CN115612902B - A kind of magnesium-based alloy micron particles that synergizes with TACE's anti-liver cancer effect and its preparation method - Google Patents

Abstract

The invention relates to a magnesium-based alloy micron particle that synergizes with TACE's anti-liver cancer effect and a preparation and application method. It uses metal magnesium as a carrier to smelt other zinc, selenium and molybdenum metal particles with anti-cancer effects, and uses a high-temperature spray method to prepare a degradation product. The micron-sized alloy particles are controllable and mild and can be stored in distilled water for long-term use. Before TACE treatment, it is locally implanted into the tumor tissue. In the early stage, the negative effects of TACE are reversed through various methods such as alkalinizing the microenvironment and anti-oxidation of the degradation products. In the final stage, the negative effects of TACE are degraded to nanoscale particles that can be endocytosed by cells and directly exert anti-cancer effects in the cells. . The present invention will provide a new anti-tumor biodegradable material for clinical practice that can reverse the negative effects of TACE and enhance the anti-liver cancer efficacy of TACE, thereby benefiting the survival of clinical malignant tumor patients.

Description

A kind of magnesium-based alloy micron particles that synergizes with TACE's anti-liver cancer effect and its preparation method

Technical field

The invention belongs to medical functional alloy materials and their preparation, and relates to a magnesium-based alloy micron particle that synergizes with the anti-liver cancer effect of TACE and its preparation and application methods. Using metal magnesium as a carrier, other beneficial metal particles such as zinc, selenium and molybdenum with anti-cancer effects are smelted. By optimizing the smelting ratio of magnesium to other metals and the particle size of the magnesium-based alloy, the degradation of magnesium-based alloys in tumor tissues is ultimately achieved. The rate is controllable. Utilize the chemical properties and degradation products of the magnesium degradation process to exert direct/indirect anti-cancer effects such as alkalinizing the tumor microenvironment and antioxidant, inhibiting the proliferation, invasion and metastasis of liver cancer, reducing the "dryness" of liver cancer, and then exerting synergistic TACE anti-liver cancer and reversing The role of TACE in treating negative effects is highly targeted in the treatment of liver cancer.

Background technique

According to the Barcelona Clinic Liver Cancer (BCLC) staging system, TACE is the preferred method for the treatment of intermediate-stage liver cancer, including unresectable multinodular liver cancer without extrahepatic spread. The BCLC system also recommends that TACE should be used when other recommended treatments are not feasible or successful in early stages of liver cancer. In Asian countries, TACE tends to be more widely recommended for a variety of clinical situations. According to different staging systems, the clinical conditions shown by TACE are slightly different, but TACE is still a mature method for the treatment of intermediate and advanced HCC. See the literature: Young Chang, Soung Won Jeong, Jae Young Jang, et al. Recent Updates of Transarterial Chemoembolilzation in Hepatocellular Carcinoma.Int.J.Mol.Sci.2020,21,8165. TACE mainly exerts its anti-liver cancer effect by blocking the blood supply of liver cancer and combining it with local injection of chemotherapy drugs (such as oxaliplatin). Although the imaging equipment and super-selection technology related to TACE treatment have been continuously improved in the past 10 years, the survival rate of liver cancer patients has not been improved. The benefits are still very limited. Multiple studies have shown that while TACE fights liver cancer, it will accelerate cancer invasion and metastasis, eventually leading to treatment failure. See the literature: Raoul JL, Gilabert M, Adhoute X: ToTACE or not to TACE? Lessons from a negative trial.Lancet GastroenterolHepatol 2017,2(8):541-543. The main reasons for the failure of TACE are: ① The hypoxic environment caused by tumor supply artery embolism will intensify the anaerobic glycolysis of cancer cells and the acidification of the microenvironment, thereby promoting cancer cell invasion and metastasis; ② Repeated chemotherapy will aggravate the drug resistance of liver cancer cells; ③ TACE The necrosis of carcinogenic tissue will also cause the infiltration of inflammatory stromal cells into the cancer tissue. This process also plays an important role in the metastasis, recurrence, and drug resistance of liver cancer.

In order to enhance clinical efficacy, TACE combination therapy has been proposed, and its efficacy and safety have been verified through a large number of clinical studies. However, studies have shown that TACE combined with radiofrequency ablation, radiotherapy, systemic therapy and other options have not brought significant benefits to patients. See the literature: Strobel O, Buchler MW: Treatment effect of liver resection vs. RFAor TACE in hepatocellular carcinoma. Chirurg 2019. The proliferation, recurrence and metastasis of liver cancer are still difficult clinical problems. Currently, there is no clinically available biodegradable material that can simultaneously reverse the negative effects of TACE and enhance the anti-liver cancer efficacy of TACE. After searching domestic and foreign journals, literature, and patents, no relevant reports or related patent authorizations were found. Therefore, seeking new strategies that can effectively improve the acidification of the microenvironment of liver cancer and reverse chemotherapy resistance for anti-liver cancer treatment in conjunction with TACE will be the key to improving the survival benefits of liver cancer patients.

Contents of the invention

Technical issues to be solved

In order to improve the shortcomings of the existing technology, the present invention proposes a magnesium-based alloy micron particle that synergizes with TACE's anti-liver cancer effect and a preparation and application method. Metal magnesium is used as a carrier to smelt other zinc, selenium and molybdenum that have anti-cancer effects. Metal particles, by smelting a certain proportion of other metals with anti-cancer effects, improve the shortcomings of the rapid degradation of micron magnesium metal particles. At the same time, the products of slow degradation in tumor tissues are used to improve the acidic microenvironment of tumors and reduce the "dryness" of tumors. ", reverse the negative effects of TACE treatment, and ultimately cooperate with TACE to exert anti-liver cancer effects. This will provide a new anti-liver cancer biodegradable material that can reverse the negative effects of TACE and enhance the anti-liver cancer efficacy of TACE, which will benefit the survival of clinical liver cancer patients.

Technical solutions

A kind of magnesium-based alloy micron particles that synergizes with TACE's anti-liver cancer effect. It is characterized by using metallic magnesium as a carrier and smelting it with other metal particles with anti-cancer effects to prepare new magnesium-based alloy micron particles; the metal magnesium and other anti-cancer effects The mass ratio of cancer-causing metal particles is 85~99:1~15.

The metal particles with anti-cancer effects include but are not limited to zinc, selenium or molybdenum metal particles.

A method for preparing the magnesium-based alloy microparticles that synergize with TACE's anti-liver cancer effect, characterized by the following steps:

Step 1: Place magnesium ingots and other metal ingots with anti-cancer effects into two pre-melting furnaces and vacuum them. Set the temperature to 300°C and purge them with high-temperature inert gas argon to remove the oxidizing atmosphere gas adsorbed on the surface. ;

Step 2: Heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C. The furnace body of other metal ingots with anti-cancer effects is heated to the melting temperature of the metal. Argon gas is used for protection to completely melt the metal. Obtain liquid magnesium and other liquid metals with anti-cancer properties;

Step 3: Pour liquid magnesium and other liquid metals with anti-cancer effects into the dish atomizer in the atomization furnace, mix the two liquid metals evenly on the dish atomizer, and then spray them into Magnesium-based alloy droplets;

Step 4: Rapidly condense the small droplets of magnesium-based alloy to form low-oxidation solid spherical magnesium-based alloy powder;

Step 5: Sieve the magnesium-based alloy powder for particle size classification, screen out 10-200 μm spherical magnesium-based alloy micron particles, and store them in distilled water/or absolute alcohol for long-term storage.

A vibrating sieve is used to classify the particle size of the alloy powder. After sieving, it passes through a 600-800 mesh sieve.

The step 2 uses an intermediate frequency heating coil to heat the furnace body.

By controlling the respective flow rates of the two liquid metals on the disc atomizer introduced into the atomization furnace in step 3, the mass ratio of the magnesium-based alloy micron particles is achieved.

In step 3, the frequency of the atomizer and the speed of the centrifuge are controlled to control the size of the metal droplets.

The atomizer frequency is 55Hz.

A method of using the magnesium-based alloy microparticles that synergize with TACE's anti-liver cancer effect, which is characterized by: injecting into the hepatic artery through the femoral artery, along with the guidewire catheter, or super-selecting into the blood supply artery of the liver segment, or the blood supply of the liver cancer lesion The artery is implanted locally into the tumor tissue and stays in the capillaries of the cancer tissue according to the particle size of the magnesium-based alloy, and cooperates with TACE therapy to exert an anti-liver cancer effect.

During use, after the magnesium-based alloy micron particles are dissolved in physiological saline, the magnesium-based alloy micron particles are locally implanted at the tumor location with the help of Taoism during TACE treatment 10 minutes before TACE treatment.

beneficial effects

The present invention proposes a magnesium-based alloy micron particle that synergizes with the anti-liver cancer effect of TACE and a preparation and application method. This medical functional alloy material takes advantage of the natural corrosion and degradation properties of magnesium metal and the anti-cancer potential of its degradation products, combined with zinc, The anti-cancer properties of molybdenum and selenium plasma are used to optimize the optimal smelting ratio of metal elements. The high-temperature spray method is used to prepare micron-sized alloy particles with a degradation rate consistent with the TACE treatment cycle, a controllable degradation rate, and a mild effect. They can be stored for a long time in Set aside in distilled water or absolute alcohol. During TACE treatment, magnesium-based alloy micron particles are implanted into tumor tissue, which exerts a synergistic anti-cancer effect and reverses the negative effects of TACE through various methods such as alkalinizing the microenvironment and anti-oxidation of its degradation products. With the terminal stage of magnesium-based micron alloy particles It is degraded into nano-sized particles in cancer tissue and can further exert a direct anti-liver cancer effect in cells through post-cytosis.

Compared with the prior art, the present invention has the following effects:

1. Local implantation of metallic magnesium achieves local sustained release and controllable degradation of liver cancer and synergistic anti-liver cancer prevention

As a medical implantable alkaline metal, magnesium is not only rich in resources, low in price, but also has good biocompatibility and degradability. It is a new type of medical functional material with great clinical application prospects. The degradation products of magnesium metal, magnesium hydroxide, hydrogen and magnesium ions, can directly/indirectly inhibit the malignant progression of liver cancer by alkalizing the microenvironment and resisting oxidation. However, their own degradation rate is fast and uneven, especially when prepared into micron particles. The shortcoming of extremely rapid degradation limits its clinical application. The present invention prepares micron-sized magnesium-based alloy particles that can be locally implanted in liver cancer tissue and have mild and controllable degradation. The synergistic anti-liver cancer effect of local implantation is difficult to achieve through inhalation, oral or intravenous injection of metal magnesium particles and their degradation products. to achieve. The anti-tumor effects of metallic magnesium in the present invention include: first, implanting magnesium metal into liver cancer tissue to degrade and release the alkaline substance magnesium hydroxide can accurately increase the local pH value of the cancer tissue and reverse the damage to tumor cells caused by the acidic microenvironment. It exerts effective anti-cancer effects by promoting cancer-promoting effects such as dedifferentiation, “stemness” enhancement, and induction of epithelial-mesenchymal transition (EMT). Second, when the oxidation reaction is enhanced and/or the antioxidant capacity is impaired, the redox reaction is destroyed and oxidative stress is generated, resulting in oxidative damage to lipids, proteins, and DNA, thereby causing/promoting the occurrence and development of tumors. . Magnesium metal is directly implanted into liver cancer tissue to degrade and slowly release hydrogen, which can accurately increase the local hydrogen concentration in liver cancer and inhibit tumor progression through antioxidant effects. The traditional hydrogen treatment strategy has very little hydrogen reaching the tumor tissue, making it difficult to exert an effective anti-tumor effect. Third, the local release of magnesium ions can accurately increase the local magnesium ion concentration in cancer tissues and inhibit the metastasis and invasion of tumor cells. Patients with radiotherapy, chemotherapy, or molecular targeted therapy are often accompanied by a decrease in serum magnesium. Appropriate supplementation of magnesium ions can enhance the anti-cancer effect of DNA-damaging therapy and act as an anti-cancer treatment sensitizer.

2. Accurately increase the safe concentration of zinc ions in liver cancer tissues, and exert anti-liver cancer effects by directly affecting the liver cancer itself and/or improving the microenvironment of liver cancer.

Zinc is involved in the composition of a variety of proteins and can be used as a signaling molecule in cells in the form of free ions to participate in regulating cell metabolism, protein kinase and phosphatase activation, etc., and plays an important role in tumor cell proliferation, apoptosis, differentiation and immune regulation. . Zinc ions play a double-edged sword role in cancer patients. Overload of zinc ions may lead to the occurrence of cachexia and even accelerate the death of patients, while insufficient concentration cannot exert an effective anti-tumor effect. Therefore, systemic medication to increase zinc ion concentration is safe for cancer patients. The windows are smaller. Therefore, only by accurately increasing the local zinc ion concentration in liver cancer tissue can the anti-tumor effect be effectively exerted. The local implantation of the present invention can maximize the killing effect of metallic zinc on liver cancer tissue and enhance the utilization rate of zinc ions in the body. The anti-liver cancer effects of metallic zinc in the present invention include: first, inhibiting the release of a large number of tumor-related inflammatory factors and increasing the expression of A20 and PPAR-a, two zinc finger proteins with anti-inflammatory effects; second, inhibiting intracellular Ras- The MAPK signaling pathway directly or cooperatively inhibits the malignant progression of tumors, including inhibiting proliferation, inducing apoptosis, or enhancing the anti-cancer efficacy of radiotherapy and chemotherapy; third, enhancing the activity of antioxidant proteins and enzymes, such as glutathione and Catalase.

3. Selenium and molybdenum metal particles integrate diagnosis and treatment, combining physical therapy with chemical therapy.

This study combines selenium and molybdenum particles with magnesium carriers to integrate diagnosis and treatment. On the one hand, molybdenum-containing compounds can be used for tumor-targeted fluorescence imaging and photothermal therapy. Through high permeability and retention effect, it accumulates at the tumor site and reaches the maximum amount, reducing damage to surrounding tissues. At the same time, its photothermal killing effect can damage the vascular endothelial cells of tumor tissue. On the other hand, it can be used as a drug carrier, a chemotherapeutic drug, and a radiotherapy sensitizing agent, and the drug efficacy can be enhanced through targeted design. The present invention not only exerts the anti-tumor effects of selenium and molybdenum metals respectively, but also improves the toxicity and stability of their component metals, improves their targeting, biocompatibility and photothermal killing effect, and reduces the immune rejection effect of the body against particles. .

Therefore, the present invention focuses on the research and development of advanced biomedical functional materials, and synthesizes magnesium-based alloy micron particles for local implantation in liver tumor tissue, solving the clinical application problems of magnesium metal being relatively active and uncontrollable degradation, and maximizing the use of magnesium. and other metal degradation products’ anti-liver cancer effects. At the same time, the present invention will provide an advanced functional material for clinical practice that can reverse the negative effects of TACE and enhance the anti-liver cancer efficacy of TACE, thereby benefiting the survival of clinical liver cancer patients.

Description of the drawings

Figure 1: Cytotoxic effects of M-6Z, M-6S, and M-6M gradient times and gradient concentrations on liver cancer cell lines, hepatic stellate cell lines, and normal liver cell lines

A: M-6Z; B: M-6S; C: M-6M

Figure 2: Effect of magnesium-based alloy microparticles on the proliferation and invasion of liver cancer cell lines

A: M-6Z, M-6S and M-6M inhibit the malignant proliferation of liver cancer cell lines;

B: M-6Z inhibits the invasion ability of liver cancer cell lines

Figure 3: Mouse liver cancer tumor growth

Figure 4: Tumor growth after liver cancer treatment in mice

Detailed ways

The present invention will now be further described with reference to the embodiments and drawings:

Magnesium-based alloy micron particles and preparation examples:

Example 1: Magnesium-zinc alloy micron particles (6% zinc), prepared by high-temperature spray method

(1) Put the magnesium ingot and zinc ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing zinc ingots is heated to 650°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid zinc into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: zinc = 94:6). Mix the magnesium liquid and the zinc liquid evenly on the disc atomizer, and then spray them into small droplets. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-zinc alloy micron particles and store them in distilled water.

Example 2: Magnesium-zinc alloy micron particles (3% zinc), prepared by high-temperature spray method

(1) Put the magnesium ingot and zinc ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing zinc ingots is heated to 650°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid zinc into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: zinc = 97:3). Mix the magnesium liquid and the zinc liquid evenly on the disc atomizer, and then spray them into small droplets. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-zinc alloy micron particles and store them in distilled water.

Example 3: Magnesium-zinc alloy micron particles (12% zinc), prepared by high-temperature spray method

(1) Put the magnesium ingot and zinc ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing zinc ingots is heated to 650°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid zinc into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: zinc = 88:12). Mix the magnesium liquid and the zinc liquid evenly on the disc atomizer, and then spray them into small droplets. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-zinc alloy micron particles and store them in distilled water.

Example 4: Magnesium-zinc alloy micron particles (15% zinc), prepared by high-temperature spray method

(1) Put the magnesium ingot and zinc ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing zinc ingots is heated to 650°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid zinc into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: zinc = 85:15). Mix the magnesium liquid and the zinc liquid evenly on the disc atomizer, and then spray them into small droplets. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-zinc alloy micron particles and store them in distilled water.

Example 5: Magnesium-selenium alloy micron particles (3% selenium), prepared by high-temperature spray method

(1) Put the magnesium ingot and selenium ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing selenium ingots is heated to 650°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid selenium into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: selenium = 97:3). Mix the magnesium liquid and selenium liquid evenly on the disc atomizer, and then spray them into small droplets. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-selenium alloy micron particles and store them in distilled water.

Example 6: Magnesium-selenium alloy micron particles (6% selenium), prepared by high-temperature spray method

(1) Put the magnesium ingot and selenium ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing selenium ingots is heated to 650°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid selenium into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: selenium = 94:6). Mix the magnesium liquid and selenium liquid evenly on the disc atomizer, and then spray them into small droplets. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-selenium alloy micron particles and store them in distilled water.

Example 7: Magnesium-selenium alloy micron particles (12% selenium), prepared by high-temperature spray method

(1) Put the magnesium ingot and selenium ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing selenium ingots is heated to 650°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid selenium into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: selenium = 88:12). Mix the magnesium liquid and selenium liquid evenly on the disc atomizer, and then spray them into small droplets. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-selenium alloy micron particles and store them in distilled water.

Example 8: Magnesium-selenium alloy micron particles (15% selenium), prepared by high-temperature spray method

(1) Put the magnesium ingot and selenium ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing selenium ingots is heated to 650°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid selenium into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: selenium = 85:15). Mix the magnesium liquid and selenium liquid evenly on the disc atomizer, and then spray them into small droplets. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-selenium alloy micron particles and store them in distilled water.

Example 9: Magnesium-molybdenum alloy micron particles (3% molybdenum), prepared by high-temperature spray method

(1) Put the magnesium ingot and molybdenum ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing molybdenum ingots is heated to 2700°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid molybdenum into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: molybdenum = 97:3). Mix the magnesium liquid and the molybdenum liquid evenly on the disc atomizer and spray them into small droplets after mixing. Control the atomizer frequency to 55Hz and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-molybdenum alloy micron particles and store them in distilled water.

Example 10: Magnesium-molybdenum alloy micron particles (6% molybdenum), prepared by high-temperature spray method

(1) Put the magnesium ingot and molybdenum ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing molybdenum ingots is heated to 2700°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid molybdenum into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: molybdenum = 94:6). Mix the magnesium liquid and the molybdenum liquid evenly on the disc atomizer and spray them into small droplets after mixing. Control the atomizer frequency to 55Hz and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-molybdenum alloy micron particles and store them in distilled water.

Example 11: Magnesium-molybdenum alloy micron particles (12% molybdenum), prepared by high-temperature spray method

(1) Put the magnesium ingot and molybdenum ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing molybdenum ingots is heated to 2700°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid molybdenum into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: molybdenum = 88:12). Mix the magnesium liquid and the molybdenum liquid evenly on the disc atomizer and spray them into small droplets after mixing. Control the atomizer frequency to 55Hz and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-molybdenum alloy micron particles and store them in distilled water.

Example 12: Magnesium-molybdenum alloy micron particles (15% molybdenum), prepared by high-temperature spray method

(1) Put the magnesium ingot and molybdenum ingot into two pre-melting furnaces respectively and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove the gas containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing molybdenum ingots is heated to 2700°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid molybdenum into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: molybdenum = 85:15). Mix the magnesium liquid and the molybdenum liquid evenly on the disc atomizer and spray them into small droplets after mixing. Control the atomizer frequency to 55Hz and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-molybdenum alloy micron particles and store them in distilled water.

Comparative Example 1: Magnesium-aluminum alloy micron particles (6% aluminum), prepared by high-temperature spray method

(1) Place magnesium ingots and aluminum ingots into two pre-melting furnaces and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove gases containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C. The furnace body containing aluminum ingots is heated to 680°C and protected by argon gas to completely melt the metal;

(3) Pour liquid magnesium and liquid aluminum into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: aluminum = 94:6). Mix the magnesium liquid and the aluminum liquid evenly on the disc atomizer, and then spray them into small droplets. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to remove 20 μm spherical magnesium-aluminum alloy micron particles and store them in distilled water.

Comparative Example 2: Magnesium-manganese alloy micron particles (6% manganese), prepared by high-temperature spray method

(1) Put magnesium ingots and manganese ingots into two pre-melting furnaces and vacuum them, set the temperature to 300°C, and use high-temperature inert gas (argon) to purge to remove gases containing oxidizing atmosphere adsorbed on the surface;

(2) Use an intermediate frequency heating ring to heat the furnace body. The furnace body containing magnesium ingots is heated to 680°C, and the furnace body containing manganese ingots is heated to 1300°C. Argon gas is used for protection to completely melt the metal;

(3) Pour liquid magnesium and liquid manganese into the disc atomizer in the atomization furnace, and control the respective flow rates to achieve a mass ratio of magnesium-based alloy micron particles (magnesium: manganese = 94:6). Mix the magnesium liquid and manganese liquid evenly on the disc atomizer, and spray them into small droplets after mixing. Control the atomizer frequency to 55Hz, and control the size of the metal droplets by controlling the centrifuge speed and other parameters;

(4) Rapidly condense the alloy droplets to form low-oxidation solid spherical magnesium-based alloy powder;

(5) Use a vibrating sieve to classify the particle size of the alloy powder. After sieving, pass it through a 600-800 mesh sieve to screen out 20 μm spherical magnesium-manganese alloy micron particles and store them in distilled water.

Table 1 Nutritional treatment plan of treatment group

1.1 Magnesium-based alloy micron particles have no direct cytotoxic effect on liver cancer cells, hepatic stellate cells and normal liver cells.

M-6Z, M-6S and M-6M were used to treat liver cancer cell lines, hepatic stellate cell lines, and normal liver cell lines with gradient times and gradient concentrations, and the expression of LDH in the cell supernatant was detected, indicating that with the increase of time and concentration, The expression of LDH in the supernatant showed an increasing trend, but the overall increase was not obvious. And M-6Z reached a plateau at 24h and 4mg. (see picture 1)

Cytotoxic effects of M-6Z, M-6S, and M-6M gradient time and gradient concentration on liver cancer cell lines, hepatic stellate cell lines, and normal liver cell lines. 1-A M-6Z; 1-B M-6S; 1-C M-6M.

1.2 Using two human liver cancer cell lines, MHCC97H and Hep3B, CCK8 detection showed that with the gradual increase in the concentration of M-6Z, M-6S and M-6M, the inhibitory effect was significantly enhanced (see Figure 2).

Matrigel was added to the Transwell chamber to construct an in vitro invasion chamber, and the M-6Z 4mg/ml system was constructed in the lower chamber. The liver cancer cell lines MHCC97H (58.33±7.51vs.24.31±6.87p=0.0044) and Hep3B (65.00±6.51 vs.40.17±8.71p =0.0095) significantly inhibited the invasion ability (see Figure 2).

Experimental Design 2 (Animal Experiment)

1.1 Experimental purpose and objects

The anti-cancer effect and safety of the combined treatment plan of the present invention and TACE are discussed.

75 8-week-old C57BL/6 healthy mice

1.2 Method

75 mice were randomly divided into 15 groups, with 5 cases in each group, corresponding to 14 treatment groups and 1 control group and given different treatment plans (see Table 2).

Construction of mouse orthotopic liver cancer model

The H22 cell line was cultured in 1640 medium containing 10% fetal calf serum, and cells in the exponential growth phase were collected. After anesthesia, the mice were fixed in the supine position. H22 liver cancer cell suspension (5×105/25μL) was injected under the capsule of the left outer lobe of the liver. A small bleb appeared at the injection site, proving that the injection was successful.

Injection of magnesium-based alloy micron particles into mice with orthotopic cancer

Mice were treated with TACE on the 30th day after inoculation with H22 cells. The skin was routinely disinfected and 5 mL of 2% lidocaine hydrochloride injection was used for local anesthesia. The first angiography determines the location of the lesion and the direction of the puncture needle, and explores the lesion in detail; the second angiography guides the puncture injection needle to enter through the guide device and the skin, through the abdomen and intercostal space, and then the magnesium-based alloy micron particles are injected after the lesion is clear ( Dissolve 1 mg in 5 ml of normal saline, mix well, and inject slowly). After the injection is completed, quickly pull out the needle and pressurize the puncture point to avoid bleeding.

Tumor ischemia surgery in mice 24 hours after local injection

Grouping: treatment group, control group

Mice were fasted 12 h before surgery and had free access to water.

Mice were anesthetized: sodium pentobarbital (100ul/20g) was injected intraperitoneally. After anesthesia, the mice were placed in a supine position, and the limbs were fixed with tape. The abdominal hair was removed with an electric razor, disinfected with 1% iodine, and covered with a sterile towel.

Make a longitudinal incision in the middle of the abdomen, cut the skin layer and muscle layer respectively, from the xiphoid process to the bladder, use Alice forceps to pull and fix the bilateral muscle layers to expose the liver, moisten the cotton swab and separate the porta hepatis. , exposing the Glisson system in the left lobe and middle lobe of the liver.

5) Use microforceps to peel off the main blood supply blood vessels of the tumor under a stereomicroscope, and use non-invasive blood vessel clips to block the blood supply of the tumor.

6) When the tumor tissue changes from reddish brown to pale, it indicates that the blood supply has been successfully blocked. After successfully blocking the blood vessels, suture and disinfect the incision layer by layer in order, place it on an electric blanket or a thermostatic board to keep warm until it wakes up, and place it in a mouse cage covered with soft shavings. Place soft food in the cage and place each mouse in it. Raise them individually and observe the living conditions of the mice.

1.1 Results

1.3.1 Growth of mice

During the experiment, the mice in both the treatment group and the control group had normal activities, their coats were shiny, they ate and drank normally, there were no abnormal changes in bowel movements and urination, and none died. Tumor tissue could be detected in all groups of mice in the liver cancer orthotopic model, and the tumor formation rate reached 100%.

1.3.2 Effect of magnesium-based alloy micron particles on tumor growth in experimental mice

An orthotopic model of human liver cancer in nude mice was constructed. Four weeks after hepatocyte injection to form tumors, magnesium-zinc alloy micron particles were injected into the liver cancer lesions at multiple points. Mouse tumor ischemia intervention was performed 24 hours after injection. One week and two weeks later, respectively The mice were sacrificed and the tumor lesions were detected, and the tumor volume was significantly reduced. After 2 weeks of treatment, the magnesium-based alloy microparticles had better anti-tumor effect, with treatment group 1 having the best effect, treatment groups 5 and 9 second, and treatment groups 13 and 14 having the worst effect. (See Figure 4).

1.3.3 Effect of magnesium-based alloy micron particles on organ coefficients of experimental mice

Compared with the control group, the organ coefficients of the organs in the treatment group were significantly decreased in treatment groups 13 and 14 (P＜0.001). Compared with the control group, the differences in the other groups were not statistically significant (P＞ 0.05). The results are shown in Table 2.

Discussion of beneficial effects:

The locally implanted magnesium-based alloy micron particles designed by the present invention to cooperate with TACE anti-liver cancer treatment take advantage of the natural corrosion degradation properties of magnesium metal and the anti-cancer potential of its degradation products, combined with the anti-cancer properties of zinc, molybdenum, selenium and other ions. Its synergistic anti-tumor effect with TACE is optimal. When local implantation of magnesium-based alloy micron particles 24 hours before TACE treatment, compared with simple TACE treatment, the overall effective rate, disease control rate, tumor volume, and survival conditions were significantly improved, and 6% magnesium-zinc alloy was the most effective. Good, there was no statistical difference in the incidence of complications between groups.

Zinc is an important trace element in the human body. It participates in the composition of various proteins. It can be used as a signaling molecule in cells in the form of free ions to participate in regulating cell metabolism, protein kinase and phosphatase activation, etc., and plays a role in tumor cell proliferation, apoptosis, Play an important role in differentiation and immune regulation. Cancer patients are often accompanied by a deficiency of zinc ions. Increasing the intake of zinc ions can reduce the incidence of tumors. Low zinc can induce the activation of NFκB signaling, thereby promoting the release of a large number of tumor-related inflammatory factors, such as TNF-a and IL-1b, while supplementing zinc ions can increase A20 and PPAR-a, two zinc finger proteins with anti-inflammatory effects. In addition, zinc ions can directly or cooperatively inhibit the malignant progression of tumors by inhibiting the intracellular Ras-MAPK signaling pathway, such as inhibiting proliferation, inducing apoptosis, or enhancing the anti-cancer efficacy of radiotherapy and chemotherapy. In the present invention, the combination of magnesium-6% zinc alloy microparticles (Mg-6Zn) and TACE has the best anti-tumor effect.

Selenium compounds inhibit metastasis of mouse melanoma cells by inducing cell cycle arrest and cell death. Additionally, selenium compounds can sensitize various cancer cells to a variety of widely used drugs. Mechanisms such as selenium inducing cell apoptosis or arresting the cell cycle, inhibiting cell proliferation, regulating redox status, detoxifying carcinogens, stimulating the immune system, and inhibiting angiogenesis have been considered important mechanisms of selenium's anti-tumor effects and are used as drugs for tumors. Treatments have been increasingly studied. However, as an organic selenium preparation, selenium compounds have high toxicity and poor targeting properties. Therefore, the present invention uses nano-selenium materials, which have higher bioavailability, stronger biological activity and lower toxicity than organic selenium and inorganic selenium. In addition, as a drug carrier, nano-selenium has the advantages of good biocompatibility, high loading rate, low toxicity, easy synthesis, and easy storage. Although nano-selenium particles can inhibit the growth of cancer cells for a long time, they cannot effectively eliminate solid tumors in a short time. Therefore, 3% magnesium-selenium alloy micron particles (Mg-3Se) are used as the optimal ratio and combined with TACE can quickly eliminate tumors and improve the therapeutic efficacy.

Molybdenum-containing compounds can be used for tumor-targeted fluorescence imaging and photothermal therapy. Through high permeability and retention effect, it accumulates at the tumor site and reaches the maximum amount, causing little damage to surrounding tissues. Its photothermal killing effect can kill vascular endothelial cells in tumor tissue.

To sum up, this special invention specifically enhances the effectiveness of TACE treatment and alleviates its drug resistance. Among them, magnesium-zinc alloy microparticles (Mg-6Zn), magnesium-selenium alloy microparticles (Mg-3Se), and magnesium-molybdenum alloy microparticles (Mg-3Mo) have good synergistic anti-tumor effects, and Mg-6Zn has the best effect. While bringing huge clinical benefits to patients, no serious adverse events occurred, and both safety and effectiveness meet the requirements.

Claims (8)

1. The magnesium-based alloy microparticles with synergistic TACE liver cancer resisting effect are characterized in that metal magnesium is used as a carrier and is smelted with other metal particles with anticancer effect to form magnesium-based alloy microparticles; the mass ratio of the magnesium metal to other metal particles with anticancer effect is 85-97:1-15;

the metal particles with anticancer effect comprise zinc, selenium or molybdenum metal particles;

the magnesium-based alloy micron particles with the synergistic TACE liver cancer resistance are prepared according to the following steps:

step 1: respectively placing magnesium ingots and other metal ingots with anticancer effect into two premelting furnaces, vacuumizing, setting the temperature to 333 ℃, and purging by adopting high-temperature inert gas argon to remove oxidizing atmosphere gas adsorbed on the surfaces;

step 2: heating the furnace body, heating the furnace body containing magnesium ingots to 683 ℃, heating the furnace body of other metal ingots with anticancer effect to the melting temperature of the metal, and adopting argon for protection to completely melt the metal to obtain liquid magnesium and other liquid metals with anticancer effect;

step 3: introducing liquid magnesium and other liquid metals with anticancer effect onto a disc atomizer in an atomizing furnace, uniformly mixing the two liquid metals on the disc atomizer, and spraying the mixed liquid metals into magnesium-based alloy droplets;

step 4: rapidly condensing the magnesium-based alloy small liquid drops to form low-oxidation solid spherical magnesium-based alloy powder;

step 5: sieving magnesium-based alloy powder, grading the granularity, and storing spherical magnesium-based alloy micron particles with 13-233um under the sieve in distilled water and/or absolute alcohol;

and (3) carrying out granularity classification on the alloy powder by adopting a vibrating screen, wherein the alloy powder is screened and then passes through a 633-833 mesh screen.

2. A method for preparing magnesium-based alloy microparticles with synergistic TACE liver cancer resistance according to claim 1, which is characterized by comprising the following steps:

step 1: respectively placing magnesium ingots and other metal ingots with anticancer effect into two premelting furnaces, vacuumizing, setting the temperature to 333 ℃, and purging by adopting high-temperature inert gas argon to remove oxidizing atmosphere gas adsorbed on the surfaces;

step 2: heating the furnace body, heating the furnace body containing magnesium ingots to 683 ℃, heating the furnace body of other metal ingots with anticancer effect to the melting temperature of the metal, and adopting argon for protection to completely melt the metal to obtain liquid magnesium and other liquid metals with anticancer effect;

step 3: introducing liquid magnesium and other liquid metals with anticancer effect onto a disc atomizer in an atomizing furnace, uniformly mixing the two liquid metals on the disc atomizer, and spraying the mixed liquid metals into magnesium-based alloy droplets;

step 4: rapidly condensing the magnesium-based alloy small liquid drops to form low-oxidation solid spherical magnesium-based alloy powder;

step 5: sieving magnesium-based alloy powder, grading the granularity, and storing spherical magnesium-based alloy micron particles with 13-233um under the sieve in distilled water and/or absolute alcohol;

and (3) carrying out granularity classification on the alloy powder by adopting a vibrating screen, wherein the alloy powder is screened and then passes through a 633-833 mesh screen.

3. The method according to claim 2, characterized in that: and step 2, heating the furnace body by using an intermediate frequency heating ring.

4. The method according to claim 2, characterized in that: and 3, controlling the flow rates of the two metal liquid states on the disc atomizer which is introduced into the atomizing furnace to realize the mass ratio of the magnesium-based alloy micron particles.

5. The method according to claim 2, characterized in that: and in the step 3, controlling the frequency of the atomizer and the rotating speed of the centrifugal machine to control the size of the metal liquid drops.

6. The method according to claim 2, characterized in that: the atomizer frequency was 55Hz.

7. A method for using magnesium-based alloy microparticles with synergistic TACE anti-liver cancer effect as claimed in claim 1, which is characterized in that: is used for synergistic TACE anti-tumor treatment.

8. The method according to claim 7, wherein: when in use, after the magnesium-based alloy microparticles are dissolved in normal saline, the magnesium-based alloy microparticles are locally implanted at the tumor position before TACE treatment for 24 hours.