A near-infrared light-responsive mitochondria-targeting drug delivery system, and a preparation method and application thereof

Abstract

The application discloses a near-infrared light response mitochondrion-targeting drug delivery system and a preparation method and application thereof, wherein the near-infrared light response mitochondrion-targeting drug delivery system is a composite nanomaterial DMON@(FeCO+Au 25 -TPP)-HA responding to glutathione and degrading, which is prepared by loading carbon monoxide releasing prodrug molecules FeCO and photosensitizer Au 25 -TPP capable of targeting mitochondria in the carrier mesoporous, and coating a targeting ligand hyaluronic acid HA on the outer layer. 25 -TPP and CO prodrug molecules FeCO in tumor tissues, achieving the effect of targeted photodynamic / CO gas synergistic anti-tumor treatment, and providing a simple, economical and safe new method for realizing efficient anti-tumor treatment.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of targeted drug delivery system, and particularly relates to a near-infrared light responsive mitochondria-targeted drug delivery system, a preparation method and application thereof. BACKGROUND

[0002] Malignant tumor is one of the global major public health problems, and is still a major killer of human health. At present, the main methods for treating malignant tumor are surgical treatment, chemotherapy and radiotherapy. These treatment methods improve the survival rate of patients, but also bring many side effects. With the development of nanomedicine, the side effects in tumor treatment are improved, but due to the special microenvironment of tumor, the number of nano-drugs reaching the tumor site is small. Studies have shown that only 0.7% of nano-particles injected intravenously eventually reach the tumor site, and only 0.0014% of them are finally taken up by tumor cells. It can be seen that exploring the improvement of the accumulation of nano-drugs in tumor sites is an effective way to improve the therapeutic effect.

[0003] Chemotherapy and radiotherapy are widely used as first-line treatment in the treatment of tumors. However, the toxicity of most existing non-specific chemotherapy and radiotherapy agents to normal tissues and blood greatly reduces the quality of life of patients. It is urgent to develop new anticancer drugs and treatment strategies with low toxicity and high efficiency. Some gas signaling molecules such as carbon monoxide (CO) and nitric oxide (NO) play an important role in various physiological and pathological metabolic activities, such as redox regulation, cell protection, and vasodilation. The physiological signaling effect of CO is closely related to its real-time concentration and the spatial location of release. CO, as a new emerging therapeutic gas molecule, can regulate cell function and tumor microenvironment, and has attracted widespread attention in cancer treatment. It can directly act on mitochondria by interfering with cytochrome c oxidase, block oxidative phosphorylation, affect the energy supply process of cell growth and proliferation, and freely cross the biological membrane and tumor gap. Therefore, researchers use its effect on the mitochondrial electron transport chain to accelerate cell respiration, consume oxygen and produce excess reactive oxygen species, clear ATP, and promote cell apoptosis for cancer treatment research.

[0004] Because of the concentration dependence of the biological effect of CO, the uncontrollable release of CO by CORMs in vivo can greatly limit the actual application of anti-tumor. Therefore, how to target delivery and precisely control the release of carbon monoxide molecules in tumor tissues has become a key problem in the application of CO treatment. It is urgent to develop a suitable nano-drug delivery system to produce CO for tumor treatment through controllable response stimulation at the tumor site, while being able to image the tumor tissue, so as to be used for imaging-guided precise treatment.

[0005] Gold-based nanoparticles as a kind of noble metal nanomaterials, due to its excellent biocompatibility, easy to surface modification and unique optical properties, as a kind of very promising disease treatment drugs has been widely concerned, among them, gold nanoclusters (AuNCs) is composed of several to hundreds of gold atoms, when their size (1-2nm) close to the fermi wavelength of electron, they show discrete energy levels. The photophysical properties of Au NCs can be adjusted by ligand to achieve strong near-infrared (more than 800nm) absorption and long lifetime of excited triplet state. Atomic level precise thiolate-protected gold clusters Au 25 (SR) 18 Due to its high stability, long lifetime of electronic excited state, the existence of triplet excited state and other characteristics, it realizes the ability to produce reactive oxygen 1 O2 by photoexcitation. Although gold nanoclusters have a very small size, but its fluorescence performance is very strong, showing more optimal CT enhanced imaging effect, and showing high sensitivity in cell fluorescence imaging.

[0006] Gold clusters Au 25 (SR) 18 The ability to generate ROS (reactive oxygen) in response to near-infrared light combined with the ability of CO prodrug CORMs to release CO under stimulation can achieve efficient photodynamic therapy while achieving controlled release of CO and used for gas synergistic therapy, enhancing the antitumor efficacy of photodynamic therapy.

[0007] In summary, it has great application prospect to develop a kind of mitochondria-targeting combined photodynamic therapy and CO gas therapy composite nanomaterial for antitumor therapy. SUMMARY

[0008] The technical problem solved by the present application is to provide a near-infrared light-responsive mitochondria-targeting drug delivery system and a preparation method thereof, which prepares a near-infrared light-responsive combined photodynamic therapy and CO gas therapy composite nanomaterial DMON@(FeCO+Au 25 -TPP)-HA, which is prepared by taking dendritic mesoporous silica DMON responsive to glutathione (GSH) as a carrier, loading carbon monoxide release prodrug molecules FeCO and mitochondria-targeting photosensitizer Au 25 (Capt) 18-m (TPP) m (simplified as Au 25 -TPP) in the mesoporous carrier, and coating the outer layer with the targeting ligand hyaluronic acid HA, which can realize the mitochondria-targeting photosensitizer Au 25The common delivery of TPP and CO prodrug molecule FeCO achieves the effect of targeted photodynamic / CO gas synergistic anti-tumor treatment, and provides a new method which is simple, economical and safe for realizing efficient anti-tumor treatment.

[0009] The application can realize that the composite nanomaterial targeting tumor tissue can generate a large amount of ROS to kill tumor cells under the irradiation of near-infrared light source 808nm laser, and the ROS stimulates the release of CO from CO release prodrug molecule FeCO, and the synergistic anti-tumor treatment is achieved, and the purpose of controllable release of CO is achieved. 25 The TPP and CO prodrug molecule FeCO are activated under the excitation of near-infrared light, and the photosensitizer Au 25 The TPP is activated to generate a large amount of ROS, and the released ROS triggers the rapid release of CO in cells, and the photosensitizer Au 25 The TPP can perform NIR-II fluorescence imaging on tumor tissue. More importantly, the released CO can sensitize ferroptosis and promote apoptosis, thereby significantly enhancing the anti-tumor effect of the photosensitizer Au 25 The TPP can perform NIR-II fluorescence imaging on tumor tissue. More importantly, the released CO can sensitize ferroptosis and promote apoptosis, thereby significantly enhancing the anti-tumor effect of the photosensitizer Au

[0010] The application adopts the following technical scheme to solve the above technical problems, a near-infrared light responsive mitochondria-targeting drug delivery system, characterized by: utilizing S-S bond bridged dendritic mesoporous silica DMON rich pore structure to load CO prodrug molecule FeCO and photosensitizer Au 25 The TPP and CO prodrug molecule FeCO are activated under the excitation of near-infrared light, and the photosensitizer Au 25 The TPP and CO prodrug molecule FeCO are activated under the excitation of near-infrared light, and the photosensitizer Au 25 The TPP and CO prodrug molecule FeCO are activated under the excitation of near-infrared light, and the photosensitizer Au 25 The TPP and CO prodrug molecule FeCO are activated under the excitation of near-infrared light, and the photosensitizer Au 1 O2 for killing tumor cells, and the active oxygen 1O2 triggers the CO prodrug molecule FeCO to release CO in cells, and the released CO can sensitize ferroptosis and promote apoptosis, thereby significantly enhancing the photosensitizer Au 25 -TPP has anti-tumor efficacy, and the released photosensitizer Au 25 -TPP can perform NIR-II fluorescence imaging on tumor tissues.

[0011] Further limited, the photosensitizer Au 25 -TPP has the ability to target mitochondria and the characteristics of photodynamic therapy at the same time, and the photosensitizer Au 25 -TPP has the chemical formula: Au 25 (SC9H 14 NO3) 18-m (C 22 H 23 Br) m , the ligand Capt represents 1-(3-mercapto-2-D-methylpropionyl)-L-proline, the ligand TPP represents 4-(triphenylphosphine)butane-1-mercapto ligand, and the chemical formula of the CO prodrug molecule FeCO is: C 11 H8FeINO5, and the chemical formula of hyaluronic acid HA is: (C 14 H 21 NO 11 ) n , and the structural formulas corresponding to the ligand Capt, the ligand TPP, the CO prodrug molecule FeCO and the hyaluronic acid HA are respectively:

[0012]

[0013] The photosensitizer Au 25 -TPP has the structural formula:

[0014] The preparation method of the near-infrared light responsive mitochondria-targeted drug delivery system provided by the application has the characteristics that the specific steps are:

[0015] Step S1, preparation of dendritic mesoporous silica DMON

[0016] Triethanolamine TEA, cetyltrimethylammonium bromide CTAB and sodium salicylate NaSal are dispersed in water and stirred and mixed uniformly, then a mixture of tetraethyl orthosilicate TEOS and bis-[gamma-(triethoxysilyl)propyl]-tetrasulfide BTES is added, and stirring reaction is carried out at room temperature to obtain DMON containing CTAB. The DMON is dissolved in an ammonium nitrate methanol solution and refluxed to react, and the reaction is repeated three times to obtain the product dendritic mesoporous silica DMON without CTAB;

[0017] Step S2, preparation of DMON@FeCO

[0018] The dendritic mesoporous silica DMON obtained in step S1 is dispersed in a mixed solvent of water and THF, and the CO prodrug molecule FeCO is added, and after stirring and mixing uniformly in the dark, centrifugal washing is performed to obtain the product DMON@FeCO loaded with the CO prodrug molecule FeCO;

[0019] Step S3, preparation of DMON@(FeCO+Au 25 -TPP)

[0020] The DMON@FeCO obtained in step S2 is dispersed in water, and an aqueous photosensitizer Au 25 -TPP solution is added, and stirring and reaction are performed in the dark, and centrifugal washing is performed to obtain the product DMON@(FeCO+Au 25 -TPP) loaded with the CO prodrug molecule FeCO and the photosensitizer Au 25 -TPP.

[0021] Step S4, preparation of DMON@(FeCO+Au 25 -TPP)-HA

[0022] The DMON@(FeCO+Au 25 -TPP) obtained in step S3 is dispersed in water, and hyaluronic acid HA is added, and stirring and reaction are performed in the dark at room temperature, and centrifugal washing is performed to obtain the near-infrared light responsive composite nanomaterial DMON@(FeCO+Au 25 -TPP)-HA for mitochondria-targeted photodynamic / CO gas synergistic anti-tumor therapy.

[0023] Further limited, the specific preparation process of the photosensitizer Au 25 -TPP is as follows: tetrachloroauric acid HAuCl4·4H2O is dissolved in methanol, and tetraoctylammonium bromide TOAB is added, and after stirring and mixing uniformly, methanol solutions of ligands captopril Capt and TPP are added, and after continuing to stir and mix uniformly, a sodium borohydride solution is added, and stirring and reaction are performed at room temperature, and the solvent is rotary evaporated to obtain a brown solid, and the solid is dissolved in anhydrous ethanol, and centrifugal washing is performed, and the supernatant is concentrated to obtain the product photosensitizer Au 25 -TPP.

[0024] The application of the near-infrared light responsive mitochondria-targeted drug delivery system in the preparation of a mitochondria-targeted photodynamic / CO gas synergistic anti-tumor therapy drug.

[0025] Compared with the prior art, the application has the following advantages and beneficial effects:

[0026] 1、The composite nanomaterial DMON@(FeCO+Au 25TPP)-HA can target tumor tissues and be degraded in response to GSH in the tumor microenvironment, thereby releasing photosensitizer Au 25 TPP and CO prodrug molecule FeCO are delivered to the tumor site together, effectively reducing the toxic side effects on normal tissues.

[0027] 2. The anti-tumor drug optimization design strategy provided by the application can effectively avoid the uncontrollable release of CO from CO prodrug molecules in CO gas treatment.

[0028] 3. Under near-infrared light, the composite nanomaterial DMON@(FeCO+Au 25 TPP)-HA can generate a large amount of active oxygen 1 O2 and CO gas are used to induce tumor cell apoptosis, achieving synergistic treatment of tumors by photodynamic / CO gas. At the same time, this may be a promising method to improve the efficiency of PDT treatment, and has good potential application prospects. BRIEF DESCRIPTION OF DRAWINGS

[0029] Figure 1 is a scanning electron microscope (A) and a transmission electron microscope (B) of the carrier dendritic mesoporous silica DMON. 25 is a preparation and application schematic diagram of the composite nanomaterial DMON@(FeCO+Au

[0030] Figure 2 is a scanning electron microscope (A) and a transmission electron microscope (B) of the carrier dendritic mesoporous silica DMON.

[0031] Figure 3 is a particle size distribution diagram (A) and a transmission electron microscope (B) of the composite nanomaterial; the picture shows that the composite nanomaterial is spherical, with a particle size of about 119 nm and a relatively narrow particle size distribution.

[0032] Figure 4 is a high-angle annular dark-field scanning transmission electron microscope (HAADF) test diagram (A) and an energy dispersive X-ray spectrometer (EDX) test diagram (B); the picture shows that Au and Fe elements are distributed in the entire material.

[0033] Figure 5 is an infrared spectrum diagram of DMON, Au 25 TPP, FeCO, DMON@FeCO, DMON@(FeCO+Au 25 TPP), DMON@(FeCO+Au 25 TPP)-HA.

[0034] Figure 6 is a ROS probe DPBF and Au 25 TPP or DMON@(FeCO+Au 25-TPP)-HA mixture is irradiated with near-infrared 808nm laser for different times to produce 1 Ultraviolet-visible spectra of O2.

[0035] Figure 7 CO probe FL-CO-1 and DMON@(FeCO+Au) 25 Fluorescence spectra of CO were generated after mixing TPP and HA and then irradiating with a near-infrared 808nm laser for different durations.

[0036] Figure 8 Different concentrations of DMON-HA, DMON@FeCO-HA, and DMON@Au 25 -TPP-HA、DMON@(FeCO+Au 25 Experiment on the cell-killing ability of 4T1 cells by 808nm laser irradiation (TPP)-HA. Detailed Implementation

[0037] The following examples further illustrate the above-described content of the present invention, but it should not be construed as limiting the scope of the subject matter of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention.

[0038] Example 1

[0039] Preparation of dendritic mesoporous silica (DMON)

[0040] 0.05 g of triethanolamine (TEA) was added to 12.5 mL of water and gently stirred magnetically for 0.5 h in an oil bath at 80 °C. Then, 200 mg of cetyltrimethylammonium bromide (CTAB) and 0.05 mg of sodium salicylate (NaSal) were added, and stirring continued for 1 h. Next, under slow stirring, 1 mL of a mixed silicon source of tetraethyl silicate (TEOS) and 1 mL of bis-[γ-(triethoxysilyl)propyl]-tetrasulfide (BTES) was added to the obtained CTAB-NaSal-TEA aqueous solution, and the reaction was continued overnight. The product was collected by high-speed centrifugation and washed three times with ethanol. To remove the CTAB template, the product was dissolved in 50 mL of a 2 wt% ammonium nitrate methanol solution, refluxed at 80 °C for 3 h, washed three times with ethanol, and refluxed three times to obtain the target product, dendritic mesoporous silica (DMON). It was dispersed in water and its concentration was calculated using the differential method.

[0041] photosensitizer Au 25 Preparation of TPP

[0042] Measure 2 mL of HAuCl4·4H2O solution into a 100 mL single-necked flask, add 3 mL of methanol solution, and stir magnetically. Then weigh 70 mg of tetraoctylammonium bromide (TOAB) and add it to the above solution. After stirring vigorously for 40 min, quickly add a methanol solution containing ligands captopril (Capt) (150 mg) and TPP (15 mg), and continue stirring for another 40 min. Then quickly add an ice-water solution of sodium borohydride (45 mg dissolved in 3 mL of ice water) to the above mixed solution, and stir at room temperature for 24 h. Rotary evaporate the solvent from the reaction solution to obtain a brown solid. Dissolve the solid in 30 mL of anhydrous ethanol, centrifuge, and concentrate the supernatant to obtain the product, the photosensitizer Au. 25 -TPP.

[0043] Synthesis of DMON@FeCO

[0044] Weigh 20 mg of the prepared dendritic mesoporous silica DMON containing SS bonds and disperse it in 1 mL of water. Weigh 20 mg of the CO prodrug molecule FeCO and place it in a centrifuge tube and dissolve it in 0.5 mL of tetrahydrofuran. Add the THF solution of FeCO to the above DMON solution containing SS bonds, stir at room temperature in the dark for 30 min, centrifuge and wash to obtain the product DMON@FeCO loaded with the CO prodrug molecule FeCO.

[0045] DMON@(FeCO+Au 25 Synthesis of -TPP

[0046] The DMON@FeCO prepared above was dispersed in 1 mL of water, and 8 mL of the photosensitizer Au prepared above was added. 25 -TPP solution was stirred in the dark for 4 hours, followed by centrifugation and washing to obtain the CO-loaded prodrug molecule FeCO and the photosensitizer Au. 25 -TPP product DMON@(FeCO+Au 25 -TPP).

[0047] DMON@(FeCO+Au 25 Synthesis of -TPP-HA

[0048] The DMON@(FeCO+Au) prepared above 25 -TPP) was dispersed in 4 mL of water, and then 20 mg of hyaluronic acid (HA) was added. The mixture was stirred at room temperature in the dark for 8 hours, and then centrifuged and washed to obtain the composite nanomaterial DMON@(FeCO+Au) 25 -TPP)-HA.

[0049] The above examples describe the basic principles, main features and advantages of the present application, and those skilled in the art should understand that the present application is not limited to the above examples, and the above examples and descriptions in the specification are only to illustrate the principles of the present application, and various changes and improvements can be made without departing from the principles of the present application, and such changes and improvements fall within the scope of the present application.

Claims

1. A near-infrared light-responsive mitochondria-targeting drug delivery system, characterized by: Utilize dendritic mesoporous silica DMON with S-S bond bridging to load CO precursor molecule FeCO and photosensitizer Au 25 -TPP, coated with hyaluronic acid HA, a targeting ligand, to obtain a near-infrared light-responsive composite nanomaterial DMON@(FeCO+Au 25 -TPP)-HA for mitochondria-targeted photodynamic / CO gas synergistic antitumor therapy 25 -TPP, which, under near-infrared light excitation, releases photosensitizer Au 25 -TPP is activated to produce a large amount of active oxygen 1 O2 is used to kill tumor cells, while active oxygen 1 O2 triggers the release of CO from CO precursor molecule FeCO in cells, and the released CO can sensitize ferroptosis and promote apoptosis, thereby significantly enhancing the antitumor efficacy of photosensitizer Au 25 -TPP, the released photosensitizer Au 25 -TPP can perform NIR-II fluorescence imaging of tumor tissues; The photosensitizer Au 25 -TPP has the ability to target mitochondria and the properties of photodynamic therapy, the photosensitizer Au 25 -TPP has the formula: Au 25 (SC9H 14 NO3) 18-m (C 22 H 23 Br) m ; the ligand Capt represents 1-(3-mercapto-2-D-methylpropionyl)-L-proline, the ligand TPP represents 4-(triphenylphosphine)butane-1-mercapto, the CO prodrug molecule FeCO has the formula: C 11 H8FeINO5, the hyaluronic acid HA has the formula: (C 14 H 21 NO 11 ) n ; the corresponding structural formulas of the ligand Capt, the ligand TPP, the CO prodrug molecule FeCO and the hyaluronic acid HA are respectively: The photosensitizer Au 25 -TPP has the structural formula: The photosensitizer Au 25 - The specific preparation process of TPP is as follows: dissolve tetrachloroauric acid HAuCl4·4H2O in methanol, then add tetraoctylammonium bromide TOAB, stir and mix uniformly, then add the methanol solution of the ligand captopril Capt and TPP, continue to stir and mix uniformly, then add sodium borohydride solution, stir and react at room temperature, rotary evaporate the solvent to obtain brown solid, dissolve the solid in anhydrous ethanol, centrifuge, take the supernatant and concentrate to obtain the product photosensitizer Au 25 - TPP.

2. A method of preparing the near-infrared light-responsive mitochondrial targeting drug delivery system according to claim 1, characterized by The specific preparation steps are as follows: Step S1, preparation of dendritic mesoporous silica DMON triethanolamine TEA, cetyltrimethylammonium bromide CTAB and sodium salicylate NaSal were dispersed in water and stirred and mixed uniformly, then a mixture of tetraethyl orthosilicate TEOS and bis-[gamma-(triethoxysilyl)propyl]-tetrasulfide BTES was added, and stirring reaction was carried out at room temperature to obtain DMON containing CTAB, and the DMON was dissolved in an ammonium nitrate methanol solution and refluxed to react for three times to obtain the product dendritic mesoporous silica DMON free of CTAB; Step S2, preparation of DMON@FeCO The dendritic mesoporous silica DMON obtained in step S1 was dispersed in a mixed solvent of water and THF with the CO prodrug molecule FeCO, and after stirring and mixing uniformly in the dark, centrifugal washing was carried out to obtain the product DMON@FeCO loaded with the CO prodrug molecule FeCO; Step S3, preparation of DMON(FeCO+Au 25 -TPP) The DMON@FeCO obtained in step S2 was dispersed in water, and a photosensitizer Au was added 25 -TPP aqueous solution, and the reaction was stirred in the dark. After centrifugation and washing, the CO prodrug molecule FeCO and the photosensitizer Au were obtained 25 -TPP) product DMON@(FeCO+Au 25 -TPP); Step S4, DMON(FeCO + Au 25 Preparation of DMON(FeCO + AuTPP)-HA DMON(FeCO+Au 25 -TPP) obtained in step S3 was dispersed in water, and hyaluronic acid HA was added. The reaction was stirred at room temperature in the dark, and centrifuged and washed to obtain near-infrared light responsive composite nanomaterial DMON(FeCO+Au 25 -TPP)-HA for mitochondria-targeted photodynamic / CO gas synergistic antitumor therapy.

3. The use of the near-infrared light responsive mitochondria-targeted drug delivery system of claim 1 in the preparation of a targeted photodynamic / CO gas synergistic anti-tumor treatment drug.

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