A photo-controlled coupled nanoprobe and preparation method thereof, photo-controlled coupled nanoparticle composition and application thereof

Abstract

The present application discloses a photo-controlled coupled nanoprobe and a preparation method thereof, a photo-controlled coupled nanoparticle composition and application, and relates to the technical field of tumor diagnosis and tumor treatment. The photo-controlled coupled nanoprobe is a metal nanoparticle coated with an amphiphilic polymer containing a furan unit and an organic photosensitizer unit; the metal nanoparticle is an Au or Pt-Au nanoparticle. The present application provides a nanoprobe with high accumulation, deep penetration and long retention time, and can achieve synergistic treatment of tumors.

Description

Technical Field

[0001] The present application relates to the technical field of tumor diagnosis and tumor treatment, and specifically to a photo-controlled coupled nanoprobe and a preparation method thereof, a photo-controlled coupled nanoparticle composition and application. Background Art

[0002] At present, traditional anti-tumor treatment still relies on surgery, radiotherapy and chemotherapy. Some chemotherapeutic drugs, such as oxaliplatin, doxorubicin (DOX) and paclitaxel (PTX), are used to treat tumors, but systemic administration also has problems such as high drug toxicity, poor selectivity, high possibility of tumor recurrence and induction of tumor cell resistance. To a certain extent, the body itself will also be damaged, causing side effects, complications, and even death.

[0003] In recent years, nanomedicines have been used in the early diagnosis and specific treatment of cancer, but the efficacy is not satisfactory. After systemic administration, only a few nanoparticles can effectively penetrate and retain in the tumor site, and most of them accumulate in the liver and spleen through nonspecific uptake, which greatly limits the development of nanomedicine. The main reason is that small-sized nanoparticles cannot accumulate in the tumor site and be pumped back into the blood; large-sized nanoparticles can accumulate in the tumor site, but cannot penetrate deeply and kill the tumor cells in the inner layer. Therefore, designing nanosystems with high accumulation, deep penetration and long retention time is particularly important to improve the therapeutic effect. Summary of the invention

[0004] In view of the shortcomings of the above-mentioned technologies, the present application provides a nanoprobe with high accumulation, deep penetration and long residence time by providing a photo-controlled coupled nanoprobe, a photo-controlled coupled nanoparticle composition and their preparation method and application, and can achieve synergistic treatment of tumors.

[0005] In order to achieve the above objectives, this application mainly provides the following technical solutions:

[0006] The present application provides a light-controlled coupled nanoprobe, wherein the light-controlled coupled nanoprobe is a metal nanoparticle coated with an amphiphilic polymer containing a furan unit and an organic photosensitizer unit;

[0007] The amphiphilic polymer containing furan units and organic photosensitizer units has a structure shown in formula (I):

[0008]

[0009] In formula (I), R1, R2, R3, and R4 are independently selected from H, methyl, ethyl, or phenyl;

[0010] S1 and S2 are both connecting groups selected from alkyl, alkoxy, amino or a combination thereof;

[0011] X is an organic photosensitizer group;

[0012] p / m=0.1-0.4; q / m=0.01-0.2; n / m=0.5-1;

[0013] The metal nanoparticles are Au or Pt-Au nanoparticles.

[0014] Preferably, the amphiphilic polymer containing furan units and organic photosensitizer units is obtained by grafting a modified furan derivative and a modified organic photosensitizer with a first copolymer;

[0015] The first copolymer has a structure shown in formula (II):

[0016]

[0017] In formula (II), R1, R2, R3, and R4 are independently selected from H, methyl, ethyl, or phenyl.

[0018] Preferably, the first copolymer is selected from at least one of poly(isobutylene-maleic anhydride), poly(styrene-maleic anhydride), and poly(ethylene-maleic anhydride).

[0019] Preferably, the organic photosensitizer is at least one of Rose Bengal or methylene blue.

[0020] Preferably, in the formula (I), X-S2-NH- has the structure of the following formula (III):

[0021]

[0022] Preferably, in the formula (I), It has the following structure:

[0023]

[0024] Preferably, the mass proportion of metal nanoparticles in the light-controlled coupling nanoprobe is 12.5%-20%.

[0025] Preferably, the particle size of the light-controlled coupling nanoprobe is 10-50 nm.

[0026] The present application also provides a method for preparing the above-mentioned light-controlled coupled nanoprobe, comprising the following steps:

[0027] (1) reacting the modified organic photosensitizer, the modified furan derivative and the first copolymer through an amide reaction to obtain an amphiphilic polymer containing a furan unit and an organic photosensitizer unit;

[0028] (2) mixing, dispersing and drying the amphiphilic polymer containing furan units and organic photosensitizer units with Au or Pt-Au nanoparticles to obtain the light-controlled coupled nanoprobe;

[0029] The modified organic photosensitizer has the following structure:

[0030] X-S2-NH2(V),

[0031] Wherein X is an organic photosensitizer group; S2 is a linking group selected from alkyl, alkoxy, amino or a combination thereof;

[0032] The modified furan derivative has the following structure:

[0033]

[0034] Wherein, S1 is a linking group selected from alkyl, alkoxy, amino or a combination thereof;

[0035] The first copolymer has the structure of the following formula (II):

[0036]

[0037] In formula (II), R1, R2, R3, and R4 are independently selected from H, methyl, ethyl, or phenyl.

[0038] Preferably, the mass ratio of the amphiphilic polymer containing furan units and organic photosensitizer units to Au or Pt-Au nanoparticles is 4:1-7:1.

[0039] Preferably, the diameter of the Au nanoparticles is 5-8 nm, and the diameter of the Pt-Au nanoparticles is 8-10 nm.

[0040] Preferably, the modified organic photosensitizer has the structure of the following formula (VII):

[0041]

[0042] Preferably, the modified organic photosensitizer is prepared by reacting Rose Bengal and 3-bromopropylamine hydrobromide.

[0043] Preferably, the modified furan derivative has the structure of the following formula (VIII):

[0044]

[0045] Preferably, the modified furan derivative is prepared by aldehyde-amine reaction of 3-furaldehyde and tert-butyl 2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate, followed by reduction and amino deprotection.

[0046] The present application also provides the application of the above-mentioned light-controlled coupled nanoprobe in tumor diagnosis and tumor treatment.

[0047] Preferably, the application process of the above-mentioned light-controlled coupled nanoprobe in tumor diagnosis and tumor treatment includes:

[0048] After the light-controlled coupled nanoprobe is intravenously injected into the patient's body, 650nm red light is used to irradiate the tumor site for 20-40 minutes.

[0049] The present application also provides a photo-controlled coupled nanoparticle composition, which comprises the above-mentioned photo-controlled coupled nanoprobe and DNA-modified Au or Pt-Au nanoparticles.

[0050] Preferably, the particle size of the DNA-modified Au or Pt-Au nanoparticles is 20-40 nm.

[0051] Preferably, the DNA sequence in the DNA-modified Au or Pt-Au nanoparticles is at least one of an adenine oligomeric chain, a guanine oligomeric chain, and a cytosine oligomeric chain.

[0052] Preferably, in the photo-controlled coupled nanoparticle composition, the molar ratio of the photo-controlled coupled nanoprobe to the DNA-modified Au or Pt-Au nanoparticles is (0.75-1.25):1.

[0053] The present application also provides the use of the above-mentioned light-controlled coupled nanoparticle composition in tumor diagnosis and tumor treatment.

[0054] Preferably, the application process of the above-mentioned light-controlled coupled nanoparticle composition in tumor diagnosis and tumor treatment includes:

[0055] After the photo-controlled coupled nanoparticle composition is intravenously injected into the patient's body, the tumor site is irradiated with 650nm red light for 20-40 minutes.

[0056] One or more technical solutions provided in the embodiments of the present application have at least the following technical effects or advantages:

[0057] The light-controlled coupling nanoprobe provided in the present application is prepared by grafting an organic photosensitizer and a furan group onto a copolymer to form an amphiphilic polymer, which is then coated with Au or Pt-Au nanoparticles. The organic photosensitizer group on the surface of the nanoprobe can be used to generate singlet oxygen under the excitation of light. After the furan group is oxidized by the singlet oxygen, it can be chemically coupled with adenine, guanine or cytosine in DNA, so that the light-controlled coupling nanoprobe can be anchored on the DNA of tumor cells, thereby enhancing the penetration and long-term retention of the nanoprobe at the tumor site; at the same time, the organic photosensitizer and the furan group are simultaneously loaded onto the surface of the light-controlled coupling nanoprobe, so that the singlet oxygen generated by the organic photosensitizer can be effectively provided to the furan group, thereby realizing in situ light-controlled coupling. Compared with the additional introduction of a photosensitizer, the effectiveness and controllability of the light-controlled coupling are greatly improved.

[0058] The light-controlled coupled nanoprobe provided in the present application uses an amphiphilic polymer to coat Au or Pt-Au nanoparticles, which can improve the biocompatibility of Au or Pt-Au nanoparticles and reduce toxic side effects; when the nanoprobe penetrates into tumor cells, the Au or Pt-Au nanoparticles coated in the middle can produce reactive oxygen to kill tumor cells, thereby achieving synergistic treatment of tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 A schematic diagram of the reaction process of light-controlled chemical coupling of the light-controlled coupling nanoprobe provided in the present application with the DNA-modified Au or Pt-Au nanoparticles;

[0060] Figure 2 TEM and particle size distribution diagram of the light-controlled coupled nanoprobe prepared in Example 1 of the present application;

[0061] Figure 3 TEM and particle size distribution diagram of DNA-modified Au nanoparticles in Example 2 of the present application;

[0062] Figure 4 This is a TEM image of the light-controlled coupled nanoparticle composition prepared in Example 2 of the present application after being illuminated;

[0063] Figure 5 TEM and particle size distribution diagram of the light-controlled coupled nanoprobe prepared in Example 3 of the present application;

[0064] Figure 6 TEM and particle size distribution diagram of DNA-modified Pt-Au nanoparticles in Example 4 of the present application;

[0065] Figure 7 This is a TEM image of the light-controlled coupled nanoparticle composition prepared in Example 4 of the present application after being illuminated;

[0066] Figure 8 This is a bright field image of cells after culturing the light-controlled coupled nanoprobe and 4T1 cells in Example 3 of the present application;

[0067] Fig. 9 This is a bright field image of cells after the light-controlled coupled nanoparticle composition and 4T1 cells are cultured in Example 4 of the present application;

[0068] Fig.10 4T1 cells were treated with Rose Bengal and the light-controlled coupled nanoparticle composition in Example 4, respectively, and the live / dead cell staining images were obtained under illumination and no illumination conditions. DETAILED DESCRIPTION

[0069] In order to further explain the technical means and effects adopted by the present application to achieve the intended application purpose, the specific implementation methods, structures, features and effects of the present application are described in detail below in combination with the accompanying drawings and preferred embodiments.

[0070] It should be noted that different "one embodiment" or "embodiment" in this application do not necessarily refer to the same embodiment. In addition, specific features, structures, or characteristics in one or more embodiments may be combined in any suitable form. It should be understood that the embodiments of this application are explanations of the scheme of this application and are not intended to limit the scope of protection of this application.

[0071] In this application, ranges may be expressed as a range from "about" one specific value and / or to "about" another specific value. When expressing such a range, examples include starting from a specific value and / or ending at another specific value. Similarly, when the antecedent "about" is used to indicate that a value is an approximation, it is understood that the specific value constitutes another aspect. It is also understood that the endpoints of each range are meaningful both in relation to the other endpoint and in relation to the other endpoint.

[0072] Unless otherwise stated, it is not intended that any method described herein be construed as requiring that its steps be performed in a specific order. Therefore, when a method claim does not actually state that its steps follow a certain order or it does not specifically indicate in any other way in the claims or description that the steps are limited to a specific order, it is not intended to imply any particular order.

[0073] Although the transitional term "comprising" may be used to disclose various features, elements, or steps of a particular embodiment, it should be understood that this implies alternative embodiments including those that may be described using the transitional terms "consisting of," "consisting essentially of." Thus, for example, implicit alternative embodiments for a method comprising A+B+C include embodiments in which the method consists of A+B+C and embodiments in which the method consists essentially of A+B+C.

[0074] The embodiment of the present application provides a light-controlled coupled nanoprobe, which is a metal nanoparticle coated with an amphiphilic polymer containing a furan unit and an organic photosensitizer unit;

[0075] The amphiphilic polymer containing furan units and organic photosensitizer units has a structure shown in formula (I):

[0076]

[0077] Formula (I), R1, R2, R3, R4 are independently selected from H, methyl, ethyl or phenyl;

[0078] S1 and S2 are both connecting groups selected from alkyl, alkoxy, amino or a combination thereof;

[0079] X is an organic photosensitizer group;

[0080] p / m=0.1-0.4; q / m=0.01-0.2; n / m=0.5-1;

[0081] The metal nanoparticles are Au or Pt-Au nanoparticles.

[0082] Specifically, the amphiphilic polymer containing furan units and organic photosensitizer units is obtained by grafting a modified furan derivative and a modified organic photosensitizer with a first copolymer;

[0083] The first copolymer has a structure shown in formula (II):

[0084]

[0085] In formula (II), R1, R2, R3, and R4 are independently selected from H, methyl, ethyl, or phenyl; n / m=0.5-1.

[0086] The light-controlled coupling nanoprobe provided in the present application is prepared by grafting an organic photosensitizer group and a furan group onto a copolymer to form an amphiphilic polymer, which is then coated with Au or Pt-Au nanoparticles. The organic photosensitizer group on the surface of the nanoprobe can be used to generate singlet oxygen under the excitation of light. After the furan group is oxidized by the singlet oxygen, it can be chemically coupled with adenine, guanine or cytosine in DNA, so that the light-controlled coupling nanoprobe can be anchored on the DNA of tumor cells, thereby enhancing the penetration and long-term retention of the nanoprobe at the tumor site; at the same time, the organic photosensitizer and the furan group are simultaneously loaded onto the surface of the light-controlled coupling nanoprobe, so that the singlet oxygen generated by the organic photosensitizer can be effectively provided to the furan group, thereby realizing in situ light-controlled coupling. Compared with the additional introduction of photosensitizers, the effectiveness and controllability of light-controlled coupling are greatly improved.

[0087] The photocontrolled coupled nanoprobe provided in the present application is coated with Au or Pt-Au nanoparticles with an amphiphilic polymer, which can enable the photocontrolled coupled nanoprobe to be stably dispersed in water, while improving the biocompatibility of Au or Pt-Au nanoparticles and reducing toxic side effects; when the nanoprobe penetrates into tumor cells, the Au or Pt-Au nanoparticles coated in the middle can produce reactive oxygen to kill tumor cells, thereby achieving synergistic treatment of tumors.

[0088] In the present application, the organic photosensitizer can be any photosensitizer that can generate singlet oxygen under light, such as rose Bengal, methylene blue, etc. Rose Bengal and methylene blue can generate singlet oxygen under red light, thereby realizing the photo-controlled coupling of nanoprobes with DNA under red light irradiation, and have good biocompatibility and water solubility, and can improve the water solubility of nanoprobes.

[0089] Specifically, when the organic photosensitizer is Rose Bengal, in formula (I), X-S2-NH- may have a structure of the following formula (III), that is, Rose Bengal is grafted onto the first copolymer via a linking group containing an amino group.

[0090]

[0091] Preferably, in the above formula (I), It has the structure of the following formula (IV), that is, the furan group is grafted onto the first copolymer through the following amino-containing connecting group.

[0092]

[0093] Specifically, the above-mentioned light-controlled coupling type nanoprobe can be prepared by the following steps:

[0094] (1) reacting the modified organic photosensitizer, the modified furan derivative and the first copolymer through an amide reaction to obtain an amphiphilic polymer containing a furan unit and an organic photosensitizer unit;

[0095] (2) mixing, dispersing and drying an amphiphilic polymer containing a furan unit and an organic photosensitizer unit with Au or Pt-Au nanoparticles to obtain a light-controlled coupled nanoprobe;

[0096] The modified organic photosensitizer has the following structure:

[0097] X-S2-NH2(V),

[0098] Wherein X is an organic photosensitizer group; S2 is a linking group selected from alkyl, alkoxy, amino or a combination thereof;

[0099] The modified furan derivative has the following structure:

[0100]

[0101] Wherein, S1 is a connecting group selected from alkyl, alkoxy, amino or a combination thereof.

[0102] In the above preparation method, the modified organic photosensitizer and the modified furan derivative are grafted by an amide reaction between the amino group and the anhydride group in the first copolymer of the following formula (II),

[0103]

[0104] In formula (II), R1, R2, R3, and R4 are independently selected from H, methyl, ethyl, or phenyl; n / m=0.5-1.

[0105] In the present application, the molecular weight of the first copolymer can be any molecular weight, preferably 5000-10000.

[0106] Preferably, the first copolymer is selected from at least one of poly(isobutylene-maleic anhydride), poly(styrene-maleic anhydride), and poly(ethylene-maleic anhydride).

[0107] In the above preparation method, specifically, by adjusting the ratio of the modified organic photosensitizer, the modified furan derivative and the first copolymer, the ratio of the grafted organic photosensitizer group and the furan group on the first copolymer can be controlled, thereby adjusting the amount of singlet oxygen generated in the light-controlled coupling nanoprobe and the anchoring ability between the light-controlled coupling nanoprobe and DNA. Since the organic photosensitizer unit and the furan unit are grafted onto the hydrophilic anhydride group of the first copolymer through an amide reaction, when the number of grafted organic photosensitizer units and furan units is too large, the hydrophobicity of the polymer increases, thereby affecting its hydrophilicity. When the number of grafted organic photosensitizer units and furan units is too small, the effectiveness of the light-controlled coupling of the nanoprobe with DNA is reduced. Specifically, the ratio of the modified organic photosensitizer, the modified furan derivative and the first copolymer is determined according to the ratio of the number of amino groups in the modified organic photosensitizer and the modified furan derivative to the maleic anhydride groups in the first copolymer, wherein the molar ratio of the amino groups in the modified organic photosensitizer to the maleic anhydride groups is preferably 0.1-0.4, and the molar ratio of the amino groups in the modified furan derivative to the maleic anhydride groups is preferably 0.01-0.2.

[0108] Preferably, the mass ratio of the amphiphilic polymer containing furan units and organic photosensitizer units to Au or Pt-Au nanoparticles is 4:1-7:1, more preferably 5:1-6:1; the mass proportion of metal nanoparticles in the prepared light-controlled coupled nanoprobe is 12.5%-20%.

[0109] Preferably, the diameter of the Au nanoparticles is 5-8 nm, and the diameter of the Pt-Au nanoparticles is 8-10 nm. Specifically, in the above preparation method, the Au or Pt-Au nanoparticles used are Au or Pt-Au nanoparticles chelated with oleylamine. The particle size of the prepared light-controlled coupled nanoprobe is 10-50 nm.

[0110] Specifically, when the organic photosensitizer is Bengal rose red, the modified organic photosensitizer may have the following structure of formula (VII):

[0111]

[0112] The modified organic photosensitizer is prepared by reacting Rose Bengal and 3-bromopropylamine hydrobromide. Specifically, the reaction process is as follows:

[0113]

[0114] Preferably, the modified furan derivative has the structure of the following formula (VIII):

[0115]

[0116] The modified furan derivative is prepared by aldehyde-amine reaction of 3-furaldehyde and tert-butyl 2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate, followed by reduction and amino deprotection. Specifically, the reaction process is as follows:

[0117]

[0118] The above-mentioned light-controlled coupled nanoprobe provided in the present application can be applied to tumor diagnosis and tumor treatment.

[0119] After the above-mentioned light-controlled coupling nanoprobe penetrates into the tumor cells, it can produce singlet oxygen under light conditions, prompting the nanoprobe to couple with the DNA in the cell, so that the nanoprobe is anchored in the cell and retained for a long time, thereby enabling fluorescence imaging. At the same time, the Au or Pt-Au nanoparticles in the nanoprobe can promote tumor cell apoptosis and play a role in tumor treatment.

[0120] Specifically, when the above-mentioned light-controlled coupled nanoprobe is used in tumor diagnosis and tumor treatment, the above-mentioned light-controlled coupled nanoprobe is injected into the patient's body intravenously, and then the tumor site is irradiated with 650nm red light for 20-40 minutes to enable the nanoprobe to couple intracellularly.

[0121] The present application also provides a photo-controlled coupled nanoparticle composition, which comprises the above-mentioned photo-controlled coupled nanoprobe and DNA-modified Au or Pt-Au nanoparticles. In the photo-controlled coupled nanoparticle composition, the photo-controlled coupled nanoprobe can chemically couple with the DNA sequence in the DNA-modified Au or Pt-Au nanoparticles under light conditions, thereby increasing the size of the nanoprobe particles, increasing its enrichment at the tumor site, and prolonging its retention time at the tumor site, thereby enhancing its tumor diagnosis effect; at the same time, the photo-controlled coupled nanoprobe and the DNA-modified Au or Pt-Au nanoparticles are photo-controlled coupled, which greatly increases the number of Au or Pt-Au nanoparticles enriched at the tumor site, which is more conducive to killing tumor cells and enhancing its tumor treatment effect.

[0122] Preferably, the particle size of the above-mentioned DNA-modified Au or Pt-Au nanoparticles is 20-40 nm.

[0123] Preferably, the DNA sequence in the above-mentioned DNA-modified Au or Pt-Au nanoparticles is at least one of an adenine oligomeric chain, a guanine oligomeric chain, and a cytosine oligomeric chain.

[0124] Preferably, in the above-mentioned photo-controlled coupled nanoparticle composition, the molar ratio of the photo-controlled coupled nanoprobe to the DNA-modified Au or Pt-Au nanoparticles is (0.75-1.25):1, more preferably (0.9-1.1):1.

[0125] When the above-mentioned light-controlled coupled nanoparticle composition is used for tumor diagnosis and tumor treatment, the light-controlled coupled nanoparticle composition is intravenously injected into the subject's body, and then 650nm red light is used to irradiate the tumor site for 20-40 minutes.

[0126] Example 1

[0127] This embodiment provides a preparation process of a light-controlled coupled nanoprobe:

[0128] (1) Preparation of an amino-modified organic photosensitizer: 1.0 g of Rose Bengal and 0.33 g of 3-bromopropionic acid ammonium hydrobromide were added to 30 mL of N,N-dimethylformamide, respectively, and the solid powder was dissolved by stirring, and the reaction was carried out at 80° C. in a dark environment for 7 h; after the reaction system was cooled to room temperature, the product was precipitated and dried with a mixed solution of ether and water (volume ratio of 1:1) to obtain an amino-modified Rose Bengal;

[0129] (2) Preparation of amino-modified furan derivatives: 104 μL of 3-furaldehyde, 0.25 g of tert-butyl 2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate, and 0.1 g of anhydrous sodium sulfate were added to 15 mL of methanol, mixed well, and reacted at 50° C. for 12 h. After the reaction system was cooled to room temperature, 0.16 g of sodium borohydride was added in an ice bath, and the mixture was stirred for 1 h while maintaining the ice bath. After the methanol was evaporated, the mixture was extracted, filtered, and rotary evaporated. The product was then dissolved in an appropriate amount of dichloromethane. Trifluoroacetic acid was slowly added dropwise in an ice bath, stirred at room temperature for 2 h, and the solvent was rotary dried to obtain an amino-modified furan derivative.

[0130] (3) Preparing an amphiphilic polymer containing a furan unit and an organic photosensitizer unit: adding 0.6 g of poly(isobutylene-maleic anhydride), 320 μL of oleylamine, the amino-modified rose Bengal obtained in step (1), and the modified furan derivative obtained in step (2) to 10 mL of N,N-dimethylformamide, respectively, and adding triethylamine to adjust the pH to 8-9, and then stirring at room temperature for 12 h; after the reaction is completed, centrifuging and washing, and finally stably dispersing the product in 5 mL of dichloromethane to obtain an amphiphilic polymer dispersion containing a furan unit and an organic photosensitizer unit;

[0131] (4) Preparation of photo-controlled coupled nanoprobes: Au nanoparticles chelated with oleylamine were uniformly dispersed in chloroform to obtain an Au nanoparticle dispersion, wherein the concentration of Au nanoparticles in the dispersion was 11 mg / mL; 100 μL of Au nanoparticle dispersion, 6 mg of amphiphilic polymer dispersion and 0.86 mL of chloroform were uniformly mixed, and then 10 mL of NaOH aqueous solution with a pH of 8.0-10.0 was added, and a pink emulsion was obtained by ultrasonic emulsification; the pink emulsion was stirred at 45°C for 3 h, and after the oil phase solvent was completely evaporated, it was naturally cooled to room temperature, and then rotary evaporated at 65°C for 15 min under vacuum conditions to obtain a photo-controlled coupled nanoprobe.

[0132] Example 2

[0133] This embodiment provides a preparation process of a photo-controlled coupled nanoparticle composition:

[0134] The photocontrolled coupled nanoprobe prepared in Example 1 is dispersed in deionized water, and the DNA-modified Au nanoparticles are dispersed in deionized water, wherein the size of the DNA-modified Au nanoparticles is 5 nm and they are prepared by conventional methods; under light-proof conditions, the photocontrolled coupled nanoprobe dispersion and the DNA-modified Au nanoparticle dispersion are mixed, wherein the molar ratio of the photocontrolled coupled nanoprobe to the DNA-modified Au nanoparticles is 1:1, to obtain a dispersion of a photocontrolled coupled nanoparticle composition.

[0135] Example 3

[0136] This embodiment provides a preparation process of a light-controlled coupled nanoprobe:

[0137] (1) Preparation of an amino-modified organic photosensitizer: 1.0 g of Rose Bengal and 0.33 g of 3-bromopropionic acid ammonium hydrobromide were added to 30 mL of N,N-dimethylformamide, respectively, and the solid powder was dissolved by stirring, and the reaction was carried out at 80° C. in a dark environment for 7 h; after the reaction system was cooled to room temperature, the product was precipitated and dried with a mixed solution of ether and water (volume ratio of 1:1) to obtain an amino-modified Rose Bengal;

[0138] (2) Preparation of amino-modified furan derivatives: 104 μL of 3-furaldehyde, 0.25 g of tert-butyl 2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate, and 0.1 g of anhydrous sodium sulfate were added to 15 mL of methanol, mixed well, and reacted at 50° C. for 12 h. After the reaction system was cooled to room temperature, 0.16 g of sodium borohydride was added in an ice bath, and the mixture was stirred for 1 h while maintaining the ice bath. After the methanol was evaporated, the mixture was extracted, filtered, and rotary evaporated. The product was then dissolved in an appropriate amount of dichloromethane. Trifluoroacetic acid was slowly added dropwise in an ice bath, stirred at room temperature for 2 h, and the solvent was rotary dried to obtain an amino-modified furan derivative.

[0139] (3) Preparing an amphiphilic polymer containing a furan unit and an organic photosensitizer unit: adding 0.6 g of poly(isobutylene-maleic anhydride), 320 μL of oleylamine, the amino-modified rose Bengal obtained in step (1), and the modified furan derivative obtained in step (2) to 10 mL of N,N-dimethylformamide, respectively, and adding triethylamine to adjust the pH to 8-9, and then stirring at room temperature for 12 h; after the reaction is completed, centrifuging and washing, and finally stably dispersing the product in 5 mL of dichloromethane to obtain an amphiphilic polymer dispersion containing a furan unit and an organic photosensitizer unit;

[0140] (4) Preparation of photo-controlled coupled nanoprobes: Pt-Au nanoparticles chelated with oleylamine were uniformly dispersed in chloroform to obtain a Pt-Au nanoparticle dispersion, wherein the concentration of Pt-Au nanoparticles in the dispersion was 6 mg / mL; 100 μL of the Pt-Au nanoparticle dispersion, 6 mg of the amphiphilic polymer dispersion and 0.86 mL of chloroform were uniformly mixed, and then 10 mL of a NaOH aqueous solution with a pH of 8.0-10.0 was added, and a pink emulsion was obtained by ultrasonic emulsification; the pink emulsion was stirred at 45°C for 3 h, and after the oil phase solvent was completely evaporated, it was naturally cooled to room temperature, and then rotary evaporated at 65°C for 15 min under vacuum conditions to obtain a photo-controlled coupled nanoprobe.

[0141] Example 4

[0142] This embodiment provides a preparation process of a photo-controlled coupled nanoparticle composition:

[0143] The photocontrolled coupled nanoprobe prepared in Example 1 is dispersed in deionized water, and the DNA-modified Pt-Au nanoparticles are dispersed in deionized water, wherein the size of the DNA-modified Pt-Au nanoparticles is 8 nm and they are prepared by conventional methods; under light-proof conditions, the photocontrolled coupled nanoprobe dispersion and the DNA-modified Pt-Au nanoparticle dispersion are mixed, wherein the molar ratio of the photocontrolled coupled nanoprobe to the DNA-modified Pt-Au nanoparticles is 1:1, to obtain a dispersion of the photocontrolled coupled nanoparticle composition.

[0144] The light-controlled coupled nanoprobe prepared in Example 1 was characterized and the following results were obtained: Figure 2 TEM and particle size distribution shown;

[0145] The DNA-modified Au nanoparticles in Example 2 were characterized to obtain Figure 3 TEM and particle size distribution diagram; the dispersion of the photo-controlled coupled nanoparticle composition in Example 2 was irradiated with a 650nm laser at 50mw / cm 2 After irradiation for 40 min, the characterization was carried out and the Figure 4 TEM images shown;

[0146] The light-controlled coupled nanoprobe prepared in Example 3 was characterized and the following results were obtained: Figure 5 TEM and particle size distribution shown;

[0147] The DNA-modified Pt-Au nanoparticles in Example 4 were characterized to obtain Figure 6 TEM and particle size distribution diagram; the dispersion of the photo-controlled coupled nanoparticle composition in Example 4 was irradiated with a 650nm laser at 50mw / cm 2 After irradiation for 40 min, the characterization was carried out and the Figure 7 TEM image shown.

[0148] from Figure 2 It can be seen that the particle size of the light-controlled coupled nanoprobe prepared in Example 1 is 13.0nm-19.8nm; Figure 3 It can be seen that the particle size of the DNA-modified Au nanoparticles in Example 2 is 23.3nm-34.3nm; Figure 4 It can be seen that when the light-controlled coupling nanoprobe prepared in Example 1 is light-controlled coupled with the DNA-modified Au nanoparticles, a coupled body with a significantly increased particle size is obtained. Figure 5 It can be seen that the particle size of the light-controlled coupled nanoprobe prepared in Example 3 is 23.0nm-42.4nm; Figure 6 It can be seen that the particle size of the DNA-modified Pt-Au nanoparticles in Example 4 is 23.3nm-32.3nm; Figure 7 It can be seen that when the light-controlled coupling nanoprobe prepared in Example 3 is light-controlled coupled with the DNA-modified Pt-Au nanoparticles, a coupled body with a significantly increased particle size is obtained.

[0149] Under light-proof conditions, the nanoprobe prepared in Example 3 was added to a culture medium (DMEM high-glucose liquid culture medium containing 10% FBS), 0.1 μM methylene blue was added, and added to 4T1 cells. After culturing for 7 h, a 650 nm laser was used to illuminate the cells at 50 mw / cm 2 Irradiate for 5 minutes at an irradiance of Figure 8 The bright field images of cells shown in Figure 1, where (a) is before laser irradiation and (b) is after laser irradiation. Figure 8 It can be seen that before laser irradiation, there is no aggregation of nanoparticles. After laser irradiation, the aggregation of nanoparticles in cells can be clearly seen (black dots in the figure), indicating that the photo-controlled coupled nanoparticle composition is coupled after laser irradiation, thereby achieving aggregation in cells.

[0150] Under light-proof conditions, the light-controlled coupled nanoparticle composition prepared in Example 4 was added to a culture medium (DMEM high-glucose liquid culture medium containing 10% FBS), 0.1 μM methylene blue was added, and added to 4T1 cells. After culturing for 7 h, a 650 nm laser was used at 50 mw / cm 2 Irradiate for 5 minutes at an irradiance of Fig. 9 Bright field image of cells shown. Fig. 9 It can be seen that after laser irradiation, the aggregation of nanoprobes (black dots in the picture) in the cells can be clearly seen.

[0151] Under light-proof conditions, Bengal rose red and the light-controlled coupled nanoparticle composition prepared in Example 4 were added to a culture medium (DMEM high-glucose liquid culture medium containing 10% FBS), and 0.1 μM methylene blue was added to each of them. Then, they were added to 4T1 cells and cultured for 7 h. The cells were divided into two groups, one of which was irradiated with laser and the other was kept in light-proof conditions. The laser irradiation conditions were: 650 nm laser at 50 mw / cm 2 After irradiation for 5 min, the cell survival was detected and the results were as follows: Fig.10 Live / dead cell staining shown, where Fig.10 a1 and Fig.10 b1 are the cell staining images of 4T1 cells treated with Bengal Rose Red before and after laser irradiation; Fig.10 a2 and Fig.10b2 is the cell staining image of 4T1 cells treated with the light-controlled coupled nanoparticle composition without laser irradiation and after laser irradiation. Fig.10 It can be seen that Fig.10 a1 and Fig.10 In b1, the number of surviving 4T1 cells treated with Rose Bengal did not change significantly under light-free and light-exposed conditions; Fig.10 a1, Fig.10 In a2, the number of surviving 4T1 cells treated with the light-controlled coupled nanoparticle composition was significantly reduced under light-free conditions. Fig.10 In b2, the number of surviving cells in 4T1 cells treated with the light-controlled coupled nanoparticle composition under illumination conditions is further reduced. This shows that the light-controlled coupled nanoparticle composition provided by the present application can penetrate into cells and kill cells, and after illumination, the light-controlled coupled nanoparticle composition can stay in cells for a long time, thereby killing more cells.

[0152] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present application and are not intended to limit it. Although the present application has been described in detail with reference to the preferred embodiments, a person of ordinary skill in the art should understand that the technical solution of the present application can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution of the present application, which should be included in the scope of the claims of the present application.

Claims

1. A light-controlled coupling nanoprobe, characterized in that: The light-controlled coupled nanoprobe is a metal nanoparticle coated with an amphiphilic polymer containing a furan unit and an organic photosensitizer unit; The amphiphilic polymer containing furan units and organic photosensitizer units has a structure shown in formula (I): In formula (I), R1, R2, R3, and R4 are independently selected from H, methyl, ethyl, or phenyl; S1 and S2 are both connecting groups selected from alkyl, alkoxy, amino or a combination thereof; X is an organic photosensitizer group; p / m=0.1-0.4; q / m=0.01-0.2; n / m=0.5-1; The metal nanoparticles are Au or Pt-Au nanoparticles.

2. The light-controlled coupling nanoprobe according to claim 1, characterized in that: The amphiphilic polymer containing furan units and organic photosensitizer units is obtained by grafting a modified furan derivative and a modified organic photosensitizer with a first copolymer; The first copolymer has a structure shown in formula (II): In formula (II), R1, R2, R3, and R4 are independently selected from H, methyl, ethyl, or phenyl; The modified organic photosensitizer has the structure of the following formula (V): X-S2-NH2(V), Wherein X is an organic photosensitizer group; S2 is a linking group selected from alkyl, alkoxy, amino or a combination thereof; The modified furan derivative has the structure of the following formula (VI): Wherein, S1 is a connecting group selected from alkyl, alkoxy, amino or a combination thereof.

3. The light-controlled coupling nanoprobe according to claim 2, characterized in that: The first copolymer is selected from at least one of poly(isobutylene-maleic anhydride), poly(styrene-maleic anhydride), and poly(ethylene-maleic anhydride).

4. The light-controlled coupling nanoprobe according to claim 1, characterized in that: The mass proportion of metal nanoparticles in the light-controlled coupled nanoprobe is 12.5%-20%.

5. The light-controlled coupling nanoprobe according to claim 1, characterized in that: The particle size of the light-controlled coupling nanoprobe is 10-50 nm.

6. The method for preparing the light-controlled coupled nanoprobe according to any one of claims 1 to 5, characterized in that: The following steps are involved: (1) reacting the modified organic photosensitizer, the modified furan derivative and the first copolymer through an amide reaction to obtain an amphiphilic polymer containing a furan unit and an organic photosensitizer unit; (2) mixing, dispersing and drying the amphiphilic polymer containing furan units and organic photosensitizer units with Au or Pt-Au nanoparticles to obtain the light-controlled coupled nanoprobe; The modified organic photosensitizer has the structure of the following formula (V): X-S2-NH2(V), Wherein X is an organic photosensitizer group; S2 is a linking group selected from alkyl, alkoxy, amino or a combination thereof; The modified furan derivative has the structure of the following formula (VI): Wherein, S1 is a linking group selected from alkyl, alkoxy, amino or a combination thereof; The first copolymer has the structure of the following formula (II): In formula (II), R1, R2, R3, and R4 are independently selected from H, methyl, ethyl, or phenyl.

7. Use of the light-controlled coupled nanoprobe according to any one of claims 1 to 5 in the preparation of tumor diagnosis and tumor treatment drugs.

8. A photo-controlled coupled nanoparticle composition, characterized in that: The photo-controlled coupled nanoparticle composition comprises the photo-controlled coupled nanoprobe according to any one of claims 1 to 5 and DNA-modified Au or Pt-Au nanoparticles.

9. The light-controlled coupled nanoparticle composition according to claim 8, characterized in that: The particle size of the DNA-modified Au or Pt-Au nanoparticles is 20-40 nm; The DNA sequence in the DNA-modified Au or Pt-Au nanoparticles is at least one of an adenine oligomeric chain, a guanine oligomeric chain, and a cytosine oligomeric chain; In the photo-controlled coupled nanoparticle composition, the molar ratio of the photo-controlled coupled nanoprobe to the DNA-modified Au or Pt-Au nanoparticles is (0.75-1.25):

1.

10. Use of the light-controlled coupled nanoparticle composition according to claim 8 in the preparation of tumor diagnosis and tumor treatment drugs.

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