Application of solanum nigrum fruit-sourced nano-vesicles in preparation of medicine for preventing and / or treating prostatic cancer

Abstract

The invention belongs to the technical field of biological medicines, and discloses application of nigrum fruit-sourced nano-vesicles in preparation of medicines / health-care products for preventing and / or treating prostatic cancer. Purified nigrum fruit source nano-vesicles are stably extracted from nigrum fruits, the nigrum fruit source nano-vesicles are natural in ingredients, free of toxic and side effects and good in biocompatibility and safety, in-vivo and in-vitro experiments prove that the nigrum fruit source nano-vesicles have strong prostatic cancer proliferation and migration inhibition effects and apoptosis promotion effects, and the tumor inhibition effect of the nigrum fruit source nano-vesicles is proved in mouse animal experiments. The solanum nigrum fruit-sourced nano-vesicles can be orally taken and absorbed, have no tissue or immunological rejection, are expected to become important materials for preparing drugs for preventing and / or treating prostatic cancer, and have wide application prospects.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and particularly relates to the use of nanovesicles derived from Solanum nigrum fruit in the preparation of drugs / health products for preventing and / or treating prostate cancer. Background Art

[0002] Prostate cancer is one of the most common malignant tumors of the male genitourinary system. Its high morbidity and mortality rates pose a serious threat to global public health. The diagnosis of prostate cancer is seriously delayed due to privacy issues, and the disease has often progressed to the middle and late stages when it is diagnosed. Currently, most prostate cancer treatment drugs have problems such as large side effects and strong drug resistance. For example, chemotherapy drugs can damage normal cells while killing tumor cells, resulting in a decrease in the patient's quality of life; and although targeted therapy drugs are highly targeted, they are prone to drug resistance after long-term use. Therefore, it is particularly important to develop new, efficient, and low-toxic drugs for the treatment of prostate cancer.

[0003] Plant-derived nanovesicles are nanoscale vesicles released by plant cells under specific conditions, which contain rich bioactive molecules such as proteins, RNA, DNA fragments and lipids. In recent years, with the rapid development of molecular biology and nanotechnology, the research on plant exosomes in the biomedical field has gradually increased, especially showing great potential in drug delivery, immune regulation and tumor treatment. However, animal exosomes have the following characteristics: immunogenicity, small quantity, difficulty in obtaining, easy degradation during storage and relatively expensive; while plant exosomes can be obtained in large quantities, have low immunogenicity, low price, high stability, small differences between different batches, but it is difficult to purify the effective substances.

[0004] Solanum nigrum Linn. belongs to the Solanaceae family and is an annual herb widely distributed throughout my country, often seen on roadsides or in fields. The whole plant of Solanum nigrum, including roots, stems, leaves and fruits, has certain medicinal value. In traditional medicine, Solanum nigrum is used to treat symptoms such as carbuncle, swelling, traumatic injury, swelling and blood circulation, and has multiple effects such as clearing away heat and detoxification, promoting blood circulation, diuresis and swelling. Although Solanum nigrum has a long history of use as a traditional Chinese medicine, and modern studies have also found that it contains a variety of bioactive ingredients (such as alkaloids, polysaccharides, non-saponin compounds, etc.), these studies mainly focus on the crude extracts of the whole plant or specific organs of Solanum nigrum, and the in-depth analysis and mechanism research of the specific components in Solanum nigrum fruit is still insufficient. In particular, there are very few studies on the emerging field of Solanum nigrum Linn.-derived nanovesicles (SDNVs).

[0005] In summary, the application of nanovesicles derived from Solanum nigrum fruit in the preparation of anti-prostate cancer drugs has broad prospects, but the current research is still in its infancy and has many defects and deficiencies. The present invention provides a new treatment option for prostate cancer patients by deeply studying the anti-tumor mechanism of plant exosomes derived from Solanum nigrum fruit and exploring its application potential in the treatment of prostate cancer. Summary of the invention

[0006] In order to overcome the above-mentioned shortcomings and deficiencies of the prior art, the primary purpose of the present invention is to provide the use of nanovesicles derived from Solanum nigrum fruit in the preparation of drugs for preventing and / or treating prostate cancer.

[0007] Another object of the present invention is to provide the use of nanovesicles derived from Solanum nigrum fruit in the preparation of health products for auxiliary prevention and / or auxiliary treatment of prostate cancer.

[0008] The purpose of the present invention is achieved through the following solutions:

[0009] Application of Solanum nigrum derived nonavesicles (SDNVs) in the preparation of drugs for preventing and / or treating prostate cancer.

[0010] In the technical solution of the present invention, the drugs are the same or different and include therapeutically effective amounts of Solanum nigrum fruit-derived nanovesicles (SDNVs).

[0011] In the technical scheme of the present invention, the drugs that are the same or different can be made into various pharmaceutical dosage forms by conventional methods, including: tablets (sugar-coated tablets, film-coated tablets, enteric-coated tablets), capsules (hard capsules, soft capsules), oral liquids, lozenges, granules, granules, pills, pills, suspensions, wine preparations, tinctures, drops and other oral dosage forms and injections and other dosage forms other than oral administration, such as injections.

[0012] In the technical solution of the present invention, the drugs described above, whether the same or different, may also contain one or more pharmaceutically acceptable carriers or excipients.

[0013] Furthermore, the carrier or auxiliary material may include a diluent, a wetting agent, a binder, a surfactant, a humectant, an adsorption carrier, a lubricant, a filler, a disintegrant, a preservative, and the like.

[0014] The present invention also provides the use of nanovesicles derived from Solanum nigrum fruit in preparing health products for auxiliary prevention and / or auxiliary treatment of prostate cancer.

[0015] The Solanum nigrum fruit-derived nanovesicles (SDNVs) of the present invention can be obtained by a nanovesicle preparation method known to those skilled in the art, such as by extraction and separation from Solanum nigrum fruit.

[0016] Furthermore, the separated Solanum nigrum fruit-derived nanovesicles (SDNVs) can be purified by sucrose solution to obtain refined SDNVs.

[0017] The present invention stably extracts purified nanovesicles from Solanum nigrum fruit, which have natural ingredients, no toxic side effects, good biocompatibility and safety, and has been proven to have a strong inhibitory effect on prostate cancer proliferation and migration and apoptosis-promoting effect through in vivo and in vitro experiments, and its tumor inhibition effect is confirmed in mouse animal experiments. Nanovesicles derived from Solanum nigrum fruit can be orally absorbed and have no tissue or immune rejection reaction, and are expected to become an important material for preparing therapeutic drugs for preventing and / or treating prostate cancer, and therefore have broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0019] Figure 1 The morphological observation of SDNVs under transmission electron microscope.

[0020] Figure 2 The CCK-8 method was used to detect the inhibitory effect of SDNVs on the proliferation of PC-3 cells and RWPE-1 cells. A. The inhibitory effect of SDNVs on the proliferation of PC-3 cells; B. SDNVs had no inhibitory effect on the proliferation of RWPE-1 cells; n=5.

[0021] Figure 3 CCK-8 method was used to detect the inhibitory effect of SDNVs and SEE on PC-3 cell proliferation. Among them, n=5.

[0022] Figure 4 The inhibition of PC-3 cell migration by SDNVs in the Transwell migration assay, qar = 200 μm, n = 3.

[0023] Figure 5 SDNVs promote apoptosis of PC-3 cells. Among them, n=3.

[0024] Figure 6 A diagram showing the anti-prostate cancer effect of SDNVs in mouse experiments.

[0025] Figure 7 The figure is a statistical chart of the prostate cancer tumor volume in each group in the mouse experiment, where n=6.

[0026] Figure 8 The figure is a statistical chart of the prostate cancer tumor weights of each group in the mouse experiment, where n=6.

[0027] In the above figures, ns>0.05, *p<0.05, **p<0.01, ***p<0.001. DETAILED DESCRIPTION

[0028] The present invention is further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto. The materials involved in the following examples can be obtained from commercial channels unless otherwise specified. The methods described are conventional methods unless otherwise specified.

[0029] In one embodiment, Solanum nigrum derived nonavesicles (SDNVs) are used in the preparation of drugs for preventing and / or treating prostate cancer.

[0030] In a specific embodiment of the present invention, the drugs are the same or different and include a therapeutically effective amount of Solanum nigrum fruit derived nanovesicles (SDNVs).

[0031] The dosage of the drug of the present invention can be prescribed in a variety of ways depending on factors such as the formulation method, administration method, patient's age, weight, gender, morbidity, diet, administration time, administration route, excretion rate and reaction sensitivity. A skilled physician can usually easily determine the prescription and the dosage that is effective for the desired treatment or prevention.

[0032] The therapeutically effective amount refers to an amount that can produce a therapeutic effect on humans and / or animals and can be accepted by humans and / or animals. For example, a therapeutically effective amount refers to the amount of the drug required to produce the desired therapeutic effect, and the therapeutic effect can be reflected by the results of clinical trials, model animal studies and / or in vitro studies. The pharmaceutically effective amount depends on several factors, including but not limited to: characteristic factors of the subject to be treated (such as the subject's height, weight, gender, age and medication history, etc.), and the severity of the disease.

[0033] In a specific embodiment of the present invention, the therapeutically effective amount refers to an amount that can produce a preventive and / or therapeutic effect on prostate cancer patients and can be accepted by the patients.

[0034] In a specific embodiment of the present invention, the administration of the drug includes, but is not limited to, oral administration, parenteral administration, administration by inhalation spray, topical administration, rectal administration, nasal administration, buccal administration, or administration by implanted drug storage device. Oral administration or injection is preferred.

[0035] In a specific embodiment of the present invention, any orally acceptable dosage form may be used, including but not limited to capsules, tablets, aqueous suspensions or solutions.

[0036] In a specific embodiment of the invention, liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs.

[0037] In a specific embodiment of the invention, solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.

[0038] In a specific embodiment of the present invention, the drug also includes a pharmaceutically acceptable carrier or adjuvant. The pharmaceutically acceptable carrier or adjuvant may contain an inert component that does not unduly inhibit the biological activity of the compound. Pharmaceutically acceptable carriers or adjuvants should be biocompatible, for example, non-toxic, non-inflammatory, non-immunogenic or no other undesirable reactions or side effects when administered to a subject. Standard pharmaceutical preparation techniques can be used.

[0039] Pharmaceutically acceptable carriers or excipients include, but are not limited to, diluents, adhesives, surfactants, wetting agents, adsorption carriers, lubricants, fillers, disintegrants, preservatives, etc. These substances are used to help the stability of the formulation or to help improve the activity or its biological effectiveness or to produce an acceptable taste or smell in the case of oral administration. The preparations that can be used in such pharmaceutical compositions can be in the form of the original compound itself or optionally in the form of a pharmaceutically acceptable salt thereof. The pharmaceutical composition thus formulated can be administered by any appropriate method known to those skilled in the art as needed.

[0040] Among them, the diluent includes but is not limited to lactose, sodium chloride, glucose, urea, starch, and water.

[0041] Binders include, but are not limited to, starch, pregelatinized starch, dextrin, maltodextrin, sucrose, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, ethylcellulose, polyvinyl alcohol, polyethylene glycol, polyvinylpyrrolidone, alginic acid and alginates, xanthan gum, hydroxypropyl cellulose and hydroxypropyl methylcellulose.

[0042] Surfactants include, but are not limited to, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, and cetyl alcohol.

[0043] Wetting agents include, but are not limited to, glycerin and starch.

[0044] Adsorption carriers include, but are not limited to, starch, lactose, bentonite, silica gel, kaolin, and bentonite.

[0045] Lubricants include, but are not limited to, zinc stearate, glyceryl monostearate, polyethylene glycol, talc, calcium and magnesium stearate, polyethylene glycol, boric acid powder, hydrogenated vegetable oil, sodium stearyl fumarate, polyoxyethylene monostearate, monolauric sucrose ester, sodium lauryl sulfate, magnesium lauryl sulfate, and magnesium dodecyl sulfate.

[0046] Fillers include, but are not limited to, mannitol (granular or powdered), xylitol, sorbitol, maltose, erythrose, microcrystalline cellulose, polymeric sugars, coupling sugars, glucose, lactose, sucrose, dextrin, starch, sodium alginate, laminarin powder, agar powder, calcium carbonate, and sodium bicarbonate.

[0047] Disintegrants include, but are not limited to, cross-linked vinyl pyrrolidone, sodium carboxymethyl starch, low-substituted hydroxypropyl methyl, cross-linked sodium carboxymethyl cellulose, and soybean polysaccharides.

[0048] In a specific embodiment of the present invention, the drug also includes other drugs for preventing and / or treating prostate cancer, that is, the Solanum nigrum fruit-derived nanovesicles can be used in combination with other compounds that can be used to prevent and / or treat prostate cancer.

[0049] Another embodiment is the use of nanovesicles derived from Solanum nigrum fruit in the preparation of health products for the auxiliary prevention and / or auxiliary treatment of prostate cancer.

[0050] In a specific embodiment of the present invention, the Solanum nigrum fruit-derived nanovesicles (SDNVs) can be obtained by a nanovesicle preparation method known to those skilled in the art, such as by extraction and separation from Solanum nigrum fruit.

[0051] In one embodiment, the extractant used in the extraction can be conventionally used water or buffer, such as an aqueous solution with added phosphate.

[0052] In one embodiment, the extraction may be performed by crushing or breaking the wall of the Solanum nigrum fruit and then soaking it in an extractant, and the obtained soaking liquid is the extract.

[0053] In one embodiment, the separation method can be to perform ultracentrifugation on the extract to separate the extract. The separated precipitate is the Solanum nigrum fruit-derived nanovesicles.

[0054] In one embodiment, the speed of the high-speed centrifugation is 100,000 g or above.

[0055] In one embodiment, the high-speed separation time is 10-100 minutes.

[0056] In one embodiment, the high-speed centrifugation time is 60-100 min.

[0057] In one embodiment, the precipitate obtained by ultracentrifugation can be resuspended in a buffer such as Tris-HCl to obtain a suspension of nanovesicles derived from Solanum nigrum fruit. One or more high-speed centrifugations can be performed to obtain further purified nanovesicles derived from Solanum nigrum fruit.

[0058] In one embodiment, pre-centrifugation is performed before high-speed centrifugation to remove impurities and precipitates.

[0059] In one embodiment, the speed of the pre-centrifugation is 10000 g or less. Pre-centrifugation can separate and remove non-target substances, thereby reducing the impact of impurities on subsequent centrifugal separation.

[0060] In one embodiment, the pre-centrifugation time is 10-60 min. The pre-centrifugation can be performed once or more.

[0061] In one embodiment, at least one pre-centrifugation is performed at a centrifugal speed of less than or equal to 10000 g.

[0062] In one embodiment, filtration is performed before the pre-centrifugation, such as using gauze to filter out insoluble sediment.

[0063] In one embodiment, the Solanum nigrum fruit-derived nanovesicle suspension is filtered through a filter before use to obtain a sterile suspension.

[0064] In one embodiment, the filter is a 0.22 μm filter.

[0065] In one embodiment, the solanum nigrum fruit-derived nanovesicles are obtained by extraction and separation of solanum nigrum fruit, and are prepared by the following steps: crushing or grinding the solanum nigrum fruit, soaking it in water, separating the supernatant, pre-centrifuging the supernatant at a centrifugal force of 10,000 g or less, taking the supernatant, and discarding the precipitate; the supernatant after pre-centrifugation is then high-speed centrifuged at a centrifugal force of 100,000 g or more, and the centrifugation time can be 10-100 min. The obtained precipitate is the solanum nigrum fruit-derived nanovesicles.

[0066] In one embodiment, the Solanum nigrum fruit is cleaned before use.

[0067] In one embodiment, the soaking time may be 1 min-1 h.

[0068] In one embodiment, stirring may be performed during the soaking process.

[0069] In one embodiment, the separated supernatant can be obtained by filtering, such as filtering the insoluble sediment using gauze.

[0070] In one embodiment, one or more pre-centrifugations may be performed; for example, the mixture may be centrifuged at 200-1000 g, 1500-2500 g, or 8000-10000 g for 10-60 min, respectively, and the supernatants are taken and the precipitates are discarded.

[0071] In one embodiment, at least one pre-centrifugation is performed at a centrifugation speed of 8000-10000 g.

[0072] In a specific embodiment of the present invention, the separated Solanum nigrum fruit-derived nanovesicles can be purified using a sucrose solution.

[0073] In one embodiment, the purification operation specifically includes adding a sample of the Solanum nigrum fruit-derived nanovesicles into a centrifuge tube, then adding sucrose solutions of different concentrations from bottom to top in order from high concentration to low concentration through the sample layer, ultracentrifuging, and taking out the Solanum nigrum fruit-derived nanovesicles between the sucrose solution layers to obtain purified Solanum nigrum fruit-derived nanovesicles.

[0074] In one embodiment, the concentrations of the sucrose solutions used are 10-20wt%, 25-35wt%, 40-50wt%, 55-60wt%, etc. The concentration gradient difference of the sucrose solutions of different concentrations used is preferably 10wt% or more.

[0075] In one embodiment, the interlayer refers to an interlayer of sucrose solution with concentrations of 10-20wt% and 25-35wt%.

[0076] In one embodiment, the sucrose solution is prepared by using pure water.

[0077] In one embodiment, the speed of ultracentrifugation can be 100,000 g or above, such as 150,000 g.

[0078] In one embodiment, the ultracentrifugation time may be 2-5 hours.

[0079] In one embodiment, the purified Solanum nigrum fruit-derived nanovesicles obtained by ultracentrifugation can be resuspended in a buffer such as Tris-HCl to obtain a suspension of Solanum nigrum fruit-derived nanovesicles. One or more high-speed centrifugations can be performed to obtain further purified Solanum nigrum fruit-derived nanovesicles.

[0080] In one embodiment, the Solanum nigrum fruit-derived nanovesicle suspension is filtered through a filter before use to obtain a sterile suspension.

[0081] In one embodiment, the filter is a 0.22 μm filter.

[0082] In one embodiment, the purification operation specifically includes the following steps: dissolving sucrose in pure water to prepare sucrose solutions with mass concentrations of 10-20wt%, 25-35wt%, 40-50wt%, and 55-60wt% (such as 15%, 30%, 45%, and 60%), respectively, and the concentration gradient difference of sucrose water of different concentrations is 10wt% or more (such as 15wt%). Take 1-2 volume parts (such as 1 volume part) of the suspension of nanovesicles derived from Solanum nigrum fruit and add them to a centrifuge tube, take 2-3 volume parts (such as 2.5 volume parts) of sucrose water of different concentrations through the sample layer and add them to the centrifuge tube from bottom to top from high concentration to low concentration, ultracentrifuge, take out the nanovesicles derived from Solanum nigrum fruit between the sucrose solution with a concentration of 10-20wt% and the sucrose solution with a concentration of 25-35wt%, resuspend them in a buffer such as Tris-HCl, and ultracentrifuge to remove the mixed sucrose water, so as to obtain the final nanovesicles derived from Solanum nigrum fruit.

[0083] In one embodiment, a transmission electron microscope (the instrument model is Tecnai G2 SpiritTwin + GATAN 832.10W) was used to perform morphological identification of Solanum nigrum fruit derived nanovesicles (SDNVs). Figure 1 As can be seen from the figure, SDNVs are round or oval vesicles, like trays, with diameters ranging from 50 to 200 nm.

[0084] In one embodiment, the particle size and particle number of SDNVs in the SDNVs suspension were measured using a particle tracking analyzer (NanoSight NS300). The results showed that the average diameter of SDNVs was 93.4±0.2nm, and the mass concentration was about 1.8mg / mL. Based on the Solanum nigrum fruit raw material, about 1.8mg SDNVs can be extracted from every 1g of Solanum nigrum fruit raw material.

[0085] In the following examples, Solanum nigrum ethanol extract was used for comparison, which was prepared by the following steps:

[0086] ① Raw material pretreatment: 20g of Chinese medicinal Solanum nigrum fruit raw material was ground into powder and passed through a 100-mesh sieve;

[0087] ② Extraction: Take the powder in ①, add an appropriate amount of 75% ethanol and extract for 24 hours, centrifuge and discard the residue to obtain the extraction clear liquid;

[0088] ③ Concentration: The extracted clear liquid in step ② is concentrated under reduced pressure to make the concentration of the concentrated liquid 10 mg / mL, a total of 8 mL, to obtain Solanum nigrum ethanol extract (SEE), which is refrigerated for later use. Based on the Solanum nigrum fruit raw material, about 4 mg SEE can be extracted from every 1 g of Solanum nigrum fruit raw material.

[0089] Example 1: SDNVs inhibit PC-3 cell proliferation activity

[0090] (1) SDNVs intervention concentration groups: 0, 12, 18, 24, 30, 36, 48 μg / mL; the treatment group with a concentration of 0 μg / mL was recorded as the blank control group (NC);

[0091] (2) P6 human prostate cancer cell line (PC-3) and human prostate epithelial cells (RWPE-1) were inoculated in 96-well plates, with 1×10 4 After the cells were completely attached to the wall, they were replaced with serum-free medium and different concentrations of SDNVs were added for 24 hours. CCK-8 reagent was added to each well at a volume ratio of 10:1, incubated at 37°C for 4 hours, and the absorbance at 450nm was detected by a multifunctional microplate reader. The results are shown in Figure 2 .

[0092] In addition, in order to compare the difference between SDNVs and SEE in the effects on PC-3 cell viability, PC-3 cells were used as the research object, 36μg / mL SDNVs was used as the experimental group, and 80μg / mL SEE was added for 24h as the positive control group (the dosage is based on the fact that 1g of Solanum nigrum fruit raw material can extract 1.8mg SDNVs or 4mg SEE, so the dose of SEE corresponding to 36μg / mL SDNVs is 80μg / mL). CCK-8 reagent was added to each well at a volume ratio of 10:1, incubated at 37℃ for 4h, and the absorbance at 450nm was detected by a multifunctional microplate reader. The results are shown in Table 1. Figure 2 and Figure 3 .

[0093] (3) Figure 2 As shown in A, with the increase of SDNVs concentration, the proliferation rate of PC-3 cells was significantly inhibited. At a concentration of 36 μg / mL, the survival rate of PC-3 cells was about 54.98%. Figure 2 As shown in B, SDNVs had no significant effect on the viability of normal prostate epithelial cells RWPE-1.

[0094] (4) Figure 3 As shown, both SDNVs and SEE can significantly inhibit the viability of PC-3 cells, but the inhibitory effect of 36 μg / mL SDNVs on PC-3 cell viability is significantly stronger than that of 80 μg / mL SEE.

[0095] Example 2: SDNVs inhibit PC-3 cell migration ability

[0096] (1) Intervention groups: blank control group NC, SDNVs, and ethanol extract group SEE;

[0097] (2) P6 PC-3 cells were inoculated in a 24-well plate, with 1×10 5 After the cell confluence reached 70%, 36 μg / mL SDNVs or 80 μg / mL SEE was added to PC-3 cells for 24 h. The cells were digested with trypsin, and 100 μL serum-free medium was added to the upper chamber of the transwell. 1.5×10 4 cells; 400 μL complete medium was added to the lower chamber. After the transwell plate was incubated in a 37°C incubator for 12 hours, the cells in the chamber were observed to see if they had crossed. After most of the cells had crossed, the medium was discarded and the cells were rinsed twice with PBS. The cells were fixed with paraformaldehyde for 10 minutes, the fixative was discarded, and the cells were washed twice with pure water. The cells were stained with 1% crystal violet (filtered with a 0.45 μm filter) for 10 minutes, and the cells were rinsed twice with PBS. The cells were photographed under an inverted microscope.

[0098] (3) The results are as follows Figure 4 As shown in the results, both SDNVs and SEE could significantly inhibit the migration ability of PC-3 cells, but SDNVs had a more significant inhibitory effect than SEE.

[0099] Example 3: Effect of SDNVs on promoting apoptosis of PC-3 cells

[0100] (1) Intervention groups: blank control group NC, SDNVs, and ethanol extract group SEE;

[0101] (2) P6 PC-3 cells were inoculated in a 12-well plate, with 2×10 5 After the cell confluence reached 70%, 36 μg / mL SDNVs or 80 μg / mL SEE was added to PC-3 cells for 24 h. The cells were digested with trypsin (without EDTA), and the digestion was terminated by pre-cooled PBS. 1-2×10 5 cells and washed twice with ice-cold PBS. TM 488 / PI double staining cell apoptosis detection kit (KTA0002, Abbkine) instructions for incubating AnnexinV-AbFluor TM 488 and PI, cells were analyzed by flow cytometry within 30 min after staining, using 491 nm and 535 nm excitation, and fluorescence was measured near 517 nm (FITC channel) and 617 nm (PI channel).

[0102] (2) The results are as follows Figure 5 As shown in the results, both SDNVs and SEE can significantly promote apoptosis of PC-3 cells, but SDNVs has a more significant apoptosis-promoting effect than SEE.

[0103] Example 4: Inhibitory effect of SDNVs on prostate cancer

[0104] (1) Experimental subjects: 8-week-old male Babl / c nude mice;

[0105] (2) Experimental groups: 6 mice in each group: ① control group (PBS gavage 100 μL / mouse); ② SDNVs gavage group (180 μg / mouse); ③ SEE gavage group (400 μg / mouse).

[0106] (3) Experimental procedures: After a one-week adaptation period, Babl / c nud mice were inoculated with PC-3 cells (1.0×10 7 cells) were subcutaneously inoculated into mice. When the tumor volume reached 20 mm 3 At 4 pm, nude mice were randomly divided into the above groups. According to the different groups, SDNVs or SEE were gavaged once a day for 4 consecutive weeks, and the control group was given an equal amount of PBS. After 4 weeks, the mice were euthanized and the tumor tissues were collected for further analysis.

[0107] (4) The results are as follows Figure 6-Figure 8 As shown, the treatment with SDNVs and SEE improved the volume and weight of the tumor in mice, and the SDNVs of the present invention had a more significant and excellent effect than SEE.

[0108] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Application of nanovesicles derived from Solanum nigrum fruit in the preparation of drugs for preventing and / or treating prostate cancer.

2. The use according to claim 1, characterized in that: The drugs are the same or different and include therapeutically effective amounts of nanovesicles derived from Solanum nigrum fruit.

3. The use according to claim 1, characterized in that: The drugs are the same or different and are made into various pharmaceutical dosage forms by conventional methods. These dosage forms include: tablets, capsules, oral liquids, lozenges, granules, granules, pills, pills, suspensions, wine preparations, tinctures, oral dosage forms of drops and injections.

4. The use according to claim 1, characterized in that: The same or different drugs are prepared into various pharmaceutical dosage forms by conventional methods, including sugar-coated tablets, film-coated tablets, enteric-coated tablets, hard capsules, soft capsules, oral liquids, lozenges, granules, granules, pills, pills, suspensions, wine preparations, tinctures, oral dosage forms of drops and injections.

5. The use according to claim 1, characterized in that: The drugs described above may be the same or different and may also contain one or more pharmaceutically acceptable carriers or excipients.

6. The use according to claim 5, characterized in that: The carrier or auxiliary material includes at least one of a diluent, a wetting agent, a binder, a surfactant, a humectant, an adsorption carrier, a lubricant, a filler, a disintegrant, and a preservative.

7. Application of nanovesicles derived from Solanum nigrum fruit in the preparation of health products for auxiliary prevention and / or auxiliary treatment of prostate cancer.

8. The use according to any one of claims 1 to 7, characterized in that: The solanum nigrum fruit-derived nanovesicles are extracted and separated from the solanum nigrum fruit.

9. The use according to claim 8, characterized in that: The separated Solanum nigrum fruit-derived nanovesicles are purified by sucrose solution to obtain refined Solanum nigrum fruit-derived nanovesicles.

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