A compound composition for resisting tumor and its preparation method

Abstract

The application discloses an anti-tumor compound composition and a preparation method thereof, and relates to the technical field of traditional Chinese medicines; the anti-tumor compound composition is composed of ferrocene formic acid grafted selenium-doped carbon nitride and a compound extract; the ferrocene formic acid grafted selenium-doped carbon nitride is used as a carrier to actively target the compound extract to tumor tissues in a mode of enhanced permeability and retention effect and responsive release, so that the bioavailability is improved, the toxic and side effects on normal tissues are reduced, and the effectiveness and safety of treatment are improved; the compound extract provides natural chemotherapy of multiple targets and multiple pathways, the ferrocene formic acid grafted selenium-doped carbon nitride prevents premature degradation of the compound extract and supplements multiple treatment pathways, further enhances the activity of the compound extract, and comprehensively enhances the anti-tumor effect.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically referring to an anti-tumor compound composition and its preparation method. Background Technology

[0002] Cancer is one of the most serious challenges facing modern medicine. Its high incidence and high mortality rate seriously threaten human health. Although strategies such as surgery, radiotherapy, chemotherapy, and immunotherapy are constantly developing, the heterogeneity of tumors, the complexity of the microenvironment, and drug resistance still greatly limit the efficacy and wide applicability of current treatment methods. In recent years, traditional Chinese medicine has received widespread attention due to its therapeutic advantages of multi-target and multi-pathway synergistic effects.

[0003] Modern Traditional Chinese Medicine (TCM) believes that tumors are often caused by a combination of factors, including congenital deficiency, invasion of external pathogens, emotional imbalance, improper diet, dysfunction of internal organs, and old age. The pathogenesis often presents a complex situation of mixed cold and heat, deficiency and excess, and disordered Qi. Among these factors, the balance of Yin and Yang is an important foundation for the body's harmony. Many physicians believe that the imbalance of Yin and Yang is one of the key pathogenesis mechanisms for the occurrence and development of malignant tumors, often manifesting as mixed cold and heat. Therefore, harmonizing Yin and Yang and balancing cold and heat are commonly used and important treatment methods in the clinical treatment of malignant tumors.

[0004] The existing technology currently has the following main problems:

[0005] Conventional Chinese medicine compositions have low bioavailability and can cause certain side effects, limiting the effectiveness and safety of tumor treatment. Summary of the Invention

[0006] In view of the above situation and to overcome the defects of the prior art, the present invention proposes an anti-tumor compound composition comprising the following components in parts by weight: 30-40 parts of selenium-doped carbon nitride grafted with ferrocene carboxylic acid, and 60-80 parts of compound extract.

[0007] The selenium-doped carbon nitride grafted with ferrocene carboxylic acid is made from the following components in parts by weight: 8-10 parts of nano-selenoglucose complex, 80-100 parts of melamine, and 0.1-0.2 parts of ferrocene carboxylic acid.

[0008] The compound extract comprises the following components in parts by weight: rhubarb 3-5 parts, aconite 3-5 parts, dried ginger 3-5 parts, ginseng 3-5 parts, and licorice 3-5 parts.

[0009] The method for preparing the selenium-doped carbon nitride grafted with ferrocene-formic acid specifically includes the following steps:

[0010] (1) Place 0.173g of sodium selenite in a 100mL round-bottom flask, add 20mL of ultrapure water, and stir at 300rpm until completely dissolved. Then add 15.0g of glucose and stir at 300rpm until completely dissolved. Heat to 70℃ while stirring at 300-500rpm. Add 0.352g of ascorbic acid to the above mixture and reflux at 70℃ for 3-4h. Cool to room temperature and concentrate under reduced pressure in a water bath at 60-70℃ using a rotary evaporator to 1 / 3 of the original volume. Dry under vacuum at 50-60℃ and grind. This process uses zero-valent nano-selenium as the functional core and hydrophilic glucose amorphous solid as the protective and dispersing matrix to form a product with good water solubility. A core-shell nanocomposite with high safety and stability was developed. In this nanocomposite, highly toxic sodium selenite is converted into low-toxicity, highly bioavailable zero-valent selenium nanoparticles. The glucose encapsulation further slows down the release of selenium atoms, avoiding acute toxic reactions such as gastrointestinal reactions caused by sudden increases in blood drug concentration, thus improving safety. The nano-selenium nanoparticles can directly inhibit tumors through multiple pathways, such as inducing tumor cell apoptosis, regulating the cell cycle, and inhibiting angiogenesis. The hydrophilic glucose shell can effectively improve the dispersibility and solubility of active ingredients in traditional Chinese medicine. The glucose matrix can also protect light-, heat-, and oxygen-sensitive traditional Chinese medicine components from degradation during storage and in vivo circulation, thereby increasing the bioavailability of the active ingredients and improving the therapeutic efficacy. This yields a nano-selenium glucose complex.

[0011] (2) After mixing the nano-selenoglucose complex described in step (1) with melamine, dry it in an oven at 60°C for 12-24 hours, then transfer it to a tube furnace and calcine it at 500-550°C for 3-4 hours under nitrogen protection at a heating rate of 5°C / min. Melamine is heated and polycondensed in an inert atmosphere, and deaminated to form a graphite-like layered structure of graphite phase carbon nitride. At the same time, the nano-selenoglucose complex decomposes, and selenium atoms enter the carbon nitride skeleton in the form of atomic doping, realizing the "carrier" and functional synergy of selenium. The carrier thus possesses the anti-tumor activity of selenium. As a basic carrier, carbon nitride provides a physical barrier to prevent the rapid release of selenium in the blood circulation. Furthermore, through the high permeability and retention effect of solid tumors, it can be enriched at the tumor site and reduce its distribution in normal tissues, thereby reducing the potential toxicity of selenium to organs such as the heart, liver, and kidneys, and obtaining selenium-doped carbon nitride.

[0012] (3) Weigh 300-400 mg of the selenoglucose-doped carbon nitride described in step (2) and disperse it in 20 mL of 25% hydrogen peroxide solution. Stir at 60-80℃ for 2-3 h, centrifuge, wash the precipitate with water until neutral, and dry to obtain pretreated selenoglucose-doped carbon nitride. Then weigh 50-100 mg of the pretreated selenoglucose-doped carbon nitride and disperse it in 20-30 mL of N,N-dimethylformamide. Under nitrogen protection and an ice-water bath at 0-5°C, ferrocene carboxylic acid, 25 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, and 2 mg of 4-dimethylaminopyridine were added sequentially. The mixture was stirred in the dark for 50-60 minutes. The ice-water bath was removed, and the temperature was raised to 60-80°C. Stirring was continued in the dark for 12-24 hours. The mixture was then centrifuged at 10000-12000 rpm for 5-10 minutes. The solid product was separated by distillation with anhydrous ethanol and ultrafiltration... The sample was washed 3-5 times with pure water, then 1-2 times with ethyl acetate, and finally vacuum dried at 60°C. The carboxyl groups introduced during the pretreatment and ferrocene carboxylic acid formed covalent bonds through amidation under the action of condensing agents and catalysts. The ferrocene groups were firmly fixed on the surface of the support material, avoiding systemic oxidative stress caused by free ferrocene molecules. This also ensured that its biological effects mainly occurred at the tumor sites enriched in the support. The dual catalytic centers formed by ferrocene carboxylic acid and selenium continuously generated reactive oxygen species, causing irreversible oxidative damage to tumor cells. Furthermore, the excellent electron conduction ability of ferrocene can act as an electron bridge to promote the rapid exchange of electrons in the catalytic reaction, making the entire selenium-iron catalytic cycle run faster and more efficiently. This accelerated local effective reaction also reduced the need for systemic high-dose drugs, reduced potential toxicity from the source, and improved the safety of treatment, resulting in selenium-doped carbon nitride grafted with ferrocene carboxylic acid.

[0013] Preferably, in step (2), the amount of nano-selenoglucose complex and melamine added is 0.8-1.0g and 8.0-10.0g, respectively. Melamine, as a precursor material of the carrier, can form a rigid porous carrier structure, thereby providing a huge specific surface area and abundant binding sites, which is conducive to loading hydrophobic Chinese medicine components through physical adsorption or chemical bonding, and can respond to the tumor microenvironment to realize intelligent control of drug release.

[0014] Preferably, in step (3), the amount of ferrocene carboxylic acid added is 10-20 mg, and the ferrocene group (Fe) 2+ / Fe 3+ The reversible redox couple can efficiently catalyze the generation of highly toxic hydroxyl radicals in the high H2O2 microenvironment of tumors, achieving direct killing. Its catalytic activity is highly dependent on the tumor-specific high H2O2 environment and is silenced in normal tissues, achieving selective attack, which improves the therapeutic effect and ensures safety.

[0015] This invention also provides a method for preparing an antitumor compound composition, specifically comprising the following steps:

[0016] S1. Rhubarb, aconite, dried ginger, ginseng, and licorice are dried and pre-treated, then pulverized and mixed evenly. 500g of the powder is placed in 6-8 times its volume of 80% ethanol solution and soaked overnight. Then, it is extracted 2-3 times under heat and pressure in a reflux condenser and rotary evaporator. The extracts are filtered, and the combined extracts are concentrated under reduced pressure to obtain an alcohol extract. This alcohol extract is then purified using an AB-8 macroporous adsorption resin. The eluent is concentrated to 1 / 3 of its original volume, then dispersed in a pH 4.8 acetate-sodium acetate buffer solution, filtered through a 0.22μm filter membrane, and freeze-dried to obtain a purified alcohol extract. Simultaneously, the residue is decocted with 3-5 times its volume of water. The resulting aqueous extract is first concentrated to 1 / 3 of its original volume, then 95% ethanol is added. The aqueous extract contains 70-75% ethanol by mass. It is then allowed to stand at 4°C for 24 hours. The precipitate is collected and washed twice with anhydrous ethanol and acetone, and finally freeze-dried to obtain refined aqueous polysaccharide. The refined alcohol extract and refined aqueous polysaccharide are then mixed at a mass ratio of 2:1. In this formula, Aconitum carmichaelii and dried ginger can warm and tonify the spleen and kidney yang to dispel cold stagnation; ginseng and licorice can greatly replenish vital energy and support the body's resistance to disease and repair; rhubarb can purge accumulations, invigorate blood circulation, and remove blood stasis to clear blockages. The combined use of these herbs embodies the tumor treatment method of "warming yang and supporting the body's resistance, combining attack and tonification." Furthermore, the direct tumor-killing mechanism of the alcohol extract is combined with the immune-boosting mechanism of the polysaccharide, forming a classic TCM treatment strategy of "simultaneously eliminating pathogens (attacking tumors) and supporting the body's resistance (enhancing immunity)," producing a safe and effective synergistic anti-tumor effect, resulting in a compound extract.

[0017] S2. Disperse 30-40 mg of ferrocene-grafted selenium-doped carbon nitride in 15 mL of pH 7.4 PBS buffer and sonicate for 40-50 min to form a suspension for later use. Then, weigh 60-80 mg of the compound extract described in step S1 and add it to 5 mL of pH 7.4 PBS buffer. Stir for 20-30 min to form a solution for later use. Then, under magnetic stirring at 300-500 rpm and in the dark, add the solution to the suspension using a constant flow pump at a rate of 1 mL / min. Place the solution in a constant temperature shaker and incubate at 120-150 rpm in the dark for 20-24 h. Then, centrifuge at 12000-15000 rpm for 10-20 min. Wash the precipitate 2-3 times with pre-cooled PBS buffer. Finally, disperse the washed precipitate in 5 mL of sterile PBS buffer. In pyrogen-free water for injection, lyophilized selenium-doped carbon nitride grafted with ferrocene carboxylic acid is used as a carrier to load the compound extract. During delivery, the extract is protected from enzymatic degradation and premature clearance, allowing more active ingredients to reach the lesion and improving bioavailability. In the slightly acidic environment of the tumor, the carboxyl groups of ferrocene carboxylic acid and the surface groups of carbon nitride undergo changes in protonation, leading to changes in the carrier structure or surface charge. This promotes the specific release of the drug at the tumor site and reduces leakage in normal tissues, effectively improving the safety of treatment. Selenium doping not only modulates the catalytic properties of the material, but also plays a role in catalytic therapy or inducing tumor cell apoptosis in the highly reactive oxygen species environment within tumor cells. It produces a synergistic anti-tumor effect with the compound extract, thereby achieving multimodal synergistic treatment and significantly improving the therapeutic effect, resulting in an anti-tumor compound composition.

[0018] Preferably, in step S1, during the purification process, water and ethanol with mass fractions of 20%, 50%, and 70% are used for stepwise elution, and the ethanol eluents with mass fractions of 50% and 70% are collected. The gradient elution method can remove water-soluble toxic components and impurities. At the same time, the main active ingredients such as saponins and flavonoids are collected, which improves the concentration and purity of the active ingredients, thereby exerting a safer and more powerful effect.

[0019] Preferably, in step S2, during the shaking incubation process, the temperature is controlled at 30-37°C. This temperature range not only increases the effective collision and contact opportunities between drug molecules and carriers, optimizing loading efficiency and drug loading, but also avoids the denaturation and inactivation of heat-sensitive active ingredients, maintaining drug activity. Furthermore, by precisely simulating the internal temperature environment of the human body, the drug and carrier can be pre-adapted to the internal environment, ensuring stability in blood circulation, significantly reducing systemic toxicity, and improving the safety of medication.

[0020] The beneficial effects achieved by this invention are as follows:

[0021] This invention utilizes ferrocene-based carboxylic acid-grafted selenium-doped carbon nitride as a carrier to actively target and deliver a compound extract to tumor tissue via enhanced penetration and retention effects and responsive release. This not only improves bioavailability but also reduces toxic side effects on normal tissues, enhancing the efficacy and safety of treatment. The compound extract provides multi-target, multi-pathway natural chemotherapy, while the ferrocene-based carboxylic acid-grafted selenium-doped carbon nitride prevents premature degradation of the extract and supplements multiple therapeutic pathways, further enhancing and protecting the activity of the extract and comprehensively strengthening its anti-tumor effect. In the ferrocene-based carboxylic acid-grafted selenium-doped carbon nitride, the ferrocene groups are covalently linked and fixed to... The carbon nitride surface doped with selenoglucose avoids the systemic oxidative stress caused by free ferrocene molecules, ensuring that the biological effects of ferrocene carboxylic acid mainly occur at the tumor site. The dual catalytic centers formed by ferrocene carboxylic acid and selenium continuously generate reactive oxygen species, causing irreversible oxidative damage to tumor cells. Furthermore, ferrocene's excellent electron conduction ability can act as an electron bridge, promoting rapid electron exchange in the catalytic reaction, making the entire selenium-iron catalytic cycle run faster and more efficiently. This accelerated local effective reaction also reduces the need for systemic high-dose drugs, reducing potential toxicity from the source and improving treatment safety. In this process, carbon nitride not only serves as a basic carrier, providing physical... The barrier prevents the rapid release of selenium into the bloodstream and can also act as a photosensitizer, generating reactive oxygen species (photodynamic therapy) or heat (photothermal therapy) under specific wavelength light at 625nm. Selenium doping and ferrocene grafting further enhance the photocatalytic efficiency of carbon nitride materials. The compound extract combines directly tumor-killing alcoholic extracts with immune-boosting polysaccharides to form a highly efficient extract that simultaneously "eliminates pathogens (attacks tumors) and strengthens the body (enhances immunity)." The refined water-extracted polysaccharides are mainly ginseng polysaccharides and licorice polysaccharides, which can activate the immune system, help the body clear tumor cells, downregulate the expression of multidrug resistance-related proteins in tumor cells, reverse or reduce the drug resistance of tumor cells, and thus... This invention enhances the killing effect of active ingredients in refined ethanol extracts on drug-resistant cells. Simultaneously, refined water-extracted polysaccharides protect the hematopoietic and immune systems, directly counteracting the risk of bone marrow cell damage present in refined ethanol extracts. They also harmonize the medicinal properties, mitigate the acute toxicity of aconite alkaloids, and reduce the intestinal irritation of rhubarb anthraquinones. Furthermore, as a hydrophilic polymer, the addition of refined water-extracted polysaccharides improves the solubility and stability of hydrophobic components in refined ethanol extracts, thereby producing a safe and effective synergistic anti-tumor effect. This invention uses ferrocene-grafted selenium-doped carbon nitride and compound extracts to prepare an anti-tumor compound composition, improving bioavailability, reducing side effects, and enhancing therapeutic efficacy and safety. Attached Figure Description

[0022] Figure 1The graph shows the cancer cell inhibition rate results of Examples 1-4 and Comparative Examples 1-3 of the present invention;

[0023] Figure 2 The graph shows the frequency of side effects in Examples 1-4 and Comparative Examples 1-3 of this invention. Detailed Implementation

[0024] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention. The preferred embodiments and materials described herein are for illustrative purposes only and do not limit the scope of this application.

[0026] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods; unless otherwise specified, the experimental materials used in the following embodiments are all purchased from commercial channels.

[0027] Example 1

[0028] This embodiment proposes an antitumor compound composition comprising the following components in parts by weight: 40 parts of selenium-doped carbon nitride grafted with ferrocene carboxylic acid, and 80 parts of compound extract.

[0029] The selenium-doped carbon nitride grafted with ferrocene carboxylic acid is made from the following components in parts by weight: 10 parts of nano-selenoglucose complex, 100 parts of melamine, and 0.2 parts of ferrocene carboxylic acid.

[0030] The compound extract contains the following components by weight: 5 parts rhubarb, 5 parts aconite, 5 parts dried ginger, 5 parts ginseng, and 5 parts licorice.

[0031] The preparation method of selenium-doped carbon nitride grafted with ferrocene-based carboxylic acid specifically includes the following steps:

[0032] (1) Place 0.173g of sodium selenite in a 100mL round-bottom flask, add 20mL of ultrapure water, and stir at 300rpm until completely dissolved. Then add 15.0g of glucose and stir at 300rpm until completely dissolved. Heat to 70℃ while stirring at 500rpm, add 0.352g of ascorbic acid to the above mixture, reflux at 70℃ for 4h, cool to room temperature, concentrate under reduced pressure in a 70℃ water bath using a rotary evaporator to 1 / 3 of the original volume, vacuum dry at 60℃, and grind. This process uses zero-valent nano-selenium as the functional core and hydrophilic glucose amorphous solid as the protective and dispersing matrix to form a solution with good water solubility and stability. In a core-shell nanocomposite, highly toxic sodium selenite is converted into low-toxicity, highly bioavailable zero-valent selenium nanoparticles. The glucose encapsulation further slows the release of selenium atoms, avoiding acute toxic reactions such as gastrointestinal reactions caused by sudden increases in blood drug concentration, thus improving safety. The nano-selenium can directly inhibit tumors through multiple pathways, such as inducing tumor cell apoptosis, regulating the cell cycle, and inhibiting angiogenesis. The hydrophilic glucose shell can effectively improve the dispersibility and solubility of the active ingredients of traditional Chinese medicine. The glucose matrix can also protect the light, heat, and oxygen-sensitive traditional Chinese medicine components from degradation during storage and in vivo circulation, thereby increasing the bioavailability of the active ingredients and improving the therapeutic efficacy, resulting in a nano-selenium glucose complex.

[0033] (2) After mixing the nano-selenoglucose complex and melamine described in step (1) evenly, the amount of nano-selenoglucose complex and melamine added is 1.0g and 10.0g, respectively. Melamine, as a precursor material for the carrier, can form a rigid porous carrier structure, thereby providing a huge specific surface area and abundant binding sites, which is conducive to loading hydrophobic Chinese medicine components through physical adsorption or chemical bonding, and can respond to the tumor microenvironment to achieve intelligent controlled release of drugs. First, dry in an oven at 60℃ for 24h, then transfer to a tube furnace, and calcine at 550℃ under nitrogen protection at a heating rate of 5℃ / min for 4 hours. h. Melamine is heated and polycondensed in an inert atmosphere, and deaminated to form a graphite-like layered structure of carbon nitride. At the same time, the nano-selenoglucose complex decomposes, and selenium atoms enter the carbon nitride framework in the form of atomic doping, realizing the "carrier" and functional synergy of selenium. The carrier thus possesses the anti-tumor activity of selenium. As a basic carrier, carbon nitride provides a physical barrier to prevent the rapid release of selenium in the blood circulation. Furthermore, through the high permeability and retention effect of solid tumors, it can be enriched at the tumor site and reduced in normal tissues, thereby reducing the potential toxicity of selenium to organs such as the heart, liver, and kidneys, and obtaining selenoglucose-doped carbon nitride.

[0034] (3) Weigh 400 mg of the selenoglucose-doped carbon nitride described in step (2) and disperse it in 20 mL of 25% hydrogen peroxide solution. Stir at 80°C for 3 h, centrifuge, wash the precipitate with water until neutral, and dry to obtain pretreated selenoglucose-doped carbon nitride. Then weigh 100 mg of the pretreated selenoglucose-doped carbon nitride and disperse it in 30 mL of N,N-dimethylformamide. Under nitrogen protection and a 5°C ice-water bath, add ferrocene carboxylic acid, 25 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, and 2 mg of 4-dimethylaminopyridine in sequence. The amount of ferrocene carboxylic acid added is 20 mg. The ferrocene group (Fe 2+ / Fe 3+ The reversible redox couple of ... With valence bonds, the ferrocene groups are firmly fixed on the surface of the carrier material, avoiding systemic oxidative stress caused by free ferrocene molecules. This also ensures that its biological effects mainly occur at the tumor sites enriched in the carrier. The dual catalytic centers formed by ferrocene carboxylic acid and selenium continuously generate reactive oxygen species, causing irreversible oxidative damage to tumor cells. Furthermore, the excellent electron conduction ability of ferrocene can act as an electron bridge, promoting the rapid exchange of electrons in the catalytic reaction, making the entire selenium-iron catalytic cycle run faster and more efficiently. This accelerated local effective reaction also reduces the need for systemic high-dose drugs, reducing potential toxicity from the source and improving the safety of treatment, resulting in selenium-doped carbon nitride grafted with ferrocene carboxylic acid.

[0035] This embodiment provides a method for preparing an antitumor compound composition, specifically including the following steps:

[0036] S1. Rhubarb, aconite, dried ginger, ginseng, and licorice were dried and pretreated, then pulverized and mixed evenly. 500g of the powder was placed in 8 times its volume of 80% ethanol solution and soaked overnight. The mixture was then extracted three times under heat and pressure in a reflux condenser and rotary evaporator. After filtration, the extracts were combined, and the collected extract was concentrated under reduced pressure to obtain an ethanol extract. This ethanol extract was then purified using AB-8 macroporous adsorption resin. During purification, water and 20%, 50%, and 70% ethanol were used for stepwise elution. The 50% and 70% ethanol eluates were collected. This gradient elution method removes highly water-soluble toxic components and impurities, while simultaneously collecting the main active ingredients such as saponins and flavonoids, increasing the concentration and purity of the active ingredients, thus resulting in a safer and more potent effect. The eluate was concentrated to 1 / 3 of its original volume, then dispersed in a pH 4.8 acetate-sodium acetate buffer solution, filtered through a 0.22μm filter membrane, and freeze-dried to obtain… The refined ethanol extract is prepared for later use. Meanwhile, the dregs are decocted with 5 times the amount of water. The resulting aqueous extract is first concentrated to 1 / 3 of its original volume, then 95% ethanol is added to bring the ethanol content to 75%. The mixture is then allowed to stand at 4°C for 24 hours. The precipitate is collected and washed twice with anhydrous ethanol and acetone, and finally freeze-dried to obtain the refined aqueous polysaccharide. Finally, the refined ethanol extract and refined aqueous polysaccharide are mixed at a mass ratio of 2:1. In this formula, aconite and dried ginger can warm and tonify the spleen and kidneys. Yang energy dissipates stagnation of Yin and cold. Ginseng and licorice can greatly replenish vital energy and support the body's resistance to disease and its ability to repair itself. Rhubarb can purge and eliminate stagnation, promote blood circulation and remove blood stasis, thus clearing blockages. The combined use of these herbs embodies the tumor treatment method of "warming Yang and supporting the body's resistance, and combining attack and replenishment." Furthermore, the direct tumor-killing mechanism of the alcohol extract is combined with the immune-boosting mechanism of polysaccharides, forming a classic TCM treatment strategy of "simultaneously eliminating pathogens (attacking tumors) and supporting the body's resistance (enhancing immunity)," producing a safe and effective synergistic anti-tumor effect, resulting in the compound extract.

[0037] S2. 40 mg of ferrocene-grafted selenium-doped carbon nitride was dispersed in 15 mL of pH 7.4 PBS buffer and sonicated for 50 min to form a suspension for later use. Then, 80 mg of the compound extract described in step S1 was weighed and added to 5 mL of pH 7.4 PBS buffer and stirred for 30 min to form a drug solution for later use. Then, under magnetic stirring at 500 rpm and in the dark, the drug solution was added to the suspension at a rate of 1 mL / min using a constant flow pump. The mixture was placed in a constant temperature shaker and incubated at 150 rpm in the dark for 24 h. During the incubation, the temperature was controlled at 37°C. This temperature range not only increases the effective collision and contact opportunities between drug molecules and the carrier, optimizing loading efficiency and drug load, but also avoids denaturation and inactivation of heat-sensitive active ingredients, maintaining drug activity. Furthermore, by precisely simulating the internal temperature environment of the human body, the drug and carrier are pre-adapted to the in vivo environment, ensuring stability in blood circulation, significantly reducing systemic toxicity, and improving the safety of medication. Centrifuge at 15,000 rpm for 20 min. Wash the precipitate three times with pre-cooled PBS buffer. Finally, disperse the washed precipitate in 5 mL of sterile, pyrogen-free water for injection and freeze-dry. Ferrocene-grafted selenium-doped carbon nitride serves as a carrier to load the compound extract, protecting it from enzymatic degradation and premature clearance during delivery, allowing more active ingredients to reach the lesion and improving bioavailability. In the slightly acidic environment of tumors, the carboxyl groups of ferrocene and the surface groups of carbon nitride undergo changes in protonation, leading to alterations in the carrier structure or surface charge, promoting specific drug release at the tumor site, and reducing leakage in normal tissues, effectively improving treatment safety. Selenium doping not only modulates the catalytic properties of the material but also plays a role in catalytic therapy or inducing tumor cell apoptosis in the highly reactive oxygen species environment within tumor cells, producing a synergistic anti-tumor effect with the compound extract. This achieves multimodal synergistic therapy, significantly improving the therapeutic effect and yielding an anti-tumor compound composition.

[0038] Example 2

[0039] This embodiment proposes an antitumor compound composition comprising the following components in parts by weight: 30 parts of selenium-doped carbon nitride grafted with ferrocene carboxylic acid, and 60 parts of compound extract.

[0040] The selenium-doped carbon nitride grafted with ferrocene carboxylic acid is made from the following components in parts by weight: 8 parts of nano-selenoglucose complex, 80 parts of melamine, and 0.1 parts of ferrocene carboxylic acid.

[0041] The compound extract contains the following components by weight: 3 parts rhubarb, 3 parts aconite, 3 parts dried ginger, 3 parts ginseng, and 3 parts licorice.

[0042] The preparation method of selenium-doped carbon nitride grafted with ferrocene-based carboxylic acid specifically includes the following steps:

[0043] (1) Place 0.173g of sodium selenite in a 100mL round-bottom flask, add 20mL of ultrapure water, and stir at 300rpm until completely dissolved. Then add 15.0g of glucose and stir at 300rpm until completely dissolved. Heat to 70℃ while stirring at 300rpm, add 0.352g of ascorbic acid to the above mixture, reflux at 70℃ for 3h, cool to room temperature, concentrate under reduced pressure in a 60℃ water bath using a rotary evaporator to 1 / 3 of the original volume, vacuum dry at 50℃, and grind. This process uses zero-valent nano-selenium as the functional core and hydrophilic glucose amorphous solid as the protective and dispersing matrix to form a solution with good water solubility and stability. In a core-shell nanocomposite, highly toxic sodium selenite is converted into low-toxicity, highly bioavailable zero-valent selenium nanoparticles. The glucose encapsulation further slows the release of selenium atoms, avoiding acute toxic reactions such as gastrointestinal reactions caused by sudden increases in blood drug concentration, thus improving safety. The nano-selenium can directly inhibit tumors through multiple pathways, such as inducing tumor cell apoptosis, regulating the cell cycle, and inhibiting angiogenesis. The hydrophilic glucose shell can effectively improve the dispersibility and solubility of the active ingredients of traditional Chinese medicine. The glucose matrix can also protect the light, heat, and oxygen-sensitive traditional Chinese medicine components from degradation during storage and in vivo circulation, thereby increasing the bioavailability of the active ingredients and improving the therapeutic efficacy, resulting in a nano-selenium glucose complex.

[0044] (2) After mixing the nano-selenoglucose complex and melamine described in step (1) evenly, the amount of nano-selenoglucose complex and melamine added is 0.8g and 8.0g, respectively. Melamine, as a precursor material for the carrier, can form a rigid porous carrier structure, thereby providing a huge specific surface area and abundant binding sites, which is conducive to loading hydrophobic Chinese medicine components through physical adsorption or chemical bonding, and can respond to the tumor microenvironment to achieve intelligent controlled release of drugs. First, dry in an oven at 60℃ for 12h, then transfer to a tube furnace, and calcine at 500℃ under nitrogen protection at a heating rate of 5℃ / min for 3 hours. h. Melamine is heated and polycondensed in an inert atmosphere, and deaminated to form a graphite-like layered structure of carbon nitride. At the same time, the nano-selenoglucose complex decomposes, and selenium atoms enter the carbon nitride framework in the form of atomic doping, realizing the "carrier" and functional synergy of selenium. The carrier thus possesses the anti-tumor activity of selenium. As a basic carrier, carbon nitride provides a physical barrier to prevent the rapid release of selenium in the blood circulation. Furthermore, through the high permeability and retention effect of solid tumors, it can be enriched at the tumor site and reduced in normal tissues, thereby reducing the potential toxicity of selenium to organs such as the heart, liver, and kidneys, and obtaining selenoglucose-doped carbon nitride.

[0045] (3) Weigh 300-400 mg of the selenoglucose-doped carbon nitride described in step (2) and disperse it in 20 mL of 25% hydrogen peroxide solution. Stir at 60°C for 2 h, centrifuge, wash the precipitate with water until neutral, and dry to obtain pretreated selenoglucose-doped carbon nitride. Then weigh 50 mg of the pretreated selenoglucose-doped carbon nitride and disperse it in 20 mL of N,N-dimethylformamide. Under nitrogen protection and 0°C ice-water bath conditions, add ferrocene carboxylic acid, 25 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2 mg of 4-dimethylaminopyridine in sequence. The amount of ferrocene carboxylic acid added is 10 mg. The ferrocene group (Fe 2+ / Fe 3+ The reversible redox couple of ferrocene acid efficiently catalyzes the generation of highly toxic hydroxyl radicals in the high-H2O2 microenvironment of tumors, achieving direct killing. Its catalytic activity is highly dependent on the tumor-specific high-H2O2 environment, remaining silent in normal tissues, thus achieving selective attack. This improves both therapeutic efficacy and ensures safety. The mixture was stirred in the dark for 50 min, then the ice-water bath was removed, the temperature was raised to 60℃, and stirring was continued in the dark for 12 h. The mixture was then centrifuged at 10000 rpm for 5 min. The solid product was washed three times alternately with anhydrous ethanol and ultrapure water, then once with ethyl acetate, and finally vacuum dried at 60℃. The carboxyl groups introduced during pretreatment and ferrocene acid formed covalent groups through an amidation reaction under the action of a condensing agent and catalyst. By bonding ferrocene groups to the surface of the carrier material, the systemic oxidative stress caused by free ferrocene molecules is prevented. This ensures that the biological effects mainly occur at the tumor sites enriched in the carrier. The dual catalytic centers formed by ferrocene carboxylic acid and selenium continuously generate reactive oxygen species, causing irreversible oxidative damage to tumor cells. Furthermore, the excellent electron conduction ability of ferrocene can act as an electron bridge, promoting the rapid exchange of electrons in the catalytic reaction, making the entire selenium-iron catalytic cycle run faster and more efficiently. This accelerated local effective reaction also reduces the need for systemic high-dose drugs, thereby reducing potential toxicity from the source and improving the safety of treatment, resulting in selenium-doped carbon nitride grafted with ferrocene carboxylic acid.

[0046] This embodiment provides a method for preparing an antitumor compound composition, specifically including the following steps:

[0047] S1. Rhubarb, aconite, dried ginger, ginseng, and licorice were dried and pretreated, then pulverized and mixed evenly. 500g of the powder was placed in 6 times its volume of 80% ethanol solution and soaked overnight. The mixture was then extracted twice under heat and pressure in a reflux condenser and rotary evaporator. After filtration, the extracts were combined, and the collected extract was concentrated under reduced pressure to obtain an alcohol extract. This alcohol extract was then purified using AB-8 macroporous adsorption resin. During purification, water and 20%, 50%, and 70% ethanol were used for stepwise elution. The 50% and 70% ethanol eluates were collected. This gradient elution method removes highly water-soluble toxic components and impurities, while simultaneously collecting the main active ingredients such as saponins and flavonoids, increasing the concentration and purity of the active ingredients, thus resulting in a safer and more potent effect. The eluate was concentrated to 1 / 3 of its original volume, then dispersed in a pH 4.8 acetate-sodium acetate buffer solution, filtered through a 0.22μm filter membrane, and freeze-dried to obtain… The refined ethanol extract is prepared for later use. Meanwhile, the dregs are decocted with three times the amount of water. The resulting aqueous extract is first concentrated to one-third of its original volume, then 95% ethanol is added to bring the ethanol content to 70%. The mixture is then allowed to stand at 4°C for 24 hours. The precipitate is collected and washed twice with anhydrous ethanol and acetone, and finally freeze-dried to obtain the refined aqueous polysaccharide. Finally, the refined ethanol extract and refined aqueous polysaccharide are mixed at a mass ratio of 2:1. In this formula, aconite and dried ginger can warm and tonify the spleen and kidneys. Yang energy dissipates stagnation of Yin and cold. Ginseng and licorice can greatly replenish vital energy and support the body's resistance to disease and its ability to repair itself. Rhubarb can purge and eliminate stagnation, promote blood circulation and remove blood stasis, thus clearing blockages. The combined use of these herbs embodies the tumor treatment method of "warming Yang and supporting the body's resistance, and combining attack and replenishment." Furthermore, the direct tumor-killing mechanism of the alcohol extract is combined with the immune-boosting mechanism of polysaccharides, forming a classic TCM treatment strategy of "simultaneously eliminating pathogens (attacking tumors) and supporting the body's resistance (enhancing immunity)," producing a safe and effective synergistic anti-tumor effect, resulting in the compound extract.

[0048] S2. Disperse 30 mg of ferrocene-grafted selenium-doped carbon nitride in 15 mL of pH 7.4 PBS buffer and sonicate for 40 min to form a suspension for later use. Then, weigh 60 mg of the compound extract from step S1 and add it to 5 mL of pH 7.4 PBS buffer, stir for 20 min to form a drug solution for later use. Then, under magnetic stirring at 300 rpm and in the dark, add the drug solution to the suspension at a rate of 1 mL / min using a constant flow pump. Place the solution in a constant temperature shaker and incubate at 120 rpm in the dark for 20 h. During the incubation, control the temperature at 30℃. This temperature range not only increases the effective collision and contact opportunities between drug molecules and carriers, optimizing loading efficiency and drug loading, but also avoids denaturation and inactivation of heat-sensitive active ingredients, maintaining drug activity. Furthermore, by accurately simulating the internal temperature environment of the human body, the drug and carrier can be pre-adapted to the in vivo environment, ensuring stability in blood circulation, significantly reducing systemic toxicity, and improving the safety of medication. Centrifuge at 12000 rpm for 10 min. Wash the precipitate twice with pre-cooled PBS buffer. Finally, disperse the washed precipitate in 5 mL of sterile, pyrogen-free water for injection and freeze-dry. Ferrocene-grafted selenium-doped carbon nitride serves as a carrier to load the compound extract, protecting it from enzymatic degradation and premature clearance during delivery, allowing more active ingredients to reach the lesion and improving bioavailability. In the slightly acidic environment of tumors, the carboxyl groups of ferrocene and the surface groups of carbon nitride undergo changes in protonation, leading to alterations in the carrier structure or surface charge, promoting specific drug release at the tumor site, and reducing leakage in normal tissues, effectively improving treatment safety. Selenium doping not only modulates the catalytic properties of the material but also plays a role in catalytic therapy or inducing tumor cell apoptosis in the highly reactive oxygen species environment within tumor cells, producing a synergistic anti-tumor effect with the compound extract. This achieves multimodal synergistic therapy, significantly improving treatment efficacy and yielding an anti-tumor compound composition.

[0049] Example 3

[0050] This embodiment proposes an antitumor compound composition comprising the following components in parts by weight: 40 parts of selenium-doped carbon nitride grafted with ferrocene carboxylic acid, and 60 parts of compound extract.

[0051] The selenium-doped carbon nitride grafted with ferrocene carboxylic acid is made from the following components in parts by weight: 9 parts of nano-selenoglucose complex, 90 parts of melamine, and 0.15 parts of ferrocene carboxylic acid.

[0052] The compound extract contains the following components by weight: 4 parts rhubarb, 4 parts aconite, 4 parts dried ginger, 4 parts ginseng, and 4 parts licorice.

[0053] The preparation method of selenium-doped carbon nitride grafted with ferrocene-based carboxylic acid specifically includes the following steps:

[0054] (1) Place 0.173g of sodium selenite in a 100mL round-bottom flask, add 20mL of ultrapure water, and stir at 300rpm until completely dissolved. Then add 15.0g of glucose and stir at 300rpm until completely dissolved. Heat to 70℃ while stirring at 400rpm. Add 0.352g of ascorbic acid to the above mixture and reflux at 70℃ for 3.5h. Cool to room temperature and concentrate under reduced pressure in a 65℃ water bath using a rotary evaporator to 1 / 3 of the original volume. Dry under vacuum at 55℃ and grind. This process uses zero-valent nano-selenium as the functional core and hydrophilic glucose amorphous solid as the protective and dispersing matrix to form a solution with good water solubility and stability. The "core-shell" nanocomposite is a nano-selenite complex in which highly toxic sodium selenite is converted into low-toxicity, highly bioavailable zero-valent selenium nanoparticles. The glucose encapsulation further slows down the release of selenium atoms, avoiding acute toxic reactions such as gastrointestinal reactions caused by a sudden increase in blood drug concentration, thus improving safety. The nano-selenium can directly inhibit tumors through multiple pathways such as inducing tumor cell apoptosis, regulating the cell cycle, and inhibiting angiogenesis. The hydrophilic glucose shell can effectively improve the dispersibility and solubility of the active ingredients of traditional Chinese medicine. The glucose matrix can also protect the light, heat, and oxygen-sensitive traditional Chinese medicine components from degradation during storage and in vivo circulation, thereby increasing the bioavailability of the active ingredients and improving the therapeutic efficacy, thus obtaining the nano-selenium glucose complex.

[0055] (2) After mixing the nano-selenoglucose complex and melamine described in step (1) evenly, the amount of nano-selenoglucose complex and melamine added is 0.9g and 9.0g, respectively. Melamine, as a precursor material of the carrier, can form a rigid porous carrier structure, thereby providing a huge specific surface area and abundant binding sites, which is conducive to loading hydrophobic Chinese medicine components through physical adsorption or chemical bonding, and can respond to the tumor microenvironment to realize intelligent control of drug release. First, dry in an oven at 60℃ for 18h, and then transfer to a tube furnace, and calcine at 525℃ under nitrogen protection at a heating rate of 5℃ / min. After 5 hours of heating and polycondensation in an inert atmosphere, melamine undergoes deamination to form a graphite-like layered structure of carbon nitride. Simultaneously, the nano-selenoglucose complex decomposes, and selenium atoms enter the carbon nitride framework in the form of atomic doping, realizing the "carrier" and functional synergy of selenium. The carrier thus possesses the anti-tumor activity of selenium. As a basic carrier, carbon nitride provides a physical barrier to prevent the rapid release of selenium in the bloodstream. Furthermore, through the high permeability and retention effect of solid tumors, it can accumulate at the tumor site and reduce its distribution in normal tissues, thereby reducing the potential toxicity of selenium to organs such as the heart, liver, and kidneys, resulting in selenoglucose-doped carbon nitride.

[0056] (3) Weigh 350 mg of the selenoglucose-doped carbon nitride described in step (2) and disperse it in 20 mL of 25% hydrogen peroxide solution. Stir at 70°C for 2.5 h, centrifuge, wash the precipitate with water until neutral, and dry to obtain pretreated selenoglucose-doped carbon nitride. Then weigh 75 mg of the pretreated selenoglucose-doped carbon nitride and disperse it in 25 mL of N,N-dimethylformamide. Under nitrogen protection and a 3°C ice-water bath, add ferrocene carboxylic acid, 25 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, and 2 mg of 4-dimethylaminopyridine in sequence. The amount of ferrocene carboxylic acid added is 15 mg. The ferrocene group (Fe 2+ / Fe 3+ The reversible redox couple of ... With valence bonds, the ferrocene groups are firmly fixed on the surface of the carrier material, avoiding systemic oxidative stress caused by free ferrocene molecules. This also ensures that its biological effects mainly occur at the tumor sites enriched in the carrier. The dual catalytic centers formed by ferrocene carboxylic acid and selenium continuously generate reactive oxygen species, causing irreversible oxidative damage to tumor cells. Furthermore, the excellent electron conduction ability of ferrocene can act as an electron bridge, promoting the rapid exchange of electrons in the catalytic reaction, making the entire selenium-iron catalytic cycle run faster and more efficiently. This accelerated local effective reaction also reduces the need for systemic high-dose drugs, reducing potential toxicity from the source and improving the safety of treatment, resulting in selenium-doped carbon nitride grafted with ferrocene carboxylic acid.

[0057] This embodiment provides a method for preparing an antitumor compound composition, specifically including the following steps:

[0058] S1. Rhubarb, aconite, dried ginger, ginseng, and licorice were dried and pretreated, then pulverized and mixed evenly. 500g of the powder was placed in 7 times its volume of 80% ethanol solution and soaked overnight. The mixture was then extracted twice under heat and pressure in a reflux condenser and a rotary evaporator. The extracts were filtered, combined, and concentrated under reduced pressure to obtain an alcohol extract. Finally, the alcohol extract was purified using AB-8 macroporous adsorption resin. During purification, water and ethanol (20% and 50% by mass) were used. Stepwise elution with 70% ethanol was performed, collecting eluates at 50% and 70% ethanol concentrations. This gradient elution method effectively removed highly water-soluble toxic components and impurities while simultaneously collecting key active ingredients such as saponins and flavonoids, increasing the concentration and purity of the active ingredients and thus resulting in a safer and more potent therapeutic effect. The eluate was concentrated to one-third of its original volume, then dispersed in a pH 4.8 acetate-sodium acetate buffer solution, filtered through a 0.22 μm filter membrane, and freeze-dried to obtain the purified product. Prepare the alcohol extract for later use. Meanwhile, add four times the amount of water to the dregs and decoct to extract the extract. Concentrate the resulting aqueous extract to one-third of its original volume, then add 95% ethanol to bring the ethanol content to 72.5%. Let it stand at 4°C for 24 hours, collect the precipitate, and wash twice with anhydrous ethanol and acetone. Finally, freeze-dry to obtain refined aqueous polysaccharide. Mix the refined alcohol extract and refined aqueous polysaccharide at a mass ratio of 2:1. In this formula, aconite and dried ginger can warm and tonify the spleen and kidneys. Yang energy dissipates stagnation of Yin and cold. Ginseng and licorice can greatly replenish vital energy and support the body's resistance to disease and its ability to repair itself. Rhubarb can purge and eliminate stagnation, promote blood circulation and remove blood stasis, thus clearing blockages. The combined use of these herbs embodies the tumor treatment method of "warming Yang and supporting the body's resistance, and combining attack and replenishment." Furthermore, the direct tumor-killing mechanism of the alcohol extract is combined with the immune-boosting mechanism of polysaccharides, forming a classic TCM treatment strategy of "simultaneously eliminating pathogens (attacking tumors) and supporting the body's resistance (enhancing immunity)," producing a safe and effective synergistic anti-tumor effect, resulting in the compound extract.

[0059] S2. 40 mg of ferrocene-grafted selenium-doped carbon nitride was dispersed in 15 mL of pH 7.4 PBS buffer and sonicated for 45 min to form a suspension for later use. Then, 60 mg of the compound extract described in step S1 was weighed and added to 5 mL of pH 7.4 PBS buffer and stirred for 25 min to form a drug solution for later use. Then, under magnetic stirring at 400 rpm and in the dark, the drug solution was added to the suspension at a rate of 1 mL / min using a constant flow pump. The mixture was placed in a constant temperature shaker and incubated at 135 rpm in the dark for 22 h. During the incubation, the temperature was controlled at 33.5℃. This temperature range not only increases the effective collision and contact opportunities between drug molecules and the carrier, optimizing loading efficiency and drug capacity, but also avoids denaturation and inactivation of heat-sensitive active ingredients, maintaining drug activity. Furthermore, by precisely simulating the internal temperature environment of the human body, the drug and carrier are pre-adapted to the in vivo environment, ensuring stability in blood circulation, significantly reducing systemic toxicity, and improving the safety of medication. Centrifuge at 13500 rpm for 15 min, wash the precipitate twice with pre-cooled PBS buffer, and finally disperse the washed precipitate in 5 mL of sterile, pyrogen-free water for injection and freeze-dry. Ferrocene-grafted selenium-doped carbon nitride serves as a carrier, loading the compound extract and protecting it from enzymatic degradation and premature removal during delivery, allowing more active ingredients to reach the lesion and improving bioavailability. Specifically, the carboxyl groups of ferrocene and the surface groups of carbon nitride undergo protonation changes in the slightly acidic environment of the tumor, leading to alterations in the carrier structure or surface charge, promoting specific drug release at the tumor site while reducing leakage in normal tissues, effectively improving treatment safety. Selenium doping not only modulates the catalytic properties of the material but also plays a catalytic therapeutic or tumor cell apoptosis-inducing role in the highly reactive oxygen species environment within tumor cells, producing a synergistic anti-tumor effect with the compound extract, thus achieving multimodal synergistic therapy and significantly improving treatment efficacy, resulting in an anti-tumor compound composition.

[0060] Example 4

[0061] This embodiment proposes an antitumor compound composition comprising the following components in parts by weight: 30 parts of selenium-doped carbon nitride grafted with ferrocene carboxylic acid, and 80 parts of compound extract.

[0062] The selenium-doped carbon nitride grafted with ferrocene carboxylic acid is made from the following components in parts by weight: 8 parts of nano-selenoglucose complex, 100 parts of melamine, and 0.1 parts of ferrocene carboxylic acid.

[0063] The compound extract contains the following components by weight: 5 parts rhubarb, 5 parts aconite, 5 parts dried ginger, 5 parts ginseng, and 5 parts licorice.

[0064] The preparation method of selenium-doped carbon nitride grafted with ferrocene-based carboxylic acid specifically includes the following steps:

[0065] (1) Place 0.173g of sodium selenite in a 100mL round-bottom flask, add 20mL of ultrapure water, and stir at 300rpm until completely dissolved. Then add 15.0g of glucose and stir at 300rpm until completely dissolved. Heat to 70℃ while stirring at 500rpm. Add 0.352g of ascorbic acid to the above mixture and reflux at 70℃ for 3h. Cool to room temperature and concentrate under reduced pressure in a 70℃ water bath using a rotary evaporator to 1 / 3 of the original volume. Dry under vacuum at 60℃ and grind. This process uses zero-valent nano-selenium as the functional core and hydrophilic glucose amorphous solid as the protective and dispersing matrix to form a solution with good water solubility and stability. In a core-shell nanocomposite, highly toxic sodium selenite is converted into low-toxicity, highly bioavailable zero-valent selenium nanoparticles. The glucose encapsulation further slows the release of selenium atoms, avoiding acute toxic reactions such as gastrointestinal reactions caused by sudden increases in blood drug concentration, thus improving safety. The nano-selenium can directly inhibit tumors through multiple pathways, such as inducing tumor cell apoptosis, regulating the cell cycle, and inhibiting angiogenesis. The hydrophilic glucose shell can effectively improve the dispersibility and solubility of the active ingredients of traditional Chinese medicine. The glucose matrix can also protect the light, heat, and oxygen-sensitive traditional Chinese medicine components from degradation during storage and in vivo circulation, thereby increasing the bioavailability of the active ingredients and improving the therapeutic efficacy, resulting in a nano-selenium glucose complex.

[0066] (2) After mixing the nano-selenoglucose complex and melamine described in step (1) evenly, the amount of nano-selenoglucose complex and melamine added is 0.8g and 10.0g, respectively. Melamine, as a precursor material for the carrier, can form a rigid porous carrier structure, thereby providing a huge specific surface area and abundant binding sites, which is conducive to loading hydrophobic Chinese medicine components through physical adsorption or chemical bonding, and can respond to the tumor microenvironment to achieve intelligent controlled release of drugs. First, dry in an oven at 60℃ for 12h, then transfer to a tube furnace, and calcine at 550℃ under nitrogen protection at a heating rate of 5℃ / min for 3 hours. h. Melamine is heated and polycondensed in an inert atmosphere, and deaminated to form a graphite-like layered structure of carbon nitride. At the same time, the nano-selenoglucose complex decomposes, and selenium atoms enter the carbon nitride framework in the form of atomic doping, realizing the "carrier" and functional synergy of selenium. The carrier thus possesses the anti-tumor activity of selenium. As a basic carrier, carbon nitride provides a physical barrier to prevent the rapid release of selenium in the blood circulation. Furthermore, through the high permeability and retention effect of solid tumors, it can be enriched at the tumor site and reduced in normal tissues, thereby reducing the potential toxicity of selenium to organs such as the heart, liver, and kidneys, and obtaining selenoglucose-doped carbon nitride.

[0067] (3) Weigh 400 mg of the selenoglucose-doped carbon nitride described in step (2) and disperse it in 20 mL of 25% hydrogen peroxide solution. Stir at 80°C for 2 h, centrifuge, wash the precipitate with water until neutral, and dry to obtain pretreated selenoglucose-doped carbon nitride. Then weigh 100 mg of the pretreated selenoglucose-doped carbon nitride and disperse it in 30 mL of N,N-dimethylformamide. Under nitrogen protection and a 5°C ice-water bath, add ferrocene carboxylic acid, 25 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, and 2 mg of 4-dimethylaminopyridine in sequence. The amount of ferrocene carboxylic acid added is 10 mg. The ferrocene group (Fe 2+ / Fe 3+ The reversible redox couple of ferrocene acid efficiently catalyzes the generation of highly toxic hydroxyl radicals in the high H2O2 microenvironment of tumors, achieving direct killing. Its catalytic activity is highly dependent on the tumor-specific high H2O2 environment, remaining silent in normal tissues, thus achieving selective attack. This improves both therapeutic efficacy and safety. The mixture was stirred in the dark for 50 min, then the ice-water bath was removed, the temperature was raised to 80℃, and stirring was continued in the dark for 12 h. The mixture was then centrifuged at 12000 rpm for 5 min. The solid product was washed five times alternately with anhydrous ethanol and ultrapure water, then twice with ethyl acetate, and finally vacuum dried at 60℃. The carboxyl groups introduced during pretreatment and ferrocene acid formed covalent groups through an amidation reaction under the action of a condensing agent and catalyst. By bonding ferrocene groups to the surface of the carrier material, the systemic oxidative stress caused by free ferrocene molecules is prevented. This ensures that the biological effects mainly occur at the tumor sites enriched in the carrier. The dual catalytic centers formed by ferrocene carboxylic acid and selenium continuously generate reactive oxygen species, causing irreversible oxidative damage to tumor cells. Furthermore, the excellent electron conduction ability of ferrocene can act as an electron bridge, promoting the rapid exchange of electrons in the catalytic reaction, making the entire selenium-iron catalytic cycle run faster and more efficiently. This accelerated local effective reaction also reduces the need for systemic high-dose drugs, thereby reducing potential toxicity from the source and improving the safety of treatment, resulting in selenium-doped carbon nitride grafted with ferrocene carboxylic acid.

[0068] This embodiment provides a method for preparing an antitumor compound composition, specifically including the following steps:

[0069] S1. Rhubarb, aconite, dried ginger, ginseng, and licorice were dried and pretreated, then pulverized and mixed evenly. 500g of the powder was placed in 8 times its volume of 80% ethanol solution and soaked overnight. The mixture was then extracted three times under heat and pressure in a reflux condenser and rotary evaporator. After filtration, the extracts were combined, and the collected extract was concentrated under reduced pressure to obtain an ethanol extract. This ethanol extract was then purified using AB-8 macroporous adsorption resin. During purification, water and 20%, 50%, and 70% ethanol were used for stepwise elution. The 50% and 70% ethanol eluates were collected. This gradient elution method removes highly water-soluble toxic components and impurities, while simultaneously collecting the main active ingredients such as saponins and flavonoids, increasing the concentration and purity of the active ingredients, thus resulting in a safer and more potent effect. The eluate was concentrated to 1 / 3 of its original volume, then dispersed in a pH 4.8 acetate-sodium acetate buffer solution, filtered through a 0.22μm filter membrane, and freeze-dried to obtain… The refined ethanol extract is prepared for later use. Meanwhile, the dregs are decocted with 5 times the amount of water. The resulting aqueous extract is first concentrated to 1 / 3 of its original volume, then 95% ethanol is added to bring the ethanol content to 75%. The mixture is then allowed to stand at 4°C for 24 hours. The precipitate is collected and washed twice with anhydrous ethanol and acetone, and finally freeze-dried to obtain the refined aqueous polysaccharide. Finally, the refined ethanol extract and refined aqueous polysaccharide are mixed at a mass ratio of 2:1. In this formula, aconite and dried ginger can warm and tonify the spleen and kidneys. Yang energy dissipates stagnation of Yin and cold. Ginseng and licorice can greatly replenish vital energy and support the body's resistance to disease and its ability to repair itself. Rhubarb can purge and eliminate stagnation, promote blood circulation and remove blood stasis, thus clearing blockages. The combined use of these herbs embodies the tumor treatment method of "warming Yang and supporting the body's resistance, and combining attack and replenishment." Furthermore, the direct tumor-killing mechanism of the alcohol extract is combined with the immune-boosting mechanism of polysaccharides, forming a classic TCM treatment strategy of "simultaneously eliminating pathogens (attacking tumors) and supporting the body's resistance (enhancing immunity)," producing a safe and effective synergistic anti-tumor effect, resulting in the compound extract.

[0070] S2. Disperse 30 mg of ferrocene-grafted selenium-doped carbon nitride in 15 mL of pH 7.4 PBS buffer and sonicate for 40 min to form a suspension for later use. Then, weigh 80 mg of the compound extract from step S1 and add it to 5 mL of pH 7.4 PBS buffer, stir for 20 min to form a drug solution for later use. Then, under magnetic stirring at 500 rpm and in the dark, add the drug solution to the suspension at a rate of 1 mL / min using a constant flow pump. Place the solution in a constant temperature shaker and incubate at 150 rpm in the dark for 20 h. During the incubation, control the temperature at 37°C. This temperature range not only increases the effective collision and contact opportunities between drug molecules and carriers, optimizing loading efficiency and drug load, but also avoids denaturation and inactivation of heat-sensitive active ingredients, maintaining drug activity. Furthermore, by accurately simulating the internal temperature environment of the human body, the drug and carrier can be pre-adapted to the in vivo environment, ensuring stability in blood circulation, significantly reducing systemic toxicity, and improving the safety of medication. Centrifuge at 15,000 rpm for 10 min. Wash the precipitate three times with pre-cooled PBS buffer. Finally, disperse the washed precipitate in 5 mL of sterile, pyrogen-free water for injection and freeze-dry. Ferrocene-grafted selenium-doped carbon nitride serves as a carrier to support the compound extract, protecting it from enzymatic degradation and premature clearance during delivery, allowing more active ingredients to reach the lesion and improving bioavailability. In the slightly acidic environment of tumors, the carboxyl groups of ferrocene and the surface groups of carbon nitride undergo changes in protonation, leading to alterations in the carrier structure or surface charge, promoting specific drug release at the tumor site, and reducing leakage in normal tissues, effectively improving treatment safety. Selenium doping not only modulates the catalytic properties of the material but also plays a role in catalytic therapy or inducing tumor cell apoptosis in the highly reactive oxygen species environment within tumor cells, producing a synergistic anti-tumor effect with the compound extract. This achieves multimodal synergistic therapy, significantly improving the therapeutic effect and yielding an anti-tumor compound composition.

[0071] Comparative Example 1

[0072] This comparative example provides an antitumor compound composition, which differs from Example 1 in that the selenium-doped carbon nitride grafted with ferrocene does not contain nano-selenoglucose complex; the preparation method of the selenium-doped carbon nitride grafted with ferrocene does not include step (1); the preparation method of the antitumor compound composition is the same as that of Example 1.

[0073] Comparative Example 2

[0074] This comparative example provides an antitumor compound composition, which differs from Example 1 in that the selenium-doped carbon nitride grafted with ferrocene does not contain ferrocene; the preparation method of the selenium-doped carbon nitride grafted with ferrocene does not include step (3); the preparation method of the antitumor compound composition is the same as that of Example 1.

[0075] Comparative Example 3

[0076] This comparative example provides an antitumor compound composition, which differs from Example 1 in that the compound extract does not contain purified water-extracted polysaccharide; the preparation method of selenium-doped carbon nitride grafted with ferrocene carboxylic acid is the same as in Example 1; the preparation method of the antitumor compound composition does not include the preparation of purified water-extracted polysaccharide in step S1, but only contains purified alcohol extract.

[0077] Experimental Example 1

[0078] Validity experiment

[0079] Test samples: Antitumor compound compositions prepared in Examples 1-4 and Comparative Examples 1-3.

[0080] Test method: Forty purebred BalB / c nude mice (half male and half female) were randomly divided into 8 groups of 5 mice each. The well-grown, milky-white ascites fluid, passaged for 5-7 days, was diluted with an appropriate amount of sterile saline to form a cancer cell suspension with a cell count of 102. 7 / mL, 0.2mL (containing 2×10⁶ cancer cells) was subcutaneously injected into the right axilla of each mouse. 6 Seven experimental groups were given 2g of the test sample, while the control group was given 0.5ml of distilled water. This treatment was administered for 10 consecutive days, with normal feeding, water, and food restrictions. The day after drug withdrawal, all mice were euthanized by cervical dislocation, and subcutaneous solid tumor masses were harvested, which were then weighed (mg). The cancer cell inhibition rate (%) was calculated using the following formula:

[0081] Cancer cell inhibition rate (%) = (average tumor weight of control group - average tumor weight of experimental group) / average tumor weight of control group × 100%.

[0082] Figure 1 The figures show the cancer cell inhibition rates of Examples 1-4 and Comparative Examples 1-3. As shown, the cancer cell inhibition rates of Examples 1-4 were 62-68%, indicating high treatment efficacy. The cancer cell inhibition rates of Comparative Examples 1-3 were 38-51%, indicating lower efficacy, suggesting moderate or poor treatment efficacy. In Comparative Example 1, the selenium-doped carbon nitride grafted with ferrocene-based acid did not contain the nano-selenoglucose complex, lacking the core precursor of selenium doping and thus failing to achieve the synergistic effect of selenium's "carrier" and catalytic function, resulting in poor treatment efficacy. In Comparative Example 2, the selenium-doped carbon nitride grafted with ferrocene-based acid did not contain ferrocene-based acid, failing to achieve responsive release and reducing the drug concentration at the tumor site, leading to poor treatment efficacy. In Comparative Example 3, the compound extract did not contain refined water-extracted polysaccharides, which is detrimental to reducing tumor cell drug resistance and consequently hinders the enhancement of the killing effect of the active ingredients in the refined alcohol extract on drug-resistant cells, resulting in moderate treatment efficacy.

[0083] Experimental Example 2

[0084] Safety Experiment

[0085] Test samples: Antitumor compound compositions prepared in Examples 1-4 and Comparative Examples 1-3.

[0086] Test method: Seventy volunteers with liver cancer (early and mid-stage) were selected and divided into 7 groups of 10 people each. They were given 2g of the test sample for drug treatment once every 2 days. After 60 days, the side effects of the drug (the most common manifestations of which are nausea, vomiting, abdominal pain, rash, dizziness and insomnia) were statistically analyzed.

[0087] Figure 2 The graph shows the frequency of side effects in Examples 1-4 and Comparative Examples 1-3. As shown, the frequency of side effects in Examples 1-4 was 5-10 times, indicating fewer side effects and higher safety. The frequency of side effects in Comparative Examples 1-3 was 12-36 times, indicating more side effects and moderate to poor safety. The ferrocene-grafted selenium-doped carbon nitride in Comparative Example 1 did not contain the nano-selenium glucose complex, lacking highly active nano-selenium. While this did not adversely affect safety, it required an increased dosage to achieve the therapeutic effect, resulting in moderate safety. The ferrocene-grafted selenium-doped carbon nitride in Comparative Example 2 did not contain ferrocene, making responsive release control impossible and increasing the risk of premature drug release in the bloodstream, leading to poor safety. The compound extract in Comparative Example 3 did not contain refined water-extracted polysaccharides, lacking the core detoxification and protective components, resulting in poor safety.

[0088] The above experimental results show that the efficacy and safety of Examples 1-4 of the present invention are significantly better than those of Comparative Examples 1-3. Among them, Example 1, which uses ferrocene-grafted selenium-doped carbon nitride and compound extract, has better efficacy and higher safety. Using ferrocene-grafted selenium-doped carbon nitride as a carrier, the compound extract is actively and targeted to tumor tissue in a manner that enhances penetration and retention effects and responsive release. This not only improves bioavailability but also reduces toxic side effects on normal tissues, thereby improving the efficacy and safety of treatment. The compound extract provides multi-target, multi-pathway natural chemotherapeutic therapy. Ferrocene-grafted selenium-doped carbon nitride not only prevents premature degradation of the compound extract but also supplements photothermal / photodynamic and catalytic therapeutic pathways, further enhancing and protecting the activity of the compound extract and comprehensively enhancing the anti-tumor effect.

[0089] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention.

[0090] The present invention and its embodiments have been described above. This description is not restrictive, and the accompanying drawings are only one embodiment of the present invention. The actual application is not limited to this. In conclusion, if those skilled in the art are inspired by this description and design similar methods and embodiments without departing from the spirit of the present invention, they should all fall within the protection scope of the present invention.

Claims

1. An antitumor compound composition, characterized in that: The antitumor compound composition comprises the following components in parts by weight: 30-40 parts of ferrocene-grafted selenium-doped carbon nitride, and 60-80 parts of compound extract; the ferrocene-grafted selenium-doped carbon nitride is made from the following components in parts by weight: 8-10 parts of nano-selenoglucose complex, 80-100 parts of melamine, and 0.1-0.2 parts of ferrocene-grafted carbon nitride; the raw materials of the compound extract include the following components in parts by weight: 3-5 parts of rhubarb, 3-5 parts of aconite, 3-5 parts of dried ginger, 3-5 parts of ginseng, and 3-5 parts of licorice.

2. A method for preparing an antitumor compound composition according to claim 1, characterized in that: Specifically, the following steps are included: S1. Rhubarb, aconite, dried ginger, ginseng, and licorice are dried and pretreated, then pulverized and mixed evenly. 500g of the powder is placed in 6-8 times its volume of 80% ethanol solution and soaked overnight. Then, it is extracted 2-3 times under heat and pressure in a reflux condenser and rotary evaporator. The extracts are filtered, and the combined extracts are concentrated under reduced pressure to obtain an alcohol extract. Finally, the alcohol extract is purified using an AB-8 macroporous adsorption resin. The resulting eluent is concentrated to 1 / 3 of its original volume and then purified using a pH 4.8 acetate-sodium acetate buffer. Dispersed, filtered through a 0.22μm filter membrane, and freeze-dried to obtain a purified alcohol extract for later use. Meanwhile, the residue was boiled with 3-5 times the amount of water for extraction. The resulting aqueous extract was first concentrated to 1 / 3 of its original volume, and then 95% ethanol was added to bring the ethanol content in the aqueous extract to 70-75%. The extract was then allowed to stand at 4°C for 24 hours. The precipitate was collected and washed twice with anhydrous ethanol and acetone. Finally, it was freeze-dried to obtain a purified aqueous polysaccharide. The purified alcohol extract and the purified aqueous polysaccharide were then mixed at a mass ratio of 2:1 to obtain a compound extract. S2. Disperse 30-40 mg of selenium-doped carbon nitride grafted with ferrocene carboxylic acid in 15 mL of PBS buffer at pH 7.4, sonicate for 40-50 min to form a suspension for later use. Then weigh 60-80 mg of the compound extract described in step S1 and add it to 5 mL of PBS buffer at pH 7.4, stir for 20-30 min to form a drug solution for later use. Then, under magnetic stirring at 300-500 rpm and in the dark, add the drug solution to the suspension at a rate of 1 mL / min using a constant flow pump, place it in a constant temperature shaker, and incubate at 120-150 rpm in the dark for 20-24 h. Then, centrifuge at 12000-15000 rpm for 10-20 min, wash the precipitate 2-3 times with pre-cooled PBS buffer, and finally, disperse the washed precipitate in 5 mL of sterile, pyrogen-free water for injection and freeze-dry to obtain the antitumor compound composition.

3. The method for preparing the antitumor compound composition according to claim 2, characterized in that: In step S1, during the purification process, water and ethanol with mass fractions of 20%, 50%, and 70% are used for stepwise elution, and ethanol eluents with mass fractions of 50% and 70% are collected.

4. The method for preparing the antitumor compound composition according to claim 3, characterized in that: In step S2, during the oscillation incubation process, the temperature is controlled at 30-37℃.

5. The method for preparing the antitumor compound composition according to claim 4, characterized in that: The method for preparing the selenium-doped carbon nitride grafted with ferrocene-formic acid specifically includes the following steps: (1) Place 0.173g sodium selenite in a 100mL round-bottom flask, add 20mL ultrapure water, stir at 300rpm until completely dissolved, then add 15.0g glucose, stir at 300rpm until completely dissolved, heat to 70℃ while stirring at 300-500rpm, add 0.352g ascorbic acid to the above mixture, reflux at 70℃ for 3-4h, cool to room temperature, concentrate under reduced pressure in a water bath at 60-70℃ using a rotary evaporator to 1 / 3 of the original volume, dry under vacuum at 50-60℃, grind to obtain nano-selenoglucose complex; (2) After mixing the nano-selenoglucose complex described in step (1) with melamine, dry it in an oven at 60°C for 12-24 hours, then transfer it to a tube furnace and calcine it at 500-550°C for 3-4 hours under nitrogen protection at a heating rate of 5°C / min to obtain carbon nitride doped with selenoglucose. (3) Weigh 300-400 mg of the selenoglucose-doped carbon nitride described in step (2) and disperse it in 20 mL of 25% hydrogen peroxide solution. Stir at 60-80℃ for 2-3 h, centrifuge, wash the precipitate with water until neutral, and dry to obtain pretreated selenoglucose-doped carbon nitride. Then weigh 50-100 mg of the pretreated selenoglucose-doped carbon nitride and disperse it in 20-30 mL of N,N-dimethylformamide. Under nitrogen protection and 0-5℃ ice-water bath conditions, add ferrocene sequentially. The mixture was prepared with 25 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2 mg of 4-dimethylaminopyridine, stirred in the dark for 50-60 min, the ice-water bath was removed, the temperature was raised to 60-80℃, and stirring was continued in the dark for 12-24 h. The mixture was then centrifuged at 10000-12000 rpm for 5-10 min. The solid product was washed 3-5 times with anhydrous ethanol and ultrapure water, then washed 1-2 times with ethyl acetate, and finally dried under vacuum at 60℃ to obtain selenium-doped carbon nitride grafted with ferrocene carbamate.

6. The method for preparing the antitumor compound composition according to claim 5, characterized in that: In step (2), the amounts of nano-selenium glucose complex and melamine added are 0.8-1.0g and 8.0-10.0g, respectively.

7. The method for preparing the antitumor compound composition according to claim 6, characterized in that: In step (3), the amount of ferrocene added is 10-20 mg.

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