A tumor-targeted photothermal / drug synergistic therapeutic material based on pillararene

Abstract

The present invention discloses a tumor-targeted photothermal / drug synergistic therapy material based on pillararenes. The present invention designs and synthesizes a copolymerized nanomaterial of pillararenes and dopamine. On the one hand, the copolymerized nanomaterial forms targeted nanoparticles through host-guest complexation between the pillararenes and the targeting molecule pyridinium conjugated folic acid (FA-Py). On the other hand, the nanoparticles are used as carriers to load doxorubicin hydrochloride (DOX) on the surface of polydopamine through physical adsorption. Under the irradiation of near-infrared laser, the polydopamine can be heated to achieve photothermal therapy, while also promoting the decomposition of the nanoparticles and the release of DOX, thereby achieving tumor-targeted photothermal / drug synergistic therapy.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a tumor-targeted photothermal / drug synergistic therapeutic material based on pillararenes. Background Art

[0002] Macrocyclic molecules hold a significant position in the supramolecular field. Through sustained and steady development, pillararenes have become a representative fifth-generation macrocyclic molecule. From synthesis and structural exploration to functional group modification and host-guest properties, pillararenes have rapidly gained a significant position in supramolecular chemistry. Pillararenes are a class of macrocyclic host molecules composed of hydroquinone or hydroquinone ethers linked para-to the benzene ring via a methylene group. Compared to previous generations of macrocyclic molecules, pillararenes not only combine the characteristics of these macrocyclic molecules, such as a highly symmetrical columnar structure and 2n substituents, but also offer amenable functional modification of the flanking groups, allowing host-guest complexation with corresponding guest molecules. In recent years, pillararenes have been widely used in the medical and biological fields. By assembling with other molecules to form nanoparticles, they are considered an excellent class of drug delivery vehicles.

[0003] Under alkaline conditions, dopamine can be converted into polydopamine through sequential oxidation, intramolecular cyclization, and oligomerization / self-assembly. Because polydopamine can effectively absorb near-infrared light and convert it into heat energy, scientists have successfully applied dopamine to treat tumor cells over the past decade. However, most of these have been done by copolymerizing inorganic materials with dopamine to form MOFs, such as ZIF-8, UiO-66, and MIL-101 to prepare stimuli-responsive multifunctional MOFs. Very few have been done by copolymerizing dopamine with pillararenes, and then using the cavities of the pillararenes to complex with targeting molecules for targeted drug transport into cells. Summary of the Invention

[0004] The purpose of the present invention is to provide a tumor-targeted photothermal / drug synergistic therapeutic material based on pillararenes.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] A dopamine-pillar aromatic copolymer nanomaterial is obtained by copolymerizing pillar aromatic hydrocarbon P[5]OH and dopamine.

[0007] The structural formula of the pillar aromatic hydrocarbon P[5]OH is shown below:

[0008]

[0009] The preparation method of the above-mentioned dopamine-pillar aromatic copolymer nanomaterial is as follows: an aqueous solution of dopamine hydrochloride is added to an aqueous solution of tris(hydroxymethylaminomethane), and then a dimethylformamide solution of pillar aromatic hydrocarbon P[5]OH is added, and the mixture is stirred and reacted at room temperature for 24 hours. The reaction solution is dialyzed to obtain dopamine-pillar aromatic copolymer particles.

[0010] Application of the above-mentioned dopamine-pillar aromatic copolymer nanomaterial in the preparation of tumor therapeutic drugs.

[0011] A tumor treatment drug comprises the above-mentioned dopamine-pillararene copolymer nanomaterial, pyridinium conjugated folic acid and a chemotherapy drug. In one embodiment of the present invention, the chemotherapy drug is doxorubicin hydrochloride.

[0012] The present invention designs and synthesizes a copolymer nanomaterial of pillararenes and dopamine. On the one hand, the copolymer nanomaterial forms targeted nanoparticles through host-guest complexation between the pillararenes and the targeting molecule pyridinium conjugated folic acid (FA-Py). On the other hand, the nanoparticles are used as carriers to load doxorubicin hydrochloride (DOX) on the surface of polydopamine through physical adsorption. Under the irradiation of near-infrared laser, the polydopamine is heated to achieve photothermal therapy, while also promoting the decomposition of the nanoparticles and the release of DOX, thereby achieving tumor-targeted photothermal / drug synergistic therapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Schematic diagram of the preparation process of the pillararenes-based tumor-targeted photothermal / drug synergistic therapeutic material of the present invention.

[0014] Figure 2 This is the H NMR spectrum of the aromatic hydrocarbon of the dopamine-modified column [5] in Example 1.

[0015] Figure 3 Infrared spectral analysis (a) of PDA, P[5]OH and PDA-P[5]OH in Example 1, scanning electron microscopy image (b) of PDA-P[5]OH and dynamic light scattering image (c) of PDA-P[5]OH.

[0016] Figure 4 、 Figure 5 、 Figure 6 This is the therapeutic effect of PDA-P[5]OH-FA-Py@DOX on tumor cells in Example 2. DETAILED DESCRIPTION

[0017] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific examples, but should not be construed as limiting the present invention. Without departing from the spirit and substance of the present invention, modifications or substitutions made to the inventive method, steps or conditions are intended to fall within the scope of the present invention. Experimental methods and reagents for which specific conditions are not specified in the examples are based on conventional conditions in the art.

[0018] Example 1

[0019] 1. Synthesis of dopamine-modified column[5] aromatics

[0020]

[0021] To a 50 mL round-bottom flask, pillararene P1 (0.4 g, 0.5 mol) and 3,4-dihydroxyphenylacetic acid (0.08 g, 0.5 mol) dissolved in 20 mL of DMF were added. O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (0.57 g, 1.5 mol) and 0.34 mL of N,N-diisopropylethylamine (DIPEA) were also added. The mixture was reacted under nitrogen for 12 hours. After completion of the reaction, excess water was added, the mixture was cooled, and the precipitated product was filtered to obtain a white solid (50%).

[0022] pass Figure 2 The successful synthesis of dopamine-modified column [5]arene (P[5]OH) was confirmed by the H NMR spectrum.

[0023] 2. Dopamine modified column [5] Preparation of copolymerized nanomaterials of aromatic hydrocarbons and dopamine

[0024] Dissolve 410 mg of tris (hydroxymethyl)aminomethane (Tris) in 20 mL of deionized water and stir at room temperature for 0.5 hours. Then, dissolve 0.04 g of dopamine hydrochloride in 0.8 mL of deionized water and add this to the reaction mixture. When the color turns dark brown, add 5 mg of P[OH] (4.86 μM) dissolved in 200 μL of DMF to the reaction mixture. Continue stirring at room temperature for 24 hours. After the reaction is complete, place the reaction mixture in a dialysis bag (molecular weight 3500) and dialyze it against deionized water for 48 hours to obtain PDA-P[5]OH nanoparticles.

[0025] 4 mg of PDA-P[5]OH was dispersed in 4 mL of deionized water, followed by 0.5 mg of FA-Py dissolved in 1 mL of deionized water. FA-Py at a concentration of 0.5 mg / mL was then added dropwise to the aqueous solution. After stirring at room temperature for 12 hours, the reaction solution was dialyzed against deionized water in a dialysis bag (molecular weight 3500) for 24 hours to obtain PDA-P[5]OH-FA-Py nanoparticles.

[0026] 5 mg of PDA-P[5]OH-FA-Py was dissolved in 5 mL of phosphate buffer (pH 7.4). 5 mL of a 5 mg / mL aqueous solution of doxorubicin hydrochloride was added dropwise to the reaction mixture. After stirring at room temperature for 24 hours, the reaction mixture was dialyzed against deionized water in a dialysis bag (molecular weight 3500) for 24 hours. During this time, free DOX was precipitated, yielding PDA-P[5]OH-FA-Py@DOX nanoparticles.

[0027] like Figure 3 As shown in the figure, the peak of C=O group which is not present in PDA appears in the spectrum of PDA-P[5]OH, indicating the formation of PDA-P[5]OH copolymer compound; the SEM image shows that the prepared PDA-P[5]OH nanoparticles are spherical, and the particle size analysis test shows that their diameter is about 130nm, and the dispersion is better than 0.4.

[0028] Example 2

[0029] Effects of PDA-P[5]OH-FA-Py@DOX nanoparticles on cervical cancer (HeLa) cells

[0030] To determine the photothermal-chemical synergistic therapeutic effect of PDA-P[5]OH-FA-Py@DOX, an MTT assay was performed. PDA-P[5]OH, PDA-P[5]OH-FA-Py, PDA-P[5]OH@DOX, and PDA-P[5]OH-FA-Py@DOX were prepared to prepare PDA-P[5]OH at concentrations of 7, 14, 28, 42, 57, and 70 μg / mL, respectively, and the corresponding DOX concentrations were 3, 6, 12, 18, 24, and 30 μg / mL, respectively. The cells were then cultured with the prepared mixture for 48 hours.

[0031] HeLa cells were stained with live / dead cell double staining reagent ( Figure 4 As shown), red represents dead cells and green represents live cells. By comparison, it was found that the treatment group containing PDA-P[5]OH-FA-Py@DOX had a stronger ability to kill cells than free DOX or other treatment groups, especially under 808nm near-infrared laser irradiation, the nanoparticles showed a stronger killing effect. And through cytotoxicity experiments ( Figure 5 The above conclusion can also be confirmed.

[0032] In order to explore the tumor therapeutic effect of PDA-P[5]OH-FA-Py@DOX, HeLa cells were first cultured in PDA-P[5]OH-FA-Py@DOX, and the fluorescence intensity of DOX in HeLa cells was recorded using a fluorescence microscope ( Figure 6 As shown in the figure, after incubation in PDA-P[5]OH-FA-Py@DOX for 1 h, the cell nucleus was stained with DAPI, emitting blue fluorescence, and obvious DOX red fluorescence appeared in the cells. As the incubation time prolonged, the red fluorescence also increased and overlapped with the blue fluorescence of the cell nucleus, indicating that PDA-P[5]OH-FA-Py@DOX had been internalized into the cells and DOX was released into the cell nucleus.

Claims

1. A dopamine-pillararene copolymer nanomaterial, characterized in that: The dopamine-pillar aromatic hydrocarbon (P[5]OH) and dopamine copolymer are prepared by the following method: adding an aqueous solution of dopamine hydrochloride and a dimethylformamide solution of pillar aromatic hydrocarbon (P[5]OH) to a 20.5 mg / mL aqueous solution of tris(hydroxymethyl)aminomethane, respectively, with the molar ratio of dopamine hydrochloride to pillar aromatic hydrocarbon (P[5]OH) being 50:1; stirring the reaction at room temperature for 24 hours; and dialyzing the reaction solution to obtain the dopamine-pillar aromatic hydrocarbon copolymer nanomaterial. The structural formula of the pillar aromatic hydrocarbon P[5]OH is shown below: 。 2. The method for preparing the dopamine-pillararene copolymer nanomaterial according to claim 1, wherein: To a 20.5 mg / mL aqueous solution of tris(hydroxymethylaminomethane), an aqueous solution of dopamine hydrochloride and a dimethylformamide solution of pillar aromatic hydrocarbon P[5]OH were added respectively, with a molar ratio of dopamine hydrochloride to pillar aromatic hydrocarbon P[5]OH of 50:

1. The reaction was stirred at room temperature for 24 hours, and the reaction solution was dialyzed to obtain dopamine-pillar aromatic hydrocarbon copolymer nanomaterials.

3. Use of the dopamine-pillararene copolymer nanomaterial according to claim 1 in the preparation of tumor therapeutic drugs.

4. A drug for treating tumors, characterized in that: The invention comprises the dopamine-pillar aromatic copolymer nanomaterial according to claim 1, pyridinium conjugated folic acid and chemotherapy drugs.

5. The tumor treatment drug according to claim 4, characterized in that: The chemotherapy drug is doxorubicin hydrochloride.

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