Pharmaceutical composition with synergistic effect for treating cancer metastasis as well as preparation method and application of pharmaceutical composition

Abstract

The invention relates to the field of medical drugs, in particular to a pharmaceutical composition with a synergistic effect for treating cancer metastasis and a preparation method and application of the pharmaceutical composition, and the pharmaceutical composition comprises Cu2-xS-PEI-Lapachonone nanoparticles wrapped by silk fibroin, and the Cu2-xS-PEI-Lapachonone nanoparticles are expressed as Cu2-xS-PEI-Lapachonone-SFNs. By adopting the technical scheme of the invention, the cancer inhibition capability is promoted through the synergistic effect of Cu2-xS, Lapacone and silk fibroin.

Description

Technical Field

[0001] The present invention relates to the field of medical drugs, and in particular to a synergistic pharmaceutical composition for treating cancer metastasis, a preparation method thereof, and an application thereof. Background Art

[0002] Cancer, also known as malignant tumor, is caused by the malignant proliferation of cells and is invasive and metastatic. The main classifications include carcinoma, sarcoma and carcinosarcoma. The cause of cancer is usually due to the long-term combined effects of chemical, physical, viral and other factors on the body, which trigger a series of abnormal genetic changes. The incidence of cancer is higher in high-risk groups, such as those with a family history of cancer and cancer-related diseases, and people over 40 years old need to be more vigilant. The clinical symptoms of cancer are related to factors such as the site of onset, the size of the lesion, and whether it has metastasized. There may be no obvious symptoms in the early stages. As the lesion progresses, local symptoms such as lumps, pain, bleeding, and obstruction may appear, as well as systemic symptoms such as fever, fatigue, anemia, and weight loss.

[0003] Synergistic effect means that when two or more anticancer drugs are used simultaneously or successively, they sometimes produce certain mutual influences, and the resulting therapeutic effect is greater than the sum of the effective rates of the two anticancer drugs alone.

[0004] Prior art, such as Patent Publication No. CN118903140A, discloses a synergistic pharmaceutical composition for treating cancer metastasis, its preparation method, and its use. Combining abemaciclib (ABE) and curcumin (Cur), this composition can boost cellular immunity. However, its efficacy in inhibiting cancer metastasis needs to be further enhanced to meet patient needs. Summary of the Invention

[0005] In order to solve the above problems, the present invention provides a synergistic pharmaceutical composition for treating cancer metastasis, a preparation method thereof and an application thereof, for use in treating cancer metastasis by Cu 2-x S, Lapachone, and silk fibroin work synergistically to promote cancer inhibition ability.

[0006] In order to achieve the above object, the technical solution of the present invention is as follows: a synergistic pharmaceutical composition for treating cancer metastasis, comprising: silk fibroin-coated Cu 2-x S-PEI-Lapachone nanoparticles, expressed as Cu 2-x S-PEI-Lapachone-SFNs.

[0007] A method for preparing a synergistic drug for treating cancer metastasis, comprising:

[0008] Step 1, preparing silk fibroin;

[0009] Step 2: Preparation of Cu 2-xS-PEI-Lapachone nanoparticles;

[0010] Step 3: Cu 2-x The S-PEI-Lapachone nanoparticle suspension was mixed in acetone solution, and the silk fibroin solution was slowly added dropwise and stirred until the acetone was completely evaporated. After centrifugation, the suspension was washed with water and ultrasonicated in an ice bath to obtain Cu 2-x S-PEI-Lapachone-SFNs.

[0011] Furthermore, in step 3, Cu 2-x The concentration of S-PEI-Lapachone nanoparticle suspension was 8 mg / mL, the concentration of silk fibroin was 5 mg / mL, and the concentration of Cu 2-x The volume ratio of S-PEI-Lapachone nanoparticle suspension, acetone solution and silk fibroin solution was 2:30:3.

[0012] Furthermore, in step 3, the centrifugation time is 10-15 minutes, the number of water washings is at least 3, and the ultrasonication time is 1-3 minutes.

[0013] Furthermore, in step 1, preparing silk fibroin includes preparing silkworm cocoon shells, washing the cocoon shells to remove the cocoon coat and impurities, soaking the cocoon shells in a Na2CO3 solution, heating and washing the cocoon shells in a water bath to obtain silk fibroin fibers, drying and degumming the silk fibroin fibers, and then soaking the silk fibroin fibers in a CaCl2-ethanol solution, heating and stirring until the silk fibroin fibers are dissolved in the CaCl2-ethanol solution, and dialyzing and centrifuging to obtain silk fibroin.

[0014] Furthermore, in step 1, the concentration of the Na2CO3 solution is 0.5%, the concentration ratio of the CaCl2 ethanol solution is CaCl2:ethanol:water is 1:2:8, and the bath ratio of the cocoon shell to the Na2CO3 solution is 1:40.

[0015] Furthermore, the number of boiling and washing in step 1 is at least 3, the product is dried at 50-60°C for 20-28 hours, and the heating and stirring temperature is 80°C.

[0016] Furthermore, in step one, water bath heating is performed using a water bath pot, a cocoon shell clamping member is provided in the water bath pot, the cocoon shell clamping member includes a Y-shaped branch rod, two branches of the Y-shaped branch rod are provided with clips, the Y-shaped branch rod is rotatably connected to the water bath pot, and a driving member for driving the Y-shaped branch rod to rotate is provided on the water bath pot.

[0017] Further, step 2 includes preparing CuCl2·2H2O, dissolving CuCl2·2H2O in PVP10 and adding NaOH dropwise, adding L-ascorbic acid dropwise after the addition of NaOH, adding Na2S after sufficient stirring, stirring in a water bath and then centrifuging and washing with water to obtain CuCl2·2H2O. 2-xS nanoparticles;

[0018] Cu 2-x S nanoparticles were added dropwise to the PEI aqueous solution, stirred thoroughly and then washed with water to obtain Cu 2-x S-PEI nanoparticles, Cu 2-x S-PEI nanoparticles and Lapachone were mixed at a mass ratio of 250:1 to obtain Cu 2-x S-PEI-Lapachone nanoparticles.

[0019] A synergistic pharmaceutical composition for treating cancer metastasis is used for breast cancer, wherein the cancer metastasis is lung metastasis.

[0020] The above scheme has the following beneficial effects:

[0021] Cu 2-x S is a nano-copper sulfide particle with good photothermal conversion efficiency and biocompatibility. It generates local high temperature under near-infrared light NIR irradiation and kills cancer cells.

[0022] PEI is polyethyleneimine, which is used to modify Cu 2-x S and silk fibroin surface, enhancing the stability and dispersibility of nanoparticles. PEI can load Lapachone molecules through electrostatic interaction or chemical bonding.

[0023] Lapachone is a quinone that induces cancer cells to produce excessive reactive oxygen species and inhibits topoisomerase, leading to cancer cell apoptosis. 2-x Combined with the photothermal effect of S, Lapachone can enhance the therapeutic effect and achieve chemical-photothermal combined therapy.

[0024] Silk protein has good drug loading capacity and controlled release properties, and can serve as a drug delivery carrier. Silk protein is a natural material with excellent biocompatibility and metabolizability, and can achieve sustained release and targeted delivery of drugs.

[0025] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Schematic diagram of the steps of the method for preparing the synergistic drug for treating cancer metastasis of the present invention;

[0027] Figure 2 This is a schematic diagram of step 1 of the method for preparing a synergistic drug for treating cancer metastasis according to the present invention;

[0028] Figure 3 This is a schematic diagram of step 2 of the method for preparing a synergistic drug for treating cancer metastasis according to the present invention;

[0029] Figure 4 This is an axonometric diagram of a water bath in the method for preparing a synergistic drug for treating cancer metastasis according to the present invention.

[0030] The reference numerals in the drawings of the specification include: 1. cocoon shell clamping member; 2. Y-shaped branch rod; 3. clamp; 4. driving member. DETAILED DESCRIPTION

[0031] The technical solution of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0032] In the description of the present invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings and are intended solely to facilitate and simplify the description of the present invention. They are not intended to indicate or imply that the devices or components referred to must have, be constructed, or operate in a specific orientation, and therefore should not be construed as limitations on the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0033] In the description of the present invention, it should be noted that, unless otherwise expressly specified or limited, the terms "mounted," "connected," and "connected" should be understood in a broad sense. For example, they may refer to fixed, detachable, or integral connections; mechanical or electrical connections; direct or indirect connections through an intermediate medium; and internal communication between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on the specific circumstances.

[0034] The following is further described in detail through specific implementation methods:

[0035] As attached Figures 1-4 Shown: A synergistic pharmaceutical composition for treating cancer metastasis, comprising: silk fibroin-wrapped Cu 2-x S-PEI-Lapachone nanoparticles, expressed as Cu 2-x S-PEI-Lapachone-SFNs.

[0036] Cu 2-xS is a nano-copper sulfide particle with good photothermal conversion efficiency and biocompatibility. It generates local high temperature under near-infrared light NIR irradiation and kills cancer cells.

[0037] PEI is polyethyleneimine, which is used to modify Cu 2-x PEI can load Lapachone molecules through electrostatic interaction or chemical bonding, and enhance the cellular uptake efficiency of nanoparticles, further improving the accumulation of drugs in cancer cells.

[0038] Lapachone is a quinone that induces cancer cells to produce excessive reactive oxygen species and inhibits topoisomerase, leading to cancer cell apoptosis. 2-x Combined with the photothermal effect of S, Lapachone can enhance the therapeutic effect and achieve chemical-photothermal combined therapy.

[0039] Silk protein has good drug loading capacity and controlled release properties, and can serve as a drug delivery carrier. Silk protein is a natural material with excellent biocompatibility and metabolizability, and can achieve sustained release and targeted delivery of drugs.

[0040] A method for preparing a synergistic drug for treating cancer metastasis, comprising:

[0041] Step 1, prepare silk cocoon shells, clean the cocoon coats and impurities of the silk cocoon shells, soak the silk cocoon shells in Na2CO3 solution, the concentration of the Na2CO3 solution is 0.5%, the bath ratio of the silk cocoon shells to the Na2CO3 solution is 1:40, heat in a water bath and boil and wash to obtain silk fibroin fibers, the number of boiling and washing is 3, dry the silk fibroin fibers at 50-60°C for 20-28h for degumming, and then soak the silk fibroin fibers in CaCl2 ethanol solution, heat and stir until the silk fibroin fibers are dissolved in the CaCl2 ethanol solution, the concentration ratio of the CaCl2 ethanol solution is CaCl2: ethanol: water is 1:2:8, the heating and stirring temperature is 80°C, and dialyze and centrifuge to obtain silk fibroin protein.

[0042] Water bath heating is carried out in a water bath pot, in which a cocoon shell clamping member 1 is provided, which includes a Y-shaped branch rod 2, on both branches of which clips 3 are installed, and the clips 3 are metal clips, and the Y-shaped branch rod 2 is rotatably connected to the water bath pot, and a driving member 4 for driving the Y-shaped branch rod 2 to rotate is provided on the water bath pot, and the driving member 4 is a motor.

[0043] Step 2: Prepare CuCl2·2H2O, dissolve CuCl2·2H2O in PVP10 and add NaOH drop by drop. After the addition of NaOH is complete, add L-ascorbic acid drop by drop. After stirring thoroughly, add Na2S. Stir in a water bath and then centrifuge and wash with water to obtain CuCl2·2H2O. 2-x S nanoparticles;

[0044] Cu 2-x S nanoparticles were added dropwise to the PEI aqueous solution, stirred thoroughly and then washed with water to obtain Cu 2-x S-PEI nanoparticles, Cu 2-x S-PEI nanoparticles and Lapachone were mixed at a mass ratio of 250:1 to obtain Cu 2-x S-PEI-Lapachone nanoparticles.

[0045] Step 3: Cu 2-x S-PEI-Lapachone nanoparticle suspension was mixed in acetone solution, Cu 2-x The concentration of S-PEI-Lapachone nanoparticle suspension was 8 mg / mL, and silk fibroin solution was slowly added dropwise. The concentration of silk fibroin was 5 mg / mL. 2-x The volume ratio of S-PEI-Lapachone nanoparticle suspension, acetone solution and silk fibroin solution is 2:30:3, stirred until the acetone is completely volatilized, centrifuged and washed with water, the centrifugation time is 10-15 minutes, and the number of water washings is at least 3. Ultrasonication is performed under ice bath conditions, the ultrasonication time is 1-3 minutes, and Cu 2-x S-PEI-Lapachone-SFNs.

[0046] Silkworm cocoon shells are natural materials, often contaminated with impurities. Therefore, cleaning is necessary for silk fibroin extraction. Using a Na₂CO₃ solution for washing effectively removes the cocoon coat and impurities, while ensuring the purity of the silk fibroin fibers through repeated washing. Using a CaCl₂ ethanol solution effectively dissolves the silk fibroin fibers, producing silk fibroin protein, which is further purified through dialysis and centrifugation to ensure high-quality silk fibroin protein.

[0047] During the water bath, the cocoon shells tend to become mixed physically, making it difficult to lay them flat before extracting the silk fibroin fibers. Therefore, a cocoon shell holder 1 is used to hold the cocoon shells during the water bath, thereby keeping them stretched. The uniform rotation of the cocoon shells improves heating uniformity and allows the water in the water bath to flow, promoting heat exchange.

[0048] Cu 2-xThe preparation process of S nanoparticles can precisely control the reaction conditions by adding the reactants NaOH and L-ascorbic acid dropwise to ensure the uniform size and morphology of the nanoparticles. 2-x S nanoparticles, enhancing their stability and biocompatibility, and further conferring their functionality by combining them with Lapachone.

[0049] Cu 2-x S-PEI-Lapachone nanoparticles were mixed with silk fibroin solution, and acetone was evaporated by slowly adding and stirring to ensure that the nanoparticles were evenly dispersed in the silk fibroin to form a stable composite drug. Subsequent ultrasonic treatment further improved the dispersibility and stability of the composite drug.

[0050] The final Cu 2-x S-PEI-Lapachone-SFNs composite drug combined with Cu 2-x The photothermal properties of S nanoparticles, the pharmaceutical activity of Lapachone, and the biocompatibility, sustained-release properties, degradability and targeting of silk fibroin have potential multifunctional application value.

[0051] A synergistic pharmaceutical composition for treating cancer metastasis is used for breast cancer, wherein the cancer metastasis is lung metastasis.

[0052] Breast cancer is a major disease that affects human health. Chemotherapeutic drugs with strong cytotoxicity often lack the ability to target metastatic breast cancer, and the treatment effect is often poor, accompanied by serious systemic side effects.

[0053] Lapachone and silk fibroin have excellent biocompatibility. 2-x After being wrapped and modified with silk fibroin, the biocompatibility of S is significantly improved, making the drug overall have low cytotoxicity and has excellent performance in the treatment of breast cancer.

[0054] Obviously, the above embodiments are merely examples for clarity of explanation and are not intended to limit the implementation methods. Those skilled in the art will readily appreciate that other variations or modifications based on the above descriptions are possible. It is not necessary and impossible to enumerate all implementation methods here. Obvious variations or modifications arising therefrom remain within the scope of protection of the present invention.

Claims

1. A synergistic pharmaceutical composition for treating cancer metastasis, characterized in that: Including, silk fibroin wrapped Cu 2-x S-PEI-Lapachone nanoparticles, expressed as Cu 2-x S-PEI-Lapachone-SFNs.

2. A method for preparing a synergistic drug for treating cancer metastasis, based on the method for preparing the pharmaceutical composition according to claim 1, characterized in that: include: Step 1, preparing silk fibroin; Step 2: Preparation of Cu 2-x S-PEI-Lapachone nanoparticles; Step 3: Cu 2-x The S-PEI-Lapachone nanoparticle suspension was mixed in acetone solution, and the silk fibroin solution was slowly added dropwise and stirred until the acetone was completely evaporated. After centrifugation, the suspension was washed with water and ultrasonicated in an ice bath to obtain Cu 2-x S-PEI-Lapachone-SFNs.

3. The method for preparing a synergistic drug for treating cancer metastasis according to claim 2, characterized in that: In step 3, Cu 2-x The concentration of S-PEI-Lapachone nanoparticle suspension was 8 mg / mL, the concentration of silk fibroin was 5 mg / mL, and the concentration of Cu 2-x The volume ratio of S-PEI-Lapachone nanoparticle suspension, acetone solution and silk fibroin solution was 2:30:

3.

4. The method for preparing a synergistic drug for treating cancer metastasis according to claim 3, characterized in that: In step 3, the centrifugation time is 10-15 minutes, the number of water washes is at least 3, and the ultrasonication time is 1-3 minutes.

5. The method for preparing a synergistic drug for treating cancer metastasis according to claim 4, characterized in that: In step 1, the preparation of silk fibroin includes preparing silk cocoon shells, cleaning the cocoon coats and impurities from the silk cocoon shells, soaking the silk cocoon shells in a Na2CO3 solution, heating them in a water bath and boiling and washing them to obtain silk fibroin fibers, drying and degumming the silk fibroin fibers, and then soaking the silk fibroin fibers in a CaCl2 ethanol solution, heating and stirring until the silk fibroin fibers are dissolved in the CaCl2 ethanol solution, and dialyzing and centrifuging to obtain silk fibroin.

6. The method for preparing a synergistic drug for treating cancer metastasis according to claim 5, characterized in that: In step 1, the concentration of the Na2CO3 solution is 0.5%, the concentration ratio of the CaCl2 ethanol solution is CaCl2:ethanol:water is 1:2:8, and the bath ratio of the cocoon shell to the Na2CO3 solution is 1:

40.

7. The method for preparing a synergistic drug for treating cancer metastasis according to claim 6, characterized in that: The number of boiling and washing in step 1 is at least 3, and the product is dried at 50-60°C for 20-28 hours, with the heating and stirring temperature at 80°C.

8. The method for preparing a synergistic drug for treating cancer metastasis according to claim 7, characterized in that: In step 1, water bath heating is performed using a water bath pot, a silkworm cocoon shell clamping member (1) is provided in the water bath pot, the silkworm cocoon shell clamping member (1) comprises a Y-shaped branch rod (2), two branches of the Y-shaped branch rod (2) are provided with a clamp (3), the Y-shaped branch rod (2) is rotatably connected to the water bath pot, and a driving member (4) for driving the Y-shaped branch rod (2) to rotate is provided on the water bath pot.

9. The method for preparing a synergistic drug for treating cancer metastasis according to claim 8, characterized in that: Step 2 includes preparing CuCl2·2H2O, dissolving CuCl2·2H2O in PVP10 and adding NaOH dropwise. After the addition of NaOH is completed, L-ascorbic acid is added dropwise. After sufficient stirring, Na2S is added. After stirring in a water bath, the mixture is centrifuged and washed with water to obtain CuCl2·2H2O. 2-x S nanoparticles; Cu 2-x S nanoparticles were added dropwise to the PEI aqueous solution, stirred thoroughly and then washed with water to obtain Cu 2-x S-PEI nanoparticles, Cu 2-x S-PEI nanoparticles and Lapachone were mixed at a mass ratio of 250:1 to obtain Cu 2-x S-PEI-Lapachone nanoparticles.

10. A synergistic pharmaceutical composition for treating cancer metastasis, the pharmaceutical composition of claim 1, characterized in that: Applied to breast cancer, where the cancer metastasizes to the lungs.

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