Multilayer structure chiral gold-cobalt nano particle for treating oral squamous cell carcinoma

Abstract

Based on chloroauric acid and a cobalt precursor, through the synergistic effect of a stabilizer and a chiral ligand, the chiral gold-cobalt nano particle with the multi-layer structure is successfully constructed. The nanoparticles show excellent physical and chemical characteristics under the selection of metal elements and the regulation and control of ligands. Through the synergistic effect of gold and cobalt, the magnetism of the material is enhanced, the material is endowed with unique optical characteristics, and meanwhile, the material has special biocompatibility and stability due to introduction of a chiral ligand. Particularly, in treatment of oral squamous cell carcinoma, by virtue of excellent magnetic and optical properties, the magnetic nano-material can effectively target a tumor site and improve the treatment effect, and a novel, efficient and safe treatment strategy is provided for treatment of oral squamous cell carcinoma. The novel chiral gold-cobalt nanoparticles are expected to play an important role in the field of cancer treatment, especially in targeted therapy and imaging diagnosis.

Description

Technical Field

[0001] The present invention relates to nanomaterials and their applications in biomedicine, and particularly to a multi-layered chiral gold-cobalt nanoparticle for the treatment of oral squamous cell carcinoma adenocarcinoma. This material combines magnetic and optical properties and realizes targeted delivery of cancer cells through the modification of chiral ligands, belonging to the fields of medical materials and cancer treatment technologies. Background Art

[0002] Oral squamous cell carcinoma is a malignant tumor originating from the oral mucosal epithelium, mainly seen in areas such as the tongue, lips, cheeks, and gums. Due to the high local invasiveness and metastasis of this cancer, treatment often faces severe challenges. Currently, the main strategies for treating oral squamous cell carcinoma include surgical resection, radiotherapy, chemotherapy, and targeted therapy, etc. Surgical resection is the preferred option for early-stage localized tumors. However, since the tumor often invades adjacent structures, such as the facial nerve and muscles related to oral functions, dysfunction is easily caused during the surgical operation, which in turn affects the patient's speech and eating abilities. In addition, the local recurrence rate after surgery is relatively high, and adjuvant treatment is often required.

[0003] As one of the treatment methods, radiotherapy can effectively reduce the tumor volume and control local lesions. However, while radiotherapy kills cancer cells, it inevitably damages the surrounding normal tissues, which may lead to side effects such as oral mucositis, dry mouth, and dysphagia. In addition, some tumors are resistant to radiotherapy, limiting the therapeutic effect; Chemotherapy is mainly applicable to advanced or metastatic oral squamous cell carcinoma, and it inhibits the proliferation and spread of cancer cells through systemic drug administration. However, due to the lack of targeting of chemotherapy drugs, their toxicity to normal cells is relatively high, often causing adverse reactions such as nausea, vomiting, hair loss, and myelosuppression; Targeted therapy aims to reduce the damage to normal cells and improve the quality of life of patients by precisely intervening in specific molecular markers of tumors. However, due to the high heterogeneity and complex molecular mechanisms of oral squamous cell carcinoma, the current efficacy of targeted therapy in clinical practice is still limited, and further research and verification are urgently needed. In recent years, with the rapid development of nanotechnology, tumor treatment methods based on nanomaterials have gradually emerged, especially the application of chiral multi-level nanomaterials in cancer treatment has attracted much attention. Due to the high compatibility and multi-functionality of chiral multi-level structures with biological systems, they have become highly potential nanotherapeutic carriers. Such materials can not only achieve specific binding to tumor cells by regulating chirality, but also realize multiple functions such as targeted delivery, drug controlled release, and imaging diagnosis by introducing magnetic and optical properties. Current research shows that assembling noble metals such as gold and transition metal elements such as cobalt with chiral ligands into complex chiral multi-level structures can not only enhance the biocompatibility of the materials, but also improve their stability in vivo and reduce side effects during the treatment process. Gold, as a noble metal, has excellent surface plasmon resonance effect, which makes it have important applications in the fields of biomedical imaging and photothermal therapy; Cobalt, as an element with significant magnetism, can achieve magnetic control treatment of tumors under the action of an external magnetic field, providing new technical support for the "diagnosis and treatment integration" treatment platform, thereby improving the treatment effect and reducing side effects.

[0004] Although nanomaterials and chiral superstructures have shown broad prospects in the field of tumor treatment, there are still some problems to be solved in current research. For example, how to achieve the efficient assembly of different functional components and how to ensure the long-term stability and low toxicity of the materials in vivo are still important challenges faced by scientists. In addition, the behavior of chiral superstructures in different biological environments and their specific interaction mechanisms with cancer cells have not been fully revealed. Therefore, based on chloroauric acid and cobalt precursors, this invention constructs a multi-level structured chiral gold-cobalt nanoparticle by the synergistic action of a stabilizer and a chiral ligand. This material combines the advantages of magnetism, optics, and chiral ligands, and exhibits excellent stability and biocompatibility in vitro and in vivo, and is expected to provide an efficient and safe treatment method for oral squamous cell carcinoma. Summary of the Invention

[0005] The present invention provides a method for preparing multi-layered chiral gold-cobalt nanoparticles for treating oral squamous cells. The multi-layered chiral gold-cobalt nanoparticles for treating oral squamous cell carcinoma are characterized in that they have a diameter of about 0.2 microns, a length of 5-10 microns, and are decorated with 100-nm nanospheres.

[0006] Step 1: Dissolve a certain amount of chloroauric acid in a solvent at 25 °C and keep it warm for 3 hours to obtain Component A.

[0007] Step 2: Add a certain amount of stabilizer and stir vigorously for 1 hour.

[0008] Step 3: Dissolve a certain amount of cobalt precursor in a solvent at 40 °C and stir at low speed for 2 hours to obtain Component B.

[0009] Step 4: Dissolve a certain amount of chiral ligand in water at 25 °C to obtain Component C.

[0010] Step 5: Mix Components A, B, and C in a certain ratio and place them in a hydrothermal reactor. React at 100-160 °C for 2-12 hours. After washing and drying, multi-layered chiral gold-cobalt nanoparticles can be obtained.

[0011] Relative to 1 gram of chloroauric acid, the amount of stabilizer used is 0.01-1 gram, the amount of cobalt precursor is 0.001-0.8 gram, and the amount of chiral ligand is 0.01-0.5 gram.

[0012] The solvent in Step 1 refers to one or a combination of several of water, ethanol, methanol, acetone, isopropanol, dichloromethane, acetonitrile, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, n-hexane, chloroform, toluene, and ethyl acetate.

[0013] The stabilizer in Step 2 refers to a combination of one or more of vitamin C, vitamin E, butylated hydroxyanisole, and butylated hydroxytoluene.

[0014] The terbium precursor in Step 3 refers to a combination of one or more of cobalt(II) chloride, cobalt sulfate, cobalt phthalocyanine, hexaaminocobalt(III) chloride, cobalt diethyldithiocarbamate, cobalt isophorone complex, cobalt tetrachloride, cobalt(III) fluoride, cobalt acetylacetonate, cobalt(II) imidazole complex, and cobalt thiocyanate.

[0015] The chiral ligand in Step 4 is one or a combination of several of tartaric acid, quinine, chiral bisphosphine ligand, chiral amine, camphorsulfonic acid, bisoxidophosphine ligand, chiral diamine, chiral amide, chiral oxazoline, chiral pyridone, alanine, serine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, histidine, aspartic acid, glutamic acid, lysine, arginine, and cystine.

[0016] In step five, the drying method is one or a combination of more than one of atmospheric drying, vacuum drying, spray drying, freeze drying, infrared drying, microwave drying, and moisture absorption drying.

[0017] Object of the Invention

[0018] The object of the present invention is to provide a multi-level structured chiral gold-cobalt nanoparticle, which realizes efficient targeted therapy for oral squamous cell carcinoma by combining magnetic, optical, and chiral properties.

[0019] Technical Solution

[0020] The preparation of the multi-level structured chiral gold-cobalt nanoparticle includes the following steps:

[0021] Step one: Dissolve chloroauric acid in a solvent at 25°C and keep it warm for 3 hours to obtain component A.

[0022] Step two: Add one or more of vitamin C, vitamin E, butylated hydroxyanisole, or butylated hydroxytoluene as a stabilizer and stir vigorously for 1 hour.

[0023] Step three: Dissolve the cobalt precursor in a solvent at 40°C and stir at a low speed for 2 hours to obtain component B.

[0024] Step four: Dissolve the chiral ligand in water at 25°C to obtain component C.

[0025] Step five: Mix components A, B, and C in a certain proportion, place them in a hydrothermal autoclave, and react at 160°C for 12 hours. After the reaction, wash and dry the product to obtain the multi-level structured chiral gold-cobalt nanoparticle.

[0026] The obtained multi-level structured chiral gold-cobalt nanoparticle has excellent properties, especially its ability to respond to the tumor microenvironment. The material undergoes cleavage under the action of reducing molecules such as high-concentration glutathione (GSH). This property ensures the precise release of the drug at the tumor site. Compared with conventional nanocarriers, this superstructure can avoid the non-specific release of the drug in the body, reduce the damage to normal tissues, and improve the specificity of treatment. At the same time, the introduction of the chiral ligand further enhances the selective binding of the material to cancer cells, making drug delivery more efficient.

[0027] In addition, the magnetic responsiveness and fluorescence characteristics of this material endow it with multiple functions in treatment and diagnosis. The cobalt element provides the ability of magnetically controlled targeted delivery and can induce apoptosis of tumor cells through the magnetic hyperthermia effect. The excellent surface plasmon resonance effect of the gold element makes it have important applications in fields such as biomedical imaging and photothermal therapy.

[0028] The size of the material is precisely controlled, with a diameter of approximately 0.2 micrometers and a length of 5 - 10 micrometers, and is accompanied by the modification of 100 - nanometer nanospheres. This structural design not only improves the stability of the material in body fluids but also enhances its dispersibility and biocompatibility in the body. The hydrothermal reaction in the preparation process ensures the high crystallinity and structural stability of the material, while various drying methods (such as freeze - drying or spray - drying) further optimize the morphology of the material and prevent particle aggregation.

[0029] In summary, the chiral gold - cobalt nanoparticles with a multi - level structure of the present invention exhibit significant advantages in terms of targeting, stability, and versatility. By responding to the tumor microenvironment to achieve cleavage and drug release, this material effectively improves the drug delivery efficiency and reduces the toxic and side effects during the treatment process. The integration of magnetic and fluorescent properties enables the material to play a role in both treatment and diagnosis, providing a new technical means for personalized treatment. Compared with traditional treatment methods, this material is simple to prepare, easy to scale up production, and has good prospects for industrial application. Brief Description of the Drawings

[0030] Figure 1 Multi - level structure chiral gold - cobalt nanoparticles synthesized by the method of Example 1

[0032] Figure 2 Comparison of the therapeutic effects and targeting efficiencies of different nanoparticles and multi - level structure chiral gold - cobalt nanoparticles on oral squamous cell carcinoma Detailed Description of the Invention

[0034] In order to make the objectives, technical solutions, and advantages of the present invention more clearly understood, the following further describes the present invention in detail with reference to the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0035] The preparation method of the multi - level structure chiral gold - cobalt nanoparticles provided by the embodiments of the present invention includes the following steps

[0036] Step 1: Dissolve a certain amount of chloroauric acid in a solvent at 25 °C and keep it warm for 3 hours to obtain Component A. The solvent refers to one or a combination of water, ethanol, methanol, acetone, isopropanol, dichloromethane, acetonitrile, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, n - hexane, chloroform, toluene, and ethyl acetate.

[0037] Step 2: Add a certain amount of stabilizer and stir vigorously for 1 hour. The stabilizer refers to a combination of one or more of vitamin C, vitamin E, butylated hydroxyanisole, and butylated hydroxytoluene.

[0038] Step 3: Dissolve a certain amount of cobalt precursor in a solvent at 40 °C and stir at a low speed for 2 hours to obtain Component B. The cobalt precursor refers to one or a combination of cobalt(II) chloride, cobalt sulfate, cobalt phthalocyanine, hexaaminecobalt(III) chloride, cobalt diethyldithiocarbamate, cobalt isophorone complex, cobalt tetrachloride, cobalt(III) fluoride, cobalt acetylacetonate, cobalt(II) imidazole complex, and cobalt thiocyanate.

[0039] Step 4: Dissolve a certain amount of chiral ligand in water at 25 °C to obtain Component C. The chiral ligand is one or a combination of tartaric acid, quinine, chiral diphosphine ligand, chiral amine, camphorsulfonic acid, bisoxidophosphine ligand, chiral diamine, chiral amide, chiral oxazoline, chiral pyridone, alanine, serine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, histidine, aspartic acid, glutamic acid, lysine, arginine, and cystine.

[0040] Step 5: Mix Components A, B, and C in a certain proportion and place them in a hydrothermal autoclave. React at 100 - 160 °C for 2 - 12 hours. After washing and drying, multi-level structured chiral gold-cobalt nanoparticles can be obtained. The drying methods include one or a combination of atmospheric drying, vacuum drying, spray drying, freeze drying, infrared drying, microwave drying, and moisture absorption drying.

[0041] Relative to 1 gram of ferric chloride, the dosage of the stabilizer is 0.01 - 1 gram, the amount of the cobalt precursor is 0.001 - 0.8 gram, and the amount of the chiral ligand is 0.01 - 0.5 gram.

[0042] The present invention will be further described below in conjunction with specific embodiments.

[0043] Example 1

[0044] Dissolve 1 gram of chloroauric acid in 100 milliliters of ethanol and keep it at 25 °C for 3 hours to obtain Component A. Add 0.1 gram of vitamin C as a stabilizer and stir vigorously for 1 hour. Take 0.1 gram of cobalt chloride and dissolve it in 50 milliliters of water, and stir at a low speed for 2 hours to obtain Component B. At the same time, dissolve 0.05 gram of tartaric acid in 10 milliliters of water to obtain Component C. Mix the three components in a ratio of 1:1:1, place them in a hydrothermal autoclave, and react at 120 °C for 4 hours. After the reaction, wash with deionized water and treat with freeze drying to obtain multi-level structured chiral gold-cobalt nanoparticles.

[0045] Example 2

[0046] Dissolve 1.5 grams of chloroauric acid in a mixed solvent of dimethyl sulfoxide (DMSO) and water (volume ratio 1:1), and keep it at 25 °C for 3 hours. Add 0.5 grams of vitamin E as a stabilizer and stir for 1 hour. Dissolve 0.2 grams of cobalt sulfate in acetonitrile and stir for 2 hours to obtain Component B. Dissolve 0.1 grams of quinine in water to obtain Component C. Mix the three components in a ratio of 2:1:1, react at 150 °C for 8 hours, and use spray drying method to process to obtain chiral multi-level nanoparticles modified with nanospheres.

[0047] Example 3

[0048] Dissolve 2 grams of chloroauric acid in a mixed solvent of methanol and acetone, keep it at 25 °C for 2 hours to obtain Component A. Add 0.05 grams of butylated hydroxyanisole as a stabilizer and stir for 1 hour. Dissolve 0.1 grams of cobalt phthalocyanine in tetrahydrofuran at 40 °C and stir for 2 hours to obtain Component B. Dissolve 0.03 grams of alanine in 5 milliliters of water to obtain Component C. Mix in a ratio of 1.5:1:0.5, place it in a hydrothermal kettle, react at 160 °C for 10 hours, and use infrared drying treatment to obtain nanostructures with a length of 7 micrometers.

[0049] Example 4

[0050] Dissolve 1 gram of chloroauric acid in 100 milliliters of dichloromethane, add 0.05 grams of butylated hydroxytoluene as a stabilizer after keeping it warm for 3 hours, and stir for 1 hour. Dissolve 0.1 grams of cobalt phthalocyanine in ethyl acetate and stir for 2 hours to obtain Component B. Dissolve 0.02 grams of camphorsulfonic acid in water to obtain Component C. Mix the three components in a ratio of 1:1:2, react at 100 °C for 6 hours to obtain chiral multi-level nanoparticles modified with nanospheres.

[0051] Example 5

[0052] Dissolve 1 gram of chloroauric acid in isopropanol, keep it warm for 3 hours to obtain Component A. Add a combined stabilizer of 0.1 grams of vitamin C and 0.05 grams of vitamin E and stir for 1 hour. Dissolve 0.2 grams of cobalt fluoride in n-hexane at 40 °C to obtain Component B. Dissolve 0.05 grams of chiral bisphosphine ligand in 10 milliliters of water to obtain Component C. Mix the three in a ratio of 1:1:1, react at 130 °C for 3 hours, and use freeze-drying method to process to prepare a material with uniform particle size.

[0053] Example 6

[0054] Dissolve chloroauric acid (1 g) in dimethylformamide (DMF), keep it at 25 °C for 3 hours to obtain Component A. Add 0.1 g of butylated hydroxyanisole as a stabilizer and stir for 1 hour. Dissolve 0.1 g of hexaaminecobalt(III) chloride complex in water and stir for 2 hours to obtain Component B. Dissolve 0.03 g of isoleucine in 5 mL of water to obtain Component C. Mix them in a ratio of 2:1:1, react at 110 °C for 5 hours, and treat by microwave drying method.

[0055] Example 7

[0056] Dissolve 0.5 g of chloroauric acid in a mixed solvent of toluene and acetonitrile (ratio 1:2), keep it warm for 3 hours. Add 0.2 g of vitamin C as a stabilizer and stir for 1 hour. Dissolve 0.2 g of cobalt isophorone complex in water at 40 °C and stir for 2 hours to obtain Component B. Dissolve 0.02 g of lysine in 10 mL of water to obtain Component C. The mixture reacts at 140 °C for 7 hours and is treated by vacuum drying to obtain a multi-level chiral material.

[0057] Example 8

[0058] Dissolve 1 g of chloroauric acid in acetone, keep it at a certain temperature for 3 hours to obtain Component A. Add 0.05 g of butylated hydroxytoluene as a stabilizer and stir for 1 hour. Dissolve 0.1 g of cobalt acetylacetonate in dimethyl sulfoxide and stir for 2 hours to obtain Component B. Dissolve 0.03 g of phenylalanine in water to obtain Component C. Mix them in a ratio of 1:2:1, react at 160 °C for 12 hours, and treat by atmospheric pressure drying method.

[0059] Example 9

[0060] Dissolve 1 g of chloroauric acid in a mixture of ethanol and water at 25 °C, keep it at a certain temperature for 3 hours to obtain Component A. Add 0.2 g of vitamin E as a stabilizer and stir for 1 hour. Dissolve 0.2 g of cobalt(II) imidazole complex in acetonitrile and stir for 2 hours to obtain Component B. Dissolve 0.02 g of histidine in water to obtain Component C. Mix the three components in a ratio of 1:1.5:1, react at 150 °C for 8 hours, and treat by moisture absorption drying method.

[0061] Example 10

[0062] Dissolve 0.8 g of chloroauric acid in ethyl acetate, keep it at a certain temperature for 3 hours to obtain Component A. Add a combined stabilizer of 0.1 g of vitamin C and 0.05 g of butylated hydroxyanisole, stir for 1 hour. Dissolve 0.15 g of cobalt thiocyanate in tetrahydrofuran and stir at low speed for 2 hours to obtain Component B. Dissolve 0.02 g of chiral oxazoline in 5 mL of water to obtain Component C. After mixing the three components, react at 120 °C for 4 hours and treat by freeze-drying method to obtain the final superstructure material.

[0063] Obviously, the above embodiments are merely examples for clear illustration and are not limitations on the implementation manners. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to enumerate all implementation manners here. And the obvious changes or modifications derived therefrom still fall within the protection scope of the present invention.

Claims

1. A multi - level structured chiral gold - cobalt nanoparticle proposed by the present invention for treating oral squamous cell carcinoma, characterized in that Preparation method of multi-level structured chiral gold-cobalt nanoparticles, comprising the following steps: Step 1, dissolve a certain amount of chloroauric acid in a solvent at 25 °C, keep warm for 3 hours to obtain component A. The solvent refers to one or a combination of several of water, ethanol, methanol, acetone, isopropanol, dichloromethane, acetonitrile, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, n-hexane, chloroform, toluene, ethyl acetate. Step 2, add a certain amount of stabilizer and stir vigorously for 1 hour. The stabilizer refers to a combination of one or more of vitamin C, vitamin E, butylated hydroxyanisole, and butylated hydroxytoluene. Step 3, dissolve a certain amount of cobalt precursor in a solvent at 40 °C, stir at low speed for 2 hours to obtain component B. The cobalt precursor refers to a combination of one or several of cobalt(II) chloride, cobalt sulfate, cobalt phthalocyanine, hexaaminocobalt(III) chloride, cobalt diethyldithiocarbamate, cobalt isophorone complex, cobalt tetrachloride, cobalt(III) fluoride, cobalt acetylacetonate, cobalt(II) imidazole complex, cobalt thiocyanate. Step 4, dissolve a certain amount of chiral ligand in water at 25 °C to obtain component C. The chiral ligand is one or a combination of several of tartaric acid, quinine, chiral diphosphine ligand, chiral amine, camphorsulfonic acid, bisoxidophosphine ligand, chiral diamine, chiral amide, chiral oxazoline, chiral pyridone, alanine, serine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, histidine, aspartic acid, glutamic acid, lysine, arginine, cystine. Step 5, mix the three components A, B, and C in a certain proportion, place them in a hydrothermal reactor, and react at 100-160 °C for 2-12 hours. After washing and drying, multi-level structured chiral gold-cobalt nanoparticles can be obtained. The drying method is one or a combination of several of atmospheric drying, vacuum drying, spray drying, freeze drying, infrared drying, microwave drying, and moisture absorption drying. Relative to 1 gram of iron chloride, the dosage of the stabilizer is 0.01-1 gram, the amount of the cobalt precursor is 0.001-0.8 gram, and the amount of the chiral ligand is 0.01-0.5 gram.

2. A multi-level structured chiral gold-cobalt nanoparticle proposed by the present invention for treating oral squamous cell carcinoma, characterized in that: About 0.2 microns in diameter, 5-10 microns in length, accompanied by 100 nm nanosphere modification.

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