CN110585207A

CN110585207A - Application of liensinine in preparation of medicine for preventing and treating brain glioma

Info

Publication number: CN110585207A
Application number: CN201910910031.3A
Authority: CN
Inventors: 付先军; 邵欣欣; 王振国; 王梦瑜; 乔利; 何珊; 陈聪; 任夏
Original assignee: Shandong University of Traditional Chinese Medicine
Current assignee: Shandong University of Traditional Chinese Medicine
Priority date: 2019-09-25
Filing date: 2019-09-25
Publication date: 2019-12-20

Abstract

The disclosure belongs to the technical field of development of brain glioma drugs, and particularly relates to application of liensinine in preparation of drugs for preventing and treating brain glioma. Brain glioma is a tumor with high harm to patients in intracranial tumors, and due to the existence of blood brain barrier, the traditional chemotherapy drugs have not ideal treatment effect on brain glioma, and the main treatment means is mainly surgery. Further developing chemical entities with inhibitory effects on brain gliomas can reduce the operative ratio well and reduce the pain of patients in treatment. The research disclosed by the disclosure provides the inhibition effect of liensinine on the in-vitro proliferation of brain glioma cells, provides the brain glioma inhibition activity of liensinine, and provides a research basis for the development of anti-brain glioma drugs.

Description

Application of liensinine in preparation of medicine for preventing and treating brain glioma

Technical Field

The disclosure belongs to the technical field of development of brain glioma drugs, and particularly relates to application of liensinine in preparation of drugs for preventing and treating brain glioma.

Background

The information in this background section is only for enhancement of understanding of the general background of the disclosure and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

Brain glioma is one of the brain tumors with the greatest harmfulness to patients, influences the cognitive function of the patients along with the progress of the disease, is easy to relapse, and seriously influences the life quality of the patients. According to WHO classification, gliomas can be classified into I-IV grades, wherein I and II are low-grade gliomas (LGG), and III and IV are high-grade gliomas (HGG), wherein the malignant degree of the glioblastoma multiforme is the highest, and the average survival time is only about 1 year; the 5-year survival rate is only 9.8 percent in research.

The current major treatment methods for brain glioma include surgical therapy, chemotherapy, radiotherapy, gene therapy, and the like. Conventional chemotherapy usually employs nitrosoureas, alkylating agents, etc. China has abundant Chinese herbal medicine data, and in recent years, researchers pay more attention to the application of natural medicine extracts and monomer compounds to the treatment of tumors. The known natural medicine monomer components comprise paclitaxel, triptolide, curcumin, resveratrol and the like, and can activate the apoptosis induction effect on brain glioma cells by influencing the action of a spindle body and inducing cell cycle arrest and regulating the expression content of apoptosis genes in cells; besides participating in pathways such as cell signal transduction of an apoptosis agent, the traditional Chinese medicine monomer components can also realize the inhibition effect on the brain glioma by regulating the immune function state of an organism: in vivo experiments show that the pulsatilla chinensis polysaccharide can reduce the levels of glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase and urea, activate superoxide dismutase and catalase in blood plasma, improve the indexes of thymus and spleen, and reduce the damage degree of liver and kidney.

Liensinine (Lie) is a dibenzyltetrahydroisoquinoline alkaloid extracted from plumula Nelumbinis of Nelumbonaccidae of Nelumbo. The existing research shows that liensinine has the activities of reducing blood pressure, resisting oxidation and the like. The researches on the tranquility show that the liensinine can inhibit the autophagy degradation process by inhibiting the fusion effect of autophagy corpuscles and lysosomes, and the research on the mechanism of the liensinine combined with various clinical chemotherapeutic drugs and acting on breast cancer cells verifies that the liensinine is expected to be used as an autophagy degradation inhibition drug and is used for synergistic treatment with conventional chemotherapeutic drugs.

Disclosure of Invention

Against the research background, the inventor considers that liensinine is expected to be developed into an anti-tumor drug for application, and further defines the action mechanism of liensinine to have important significance in defining the usage of liensinine. The in vitro cell activity experiment proves that the liensinine has an inhibiting effect on cell proliferation of brain glioma, and suggests that the liensinine (CAS: 2586-96-1) is expected to have a therapeutic effect on brain glioma.

Aiming at the research result, the disclosure provides the following technical scheme:

in a first aspect of the disclosure, an application of liensinine in preparing a medicament for preventing and treating brain glioma is provided.

In some embodiments of the disclosure, the disclosure demonstrates that liensinine has an inhibitory effect on the proliferation of brain glioma cells by an in vitro cytostatic assay. The method proves that the inhibition effect of liensinine on brain glioma has certain concentration dependence by setting a series of concentration gradients of liensinine. The liensinine of more than 10ug/ml shows an inhibition effect on brain glioma cells, when the liensinine concentration reaches 40ug/ml, the inhibition effect on the brain glioma cells can reach more than 80%, and the inhibition effect is obvious. The good in-vitro inhibition effect of the brain glioma cells suggests that the liensinine can be used as an active ingredient to be applied to an anti-brain glioma preparation.

In a second aspect of the present disclosure, an anti-brain glioma formulation comprising liensinine as an active ingredient.

Preferably, the preparation comprises a brain glioma inhibition model drug and an anti-brain glioma drug.

Preferably, the liensinine in the preparation is used as an active ingredient and accounts for 1-99% of the total weight of the preparation.

Preferably, the preparation also comprises a pharmaceutically acceptable carrier. And conventional adjuvants such as correctant can be further included.

Preferably, the formulations may be in the form of tablets, capsules, powders, syrups, solutions, suspensions, aerosols, etc., and may be presented in suitable solid or liquid carriers or diluents and in suitable sterile devices for injection or instillation, and prepared according to conventional methods of preparation in the pharmaceutical field.

In a third aspect of the present disclosure, there is provided a method of treatment of brain glioma comprising treatment with the anti-brain glioma formulation of the second aspect.

Compared with the prior art, the beneficial effect of this disclosure is:

brain glioma is a brain tumor disease with great harmfulness, and due to the existence of blood brain barrier, the traditional chemotherapy drugs have not ideal inhibitory effect on brain glioma. According to the application, the cell activity experiment shows that the liensinine can obviously inhibit the proliferation of the brain glioma cells, has good in-vitro inhibitory activity on the brain glioma cells, suggests that the liensinine is expected to be developed as an anti-brain glioma drug, and further develops the research and development progress of the anti-brain glioma drug.

Drawings

The accompanying drawings, which are included to provide a further understanding of the disclosure, illustrate embodiments of the disclosure and together with the description serve to explain the disclosure and are not to limit the disclosure.

FIG. 1 is a line graph showing the results of the cell inhibition experiment in example 1;

FIG. 2 is a graph showing the results of immunofluorescence observation in example 1.

Detailed Description

It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. 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 disclosure belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present disclosure. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.

As described in the background art, brain glioma is a serious intracranial tumor, and the development of a chemical monomer capable of penetrating the blood brain barrier and having good inhibitory activity is of great significance for the treatment of brain glioma. The present disclosure provides the use of liensinine as an active ingredient in the preparation of a medicament for the prevention and treatment of brain glioma.

In order to make the technical solutions of the present disclosure more clearly understood by those skilled in the art, the technical solutions of the present disclosure will be described in detail below with reference to specific examples and comparative examples.

Example 1

1) Cell inhibition assay

In this example, brain glioma U87MG was used as a research model to study the effect of liensinine on the activity of brain glioma cells. Liensinine, mother liquor 40mg/ml, cell culture was performed by diluting the solution with medium to 40ug/ml, 20ug/ml, 10ug/ml, 5ug/ml, 2.5ug/ml, (DMSO controlled at 0.1%), culturing was performed for 24 hours, MTT was measured, and liensinine was observed to inhibit cell proliferation (as shown in table 1 and fig. 1):

TABLE 1

As can be seen from the results in table 1, the brain glioma inhibitory rate gradually increased as the liensinine concentration increased. When the concentration of the liensinine reaches 40ug/ml, the inhibition rate of the tumor cells can reach more than 80 percent, and the inhibition effect is good.

2) Immunofluorescence Observation

When the cells grew to cover 60-80% of the coverslip, the experiment was started

(1) Sample adding: discarding the culture medium, adding 0.1% Lie working solution (culture medium dilution), and incubating for 2min, 4min, and 8 min; 500 ul/well; discard, add PBS and wash 1 time.

(2) Fixing: discarding PBS, adding 4% paraformaldehyde, and fixing at room temperature for 15 min; discarded and washed 1 time with PBS for 5min each (longer immobilization will affect the immunogenicity of the protein).

(3) Dyeing: alexa fluor 546 pharoids were diluted with 1% BSA and incubated at room temperature for 30min, 200ul per well; washing with PBS for 5min for 2 times; care was taken to avoid light.

(4) DAPI mounting: when DAPI is added, the cell is placed in contact with DAPI by mounting the cell-bearing plate facing down.

(5) And (4) observing results: and (5) observing and photographing under a fluorescence microscope, and recording the experimental result.

The results show (FIG. 2) that the glial cytoskeleton morphology is significantly altered and that liensinine distributes F-actin in lamellipodia and invasion pseudopodia. Liensinine was shown to inhibit migration by affecting the skeletal morphology of the cells.

The above description is only a preferred embodiment of the present disclosure and is not intended to limit the present disclosure, and various modifications and changes may be made to the present disclosure by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present disclosure should be included in the protection scope of the present disclosure.

Claims

1. Use of liensinine in preparing medicine for preventing and treating brain glioma is provided.

2. An anti-glioma formulation comprising liensinine.

3. The anti-glioma formulation of claim 2 wherein the liensinine is an active ingredient.

4. The anti-glioma formulation of claim 2 wherein the formulation comprises a brain glioma-inhibiting model drug.

5. The anti-glioma formulation of claim 2 wherein the formulation comprises an anti-glioma drug.

6. The anti-glioma formulation of claim 2 wherein the liensinine is present as an active ingredient in an amount of 1-99% by weight of the total formulation.

7. The anti-glioma formulation of claim 2 further comprising a pharmaceutically acceptable carrier.

8. The anti-glioma formulation of claim 7 further comprising a flavoring agent.

9. The anti-glioma formulation of claim 7 wherein the formulation is a tablet, capsule, powder, syrup, solution, suspension, or aerosol.

10. A method of treatment of brain glioma comprising treatment with an anti-brain glioma formulation according to any one of claims 2 to 9.