CN113952373B - Application of gambogic acid in preparation of medicine for inhibiting enterococcus faecalis - Google Patents
Application of gambogic acid in preparation of medicine for inhibiting enterococcus faecalis Download PDFInfo
- Publication number
- CN113952373B CN113952373B CN202111493469.XA CN202111493469A CN113952373B CN 113952373 B CN113952373 B CN 113952373B CN 202111493469 A CN202111493469 A CN 202111493469A CN 113952373 B CN113952373 B CN 113952373B
- Authority
- CN
- China
- Prior art keywords
- acid
- gambogic acid
- enterococcus faecalis
- staphylococcus aureus
- upp
- 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
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/38—Clusiaceae, Hypericaceae or Guttiferae (Hypericum or Mangosteen family), e.g. common St. Johnswort
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Communicable Diseases (AREA)
- Organic Chemistry (AREA)
- Alternative & Traditional Medicine (AREA)
- Oncology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses antibacterial pharmacological activity of gamboge extract, which is developed through a large number of experimental screening, and the gamboge extract comprising neogambogic acid and gambogic acid shows good antibacterial activity to gram positive bacteria such as staphylococcus aureus, enterococcus faecalis, methicillin-resistant staphylococcus aureus and the like through combining with UPP synthetase and reducing the enzyme activity of the UPP synthetase. Is expected to be developed into a new antibacterial natural product, and has important application value.
Description
Technical Field
The invention belongs to the field of biological medicine, and in particular relates to application of active components of gambogic acid and gambogic acid in gamboge extract as a UPP synthetase inhibitor of targeted Yu Gelan positive bacteria.
Background
In recent years, the appearance of drug-resistant or multi-drug-resistant strains of various pathogenic bacteria such as staphylococcus aureus, enterococcus faecalis and the like seriously endangers the physical health of people, and the development of novel drug-resistant bacteria infection resistant drugs has become an urgent requirement in the field of public health. The traditional Chinese medicine is a unique resource in China, and is worthy of inheritance, development and utilization. The Chinese medicinal material gamboge is a gamboge plant secretion resin, has the effects of breaking blood and resolving hard mass, attacking toxin and corroding sore, is applied to the aspects of removing putrefaction and promoting wound healing, attacking toxin and detumescence, stopping bleeding, killing insects and the like, and is clinically used for mainly treating ulcers, wet sores, tumors, carbuncles and swelling toxins, stubborn tinea, traumatic injuries, wound bleeding and scalds. Research has reported that gambogic acid (gambogic acid) and neogambogic acid (neogambogic acid) as main components in gamboge have remarkable effects of resisting tumor-induced tumor cell differentiation and apoptosis, resisting inflammation and resisting viruses, and have the characteristics of wide antitumor spectrum and low toxicity.
The bacterial cell wall plays an important biological function in the aspects of maintaining the cell morphology, controlling the cell proliferation, resisting external damage and the like. Peptidoglycans are the main component of bacterial cell walls. In the early stages of peptidoglycan synthesis, the synthesis of the peptidoglycan precursor substance C55 isoprenoid pyrophosphoric acid is dependent on UPP synthase (undecaprenyl pyrophosphate synthase). UPP synthase catalyzes the condensation of 8 molecules of isopentenyl pyrophosphate (IPP) containing five carbon units and 1 molecule of farnesyl pyrophosphate (FPP) containing fifteen carbon units to form C55 isoprenoid pyrophosphate. The C55 isoprenoid pyrophosphoric acid participates in synthesis and transportation of cell wall peptidoglycan, also participates in synthesis of other cell wall polysaccharide, and plays an important role in growth and reproduction of bacteria. UPP synthase is therefore essential for bacterial growth. Because the genome of eukaryotes such as human beings does not contain UPP synthetase homologs, the genome is a potential drug target.
At present, no report about the application of neogambogic acid and gambogic acid with UPP enzyme activity inhibition effect on gram-positive bacteria is available, and particularly, the report about the antibacterial effect on methicillin-resistant staphylococcus aureus with drug resistance is available.
Disclosure of Invention
The invention aims to screen out new clinical application of gamboge based on the existing pharmacological activity of gamboge through a large number of experiments, and develop that gamboge extracts including gamboge acid and gamboge acid show good antibacterial activity to gram-positive bacteria such as staphylococcus aureus, enterococcus faecalis, methicillin-resistant staphylococcus aureus and the like through combining with UPP synthetase and reducing the enzyme activity of UPP synthetase.
It has been found herein that neogambogic acid and gambogic acid bind to and effectively inhibit the activity of UPP synthase, which has important biological functions on bacterial cell wall integrity. Thus, the present application provides two types of gamboge extracts which bind to the UPP synthase and effectively inhibit the activity thereof to exert antibacterial activity, and thus, the present invention screens neogambogic acid, gambogic acid and their derivatives for the preparation of inhibitors of UPP enzyme activity.
Preferably, the gamboge extract or gambogic acid and gambogic acid are prepared into tablets, granules, capsules, pills, powder, ointment, oral liquid and injection with pharmaceutically acceptable carriers.
The invention discovers that the minimum inhibitory concentration MIC of the neogambogic acid on staphylococcus aureus, enterococcus faecalis and methicillin-resistant staphylococcus aureus is 4 mug/ml, 2 mug/ml and 4 mug/ml respectively. The minimum inhibitory concentrations MIC of gambogic acid on staphylococcus aureus, enterococcus faecalis and methicillin-resistant staphylococcus aureus are 2 mug/ml, 2 mug/ml and 4 mug/ml respectively. The novel gambogic acid and gambogic acid have good antibacterial effect, and particularly have effective antibacterial activity on drug-resistant staphylococcus aureus. Staphylococcus aureus, enterococcus faecalis and methicillin-resistant staphylococcus aureus are all important pathogenic bacteria for causing clinical bacterial infection, and the invention has important clinical significance.
Drawings
FIG. 1 is a diagram showing the IC of neogambogic acid and gambogic acid against enterococcus faecalis UPP synthase 50 。
FIG. 2 is the interaction of neogambogic acid and gambogic acid with enterococcus faecalis UPP synthase.
Detailed Description
Through a large number of experimental screening, the invention discovers that the neogambogic acid and gambogic acid can inhibit the growth of staphylococcus aureus, enterococcus faecalis and methicillin-resistant staphylococcus aureus in vitro. The invention is further illustrated below in connection with specific embodiments, but these examples should not be construed as limiting the invention.
Example 1 in vitro antibacterial Activity of neogambogic acid and gambogic acid against gram-positive pathogens
Compounds tested: neogambogic acid and gambogic acid.
MIC refers to the minimum inhibitory concentration, which is an index for measuring the antibacterial activity of an antibacterial drug, and refers to the minimum drug concentration that can inhibit the growth of pathogenic bacteria in a culture medium after bacteria are cultured in vitro for 18 to 24 hours.
MIC determination method: the bacteria selected for measurement comprise staphylococcus aureus, enterococcus faecalis and methicillin-resistant staphylococcus aureus. Coli served as negative control bacteria and tetracycline and ampicillin served as positive control drugs. The test strain is from China center for type culture Collection, and the medicine is from biological engineering Co., ltd. The antibacterial activity and the minimum antibacterial concentration of the gambogic acid are measured by adopting a trace broth dilution method. And (3) picking a monoclonal colony in 5ml of LB culture medium, shaking for 12 hours at 200rpm, and continuously diluting 10000 times to obtain an experimental bacterial liquid for later use. The 96-well plate is adopted, the highest concentration of the compound to be tested (neogambogic acid or gambogic acid) in the first well is 16 mug/ml, the concentrations are sequentially diluted by times, the lowest concentration of the compound to be tested in the 6 th well is 0.5 mug/ml, the 7 th well is the bacterial liquid as a positive control, the 8 th well is added with LB culture medium as a blank negative control, and the culture is carried out for 18 hours at 37 ℃, wherein the lowest compound concentration of the bacterial liquid which is completely clear can be regarded as the lowest antibacterial concentration. The minimum inhibitory concentrations MIC of the neogambogic acid on staphylococcus aureus, enterococcus faecalis and methicillin-resistant staphylococcus aureus are 4 mug/ml, 2 mug/ml and 4 mug/ml respectively. The minimum inhibitory concentrations MIC of gambogic acid on staphylococcus aureus, enterococcus faecalis and methicillin-resistant staphylococcus aureus are 2 mug/ml, 2 mug/ml and 4 mug/ml respectively. The specific experimental results are shown in tables 1 and 2.
TABLE 1 antibacterial Activity of neogambogic acid
TABLE 2 antibacterial Activity of gambogic acid
EXAMPLE 2 Neogranic acid and gambogic acid IC for enterococcus faecalis UPP synthase 50 。
By measuring the minimum inhibitory concentration, both neogambogic acid and gambogic acid showed good antibacterial activity. Thus, further analysis of its inhibition of the target enzyme UPP synthetase activity IC 50 。
The experimental principle equation is as follows:
the upper side is the reaction formula of UPP synthetase, which acts on two substrates of isopentyl diphosphate and farnesyl diphosphate, and the binding of inhibitor to UPP synthetase affects the production of pyrophosphoric acid, and the inhibition ability can be seen by calculating the fluorescence percentage.
The specific method comprises the following steps:
the UPP synthase reaction was performed on a black 96-well microtiter plate (Corning, NY, USA). The purified UPP synthetase was added to a solution containing 50. Mu.l of the reaction mixture (100 mM Tris-HCl (pH 7.5), 0.5mM MgCl2,50mM KCl,35 which inhibits isopentyl diphosphate (IPP), 5. Mu.M farnesyl diphosphate (FPP) and 0.005% (w/v) Triton X-100) with 9mM, 8mM, 6mM, 5mM, 4mM, 2.5mM, 2mM, 1.5mM, 1.25mM, 0.4mM, 0.25mM, 0.2mM of neogambogic acid or gambogic acid, tetracycline (as a negative control). The reaction was carried out at 37℃and terminated after 30min by adding 10. Mu.l of 0.5M EDTA solution. Mu.l of the reaction mixture was transferred to a new 96-well microwell plate well and quenched with 50. Mu.l of Master reaction Mix from pyrophosphate detection kit (Sigma-Aldrich, st Louis, USA). After 30 minutes incubation at room temperature, fluorescence of the reaction mixture at 316 and 456nm was measured with FlexStation 3 (MD, USA). Calculated percent fluorescence was plotted against mixture concentration using GraphPad Prism (La Jolla, usa). Experiments were repeated at least 3 times independently. The experimental results are shown in FIG. 1.
Example 3 interaction of neogambogic acid and gambogic acid with enterococcus faecalis UPP synthase.
Using Monolith NT TM Protein labeling kit Monolith TM His-Tag Labeling Kit RED-tris-NTA2nd Generation (NanoTemper Technologies) His-tagged UPP was tagged with the dye RED-tris-NTA2nd Generation dye. Serial dilutions of unlabeled compound and 10nM of labeled UPP were mixed in binding buffer (1.8mM KH2PO4,10mM Na2HPO4,137mM NaCl,2.7mM KCl,0.05%Tween-20, ph 7.8) to a final volume of 10 μl. Subsequently, the sample was loaded into nt.115 quality coated capillary (NanoTemper Technologies). The combined experiments were performed using a Monolith NT.115Pico device (NanoTemper Technologies) with parameters of LED power 5% and MST power high. The result was obtained using MO Control software version 1.6.MO affinity analysis software version 2.3 was used to determine the fraction of complex formed. Using GraphPad Prism 7 software, assuming a specific binding site, the apparent dissociation constant (Kd) was calculated using a nonlinear fit as follows: y=bmax X/kd+x (where BMax is the maximum theoretical specific binding, where bmax=1). The experimental results are shown in FIG. 2.
Claims (2)
1. Application of gambogic acid in preparing medicine for inhibiting enterococcus faecalis is provided.
2. The use according to claim 1, wherein gambogic acid is formulated with a pharmaceutically acceptable carrier into a tablet, granule, capsule, pill, powder, paste, oral liquid or injection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111493469.XA CN113952373B (en) | 2021-12-08 | 2021-12-08 | Application of gambogic acid in preparation of medicine for inhibiting enterococcus faecalis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111493469.XA CN113952373B (en) | 2021-12-08 | 2021-12-08 | Application of gambogic acid in preparation of medicine for inhibiting enterococcus faecalis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113952373A CN113952373A (en) | 2022-01-21 |
| CN113952373B true CN113952373B (en) | 2023-04-21 |
Family
ID=79473027
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111493469.XA Active CN113952373B (en) | 2021-12-08 | 2021-12-08 | Application of gambogic acid in preparation of medicine for inhibiting enterococcus faecalis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113952373B (en) |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107648217B (en) * | 2017-09-29 | 2020-08-18 | 中国农业科学院哈尔滨兽医研究所 | Use of neogambogic acid or its derivative for preparing medicine for preventing and/or treating relevant diseases caused by bacteria |
| US20210113615A1 (en) * | 2018-04-13 | 2021-04-22 | Ludwig Institute For Cancer Research Ltd. | Heterodimeric inactivatable chimeric antigen receptors |
| WO2021168168A1 (en) * | 2020-02-20 | 2021-08-26 | University Of Florida Research Foundation, Incorporated | Identification of host-targeting modulators of bacterial uptake and assay for quantifying intracellular bacteria |
-
2021
- 2021-12-08 CN CN202111493469.XA patent/CN113952373B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN113952373A (en) | 2022-01-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Eldeen et al. | Cyclooxygenase inhibition and antimycobacterial effects of extracts from Sudanese medicinal plants | |
| CN113072611B (en) | Preparation method of glycyrrhetinic acid modified polypyridine ruthenium complex antibacterial agent | |
| KR20080064177A (en) | Method of treating clostridium difficile-associated diarrhea | |
| US11654174B2 (en) | Method of chinese herbal medicine extract used for treating multiple diseases caused by drug resistant bacteria infection | |
| CN113952373B (en) | Application of gambogic acid in preparation of medicine for inhibiting enterococcus faecalis | |
| CN101543499B (en) | Application of natural compound ursolic acid on antibiosis | |
| Ibrahim et al. | Antimicrobial Property of Water and Ethanol Extract Chlorella vulgaris: A Value-Added Advantage for a New Wound Dressing Material. | |
| CN110151752A (en) | A kind of tea polyphenols composition and its preparing the application in anti-streptococcus suis drug | |
| CN111419829B (en) | Application of honokiol in inhibiting streptococcus suis or biofilm thereof | |
| Yang et al. | Antioxidant and antimicrobial activities of the extracts from Sophora flavescens | |
| CN110870865B (en) | Application of sanguinarine in inhibition and removal of multiple-drug-resistant providencia rettgeri biofilm | |
| Hamma et al. | Phytochemical screening and antibacterial activity of the crude extract of scent leaf (Ocimum gratissimum) on Escherichia coli and Staphylococcus aureus | |
| KR102504101B1 (en) | Chrysin Derivatives and Antibacterial compositions comprising the Same | |
| CN110960545A (en) | Application of punicalagin in inhibiting growth of multidrug-resistant klebsiella pneumoniae | |
| CN104988083A (en) | Streptomyces platensis and applications of Streptomyces platensis in production of platensimycin and platencin | |
| CN116211879B (en) | Sabina alcohol and clindamycin composition and application thereof in bacteria inhibition | |
| Regmi et al. | Antidiabetic and Antimicrobial Properties of Some High Altitude Medicinal Plants of Nepal: Antidiabetic and Antimicrobial Properties of High Altitude Plants of Nepal | |
| JP2015013825A (en) | Novel compound quinofuracins, method for producing same, and application thereof, and novel microorganisms | |
| Khalaf et al. | Inhibitory effect of Cepressus sempervirens L. and Ricinus communis L. extracts against growth of clinical Staphylococcus aureus and Pseudomonas aeruginosa isolated from cutaneous infections. | |
| RU2444366C1 (en) | Antibiotic resistant therapeutic biological product for bacterial and mycotic infections | |
| CN110215446B (en) | Application of vanillic acid in inhibiting growth of multiple drug-resistant enterobacter cloacae | |
| WO2022188162A1 (en) | Endophytic fungus in ginkgo biloba and anti-bacterial and antitumor application thereof | |
| CN106673989A (en) | Pharmaceutical composition as well as preparation method and application thereof | |
| CN116211879A (en) | Sabina alcohol and clindamycin composition and application thereof in bacteria inhibition | |
| CN116196324A (en) | Composition of pyrolin and clindamycin and application thereof in bacteria inhibition |
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 |