CN112245422A

CN112245422A - Novel application of acetyl valerian

Info

Publication number: CN112245422A
Application number: CN202010968464.7A
Authority: CN
Inventors: 毛新良; 孙彤; 徐宇嘉
Original assignee: Guangzhou Medical University
Current assignee: Guangzhou Medical University
Priority date: 2020-09-15
Filing date: 2020-09-15
Publication date: 2021-01-22
Anticipated expiration: 2040-09-15
Also published as: CN112245422B

Abstract

The invention provides a new application of acevalerian, and particularly relates to an application of acevalerian in preparing a medicament for preventing and/or treating tumors. The invention discovers that the acevalosin has obvious inhibition effect on tumor cells, and the inhibition effect is verified on cells and animal living bodies. After verification: the acevalosin can induce the degradation of c-Maf, CCND1 and JAK2 oncoproteins, and inhibit Otub1, c-Maf, USP10 and STAT3 signal channels, thereby effectively controlling the proliferation of tumor cells, inducing the apoptosis of the tumor cells and having the effects of inhibiting and treating tumors. Meanwhile, as a plant extract, the acevalosin also has the advantage of low toxicity.

Description

Novel application of acetyl valerian

Technical Field

The invention belongs to the field of pharmacology, and particularly relates to a new application of a compound of acevalerian.

Background

Cancer or tumor, which is a malignant lesion originated from human epithelial or leaf tissue, has biological characteristics of abnormal cell differentiation and proliferation, growth loss control, infiltration and metastasis, and the like, and the occurrence of the cancer is a complex process with multiple factors and multiple steps. Tumors can originate from almost all tissue and organs and can therefore form common different tumors, such as lung cancer, breast cancer, brain cancer, liver cancer, prostate cancer, kidney cancer, blood cancer, and the like. The malignant tumors are serious diseases seriously threatening the physical and mental health of people in China, and the investigation of main death reasons of urban and rural residents published by the ministry of health in China in 2006 shows that: the number of people who die of malignant tumors in China is nearly 200 thousands, the malignant tumors become the first cause of death, but most of tumors have no effective cure measures at present. The research of malignant tumor medicine with good curative effect and low toxic and side effect is one of the important contents of the national medium and long term scientific and technical development planning compendium.

While the occurrence of malignant tumors is closely related to the changes in various molecular events. For example, Multiple Myeloma (MM) is a terminally differentiated malignant B-cell tumor characterized by abnormal proliferation of plasma cells in the bone marrow, secretion of monoclonal immunoglobulins or fragments thereof, and induction of tissue or organ damage. The vast majority of B cell tumors are mainly characterized by immunoglobulin heavy chain gene (IgH) switch region rearrangement, antigen selection, high frequency somatic mutation in B cells at the germinal center or the postgerminal center, and immunoglobulin gene regulation, which in turn will selectively act on VDJ and switch region sequences of immunoglobulin (Ig) genes to generate IgH translocation. Multiple myeloma IgH translocations are also caused by errors in IgH switch domain recombination or by high frequency somatic mutations during B cell maturation at the germinal center, with a maximum of 5 associated chromosomal sites: 4p16 (histone methyltransferase-multiple myeloma SET domain protein, MMSET and fibroblast growth factor receptor 3, FGFR3), 6p21 (cyclin D3, CCND3), 11q13 (cyclin D1, CCND1), 16q23(MAF), 20q11 (MAFB). However, in the MGUS stage (monoclonal immunoglobulin disease of unknown significance, the pre-stage of multiple myeloma), IgH translocations accumulate rarely 4p16 t (4,14) and 16q23t (14,16), which indicates that the translocations may promote the development of pathological states such as MGUS and SMM, which may eventually lead to the occurrence of multiple myeloma, or have a certain relationship with the rapid development of MGUS into multiple myeloma.

Valerian is mainly distributed in Europe, Asia, northeast China, etc., has mind tranquilizing effect, and is mainly used for treating uneasiness, cardiopalmus and insomnia, mania, noise, rheumatalgia, dysmenorrheaIt can be used for treating amenorrhea, traumatic injury, etc., and has pharmacological effects in strengthening cerebral cortex inhibiting process, reducing reflex excitability, relieving smooth muscle spasm, resisting bacteria, and promoting urination. The acevalant has a structure shown as the following formula I and a molecular formula C₂₄H₃₂O₁₀And the molecular weight of 480.50, is a natural extract of valerian, and has effects of tranquilizing and relieving pain.

Disclosure of Invention

Based on the situation, the main purpose of the invention is to provide a new application of the acevalol, in particular to a medicine prepared from the acevalol and used for preventing and/or treating tumors.

Application of acevalerian in preparing medicine for preventing and/or treating tumor is disclosed.

In one embodiment, the tumor is myeloma, brain tumor, leukemia, hodgkin's disease, non-hodgkin's lymphoma, B cell lymphoma, T cell lymphoma, fahrenheit macroglobulinemia, myeloproliferative syndrome, myelodysplastic syndrome, lung cancer, liver cancer, prostate cancer, oral cancer, or breast cancer.

In one embodiment, the myeloma is multiple myeloma.

In one embodiment, the brain tumor is a glioma.

In one embodiment, the medicament comprises the acevalant and pharmaceutically acceptable excipients.

In one embodiment, the medicament comprises from 0.1 wt% to 99.5 wt% of the acevalant.

In one embodiment, the excipient is selected from at least one of diluents, wetting agents, binders, disintegrants, lubricants, color, flavor modifiers, solvents, solubilizers, suspending agents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, local analgesics, pH modifiers, and isotonic or isotonic modifiers.

In one embodiment, the diluent is selected from at least one of starches, sugars, celluloses and inorganic salts; or/and the wetting agent is selected from at least one of water and ethanol; or/and the adhesive is selected from at least one of starch slurry, dextrin, sugar, cellulose derivative, gelatin, povidone and polyethylene glycol; or/and the disintegrant is selected from at least one of starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, croscarmellose sodium, crospovidone, surfactant and effervescent disintegrant; or/and the lubricant is selected from at least one of talcum powder, calcium stearate, magnesium lauryl sulfate, superfine silica powder and polyethylene glycol; or/and the color, aroma and taste regulator is selected from at least one of pigment, spice, sweetener, mucilage and flavoring agent; or/and the solvent is at least one selected from water, oil, ethanol, glycerol, propylene glycol, polyethylene glycol, dimethyl sulfoxide, liquid paraffin, fatty oil and ethyl acetate; or/and the solubilizer is selected from at least one of tween, maize, polyoxyethylene fatty alcohol ether, soap, sulfate and sulfonate; or/and the cosolvent is at least one selected from organic acids and salts thereof, amides and amine compounds, inorganic salts, polyethylene glycol, povidone and glycerol; or/and the emulsifier is selected from at least one of span, tween, maize, beneze, glycerin fatty acid ester, higher fatty acid salt, sulfate, sulfonate, acacia, tragacanth, gelatin, pectin, phospholipid, agar, sodium alginate, hydroxide, silicon dioxide and bentonite; or/and the suspending agent is at least one selected from glycerol, syrup, acacia gum, tragacanth gum, agar, sodium alginate, cellulose derivatives, povidone, carbopol, polyvinyl alcohol and thixotrope; and/or, the antioxidant is at least one selected from sulfite, pyrosulfite, bisulfite, ascorbic acid, gallic acid and esters thereof; or/and the metal complexing agent is selected from one of disodium ethylene diamine tetraacetate and polycarboxylic acid compound; or/and the inert gas is selected from one of nitrogen and carbon dioxide; or/and the preservative is at least one of nipagin, organic acid and salt thereof, quaternary ammonium compound, chlorhexidine acetate, alcohol, phenol and volatile oil; or/and the local analgesic is selected from at least one of benzyl alcohol, chlorobutanol, lidocaine and procaine; or/and the pH regulator is at least one selected from hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, tartaric acid, acetic acid, sodium hydroxide, sodium bicarbonate, ethylenediamine, meglumine, phosphate, acetate and citrate; and/or, the isotonic or isotonic regulator is at least one selected from glucose, sodium chloride, sodium citrate, sorbitol and xylitol.

In one embodiment, the dosage form of the medicament is tablets, granules, pills, powder, capsules, injections, oral liquid, ophthalmic preparations or external preparations.

In one embodiment, the route of administration of the drug includes intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration, or transdermal administration.

A combined medicine composition for preventing and/or treating tumors comprises active ingredients of at least one of an anti-tumor medicine and an immunomodulator and acetyl valerianin.

In one embodiment, the antineoplastic agent is selected from at least one of cisplatin and doxorubicin; or/and the immunomodulator is ranitidine.

The invention has the following beneficial effects:

the invention discovers that the acevalosin has obvious inhibition effect on tumor cells, mainly embodies that the acevalosin regulates and controls various factors related to tumors, and the regulation and control effect is verified on cells and animal living bodies. After verification: the acevalosin can induce the degradation of c-Maf, CCND1 and JAK2 oncoproteins, and inhibit Otub1, c-Maf, USP10 and STAT3 signal channels, thereby effectively controlling the proliferation of tumor cells, inducing the apoptosis of the tumor cells and having the effects of inhibiting and treating tumors. Meanwhile, as a plant source component, the acetovalerian also has the advantage of low toxicity.

Drawings

FIG. 1 is a graph showing the results of preliminary screening of acevalosin for tumors in example 1.

FIG. 2 is a graph showing the results of Western blotting (Western Blot) and RT-PCR analysis and luciferase activity analysis of the inhibitory effect of acevalosin, acevalant against c-Maf, which is a cancer transcription factor, in example 2.

FIG. 3 is a graph showing the results of luciferase activity analysis and Western immunoblotting (Western Blot) of the inhibitory effect of acetylvalerian on the cancer transcription factor STAT3 in example 3.

FIG. 4 is a graph of Co-immunoprecipitation (Co-IP) and immunoblots for degradation of oncoproteins c-Maf and CCND1 following treatment of various cell lines with acetovalosin in example 4.

FIG. 5 is a graph showing flow analysis of apoptosis of different strains of multiple myeloma treated with Val-acetyl in example 5.

FIG. 6 is a graph showing the flow analysis of apoptosis of individual cell lines of brain tumors treated with acetovalosin in example 6.

FIG. 7 is a graph showing the change in tumor volume and the change in body weight of the mice treated with acetovalosin of example 7; FIG. 7A is a graph showing the relationship between the tumor volume and the number of days of administration in mice, and FIG. 7B is a graph showing the relationship between the body weight and the number of days of administration in mice.

FIG. 8 is a graph showing the analysis of the proliferation of bone marrow cells of a patient with acetovalacyclovir-treated myeloma in example 8.

FIG. 9 is a Western Blot analysis of the tumor apoptosis promoting proteins of the combination of acevalosin and various drugs in example 9.

Detailed Description

In order that the invention may be more fully understood, reference will now be made to the following description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Experimental procedures without specific conditions noted in the following examples, generally followed by conventional conditions, such as Sambrook et al, molecular cloning: the conditions described in the Laboratory Manual (New York: Cold Spring Harbor Laboratory Press,1989), or according to the manufacturer's recommendations. The various chemicals used in the examples are commercially available.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The tumor types in the embodiments of the present invention are not particularly limited, and include, but are not limited to: myeloma, brain tumor, leukemia, Hodgkin's disease, non-Hodgkin's lymphoma, B cell lymphoma, T cell lymphoma, Waldenstrom's macroglobulinemia, myeloproliferative syndrome, myelodysplastic syndrome, lung cancer, liver cancer, prostate cancer, oral cancer, breast cancer, etc.

The leukemia in the embodiment of the invention is acute myeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia or chronic lymphocytic leukemia.

The myeloma according to the embodiment of the present invention is multiple myeloma.

The brain tumor in the embodiment of the invention is glioma.

It is understood that the medicament of the embodiment of the invention comprises the acevalant and pharmaceutically acceptable auxiliary materials.

Preferably, the medicament comprises from 0.1 wt% to 99.5 wt% of the acevalant.

It is understood that the excipients of the embodiments of the present invention may be any pharmaceutically acceptable excipient, including, but not limited to, diluents, wetting agents, binders, disintegrants, lubricants, color, flavor modifiers, solvents, solubilizers, solubilizing agents, suspending agents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, topical analgesics, pH adjusting agents, isotonic or isotonic adjusting agents, and the like. Further, including but not limited to: the diluent is selected from starch, saccharide, cellulose and inorganic salt; the wetting agent is selected from water, ethanol and the like; the adhesive is selected from starch slurry, dextrin, sugar, cellulose derivatives, gelatin, polyvidone, polyethylene glycol and the like; the disintegrant is selected from starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, croscarmellose sodium, crospovidone, surfactant, effervescent disintegrant and the like; the lubricant is selected from talcum powder, calcium stearate, magnesium dodecyl sulfate, superfine silica gel powder, polyethylene glycol and the like; the color, aroma and taste regulator is selected from pigments, spices, sweeteners, mucilage agents, flavoring agents and the like; the solvent is selected from water, oil, ethanol, glycerol, propylene glycol, polyethylene glycol, dimethyl sulfoxide, liquid paraffin, fatty oil, ethyl acetate and the like; the solubilizer is selected from tween, maize, polyoxyethylene fatty alcohol ether, soap, sulfate, sulfonate and the like; the cosolvent is selected from organic acid and salts thereof, amide and amine compounds, inorganic salt, polyethylene glycol, polyvidone, glycerol and the like; the emulsifier is selected from span, Tween, Brix, glycerol fatty acid ester, higher fatty acid salt, sulfate, sulfonate, gum arabic, tragacanth, gelatin, pectin, phospholipid, agar, sodium alginate, hydroxide, silicon dioxide, bentonite, etc.; the suspending agent is selected from glycerol, syrup, acacia, tragacanth, agar, sodium alginate, cellulose derivatives, polyvidone, carbopol, polyvinyl alcohol, and thixotrope; the antioxidant is selected from sulfite, pyrosulfite, bisulfite, ascorbic acid, gallic acid and esters thereof; the metal complexing agent is selected from disodium ethylene diamine tetraacetate, polycarboxylic acid compounds and the like; the inert gas is selected from one of nitrogen and carbon dioxide; the preservative is selected from parabens, organic acids and salts thereof, quaternary ammonium compounds, chlorhexidine acetate, alcohols, phenols, volatile oil and the like; the local analgesic is selected from benzyl alcohol, chlorobutanol, lidocaine, procaine and the like; the pH regulator is selected from hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, tartaric acid, acetic acid, sodium hydroxide, sodium bicarbonate, ethylenediamine, meglumine, phosphate, acetate, citrate and the like; the isotonic or isotonic regulator is selected from glucose, sodium chloride, sodium citrate, sorbitol, xylitol, etc.

It is understood that the dosage form of the drug is not particularly limited in the examples of the present invention, and includes, but is not limited to: tablet, granule, pill, powder, capsule, injection, oral liquid, ophthalmic preparation or external preparation, etc.

It is to be understood that the administration route of the drug is not particularly limited in the examples of the present invention, and includes, but is not limited to: intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration, transdermal administration, etc.

The embodiment of the invention also provides a combined medicine composition for preventing and/or treating tumors, and the active ingredients of the combined medicine composition comprise at least one of an anti-tumor medicine and an immunomodulator and the acevalacilin.

Preferably, the antineoplastic agent is cisplatin, adriamycin or the like.

Preferably, the immunomodulator is, for example, ranitidine or the like.

The acevalacilin of the present invention is commercially available.

Example 1

Otub1, HA-c-Maf and c-Maf specific recognition unit (pMARE) driven luciferase, luci. plasmid pMARE. luci, were transfected into HEK293T cells, and after 24 hours of culture, the cells were seeded into 96 well cell culture plates at 10000 cells per well, and after cell attachment, different native compounds were added separately (all compounds were dissolved in dimethyl sulfoxide (DMSO) and diluted to application concentrations; cells were collected for luciferase activity determination after 24 hours of continued culture at 37 ℃. the relative luciferase activity (RLU) after treatment of the cells with each compound was plotted as the Log2 value of luciferase in the treated sample divided by DMSO control cells.

Results referring to FIG. 1, according to FIG. 1, acevalosin significantly inhibited c-Maf-driven luciferase activity.

Example 2

To further analyze the inhibitory effect of Acetovalosin (AVT) on the oncogene transcription factor c-Maf, the present invention first transfected Otub1, HA-c-Maf, pMARE.Luci. plasmid and the internal control β -Gal in HEK293T cells. After 24 hours, the cells are inoculated into a 96-well cell culture plate at the density of 10000 cells per well, after the cells are attached to the wall, the acetovalacilin with different concentrations is added respectively, the cells are collected after being cultured for 24 hours at 37 ℃ for luciferase activity determination, and the activity of the acetovalacilin for inhibiting luciferase is analyzed.

Furthermore, after the multiple myeloma cell strains (RPMI-8226 and LP1) are treated by different concentrations of acevalosin (0 mu M, 1 mu M, 2 mu M and 4 mu M, medicines are prepared by 100% DMSO and then diluted in an IMDM culture medium without serum, and the final concentration of DMSO is less than 0.1%), the total cell protein is extracted, and the protein level of c-Maf and the protein level of the regulated target gene thereof are detected.

Meanwhile, the AVT is used for treating cells, total RNA of the cells is extracted, and the mRNA levels of the c-Maf downstream target genes CCND2 and Intergin beta 7 are detected by a PCR instrument after the cells are subjected to reverse transcription to form cDNA.

Results referring to FIG. 2, FIG. 2 shows that the concentration-dependent inhibition of the c-Maf-controlled luciferase activity by the acevaloyl triester, acevaloyl valerianin, inhibits the level of RNA and protein transcribed from its downstream target genes. Specifically, the method comprises the following steps: FIG. 2A shows that concentration-dependent inhibition of c-Maf luciferase activity by Valerianin; FIG. 2B shows that the concentration-dependent inhibition of the protein expression of the c-Maf target gene by Valerianin; FIG. 2C shows that the concentration-dependent inhibition of transcription of the target gene of C-Maf by acetovalacilin.

Example 3

To analyze the inhibitory effect of Acetovalosin (AVT) on the oncogene transcription factor STAT3, the STAT 3-specific recognition unit-driven luciferase plasmid (pSTAT3-Luc) and the internal control β -Gal plasmid were transfected in HEK293T cells. After 24 hours, the cells are inoculated into a 96-well cell culture plate at the density of 10000 cells per well, and after the cells are attached to the wall, the acetovalacilin with different concentrations is added respectively. After further incubation at 37 ℃ for 24 hours, the cells were harvested for luciferase activity assay and the activity of the luciferase was inhibited by the acetovalosin. In addition, after the multiple myeloma cell line LP1 was treated with different concentrations of acevalosin (0, 1, 2, 4. mu.M) or with interleukin 6(IL-6) or STATIC, an inhibitor STATIC 3, for 24 hours, total cell protein was extracted, and the JAK2, p-STAT3 and c-Maf protein levels were detected using Western blot.

The results are shown in FIG. 3, specifically: the results show that the acevalosin significantly inhibits STAT 3-driven luciferase activity and inhibits the JAK2/STAT3 signaling pathway. From FIG. 3A, it can be seen that the concentration-dependent inhibition of the concentration of valacilin by the luciferase activity of the transcription factor STAT 3; from FIG. 3B, it is seen that acetovalacilin significantly inhibited the JAK2/STAT3 signaling pathway.

Example 4

In order to elucidate the mechanism of the acevalosin to inhibit the c-Maf cancer transcriptional activity, the invention firstly treats multiple myeloma LP1 and RPMI-8226 cells with acevalosin with different concentration gradients (0 muM, 1 muM and 2 muM) for 24 hours, then collects the cells, adds c-Maf antibody and Protein A + G agarose beads in sequence for incubation so as to carry out immunoprecipitation on c-Maf, then carries out elution to remove non-specific binding Protein, and finally determines the polyubiquitination level of c-Maf by Western blot.

Further, the present invention treated brain tumor U251 and HS683 cells with different concentrations of acetovalosin concentration gradient (0. mu.M, 1. mu.M, 2. mu.M) for 24 hours. Then collecting cells, adding a cyclin CCND1 antibody and Protein A + G agarose beads in sequence for incubation to carry out immunoprecipitation on CCND1, then eluting to remove non-specific binding Protein, and finally determining the polyubiquitination level of CCND1 by Western blot.

In addition, the invention also analyzes the effect of the acevalant on the stability of the c-Maf and CCND1 proteins. Specifically, LP1 and RPMI-8226 cells are respectively treated with acetyl valerian (2 mu M) for 4 hours, then are treated with protein synthesis inhibitor cycloheximide (CHX, 100 mu g/mL), the cells are collected at 0 hour, 2 hours, 4 hours and 6 hours, the total cell protein is extracted, and the expression level of c-Maf is detected by Western blot.

Meanwhile, the invention treats the cells of the brain tumor U251 and the HS683 with the acetovalin (0 mu M, 2 mu M and 4 mu M) for 24 hours respectively, then collects the cells, extracts the total protein of the cells, and detects the protein level of the CCND1 by using Western Blot.

The results are shown in fig. 4, specifically: as shown in FIG. 4A, Val-acetyl significantly increased the polyubiquitination level of c-Maf and down-regulated c-Maf protein; from FIG. 4B, it can be seen that valacitin significantly increased the polyubiquitination level of CCND1 and down-regulated the CCND1 protein; as shown in FIG. 4C, it can be seen that the acevalosin significantly shortens the half-life of the C-Maf protein and promotes its degradation; from FIG. 4D, it can be seen that the concentration dependence of valacilin down-regulates the protein level of CCND 1.

Example 5

To analyze the ability of acetovalosin to induce apoptosis in multiple myeloma cells, different myeloma cells (LP1, RPMI-8226, OPM2) were plated at 50000 cells per well in 96-well plates and treated with acetovalosin (0. mu.M, 2. mu.M, 4. mu.M) for 24 hours. Then cells are collected and double-stained by Annexin V-FITC and Propidium Iodide (PI), and the proportion of Annexin V positive cells is detected by a flow cytometer after 5 minutes.

The results are shown in fig. 5, specifically: the concentration dependence of valacilin promotes apoptosis of multiple myeloma cells.

Example 6

In order to analyze the ability of the acetovalerian to induce the apoptosis of brain tumor cells, different glioma cells (HS683, U251) in logarithmic growth phase are taken, 50000 cells are paved on a 96-well plate in each well, the acetovalerian is added after the cells are attached to the wall, the cells are collected after incubation and culture for 24 hours at 37 ℃, the cells are double-stained by Annexin V-FITC and Propidium Iodide (PI), and the proportion of Annexin V positive cells is detected by a flow cytometer.

Results see fig. 6, specifically: the concentration dependence of acetovalosin promotes apoptotic cells of glioma cells.

Example 7

To further analyze the anti-tumor ability of the acevalosin in vivo, 5-6-week-old female Balb/C nude mice (provided by Shanghai Slac animal center) were selected, and after one week of quarantine and quarantine, subcutaneous myeloma cells LP1 and RPMI-8226 (3X 10) were performed at the medial proximal vessel of the left hind limb⁷Cell/cell). When the tumor grew to a palpable state (about 5-10mm in diameter), the tumorigenic nude mice were randomly divided into two groups (control group, administration group). The experimental group was administered 50mg/kg of acetylvaleric (AVT, solvent dissolved in PBS and sesame oil at a volume ratio of 1: 1) daily, the control group was administered the same volume of solvent continuously for 15 days, and changes in tumor volume were measured daily while monitoring changes in body weight of the mice.

The results are shown in fig. 7, specifically: FIG. 7A is a graph showing the relationship between the tumor volume and the number of days of administration in mice, and FIG. 7B is a graph showing the relationship between the body weight and the number of days of administration in mice. As can be seen from fig. 7: the body weights of the mice of the experimental group and the control group are not obviously changed; the Valerianin significantly inhibited the growth of tumors in two tumor-bearing mice, and the differences of the tumor volumes of the Valerianin intervention group compared with the control group were statistically significant (p < 0.01;. p < 0.001).

Example 8

To further evaluate the effect of acetovalgenin on tumor cells in patients, bone marrow cells from healthy and myeloma patients were collected, mononuclear cells were extracted from blood after isolation from lymphocyte isolates, cells were seeded in soft agar with the addition of acetovalgenin, and colonies of greater than 50 cells were counted microscopically after 1-2 weeks of culture.

The results are shown in FIG. 8, and it can be seen from FIG. 8 that the number of colonies in healthy persons and myeloma patients is substantially unchanged and the number of colonies in myeloma patients is significantly reduced after the addition of acevalosin. The acetovalerian is suggested to be capable of effectively inhibiting the proliferation and cloning of myeloma cells of myeloma patients.

Example 9

Collecting and culturing lung cancer cells (A549 and H1299), prostate cancer cells (PC3 and DU145) and oral cancer cells (SCC9 and Cal27), treating the cells with acetovalerian and/or cisplatin for 24 hours, collecting the cells, extracting proteins, and analyzing the apoptosis proteins of the cells by Western Blot.

The results are shown in FIG. 9. Specifically, the method comprises the following steps: under the condition of lower dosage, the combined use of the acevalosin and the cisplatin can obviously promote the expression of apoptosis protein of tumor cells, and the apoptosis level of the cells is greatly improved.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. Application of acevalerian in preparing medicine for preventing and/or treating tumor is disclosed.

2. The use of claim 1, wherein the tumor is myeloma, brain tumor, leukemia, hodgkin's disease, non-hodgkin's lymphoma, B-cell lymphoma, T-cell lymphoma, fahrenheit macroglobulinemia, myeloproliferative syndrome, myelodysplastic syndrome, lung cancer, liver cancer, prostate cancer, oral cancer, or breast cancer.

3. The use of claim 2, wherein the myeloma is multiple myeloma; or/and the brain tumor is glioma.

4. The use according to any one of claims 1 to 3, wherein the medicament comprises the acevalosin and a pharmaceutically acceptable excipient.

5. The use according to claim 4, wherein the medicament comprises from 0.1% to 99.5% by weight of the acevalant.

6. The use according to claim 4, wherein the adjuvant is selected from at least one of diluents, wetting agents, binders, disintegrants, lubricants, color and flavor modifiers, solvents, solubilizers, cosolvents, suspending agents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, topical analgesics, pH modifiers, and isotonic or isotonic modifiers.

7. The use according to claim 6, wherein the diluent is at least one selected from the group consisting of starches, sugars, celluloses, and inorganic salts; or/and the wetting agent is selected from at least one of water and ethanol; or/and the adhesive is selected from at least one of starch slurry, dextrin, sugar, cellulose derivative, gelatin, povidone and polyethylene glycol; or/and the disintegrant is selected from at least one of starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, croscarmellose sodium, crospovidone, surfactant and effervescent disintegrant; or/and the lubricant is selected from at least one of talcum powder, calcium stearate, magnesium lauryl sulfate, superfine silica powder and polyethylene glycol; or/and the color, aroma and taste regulator is selected from at least one of pigment, spice, sweetener, mucilage and flavoring agent; or/and the solvent is at least one selected from water, oil, ethanol, glycerol, propylene glycol, polyethylene glycol, dimethyl sulfoxide, liquid paraffin, fatty oil and ethyl acetate; or/and the solubilizer is selected from at least one of tween, maize, polyoxyethylene fatty alcohol ether, soap, sulfate and sulfonate; or/and the cosolvent is at least one selected from organic acids and salts thereof, amides and amine compounds, inorganic salts, polyethylene glycol, povidone and glycerol; or/and the emulsifier is selected from at least one of span, tween, maize, beneze, glycerin fatty acid ester, higher fatty acid salt, sulfate, sulfonate, acacia, tragacanth, gelatin, pectin, phospholipid, agar, sodium alginate, hydroxide, silicon dioxide and bentonite; or/and the suspending agent is at least one selected from glycerol, syrup, acacia gum, tragacanth gum, agar, sodium alginate, cellulose derivatives, povidone, carbopol, polyvinyl alcohol and thixotrope; and/or, the antioxidant is at least one selected from sulfite, pyrosulfite, bisulfite, ascorbic acid, gallic acid and esters thereof; or/and the metal complexing agent is selected from at least one of disodium ethylene diamine tetraacetate and polycarboxylic acid compound; or/and the inert gas is selected from at least one of nitrogen and carbon dioxide; or/and the preservative is at least one of nipagin, organic acid and salt thereof, quaternary ammonium compound, chlorhexidine acetate, alcohol, phenol and volatile oil; or/and the local analgesic is selected from at least one of benzyl alcohol, chlorobutanol, lidocaine and procaine; or/and the pH regulator is at least one selected from hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, tartaric acid, acetic acid, sodium hydroxide, sodium bicarbonate, ethylenediamine, meglumine, phosphate, acetate and citrate; and/or, the isotonic or isotonic regulator is at least one selected from glucose, sodium chloride, sodium citrate, sorbitol and xylitol.

8. The use according to any one of claims 1 to 3 and 5 to 7, wherein the medicament is in the form of tablets, granules, pills, powders, capsules, injections, oral liquids, ophthalmic preparations or external preparations; and/or the first and/or second light sources,

the administration route of the medicine comprises intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration or transdermal administration.

9. A combined medicine composition for preventing and/or treating tumors is characterized in that the active ingredients of the combined medicine composition comprise at least one of an anti-tumor medicine and an immunomodulator and acetovalerian.

10. The combination composition for the prevention and/or treatment of tumor according to claim 9, wherein the antitumor drug is at least one selected from cisplatin and adriamycin; or/and the immunomodulator is ranitidine.