Vesicle-like nano-particles derived from traditional Chinese medicine and application of vesicle-like nano-particles
By preparing the Yifei Sanjie formula into vesicle-like nanoparticles with a particle size of 100-150nm, the problem of low oral administration efficiency of the current Yifei Sanjie formula is solved, intravenous administration is achieved, and its application in lung cancer treatment is expanded, especially for patients who cannot take the medicine orally.
Patent Information
- Application Number
- CN202510972088.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-15
- Publication Date
- 2025-09-26
AI Technical Summary
Currently, Yifei Sanjie prescription mainly exists in the form of decoction. Oral administration is affected by gastrointestinal absorption efficiency and the first-pass effect of the liver, has low bioavailability, and cannot be administered by injection, which limits its application in the treatment of lung cancer, especially for patients who cannot take the medicine orally.
The Fritillaria thunbergii, Herba Cat's Claw, Bombyx Batryticatus, Sarcandra scabra, Trillium gracile, Pinellia ternata, Ganoderma lucidum and American ginseng in the Yifei Sanjie formula are prepared into vesicle-like nanoparticles with a particle size of 100-150nm and used through intravenous injection, simplifying the route of administration.
Excellent lung cancer treatment effects have been achieved. Vesicle-like nanoparticles can be safely used through intravenous injection, which simplifies the material basis, expands the route of administration, and is suitable for patients who cannot take the drug orally.
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Figure CN120695111A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to vesicle-like nanoparticles derived from traditional Chinese medicine and applications thereof. Background Art
[0002] The Yifei Sanjie Recipe, once known as the Qi-invigorating and Phlegm-removing Recipe, contains ingredients including Fritillaria thunbergii, Cat's Claw, Bombyx batryticatus, Sarcandra scabra, Trillium gracile, Pinellia ternata, Ganoderma lucidum, and American ginseng. It boasts Qi-invigorating and phlegm-removing, anti-cancer, and detoxifying properties, and has been widely used in the adjuvant treatment of lung cancer patients. Previous clinical studies have shown that lung cancer patients can benefit from the Yifei Sanjie Recipe. It can be used alone for elderly lung cancer, as a maintenance treatment after chemotherapy, and for patients who cannot tolerate or are unwilling to undergo chemotherapy. It is also widely used in the treatment of lung cancer at all stages, in combination with Western medical treatments such as surgery, chemotherapy, radiotherapy, targeted therapy, and immunotherapy. Basic research has also shown that the Yifei Sanjie Recipe can reduce toxicity and enhance efficacy in multiple tumor-bearing mouse models, and its effectiveness in reversing immune escape and improving EGFR-TKI resistance has been validated.
[0003] However, the Yifei Sanjie formula currently available on the market is primarily in the form of a decoction, limited to oral administration. Oral administration is not only affected by factors such as gastrointestinal absorption efficiency and the first-pass effect in the liver, resulting in low bioavailability, but the decoction also contains a large number of complex ingredients, including multiple chemical components and impurities. The material basis of its anti-lung cancer effect is unclear, hindering in-depth research, quality control, and clinical application promotion of this formula.
[0004] At the same time, due to the dosage form characteristics of decoctions, injection is not possible, which greatly limits their application scenarios in the treatment of lung cancer. It is difficult to meet the treatment needs of patients with different conditions, especially for lung cancer patients who cannot take the medicine orally, and their therapeutic effect cannot be exerted.
[0005] Therefore, how to effectively process the Yifei Sanjie prescription, simplify its material basis while retaining its anti-lung cancer effect, and expand its route of administration to promote the further promotion and application of the Yifei Sanjie prescription has become one of the technical problems that need to be solved urgently. Summary of the Invention
[0006] To address the shortcomings of the existing technology, the present invention provides vesicle-like nanoparticles derived from traditional Chinese medicine and their applications. These vesicle-like nanoparticles are prepared from Fritillaria thunbergii, Herba Lycopodii, Bombyx batryticatus, Sarcandra scabra, Trillium gracile, Pinellia ternata, Ganoderma lucidum, and American ginseng. These vesicle-like nanoparticles can be safely administered via intravenous injection, simplifying the complex system of existing compound formulas and achieving excellent therapeutic effects for lung cancer.
[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:
[0008] In a first aspect, the present invention provides a vesicle-like nanoparticle derived from traditional Chinese medicine, wherein the raw materials for preparing the vesicle-like nanoparticle include fritillary bulb of thunbergii, cat's claw, bombyx batryticatus, scutellaria baicalensis, trillium, pinellia ternata, ganoderma lucidum and American ginseng.
[0009] This invention creatively prepares vesicle-like nanoparticles using the same ingredients as the Yifei Sanjie formula: Fritillaria thunbergii, Herba Lycopodii, Bombyx batryticatus, Sarcandra scabra, Trillium gracile, Pinellia ternata, Ganoderma lucidum, and American ginseng. Experiments have demonstrated that these vesicle-like nanoparticles achieve excellent lung cancer treatment efficacy and can be safely administered via intravenous injection.
[0010] Preferably, the raw materials for preparing the vesicle-like nanoparticles include, by mass, 5-15 parts of Fritillaria thunbergii, 25-35 parts of Radix Ranunculi, 5-15 parts of Bombyx batryticatus, 10-20 parts of Sarcandra scutellariae, 7-12 parts of Trillium gracile, 7-12 parts of Pinellia ternata, 10-15 parts of Ganoderma lucidum and 3-8 parts of American ginseng.
[0011] Among them, the specific point values in 5-15 portions can be selected from 5, 7, 9, 11, 13, 15, etc., the specific point values in 25-35 portions can be selected from 25, 27, 29, 31, 33, 35, etc., the specific point values in 10-20 portions can be selected from 10, 12, 14, 16, 18, 20, etc., the specific point values in 7-12 portions can be selected from 7, 8, 9, 10, 11, 12, etc., the specific point values in 10-15 portions can be selected from 10, 11, 12, 13, 14, 15, etc., and the specific point values in 3-8 portions can be selected from 3, 4, 5, 6, 7, 8, etc.
[0012] Preferably, the raw materials for preparing the vesicle-like nanoparticles include, by mass, 10 parts of Fritillaria thunbergii, 30 parts of Herba Lycopodii, 10 parts of Bombyx batryticatus, 15 parts of Radix Sarcodiformis, 9 parts of Trillium, 9 parts of Rhizoma Pinelliae, 12 parts of Ganoderma lucidum and 6 parts of American ginseng.
[0013] Preferably, the particle size of the vesicle-like nanoparticles is 100-150 nm, for example, 100 nm, 110 nm, 120 nm, 130 nm, 140 nm, 150 nm, etc.
[0014] Preferably, the PDI of the vesicle-like nanoparticles is ≤0.4, for example, it may be 0.4, 0.3, 0.2, 0.1, etc.
[0015] Preferably, the Zeta potential of the vesicle-like nanoparticles is ≤0 mV, for example, it can be 0 mV, -2 mV, -4 mV, -6 mV, -8 mV, -10 mV, etc.
[0016] Preferably, the vesicle-like nanoparticles are prepared by a preparation method comprising the following steps:
[0017] (1) mixing the raw materials for preparing vesicle-like nanoparticles with water, soaking, boiling, and centrifuging to collect the supernatant I;
[0018] (2) The supernatant I collected in step (1) is centrifuged again, and the supernatant II is collected to obtain vesicle-like nanoparticles.
[0019] Preferably, the mass volume ratio of the raw materials for preparing the vesicle-like nanoparticles in step (1) to water is 1:(8-12) g / mL, for example, it can be 1:8 g / mL, 1:9 g / mL, 1:10 g / mL, 1:11 g / mL, 1:12 g / mL, etc.
[0020] Preferably, the soaking temperature in step (1) is 2-10° C., and the soaking time is 30-90 min.
[0021] Preferably, the decocting time in step (1) is 30-90 minutes.
[0022] Preferably, after the decoction in step (1), the decoction liquid is cooled to 20-30°C.
[0023] Preferably, the centrifugal speed in step (1) is 500-2000 rpm, and the centrifugal time is 10-50 min.
[0024] Among them, the specific point values in 2-10℃ can be selected from 2℃, 4℃, 6℃, 8℃, 10℃, etc., the specific point values in 30-90min can be selected from 30min, 50min, 70min, 90min, etc., the specific point values in 20-30℃ can be selected from 20℃, 22℃, 24℃, 26℃, 28℃, 30℃, etc., the specific point values in 500-2000rpm can be selected from 500rpm, 1000rpm, 1500rpm, 2000rpm, etc., and the specific point values in 10-50min can be selected from 10min, 20min, 30min, 40min, 50min, etc.
[0025] Preferably, the centrifugal speed in step (2) is 8000-15000 rpm, and the centrifugal time is 10-50 min.
[0026] Among them, the specific point values in 8000-15000rpm can be selected as 8000rpm, 9000rpm, 10000rpm, 11000rpm, 12000rpm, 13000rpm, 14000rpm, 15000rpm, etc., and the specific point values in 10-50min can be selected as 10min, 20min, 30min, 40min, 50min, etc.
[0027] Preferably, after collecting the supernatant II in step (2), the method further comprises dialyzing the supernatant II.
[0028] Preferably, the dialysate of the dialysis treatment comprises water.
[0029] Preferably, the dialysis treatment time is 20-30 hours, for example, it can be 20 hours, 22 hours, 24 hours, 26 hours, 28 hours, 30 hours, etc.
[0030] Preferably, the molecular weight cut-off of the dialysis bag used for the dialysis treatment is 3-4 KDa, for example, 3 KDa, 3.2 KDa, 3.4 KDa, 3.6 KDa, 3.8 KDa, 4 KDa, etc.
[0031] Preferably, the dialysis bag used in the dialysis treatment is of the MD44 type.
[0032] In a second aspect, the present invention provides a use of the vesicle-like nanoparticles as described in the first aspect in the preparation of anti-tumor drugs.
[0033] Preferably, the tumor comprises lung cancer.
[0034] Preferably, the dosage form of the drug includes tablets, capsules, granules, powders, injections or sprays.
[0035] In a third aspect, the present invention provides an injection comprising the vesicle-like nanoparticles derived from traditional Chinese medicine as described in the first aspect and water for injection.
[0036] The numerical range described in the present invention includes not only the point values listed above, but also any point values between the above numerical ranges that are not listed. Due to space limitations and for the sake of simplicity, the present invention no longer exhaustively lists the specific point values included in the range.
[0037] Compared with the prior art, the present invention has the following beneficial effects:
[0038] This study creatively developed a vesicle-like nanoparticle derived from traditional Chinese medicine, which has the same anti-tumor therapeutic effect as the Yifei Sanjie formula. The vesicle-like nanoparticle can achieve excellent lung cancer treatment effects and can be safely administered via intravenous injection.
[0039] Furthermore, the present invention utilizes a decoction of the original formula, which is time-efficient, low-cost, and requires no toxic reagents. High-speed centrifugation and deionized water dialysis yield clean, pollution-free vesicle-like nanoparticles, without the addition of any artificial excipients. These vesicle-like nanoparticles offer excellent compound alternatives and biosafety, and could be further developed into a new injectable formulation for the treatment of lung cancer. BRIEF DESCRIPTION OF THE DRAWINGS
[0040] Figure 1 This is a comparison chart of the particle sizes of YFSJF-O and YFSJF-N in Verification Example 1.
[0041] Figure 2 This is a comparison chart of the Zeta potential of YFSJF-O and YFSJF-N in Verification Example 1.
[0042] Figure 3 This is the transmission electron microscope image of YFSJF-N in Verification Example 1.
[0043] Figure 4 This is a comparison chart of the cytotoxic effects of YFSJF-O and YFSJF-N in Verification Example 2.
[0044] Figure 5 This is a graph showing the tumor growth curve of the Lewis lung cancer mouse model in Verification Example 3.
[0045] Figure 6 This is a comparison chart of tumor fluorescence intensity in the Lewis lung cancer mouse model in Verification Example 3.
[0046] Figure 7 This is a comparison chart of biochemical indicators of the Lewis lung cancer mouse model in Verification Example 3. DETAILED DESCRIPTION
[0047] The technical solution of the present invention is further described below by way of specific embodiments. It should be understood by those skilled in the art that the embodiments are merely to help understand the present invention and should not be regarded as specific limitations of the present invention.
[0048] Example 1
[0049] Vesicle-like nanoparticles and Chinese medicine decoctions derived from traditional Chinese medicine were prepared respectively.
[0050] (1) Add Yifei Sanjie prescription (including: 10g of Fritillaria thunbergii, 30g of Herba Cat's Claw, 10g of Bombyx batryticatus, 15g of Radix Sarcocarpa, 9g of Trillium, 9g of Pinellia ternata, 12g of Ganoderma lucidum, and 6g of American ginseng, hereinafter referred to as YFSJF) to 1000mL of cold water, soak at 4℃ for 60min, and boil for 60min. Then filter the extract with gauze and let it cool to room temperature. Centrifuge at 1000rpm at 4℃ for 30min using a high-speed centrifuge, discard the precipitate and collect the supernatant to obtain a Chinese medicine decoction (hereinafter referred to as YFSJF-O), which is stored at 4℃.
[0051] (2) The Chinese herbal medicine decoction was centrifuged at 10,000 rpm and 4°C for 30 min, then placed in an MD44-3.5 kDa dialysis bag and dialyzed in double-distilled water for 24 h to obtain vesicle-like nanoparticles (hereinafter abbreviated as YFSJF-N), which were then stored at 4°C.
[0052] Verification Example 1
[0053] The structure of vesicle-like nanoparticles was characterized.
[0054] Dynamic light scattering (Malvern zeta sizer, Malvern) was used to measure the size distribution and Zeta potential of YFSJF-O and YFSJF-N. The test results are as follows: Figure 1 and Figure 2 shown.
[0055] The results showed that the particle size of YFSJF-O was approximately 264 nm, with a PDI of approximately 0.55 and a potential of -20 mV, while the extracted vesicle-like nanoparticles (YFSJF-N) had a particle size of approximately 119 nm, a PDI of approximately 0.38, and a potential of -9 mV. The extracted vesicle-like nanoparticles were smaller and more uniform in size, making them more suitable for intravenous administration.
[0056] The morphology of YFSJF-N was characterized using a transmission electron microscope (TEM, Hitachi-HT7700). YFSJF-N was diluted 5 times, and 10 μL of the solution was dripped onto a copper grid coated with a carbon film. The excess liquid around it was absorbed with filter paper, and the sample was then immersed in lead citrate for staining. After staining for 2 minutes, the copper grid was dried and transferred to a sample holder. YFJSF-N was observed under a transmission electron microscope. The results are as follows: Figure 3 As shown, it can be seen that the extracted vesicle-like nanoparticles are indeed irregular vesicle-like structures.
[0057] Verification Example 2
[0058] The anti-tumor effect of vesicle-like nanoparticles was verified at the cellular level.
[0059] 100 μL of 3000-7000 A549 cells / well were seeded on a 96-well plate, 100 μL of DMEM medium was added, and cultured at 37° C., 5% CO 2 , and 90% humidity for 24 h. On the second day, the cell growth state and density were observed under a microscope. Wells with good growth state, uniform cell distribution and density were selected for the experiment. Gradient concentrations of YFSJF-O and YFSJF-N solutions were added (the concentration gradient was 0.0625 mg / mL, 0.125 mg / mL, 0.25 mg / mL, 0.5 mg / mL, 0.75 mg / mL, 1 mg / mL, 1.5 mg / mL, and 2 mg / mL), respectively. The cells were cultured at 37°C, 5% CO2, and 90% humidity for 72 hours, and the test was performed at 72 hours. 10 μL of CCK-8 solution was added to each well. After incubation for 30 minutes, the absorbance was measured at 450 nm using a microplate reader. The IC50 value of the cell viability at different concentration gradients was calculated. The results are shown in Figure 5. Figure 4 shown.
[0060] Verification Example 3
[0061] The anti-tumor effect of vesicle-like nanoparticles was verified at the animal level.
[0062] (1) Construction of C57 BL / 6 Lewis subcutaneous implant cancer model:
[0063] C57 BL / 6J mice (5-6 weeks old, 18-21 g, purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.) were adaptively fed for 1 week. Luciferase-labeled Lewis lung cancer cells in the logarithmic growth phase were digested with 0.25% trypsin to remove the cell wall. The cells were centrifuged at 1000 rpm for 3 min in normal saline and washed twice to remove trypsin and serum in the culture medium. The tumor cells were diluted with normal saline to a concentration of 2 × 10 7 Cell suspension (containing 4 × 10 cells / mL) was inoculated into the right armpit of each mouse using a 1 mL syringe. 6 Lewis lung cancer cells were used to establish a C57 BL / 6 Lewis subcutaneous implantation cancer model.
[0064] (2) Trial grouping and intervention methods:
[0065] On the second day after establishing the C57 BL / 6 Lewis subcutaneous implant cancer model, the mice were randomly divided into four groups, with six mice in each group. The mice in each group were treated as follows:
[0066] ① Model group: Normal feeding, drug administration once every 2 days, 0.9% sodium chloride solution was given orally each time, 0.5 mL / animal, for 12 consecutive days;
[0067] ②YFSJF-O gavage group (YFSJF-O-IG): Normal feeding, administration once every 2 days, 21.6 mg / mL YFSJF-O per gavage, 0.5 mL / mouse, for 12 consecutive days;
[0068] ③YFSJF-N gavage group (YFSJF-N-IG): Normal feeding, administration once every 2 days, 4.2 mg / mL YFSJF-N, 0.5 mL / mouse per gavage, for 12 consecutive days;
[0069] ④YFSJF-N intravenous injection group (YFSJF-N-IV): Normal feeding, administration once every 2 days, each intravenous injection of 4.2 mg / mL YFSJF-N, 0.2 mL / mouse, for 12 consecutive days.
[0070] (3) Test the anti-tumor effect of each experimental group:
[0071] (3.1) From the start of drug administration, the size of the tumor was measured daily using a vernier caliper. The volume was calculated using the following formula: Volume = (length × width / 2) 2 , and obtain the tumor growth curve according to the tumor volume. The test results are as follows Figure 5 As shown (where the horizontal axis is the number of days from modeling, and the second day of modeling is the initial day of drug administration). The results showed that the oral YFSJF-N-IG group had a better tumor inhibition effect, while there was no significant difference between the YFSJF-N-IG group and the YFSJF-Y-IG group.
[0072] (3.2) After the administration, D-luciferin potassium salt was dissolved in ultrapure water to prepare a 4 mg / mL working solution. 100 μL of the solution was injected intraperitoneally into each mouse. After 10 minutes, the labeled tumors began to fluoresce under the conditions of an excitation wavelength of 420 nm and an emission wavelength of 560 nm in a small animal 3D in vivo imaging system (Perkin Elmer). The fluorescence intensity values were obtained in the software. Figure 6 As shown in the fluorescence images of tumor-bearing mice, it can also be observed that the tumor fluorescence intensity of mice receiving YFSJF-N-IG is lower, but there is no significant difference between YFSJF-N-IG, YFSJF-Y-IG, and YFSJF-N-IV, further illustrating the equivalent relationship between the two.
[0073] (3.3) After the end of the experiment, 1 mL of blood was collected from the mouse eyeballs and the mice were killed by cervical dislocation. The red blood cells (RBC), hemoglobin (HGB), platelets (PLT), and lymphocytes (LYM) were tested using an automatic blood analyzer (Shenzhen Mindray-BC-5180CRP). The aspartate aminotransferase (AST), alanine aminotransferase (ALT), and serum creatinine (CREA) were analyzed using a fully automatic biochemical analyzer (Toshiba-TBA40FR, Japan) to verify the safety of YFSJF-N. The results are as follows: Figure 7 In the safety evaluation, no anemia, bone marrow suppression, or liver and kidney function damage were observed in hematological examinations, regardless of whether YFSJF-N was administered orally or through the tail vein. There were no statistically significant differences, indicating a high safety profile.
[0074] In summary, these experiments have demonstrated, at both the cellular and animal levels, that the vesicle-like nanoparticles extracted from this invention can exert anti-tumor effects similar to those of YFSJF. These vesicle-like nanoparticles further simplify the composition of traditional Chinese medicine formulas and can be formulated into injectable formulations to exert anti-tumor effects, providing a new approach for the treatment of lung cancer.
[0075] The applicant declares that the present invention is illustrated by the above-described embodiments, but the present invention is not limited to the above-described embodiments. This does not mean that the present invention must rely on the above-described embodiments in order to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent replacements for raw materials in the present invention, additions of auxiliary ingredients, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present invention.
[0076] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.
[0077] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.
Claims
1. A vesicle-like nanoparticle derived from traditional Chinese medicine, characterized in that: The raw materials for preparing the vesicle-like nanoparticles include fritillary bulb of thunbergii, cat's claw, fried bombyx batryticatus, scutellaria baicalensis, trillium, pinellia ternata, ganoderma lucidum and American ginseng.
2. The vesicle-like nanoparticle according to claim 1, characterized in that The raw materials for preparing the vesicle-like nanoparticles include, by weight, 5-15 parts of Fritillaria thunbergii, 25-35 parts of Herba Lycopodii, 5-15 parts of Bombyx batryticatus, 10-20 parts of Radix Sarcodiformis, 7-12 parts of Psoralea corylifolia, 7-12 parts of Pinellia ternata, 10-15 parts of Ganoderma lucidum, and 3-8 parts of American ginseng. Preferably, the raw materials for preparing the vesicle-like nanoparticles include, by mass, 10 parts of Fritillaria thunbergii, 30 parts of Herba Lycopodii, 10 parts of Bombyx batryticatus, 15 parts of Radix Sarcodiformis, 9 parts of Trillium, 9 parts of Rhizoma Pinelliae, 12 parts of Ganoderma lucidum and 6 parts of American ginseng.
3. The vesicle-like nanoparticle according to claim 1 or 2, characterized in that The particle size of the vesicle-like nanoparticles is 100-150 nm.
4. The vesicle-like nanoparticle according to any one of claims 1 to 3, characterized in that The vesicle-like nanoparticles are prepared by a preparation method comprising the following steps: (1) mixing the raw materials for preparing vesicle-like nanoparticles with water, soaking, boiling, and centrifuging to collect the supernatant I; (2) The supernatant I collected in step (1) is centrifuged again, and the supernatant II is collected to obtain vesicle-like nanoparticles.
5. The vesicle-like nanoparticle according to claim 4, characterized in that The mass volume ratio of the raw materials for preparing the vesicle-like nanoparticles in step (1) to water is 1:(8-12) g / mL; Preferably, the soaking temperature in step (1) is 2-10° C., and the soaking time is 30-90 min; Preferably, the decocting time in step (1) is 30-90 minutes; Preferably, the centrifugal speed in step (1) is 500-2000 rpm, and the centrifugal time is 10-50 min.
6. The vesicle-like nanoparticle according to claim 4 or 5, characterized in that The centrifugal speed in step (2) is 8000-15000 rpm, and the centrifugal time is 10-50 min.
7. The vesicle-like nanoparticle according to any one of claims 4 to 6, characterized in that After collecting the supernatant II in step (2), the method further comprises dialyzing the supernatant II; Preferably, the dialysate of the dialysis treatment comprises water; Preferably, the dialysis treatment time is 20-30 hours; Preferably, the molecular weight cut-off of the dialysis bag used for the dialysis treatment is 3-4 KDa; Preferably, the dialysis bag used in the dialysis treatment is of the MD44 type.
8. Use of the vesicle-like nanoparticles according to any one of claims 1 to 7 in the preparation of anti-tumor drugs; Preferably, the tumor comprises lung cancer.
9. The use according to claim 8, characterized in that The dosage forms of the drug include tablets, capsules, granules, powders, injections or sprays.
10. An injection, characterized in that: The injection comprises the vesicle-like nanoparticles derived from traditional Chinese medicine according to any one of claims 1 to 8 and water for injection.