CN111494391B - Application of buxine in preparation of autophagy inducer - Google Patents
Application of buxine in preparation of autophagy inducer Download PDFInfo
- Publication number
- CN111494391B CN111494391B CN202010554875.1A CN202010554875A CN111494391B CN 111494391 B CN111494391 B CN 111494391B CN 202010554875 A CN202010554875 A CN 202010554875A CN 111494391 B CN111494391 B CN 111494391B
- Authority
- CN
- China
- Prior art keywords
- autophagy
- buxine
- preparation
- application
- cancer cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses application of buxine in preparation of an autophagy inducer, which is researched and found that the buxine can induce autophagy of cancer cells and block autophagy flux in lysosomes, and further research and found that both lysosome autophagy and autophagy blocking induced by the buxine can be recovered through a BAFA 1V-ATPase inhibitor, so that the buxine can be used for autophagy-related diseases or building autophagy models.
Description
Technical Field
The invention relates to the field of biological medicines, in particular to application of buxine in preparation of autophagy inducers.
Background
Autophagy, or Autophagy, is a process involving the breakdown of the cell's own structure by lysosomal mechanisms, which is tightly regulated and helps maintain a state of equilibrium between cellular products during synthesis, degradation, and subsequent cycling. After the cells receive the autophagy-inducing signal, a flat "liposome" -like membrane structure called Phagophore is formed somewhere in the cytoplasm. With the extension of Phagophore, the components such as organelles in the cytoplasm are completely wrapped to form a closed structure, which is called "autophagosome (autophagosome)". After autophagosome formation, it can fuse with lysosomes, the contents of autophagosome are degraded, the products (amino acids, fatty acids, etc.) are transported into cytoplasm for cell reuse, and the residues are either discharged outside the cell or retained in the cytoplasm.
Researchers have focused on the regulation of autophagy and various vital activities, in addition to exploring the molecular mechanisms of autophagy initiation and termination. It has been found that autophagy is closely associated with tumors, immunotherapy, modulation of inflammatory signaling pathways, neurodegenerative diseases, immunochemotherapy and radiotherapy, epigenetics, drug therapy and drug resistance. Autophagy plays an important role in various life activities, such as studying how autophagy participates in the development of tumors, how autophagy participates in the drug resistance and recurrence metastasis of tumors, how autophagy participates in the effect of tumor immunotherapy, how autophagy participates in inflammatory reaction, how autophagy participates in the development and treatment of autism and alzheimer's disease, and the like. The following autophagy inducers and autophagy blockers are widely used in the market:
1. autophagy inducer: (a) Bredeldin A/Thapsiggin/Tunicamycin: simulating endoplasmic reticulum stress; (b) Carbamazepine/L-690, 330/Lithium Chloride: an IMPase inhibitor (i.e., inositol monophosphatase); (c) Earle's balanced salt solution: making hunger; (D) N-Acetyl-D-sphingosine (C2-ceramide): class I PI3K Pathway inhibitors; (e) Rapamycin: an mTOR inhibitor; f) Xestospongin B/C: IP3R blockers, and the like.
2. Autophagy blocking agent: (a) 3-Methylladenine (3-MA): (Class III PI 3K) hVps34 inhibitors; (b) Bafilomycin A1: a proton pump inhibitor; (c) And lysosomal inhibitors Hydroxychloroquine (Hydroxychloroquine), and the like. The above agents exert different functions through different target sites at different autophagy stages (autophagosome formation stage, autophagosome and lysosome fusion stage, autophagososome degradation stage) in the autophagy pathway. However, the autophagy pathway is extremely complex, and inhibitors with more definite action periods and target sites are required to be further developed and applied.
The treatment of traditional Chinese medicine is always an attractive subject in the medical field. The Chinese medicine monomer has the advantages of clear molecular formula, clear action mechanism and the like, and gradually becomes a new favorite of medicines for treating various diseases. The buxine as a common medicament for human bodies has small toxic and side effects and is widely applied to the treatment of cardiovascular diseases. However, no report is found on the study of buxus sinica base on lysosome autophagy.
Disclosure of Invention
In view of the above, the present invention aims to provide buxine as an autophagy inducer; the second purpose of the invention is to provide the application of buxine in the preparation of drugs for treating autophagy-related diseases; the invention also aims to provide the application of buxine in the preparation of medicaments for enhancing the activity of V-ATPase; the fourth purpose of the invention is to provide the application of buxine in the preparation of drugs for constructing cell autophagy models; the fifth purpose of the invention is to provide the application of buxine in the preparation of drugs for inducing autophagy block of cancer cells by autophagy blocker.
In order to achieve the purpose, the invention provides the following technical scheme:
1. application of buxine in preparing cell autophagy inducer is provided.
Preferably, the autophagy-inducing agent is a lysosomal autophagy-inducing agent.
Preferably, the cell is a cancer cell.
Preferably, the cancer cell is a skin cancer cell, a liver cancer cell, a glioma cell, a colorectal cancer cell or a gastric cancer cell.
2. Application of buxine in preparing medicine for treating autophagy related diseases is provided. The autophagy-related diseases comprise gastric cancer, skin cancer, liver cancer, glioma and colorectal cancer.
3. Application of buxine in preparing medicine for enhancing activity of V-ATPase is provided.
4. Application of buxine in preparation of drugs for constructing autophagy model of cells.
5. Application of buxine in preparing medicine for inducing autophagy of cancer cells by autophagy blocker.
Preferably, the cancer cell is a skin cancer cell, a liver cancer cell, a glioma cell, a colorectal cancer cell or a gastric cancer cell.
The invention has the beneficial effects that: the invention proves that buxine can induce cancer cells to generate lysosome autophagy and induce late autophagy blocking for the first time, and the blocking mechanism of buxine promotes the activity of V-ATPase to cause the pH value of lysosome to be reduced and lose the capacity of degrading autophagosomes. The invention also verifies the target molecule of buxine, the buxine specificity promotes the activity of V-ATPase, and the above results have important significance for clinical medication.
Drawings
In order to make the object, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:
FIG. 1 shows the drug effect test (A: buxine structural formula; B: morphological analysis of buxine added with different concentrations in gastric cancer cells MKN-45 and SGC-7901).
FIG. 2 is an electron micrograph of gastric cancer cells treated with buxine 30 μ M for 12 hours; arrows show autophagy vesicles (autophagosomes).
FIG. 3 shows autophagy flow detection (A: lentivirus autophagy double-standard detection result; B: adenovirus autophagy double-standard detection result; C: immunofluorescence LC3B antibody detection result; and D: adenovirus autophagy single-standard detection result).
FIG. 4 shows vacuole detection-immunofluorescence lysosomal membrane protein LAMP1 antibody detection.
FIG. 5 shows the autophagy type assay (A: lysomal. Beta. -Galactosidase assay; B: immunofluorescent lysosomal Galectin3 (Galectin 3) assay).
FIG. 6 shows the lysosome pH assay (A: lysoSensor DND-160 lysosome acidity assay; B: oregon Green 488 dextran (Invitrogen, D7170) lysosome pH assay).
FIG. 7 shows that drug target detection BAFA1 (V-ATPase inhibitor) and CVB-D effect can counteract each other.
FIG. 8 shows the effect of buxine on autophagic flux of other cancer cells such as (liver, skin, glioma, colon).
Detailed Description
The present invention is further described with reference to the following drawings and specific examples so that those skilled in the art can better understand the present invention and can practice the present invention, but the examples are not intended to limit the present invention.
Example 1 Effect of buxine on gastric cancer cell autophagy
The molecular structural formula of buxine is shown as A in figure 1. Then, the cell morphology of the gastric cancer cell lines MKN-45 and SGC-7901 after treatment with buxine at concentrations of 0. Mu.M, 20. Mu.M, 40. Mu.M, 80. Mu.M and 120. Mu.M was observed, and the results are shown as B in FIG. 1. According to morphological observation, a large amount of vacuoles are generated, and buxun base is supposed to induce autophagy.
The results of electron microscopy of gastric cancer cells treated with buxine 30 μ M for 12 hours are shown in fig. 2. The results show that buxine can induce the generation of gastric cancer cell autophagy.
To further investigate the type of autophagy, immunofluorescence experiments were performed. The detection results are shown in fig. 3, which are respectively detected by a lentivirus dual-label system, an adenovirus dual-label system, an immunofluorescence LC3B antibody and an adenovirus single-label system. The results indicate that buxine induces autophagy in gastric cancer cell lines. Vacuole detection-immunofluorescence lysosome membrane protein LAMP1 antibody detection results are shown in FIG. 4, and show that the autophagy flow is blocked by late autophagy.
Example 2 investigation of Buxus sinica Diels alkali-induced autophagy types
The results according to figure 4 show that the autophagosomes are highly coincident with the lysosomal membrane and that the vacuoles present in the cells are mostly able to fuse with the lysosomal membrane. Therefore, it is assumed that the lysozymes are changed, which in turn leads to lysosome autophagy. The lysosomal galactosidase was then subsequently tested and the results showed a significant decrease in galactosidase activity on the lysosomes, as shown in a in figure 5. We subsequently detected lactoglobulin 3 by immunofluorescence and the results are shown in fig. 5B. The results show that the buxus base-induced autophagy type is lysosome autophagy.
Example 3 study of Buxus sinica alkali-induced autophagy blockade mechanism
In order to explore the autophagy blocking mechanism, the emphasis is focused on lysosomes, and through the detection of the pH value of the lysosomes, buxun alkali is found to be capable of inducing the pH value to be reduced, and the result is shown as A in figure 6. Further we examined the activity of the proton pump V-ATPase of lysosomes. The activity was found to be enhanced, and the results are shown in FIG. 6, panel B. Further we used the V-ATPase inhibitor BAFA1, and found that both inhibitors were able to counteract each other their separately induced autophagy phenomena, the results are shown in fig. 7. The above results further demonstrate that buxine is induced by lysosomal autophagy blockade via increased activity of V-ATPase.
Example 4 Effect of buxine on autophagy of other tumors
In order to investigate the effect of buxine on autophagy of other tumors, we tested autophagy flux of liver cancer Huh-7, skin cancer a375, glioma LN229 and colon cancer HCT-116 using a lentivirus dual-marker system, and found that buxine (30 μ M) can induce autophagy flux blockade in other cancer cells as well, and the results are shown in fig. 8.
In conclusion, the research shows that buxine can be used as a broad-spectrum autophagy inducer to induce autophagy. Meanwhile, buxine can be used as an autophagic flow blocker after autophagy occurs, so that the degradation pathway of autophagosomes is cut off, and autophagy blocking of cancer cells is induced.
The above-mentioned embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of the present invention is not limited thereto. The equivalent substitution or change made by the technical personnel in the technical field on the basis of the invention is all within the protection scope of the invention. The protection scope of the invention is subject to the claims.
Claims (1)
1. The application of buxine in the preparation of drugs for inducing autophagy block of cancer cells by autophagy flow blocker is characterized in that: the cancer cell is skin cancer cell, and the buxine has a structural formula
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010554875.1A CN111494391B (en) | 2020-06-17 | 2020-06-17 | Application of buxine in preparation of autophagy inducer |
| PCT/CN2021/111154 WO2021254537A1 (en) | 2020-06-17 | 2021-08-06 | Use of cyclovirobuxine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010554875.1A CN111494391B (en) | 2020-06-17 | 2020-06-17 | Application of buxine in preparation of autophagy inducer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111494391A CN111494391A (en) | 2020-08-07 |
| CN111494391B true CN111494391B (en) | 2023-03-17 |
Family
ID=71865583
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010554875.1A Active CN111494391B (en) | 2020-06-17 | 2020-06-17 | Application of buxine in preparation of autophagy inducer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111494391B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021254537A1 (en) * | 2020-06-17 | 2021-12-23 | 西南大学 | Use of cyclovirobuxine |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103450314A (en) * | 2013-09-03 | 2013-12-18 | 沈阳药科大学 | Plant extract with anti-tumor activity and active ingredients thereof |
| CN104906111A (en) * | 2015-06-25 | 2015-09-16 | 黑龙江中医药大学 | Pharmaceutical composition prepared from hydroxytyrosol for treating liver injury and preparation process of pharmaceutical composition |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE20214753U1 (en) * | 2002-09-24 | 2002-12-12 | Hando Handels Ges M B H | Pharmaceutical composition for the treatment and / or prevention of light eruption |
| CN103288912A (en) * | 2012-03-04 | 2013-09-11 | 上海壹志医药科技有限公司 | Salts of buxus microphylla base derivative |
| CN104877127B (en) * | 2015-06-23 | 2017-11-10 | 厦门赛诺邦格生物科技股份有限公司 | A kind of eight arms polyethyleneglycol derivative, preparation method and its bio-related substance of modification |
-
2020
- 2020-06-17 CN CN202010554875.1A patent/CN111494391B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103450314A (en) * | 2013-09-03 | 2013-12-18 | 沈阳药科大学 | Plant extract with anti-tumor activity and active ingredients thereof |
| CN104906111A (en) * | 2015-06-25 | 2015-09-16 | 黑龙江中医药大学 | Pharmaceutical composition prepared from hydroxytyrosol for treating liver injury and preparation process of pharmaceutical composition |
Non-Patent Citations (3)
| Title |
|---|
| Cyclovirobuxine D Inhibits Cell Proliferation and Induces Mitochondria-Mediated Apoptosis in Human Gastric Cancer Cells;Jie Wu et al.，;《Molecules》;20151119;第20卷(第11期);第20659-20668页 * |
| 新型天然生物碱抗癌机制研究进展;李滢等;《天然产物研究与开发》;20161231(第11期);第1850-1855页 * |
| 环维黄杨星D药理研究概况;周玖瑶;《广州中医药大学学报》;20050131;第22卷(第1期);第70-74页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111494391A (en) | 2020-08-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kozma et al. | Pseudo-anaphylaxis to polyethylene glycol (PEG)-coated liposomes: roles of anti-PEG IgM and complement activation in a porcine model of human infusion reactions | |
| Wu et al. | p53 promotes ferroptosis in macrophages treated with Fe3O4 nanoparticles | |
| Obaid et al. | Remediating desmoplasia with EGFR‐targeted photoactivable multi‐inhibitor liposomes doubles overall survival in pancreatic cancer | |
| Zhong et al. | “Plug and play” functionalized erythrocyte nanoplatform for target atherosclerosis management | |
| CN111494391B (en) | Application of buxine in preparation of autophagy inducer | |
| Meng et al. | Ferroptosis‐Enhanced Immunotherapy with an Injectable Dextran‐Chitosan Hydrogel for the Treatment of Malignant Ascites in Hepatocellular Carcinoma | |
| Wang et al. | Boosting nutrient starvation-dominated cancer therapy through curcumin-augmented mitochondrial Ca2+ overload and obatoclax-mediated autophagy inhibition as supported by a novel nano-modulator GO-Alg@ CaP/CO | |
| Chung et al. | Potentiation of anti-angiogenic activity of heparin by blocking the ATIII-interacting pentasaccharide unit and increasing net anionic charge | |
| Zhang et al. | In situ autophagy disruption generator for cancer theranostics | |
| Kudo | Changing the treatment paradigm for hepatocellular carcinoma using atezolizumab plus bevacizumab combination therapy | |
| Li et al. | Cytotoxic effect of protoporphyrin IX to human Leukemia U937 cells under ultrasonic irradiation | |
| Yu et al. | Study on glyco-modification of endostatin-derived synthetic peptide endostatin2 (ES2) by soluble chitooligosaccharide | |
| Liu et al. | Promoting cardiac repair through simple engineering of nanoparticles with exclusive targeting capability toward myocardial reperfusion injury by thermal resistant microfluidic platform | |
| TW201434497A (en) | Pharmaceutical formulation containing glycosaminoglycan | |
| Deng et al. | Metal polyphenol network/cerium oxide artificial enzymes therapeutic nanoplatform for MRI/CT-aided intestinal inflammation management | |
| Banach et al. | Synthesis and hemostatic activity of new amide derivatives | |
| Hur et al. | Multicomponent metal-organic framework nanocomposites for tumor-responsive synergistic therapy | |
| Liu et al. | New fungal protein from Pleurotus ferulae lanzi induces AMPK-mediated autophagy and G1-phase cell cycle arrest in A549 lung cancer cells | |
| Yao et al. | Modulation of glucose metabolism through macrophage-membrane-coated metal-organic framework nanoparticles for triple-negative breast cancer therapy | |
| Han et al. | Current trends and future perspectives of stroke management through integrating health care team and nanodrug delivery strategy | |
| Yang et al. | Evaluation of a tumor-targeting oligosaccharide nanosystem in BNCT on an orthotopic hepatocellular carcinoma model | |
| Hori et al. | Medicinal electronomics bricolage design of hypoxia-targeting antineoplastic drugs and invention of boron tracedrugs as innovative future-architectural drugs | |
| Bin et al. | Biodegradable silk fibroin nanocarriers to modulate hypoxia tumor microenvironment favoring enhanced chemotherapy | |
| Wen et al. | Slc9a1 plays a vital role in chitosan oligosaccharide transport across the intestinal mucosa of mice | |
| Song et al. | Improvement of TNBC immune checkpoint blockade with a microwave-controlled ozone release nanosystem |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |