1% penicillin-streptomycin mixture). After two passages, start the experiment.
[0090] (2) When the cells reached the logarithmic growth phase, they were seeded into two 96-well plates at a density of 5,000 cells / well (100 μL of culture medium was used to culture cells in each well, one plate was for the light control group and the other was for the dark control group), and cultured in a 37°C, 5% CO2 incubator.
[0091] (3) After the cells adhere to the wall, the original culture medium was aspirated and 100 μL of ruthenium complexes of seven concentrations, 100, 50, 10, 1, 0.1, 0.01, and 0.001 μM, were added to each well. The cells were gently shaken and incubated in a carbon dioxide incubator (37°C, 5% CO2) in the dark.
[0092] (4) After incubation for 6 h, the cell culture plates in the illumination group were placed under white light for 20 min (light dose of 52.56 J / cm 2 ), and then returned to the incubator for incubation in the dark for 42 h (the cells in the dark control group were kept in the incubator incubated in the dark).
[0093] (5) After incubation for 48 h, the culture medium was discarded from each well, and 80 μL of resazurin (100 μg / mL) was added to each well. After further incubation in a 37°C incubator for 4 h, EX540 / EM590 was detected using the fluorescence plate of an enzyme-linked immunosorbent assay (ELISA) to calculate the cell proliferation inhibition rate and obtain the IC 50 value (drug concentration when the inhibition rate is equal to 50%).
[0094] like Figure 6 As shown in the figure, the resazurin method was used to detect the killing effect of different concentrations of ruthenium complexes on cisplatin-resistant human non-small cell lung cancer cell line (A549 / DDP cells) under dark and light treatment conditions. It can be seen that in the absence of light, the IC 50 >100μM, IC against cisplatin-resistant human non-small cell lung cancer cell lines under light conditions 50 It is 0.08 μM, indicating that the ruthenium complex of the present invention has a strong photodynamic therapy effect.
[0095] 6. Dark toxicity and phototoxicity of binuclear ruthenium complexes against Staphylococcus aureus
[0096] The typical Gram-positive bacteria Staphylococcus aureus was used as the research object to study the photodynamic antibacterial effect of binuclear ruthenium complexes. Staphylococcus aureus was inoculated in fresh LB medium and shaken at 150 rpm at 37°C until the exponential phase. The bacterial concentration was then determined by measuring the optical density of the bacteria at 600 nm. The specific experimental method is as follows:
[0097] The overnight cultured Staphylococcus aureus was diluted to 2×10 6 CFU / mL, and then a series of drug-containing bacterial solutions with gradient concentrations were prepared by the half-dilution method. That is, binuclear ruthenium (50μM) was added to the diluted bacterial solution for gradient dilution, and 7 concentration gradients (0.78, 1.56, 3.13, 6.25, 12.5, 25, 50μM) were prepared. The prepared bacterial suspension (100μL) was added to a 96-well plate in sequence, and 100μL of sterile and drug-free LB culture medium was added to the negative control group, and drug-free bacterial solution was added to the positive control group. The dark group was placed in a 37℃ static incubator for 18h, and the absorbance at OD=600nm was read. After the light group was statically incubated for 4h, it was irradiated with a white light source for 0, 20, and 40min (30mW / cm 2 ), and then put it back in the incubator for incubation for 14 hours. Observe whether the negative control group is clear, and clarity proves that the data is reliable. Use an enzyme marker to measure the absorbance at 600nm, and determine the light and dark toxicity of the binuclear metal ruthenium complex according to the following formula. The drug concentration at which the bacterial survival rate is <10% is the MIC value.
[0098] Survival rate (%) = (C experimental group - C negative control group) / (C positive control group - C negative control group) × 100%.
[0099] like Figure 7 As shown in the figure, different concentrations of binuclear ruthenium complexes have different killing effects on Staphylococcus aureus under dark and light treatment conditions. The binuclear ruthenium complex in the experiment has low toxicity to Staphylococcus aureus in the absence of light, but has a strong growth inhibition ability on Staphylococcus aureus under light conditions (light MIC 90 =6.25 μM).
[0100] In summary, the binuclear ruthenium complex of the present invention has a photocatalytic therapeutic effect on cisplatin-resistant human non-small cell lung cancer strain (A549 / DDP) and Staphylococcus aureus (S. aureus). Under light conditions, it has a proliferation inhibition ability (IC 50 0.08 μM), and has the ability to inhibit the proliferation of Staphylococcus aureus (MIC 90 6.25μM), and can produce superoxide anions and singlet oxygen under light conditions, and has photocatalytic oxidation ability for NADH / NADPH of cisplatin-resistant human non-small cell lung cancer strain (A549 / DDP) and Staphylococcus aureus (S.aureus), which is of great significance for the study of binuclear metal ruthenium photosensitizers for anti-tumor and antibacterial drugs, and provides a new idea for the clinical development of anti-tumor and antibacterial metal photosensitizers.
[0101] The embodiments of the present invention are described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions and variations of these embodiments are made without departing from the principles and spirit of the present invention, and still fall within the protection scope of the present invention.
Claims
1. A binuclear ruthenium complex with full spectrum absorption, characterized in that: The binuclear metal ruthenium complex has a structure as shown in formula (I):
2. The method for preparing the binuclear ruthenium complex according to claim 1, characterized in that: The preparation of the binuclear ruthenium complex comprises the following steps: S1, generating a precursor compound Ru(tpy-Br)Cl3 by reacting 4'-bromo-2,2':6',2"-terpyridine with ruthenium(III) chloride hydrate; S2, reacting the precursor compound Ru(tpy-Br)Cl3 of step S1 with 2,6-di(quinolin-8-yl)pyridine to generate a ruthenium complex [Ru(dqp)(tpy-Br)](PF6)2 after ion replacement with ammonium hexafluorophosphate; S3. The binuclear metal ruthenium complex [Ru(dqp)(tpy-Br)](PF6)2 of step S2 is reacted with 2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophene-2-yl)pyrrolo[3,4-c]pyrrole-1,4-(2H,5H)-dione to obtain the binuclear metal ruthenium complex.
3. The method for preparing a binuclear ruthenium complex according to claim 2, characterized in that: Step S1 is specifically as follows: 4'-bromo-2,2':6',2"-terpyridine and ruthenium (III) chloride hydrate are dissolved in an ethanol solution, and then refluxed at 75-95°C for 8-16h, and then cooled to room temperature.
4. The method for preparing a binuclear ruthenium complex according to claim 2, characterized in that: Step S2 is specifically as follows: the precursor Ru(tpy-Br)Cl3 and 2,6-di(quinolin-8-yl)pyridine are dissolved in ethylene glycol solution, refluxed at 180-220°C for 10-30 minutes and then cooled to room temperature, and then ammonium hexafluorophosphate is added for ion replacement, and purified by filtering, washing, drying and purification.
5. The method for preparing a binuclear ruthenium complex according to claim 2, characterized in that: Step S3 is specifically as follows: under argon protection, [Ru(dqp)(tpy-Br)](PF6)2,2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophene-2-yl)pyrrolo[3,4-c]pyrrole-1,4-(2H,5H)-dione and tetrakis(triphenylphosphine)palladium are dissolved in toluene / N,N-dimethylformamide, and stirred at 100-135°C for 18-24h; after the reaction is completed, the mixture is cooled to room temperature, saturated brine is added, the precipitate is collected by filtration, and the precipitate is washed with water and toluene and dried.
6. Use of the binuclear ruthenium complex according to claim 1 in the preparation of anti-tumor photocatalytic drugs, characterized in that: The tumor is lung cancer.
7. Use of the binuclear ruthenium complex according to claim 1 in the preparation of a drug for inhibiting the proliferation of cisplatin-resistant strains of human non-small cell lung cancer, characterized in that: The human non-small cell lung cancer cisplatin-resistant strain is A549 / DDP cell.