Saikosaponin D self-assembled nano-drug as well as preparation method and application thereof

Abstract

The invention discloses a saikoside D self-assembled nano-drug as well as a preparation method and application thereof, saikoside D is self-assembled to form the saikoside D nano-drug (SSD-NPs), and the prepared SSD-NPs has remarkable anti-cancer activity. An in-vitro experiment shows that the SSD-NPs can be used for remarkably inhibiting the activity of HCT-116 and HT-29 colon cancer cells. In addition, the SSD-NPs can also inhibit the cell activity of the hepatoma carcinoma cell Hepg2. Then, an HCT-116 colorectal cancer cell line is taken as an example, the anticancer efficacy of the SSD-NPs is verified in a subcutaneous tumor-bearing mouse model, and the SSD-NPs shows a remarkable anti-colorectal cancer effect in the aspect of improving the size and volume of a subcutaneous tumor-bearing of a CRC mouse. Therefore, the saikoside D nano-drug (SSD-NPs) provided by the invention provides a new candidate compound for cancer treatment.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a saikosaponin D self-assembly nanomedicine and applications thereof. Background Art

[0002] Colorectal cancer is one of the malignant tumors with the highest morbidity and mortality rates worldwide. Its high recurrence rate and chemotherapy resistance have always been the main challenges in clinical treatment. At present, the clinical treatments for colorectal cancer mainly include surgical resection, chemotherapy, radiotherapy, targeted therapy and immunotherapy, among which chemotherapy is still the core treatment for advanced and metastatic colorectal cancer. However, traditional chemotherapy drugs (such as 5-fluorouracil, oxaliplatin, irinotecan, etc.) have problems such as poor targeting, low bioavailability, and strong toxic side effects (such as gastrointestinal reactions, bone marrow suppression, and neurotoxicity), which seriously affect patients' treatment compliance and quality of life.

[0003] Self-assembling nanomedicines have emerged as an emerging research direction in recent years. They spontaneously form ordered nanostructures through the material's own intermolecular forces, eliminating the need for complex exogenous modification or processing. They offer advantages such as ease of preparation, high stability, and excellent biocompatibility. Naturally derived bioactive ingredients (such as polysaccharides, flavonoids, and alkaloids from plant extracts) are ideal raw materials for self-assembling nanomedicines due to their inherent biocompatibility, low toxicity, and potential pharmacological activity.

[0004] Bupleurum (Bupleurum Chinese DC), a plant of the Umbelliferae family, was first mentioned in Shennong's Herbal Classic and was listed as a top-grade herb. It has been included in various herbal remedies throughout the dynasties. It has the functions of dispersing and reducing fever, relieving liver depression, raising Yang Qi, clearing the gallbladder, and preventing malaria. It is commonly used to treat colds, fevers, alternating chills and fever, chest and flank pain, menstrual irregularities, uterine prolapse, and rectal prolapse. The most common component in Bupleurum is saikosaponin, which is primarily a pentacyclic triterpenoid oleanane derivative. The most representative of these is saikosaponin D (SSD), the structure of which is shown below:

[0005]

[0006] Similar to steroids, the chemical formula is C 42 H 68 O 13 , molecular weight is 780.98, and it has the pharmacological effects of antipyretic, sedative, anti-inflammatory, antibacterial, liver-protecting, anti-nephritis, and immune regulation.

[0007] Extensive research has been conducted on SSD. In 2019, Ningxia Medical University published a patent titled "Application of Bupleurum Saponin Compounds in the Preparation of Drugs for the Treatment of Neurodegenerative Diseases" [CN105079016B]. In a nesting experiment, the patent highlighted that SSA significantly enhanced the nesting ability of mice with LPS-induced learning and memory impairment. In 2022, Liaoning China Resources Benxi Sanyao Co., Ltd. published a patent titled "An Analgesic and Anti-inflammatory Pharmaceutical Composition, Its Preparation Method, and Use" [CN107308177B]. This patent, in the field of pharmaceuticals or health products, specifically relates to an analgesic and anti-inflammatory pharmaceutical composition, its preparation method, and its use. The patent involves quantitatively selecting and uniformly mixing Bupleurum Saponin D, paeoniflorin, feroxamine, and glycyrrhizic acid. Long Hongping publicly applied for "An antidepressant Chinese medicine composition and its application" [CN119055665A]. The present invention is composed of saikosaponin D, paeoniflorin, atractylodes lactone III, and formononetin, with the optimal ratio being 2.0:3.0:1.0:1.0.

[0008] Although the prior art discloses that saikosaponin D has multiple pharmacological effects such as antidepressant, anti-epileptic, anti-inflammatory, liver protection, and antipyretic, there are currently no reports on the preparation of saikosaponin D into nanomedicines through self-assembly and its application in the treatment of colorectal cancer. Summary of the Invention

[0009] In view of this, the object of the present invention is to provide a saikosaponin D self-assembled nanomedicine and application thereof.

[0010] To achieve the above object, the technical solution of the present invention is as follows:

[0011] In a first aspect, the present invention provides a saikosaponin D self-assembled nanomedicine, characterized in that the nanomedicine is self-assembled from saikosaponin D, the particle size of the nanomedicine is 151.2±3.85 nm, and the zeta potential of the nanomedicine is -20.03±1.09 mV;

[0012] The structure of saikosaponin D is shown below:

[0013]

[0014] In a second aspect, the present invention provides a method for preparing the aforementioned saikosaponin D self-assembled nanomedicine, comprising the following steps:

[0015] Saikosaponin D was used as a raw material, dissolved in ethanol, and the ethanol was removed by rotary evaporation. DMSO was added as an aqueous solvent, and the mixture was stirred and then ultrasonicated using a cell probe disruptor to obtain a Saikosaponin D self-assembled nanomedicine.

[0016] Furthermore, the rotary evaporation temperature is 40-50° C., the proportion of DMSO in the DMSO aqueous solvent is 0.5-2%, and the stirring time is 6-10 h.

[0017] Furthermore, the mass-to-volume ratio of the saikosaponin D to the ethanol is 1 mg: 0.2-0.5 mL; the mass-to-volume ratio of the saikosaponin D to the DMSO aqueous solvent is 1 mg: 1.5-3 mL.

[0018] Furthermore, during the ultrasonic treatment, the power is 40-60W, the ultrasonic treatment is performed for 4-6 seconds and then stopped for 1-3 seconds, and the total ultrasonic treatment time is 10-15 minutes.

[0019] In a third aspect, the present invention provides use of the aforementioned saikosaponin D self-assembled nanomedicine in the preparation of a drug for treating colon cancer or liver cancer.

[0020] Furthermore, the colon cancer cells are HCT-116 cells and HT-29 cells; the liver cancer cells are HepG2 cells.

[0021] The beneficial effects of the present invention include at least:

[0022] The present invention prepares saikosaponin D nanopharmaceuticals (SSD-NPs). This method, which nanosizes traditional small-molecule Chinese medicines to form nanopharmaceutical preparations, is an emerging discipline in modern pharmaceutical innovation research. The isolation, extraction, and preparation of certain delivery vehicles (such as extracellular vesicles) are challenging, including complex processes and obstacles to large-scale, cost-effective production. In contrast, the preparation of SSD-NPs is very simple, forming supramolecular nanopharmaceuticals in a self-assembled manner with a high drug loading rate, which increases the efficiency of Chinese medicine delivery in the body and the therapeutic effect. It has the potential to promote low-cost, large-scale production, thereby advancing its future practical clinical application. The nanomedicine provides a new treatment for colorectal cancer patients. The CCK-8 assay was used to verify the inhibitory effect of SSD-NPs on colon cancer cell lines HCT-116 and HT-29, as well as the inhibitory effect of SSD-NPs on liver cancer cells Hepg2, in vitro. This demonstrated the good in vitro anti-tumor effect of SSD-NPs. The HCT-116 cell scratch assay further confirmed the anti-tumor effect of SSD-NPs. As the concentration of SSD-NPs increased, the migration rate of HCT-116 cancer cells gradually decreased. In vivo experiments, there was no difference in tumor size between the low-dose SSD-NPs (5 mg / kg) and model groups. However, the medium-dose SSD-NPs (10 mg / kg) and high-dose SSD-NPs (20 mg / kg) showed better tumor growth inhibition as the dose increased. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1This is a real picture of the prepared saikosaponin D nanomedicine. A: Water is used as a negative control on the left side of the figure. B: The experimental results show that the saikosaponin D nanomedicine exhibits a stable light path.

[0024] Figure 2 This is the characterization diagram of the saikosaponin D nanomedicine, including the particle size of 151.2±3.85nm, the polydispersity index (Pdi) of 0.121±0.02, and the zeta potential of -20.3±1.09mV.

[0025] Figure 3 Figure 3. Stability test results of saikosaponin D nanoparticles (SSD-NPs) at different pH conditions. A: Particle size at pH 5 was 150.8 nm, at pH 6 was 152.5 nm, at pH 7 was 150.1.2 nm, at pH 8 was 169.7 nm, and at pH 9 was 162 nm. B: Zeta potential at pH 5 was -19.37 mV, at pH 6 was -19.69 mV, at pH 7 was -20.03 mV, at pH 8 was -18.73 mV, and at pH 9 was -20.23 mV.

[0026] Figure 4 The inhibitory effect on human colon cancer cell lines; A: Statistical graph of saikosaponin D nanomedicine on HCT-116 cell viability; B: IC of saikosaponin D nanomedicine on HCT-116 cell viability 50 picture.

[0027] Figure 5 The inhibitory effect of saikosaponin D nanomedicine on HT-29 colorectal cancer cells; A: Statistical graph of saikosaponin D nanomedicine on HT-29 cell viability; B: IC of saikosaponin D nanomedicine on HT-29 cell viability inhibition 50 picture.

[0028] Figure 6 The inhibitory effect of saikosaponin D nanomedicine on liver cancer cells; A: Statistical graph of saikosaponin D nanomedicine on Hepg2 cell viability; B: IC of saikosaponin D nanomedicine on Hepg2 cell viability 50 picture.

[0029] Figure 7 This is the scratch test of saikosaponin D nanomedicine on HCT-116 cells (**P<0.01; vs.Con).

[0030] Figure 8This is an evaluation of the inhibition of tumor growth using saikosaponin D nanomedicine in a mouse colon cancer model; A: a physical picture of the subcutaneous tumor peeled off after the end of the tumor administration cycle; B: a statistical graph of the tumor weight of the subcutaneous tumor peeled off after the end of the administration cycle (*P < 0.05; ***P < 0.001 vs. Mod); C: a graph of the tumor growth trend after the end of the administration cycle; D: a statistical graph of the tumor volume of the subcutaneous tumor peeled off (*P < 0.05; ***P < 0.001 vs. Mod); E: an H&E staining image of the peeled tumor section. DETAILED DESCRIPTION

[0031] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of them. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0032] In addition, the technical solutions between the various embodiments of the present invention can be combined with each other, but it must be based on the fact that ordinary technicians in this field can implement it. When the combination of technical solutions is mutually contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0033] In the experiments of the present invention, Bupleurum chinense and saikosaponin D (CAS: 20874-52-6) were purchased from Beijing Biotech Co., Ltd., product number CFN9989, and HCT116 cell line (product number FH0027) and Hepg2 cell line (product number FH0076) were purchased from Shanghai Fuheng Biotechnology Co., Ltd.

[0034] The solution proposed by the present invention is described in detail below through specific embodiments:

[0035] Example 1: Preparation of saikosaponin D nanoparticles (SSD-NPs)

[0036] First, 5 mg of the monomer compound saikosaponin D was dissolved in 2 mL of ethanol. The ethanol was dried using a rotary evaporator at 45°C, and 10 mL of 1% DMSO aqueous solvent was added. Ultrapure water was then added for re-dissolution. The mixture was then placed in a magnetic stirrer at 50°C and 1000 rpm for 8 hours. Finally, a cell probe disruptor was used with a power of 50 W and ultrasonication for 10 minutes with an interval of 5 seconds for 2 seconds. The preparation of saikosaponin D nanomedicine was completed. The results are shown in FIG. Figure 1 A and 1B, the use of infrared laser can produce a stable light path, which is the Tyndall effect, indicating the formation of nanoparticles, that is, the formation of saikosaponin D nanomedicine.

[0037] Example 2: Characterization of saikosaponin D nanoparticles (SSD-NPs)

[0038] The particle size, zeta potential, and polydispersity index (Pdi) of SSD-NPs were measured using a dynamic light scattering (DLS) instrument (Malvern Zetasizer ZS-90, Malvern, UK) at room temperature of 25°C. The particle size was 151.2 ± 3.85 nm, the Pdi was 0.121 ± 0.02, and the zeta potential was -20.03 ± 1.09 mV. Figure 2 .

[0039] Example 3: Stability of saikosaponin D nanoparticles (SSD-NPs) under different pH conditions

[0040] The particle size of SSD-NPs was tested under pH conditions of 5, 6, 7, 8, and 9. The results are as follows: Figure 3 The results show that the particle sizes of SSD-NPs under different pH conditions are not much different, namely 150.8nm, 152.5nm, 151.2nm, 169.7nm, and 162nm, respectively. This indicates that the stability of SSD-NPs can adapt to the physiological conditions in the body and the acidic tumor microenvironment. The Zeta potential is also slightly affected, namely -19.37mV, -19.69mV, -20.03mV, -18.73mV, and -20.23mV, respectively.

[0041] Example 4: Cell viability detection of HCT-116 cells by saikosaponin D nanoparticles (SSD-NPs)

[0042] HCT116 cells were cultured in DMEM high-glucose medium (containing 1% penicillin and streptomycin) supplemented with 10% fetal bovine serum in a CO2 incubator maintained at 37°C with a continuous supply of 5% CO2 and saturated humidity.

[0043] Cell recovery: sterilize the clean bench with ultraviolet light for 30 minutes, and wipe the table with 75% medical alcohol for later use; preheat the complete culture medium in a 37°C constant temperature water bath, take 4 mL and add it to a 15 mL centrifuge tube; remove the frozen cells from liquid nitrogen, quickly place them in a 37°C constant temperature water bath, and shake them gently until they are completely thawed. Use a pipette to aspirate the 1 mL of thawed cell suspension into the previously prepared 4 mL of complete culture medium in the clean bench; centrifuge at 1000 rpm for 5 minutes, and carefully discard the supernatant; resuspend the cells in complete culture medium and inoculate them into a cell culture flask. After tightening the bottle cap, shake the bottle gently to spread the cell suspension evenly on the bottom of the bottle; place the cell culture flask in a 5% CO2 incubator and culture at 37°C. Observe the cell adhesion overnight and change the complete culture medium every 2 days.

[0044] Cell passaging: When the cell density is about 80% as observed under a microscope, the cells can be passaged. The operation is carried out in a clean bench. Aspirate the culture medium in the cell culture flask, add 2 mL of PBS, and gently shake the bottle to allow the PBS to fully wash the cells and aspirate it. Wash twice in total. After removing the PBS, add 1 mL of trypsin to the culture flask, and place the culture flask in a 37°C incubator. When the cells become round and have not fallen off under a microscope, aspirate the trypsin, add 3 mL of complete culture medium to the culture flask to terminate the digestion, and gently blow the cells off from the bottom of the flask with a pipette. Transfer the digested cell suspension to a 15 mL centrifuge tube, centrifuge at 1000 rpm for 5 minutes, and carefully aspirate the supernatant. Add 2 mL of complete culture medium to the cell pellet and gently resuspend the cells with a pipette. Pipette 10 μL of cell suspension, count it in a cell counter, and divide the cells to be passaged into new cell culture flasks at a ratio of 1:3 according to the experimental needs. Mark the date and ratio and place them in the incubator for culture.

[0045] Cell freezing: Use non-programmed cell freezing solution to digest the cells according to the previous method, centrifuge at 1000 rpm for 5 minutes, aspirate the supernatant, resuspend the cells in non-programmed cell freezing solution, transfer 1 mL of cell suspension to a cryovial, label the tube and cap with the cell name and freezing date, place the cryovial in a freezing box and transfer to a -80°C freezer. If cells need to be stored long-term, transfer the cells to liquid nitrogen after overnight storage at -80°C.

[0046] Cell plating: All cells used in this experiment were passage 2 to 7. HCT116 cells in the logarithmic growth phase were digested, centrifuged, and resuspended. Cells were counted using a cell counter. 5000 cells / well were seeded into 96-well plates and cultured overnight in a 37°C, 5% CO2 incubator.

[0047] Drug treatment: After the cells adhered overnight, several drug treatment groups and control groups were set up according to the drug concentration gradient of the experimental scheme, with 5 replicate wells in each group. After adding the drug, the cells were placed in 37°C and 5% CO2 for 24 hours. CCK-8 assay: 1:10 CCK-8 reagent was added to each well of the 96-well plate and incubated for 1.5 hours. Finally, the absorbance of each test well at 450nm was read using a microplate reader and the OD value was recorded. The cell viability results are as follows: Figure 4 A, shows that the inhibitory effect of saikosaponin D nanoparticles (SSD-NPs) on HCT116 cells becomes more significant as the concentration increases. 50 =4.347μM, the results are as follows Figure 4 B. The formula for cell survival rate is: Cell survival rate (%) = (OD value of drug-treated group / OD value of control group) × 100%. All statistical charts in this experiment were drawn using Graphpadprism 10.1.2 statistical software.

[0048] Example 5: Detection of HT-29 cell viability by saikosaponin D nanoparticles (SSD-NPs)

[0049] Cell culture and plating are as in Implementation Case 4. Drug treatment: After the cells adhere to the wall overnight, several drug treatment groups are set up according to the drug concentration gradient of the experimental plan, and a control group is set up, with 5 replicate wells in each group. After adding the drug, place it in 37°C and 5% CO2 for 24 hours. CCK-8 assay: Add 1:10 CCK-8 reagent to each well of the 96-well plate and incubate for 1.5 hours. Finally, the microplate reader reads the absorbance of each test well at 450nm and records the OD value. The cell viability results are as follows Figure 5 A, indicating that the inhibitory effect of saikosaponin D nanoparticles (SSD-NPs) on HT-29 cells becomes more significant as the concentration increases. 50 =5.14μM, the results are as follows Figure 5 B. The formula for cell survival rate is: Cell survival rate (%) = (OD value of drug-treated group / OD value of control group) × 100%.

[0050] Example 6: Detection of Hepg2 cell viability by saikosaponin D nanoparticles (SSD-NPs)

[0051] Cell culture and plating were performed as in Example 4. HepG2 cells were cultured in MEM medium containing 10% fetal bovine serum (1% penicillin and streptomycin). The CO2 incubator was maintained at a constant temperature of 37°C, with a continuous supply of 5% CO2 and saturated humidity. Cell viability results were as follows: Figure 6 A. It shows that the inhibitory effect of saikosaponin D nanoparticles (SSD-NPs) on Hepg2 cells becomes more obvious with the increase of concentration, IC 50 =5.299μM The results are as follows Figure 6 B. The formula for cell survival rate is: Cell survival rate (%) = (OD value of drug-treated group / OD value of control group) × 100%.

[0052] Example 7: Scratch test of saikosaponin D nanoparticles (SSD-NPs) on HCT-116 cells

[0053] Cell culture was performed as in Example 4. HCT116 cells in the logarithmic growth phase were digested, centrifuged, and resuspended. The cells were counted using a cell counter. 300,000 cells / well were seeded into 6-well plates and placed in a 37°C, 5% CO2 incubator for overnight culture. The next day, a 10 μL pipette tip was used to scratch the 6-well plate evenly to simulate a wound healing experiment. A microscope photo was taken and recorded as 0h. After 24h, the 6-well plate was washed with PBS. The results are shown in Figure 2. Figure 7The results show that the cells in the non-drug group (Con) migrated quickly within 24 hours, and the positive drug cisplatin (CDDP) inhibited the growth and migration of the cells. As the concentration of SSD-NPs increased, the growth and migration of tumor cells were inhibited. The migration rate of the blank group was 39.23%. Compared with the blank group, the average migration inhibition rate of the positive drug cisplatin (CDDP) group was 28.69% (**P < 0.01), the average migration inhibition rate of the low-dose group was 37.97%, the average migration inhibition rate of the medium-dose group was 19.81% (**P < 0.01), and the average migration inhibition rate of the high-dose group was 16.53% (**P < 0.01).

[0054] Example 8: Inhibitory effect of saikosaponin D nanoparticles (SSD-NPs) on colon cancer in vivo

[0055] Male nude mice were BALB / c-nu, aged 4-6 weeks, weighing 16-18 g. They were fed adaptively for one week until their weight grew to about 20 g. Colon cancer cell HCT116 cells were cultured in large quantities. When the cells were in the logarithmic growth phase, they were digested with trypsin, collected, and prepared into 1×10 7 A cell suspension of 500 mg / mL was subcutaneously injected into the right armpit of male nude mice, 0.2 mL per mouse. After successful inoculation, the condition of the mice was observed daily, and the tumor volume was measured every 3 days. About a week after the tumor was formed, the animals were randomly divided into 4 groups, with 6 mice in each group. The specific groupings were model group, SSD-NPs low-dose group (5 mg / kg), SSD-NPs medium-dose group (10 mg / kg), and SSD-NPs high-dose group (20 mg / kg). Intraperitoneal administration was used, 0.1 mL / mouse / day. After continuous administration for 14 days, the mice were killed by cervical dislocation, and the tumors were removed and weighed.

[0056] Analysis of tumor material removed after tumor administration Figure 8 A; tumor weight Figure 8 B. The results showed that compared with the control group, the average tumor weight in the SSD-NPs low-dose group was 1.3454 g, the average tumor weight in the SSD-NPs medium-dose group was 0.6288 g (*P < 0.05), and the average tumor weight in the SSD-NPs high-dose group was 0.4433 g (***P < 0.001). Figure 8 C, Final tumor volume statistical analysis as shown Figure 8 D. The results showed that the average tumor volume of the model group was 1419.419 mm 3 Compared with the model group, the average tumor volume in the low-dose SSD-NPs group was 1421.775 mm 3The average volume of the SSD-NPs medium-dose group was 813.737 mm 3 (*P<0.05), the average volume of the SSD-NPs high-dose group was 438.108 mm 3 (***P<0.001), it can be seen intuitively that medium-dose and high-dose SSD-NPs have a significant inhibitory effect on colon cancer.

[0057] After embedding and dehydration, the excised tumor was stained with H&E. Figure 8 The tumor necrosis area in the E model group is a pathophysiological manifestation caused by insufficient nutrients, which is common in rapidly growing tumors. After drug intervention, the area of ​​the tumor necrosis area tends to decrease, which means that under the intervention of the drug, the tumor growth rate decreases.

[0058] In summary, the present invention prepares saikosaponin D nanomedicines (SSD-NPs). This method nanosizes traditional Chinese medicine small molecule drugs to form nanomedicine preparations, which is an emerging discipline in modern drug innovation research. The separation, extraction and preparation of certain delivery carriers (such as extracellular vesicles) will encounter challenges, including complex processes and obstacles to large-scale, cost-effective production. On the contrary, the preparation of SSD-NPs is very simple, forming supramolecular nanomedicines in a self-assembled manner, and has a high drug loading rate, which increases the efficiency of Chinese medicine delivery in vivo and the therapeutic effect. It has the potential to promote low-cost, large-scale production, thereby advancing its future practical clinical application. And its nanomedicine provides a new treatment method for patients with colorectal cancer. The inhibitory effect of SSD-NPs on colon cancer cell HCT-116 and HT-29 cell lines, as well as the inhibitory effect of SSD-NPs on liver cancer cell Hepg2, were verified in vitro by the CCK-8 method, proving that SSD-NPs have good anti-tumor effects in vitro, and the HCT-116 cell scratch experiment further confirmed it.

[0059] The anti-tumor effect of SSD-NPs was demonstrated. As the concentration of SSD-NPs increased, the migration rate of HCT-116 cancer cells gradually decreased. In in vivo experiments, there was no difference in tumor size between the low-dose SSD-NPs (5 mg / kg) and the model group. However, increasing the dose of SSD-NPs (10 mg / kg) and the high-dose SSD-NPs (20 mg / kg) showed a greater inhibitory effect on tumor growth.

[0060] It should be noted that, in this document, the terms "comprises," "includes," or any other variations thereof are intended to encompass non-exclusive inclusion, such that a process, method, article, or apparatus comprising a series of elements includes not only those elements but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus. In the absence of further limitations, an element defined by the phrase "comprising a ..." does not exclude the presence of other identical elements in the process, method, article, or apparatus comprising the element.

[0061] The serial numbers of the above embodiments of the present invention are for description only and do not represent the advantages or disadvantages of the embodiments.

[0062] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, and all of these should be included in the scope of the claims of the present invention.

Claims

1. A saikosaponin D self-assembled nanomedicine, characterized in that: The nano drug is self-assembled by saikosaponin D, the particle size of the nano drug is 151.2±3.85 nm, and the Zeta potential of the nano drug is -20.03±1.09 mV.

2. The method for preparing the saikosaponin D self-assembled nanomedicine according to claim 1, characterized in that: The following steps are involved: Saikosaponin D was used as a raw material, dissolved in ethanol, and the ethanol was removed by rotary evaporation. DMSO was added as an aqueous solvent, and the mixture was stirred and then ultrasonicated using a cell probe disruptor to obtain a Saikosaponin D self-assembled nanomedicine.

3. The preparation method according to claim 2, characterized in that The rotary evaporation temperature is 40-50° C., the proportion of DMSO in the DMSO aqueous solvent is 0.5-2%, and the stirring time is 6-10 hours.

4. The preparation method according to claim 2, characterized in that The mass volume ratio of the saikosaponin D to the ethanol is 1 mg: 0.2-0.5 mL; the mass volume ratio of the saikosaponin D to the DMSO aqueous solvent is 1 mg: 1.5-3 mL.

5. The preparation method according to claim 2, characterized in that During the ultrasonic treatment, the power is 40-60W, the ultrasonic treatment is performed for 4-6 seconds and the ultrasonic treatment is stopped for 1-3 seconds, and the total ultrasonic treatment time is 10-15 minutes.

6. Use of the saikosaponin D self-assembled nanomedicine according to claim 1 in the preparation of a drug for treating colon cancer or liver cancer.

7. The use according to claim 6, characterized in that The colon cancer cells are HCT-116 cells and HT-29 cells; the liver cancer cells are HepG2 cells.

8. The use according to claim 6, characterized in that The effective concentration of the saikosaponin D self-assembled nanomedicine is 10 mg / kg to 20 mg / kg.

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