CN105837660A - Preparation method and anticancer use of novel antibacterial peptide - Google Patents
Preparation method and anticancer use of novel antibacterial peptide Download PDFInfo
- Publication number
- CN105837660A CN105837660A CN201510057977.1A CN201510057977A CN105837660A CN 105837660 A CN105837660 A CN 105837660A CN 201510057977 A CN201510057977 A CN 201510057977A CN 105837660 A CN105837660 A CN 105837660A
- Authority
- CN
- China
- Prior art keywords
- compound
- formula
- preparation
- formulas
- antibacterial peptide
- 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.)
- Pending
Links
Abstract
The invention discloses a novel antibacterial peptide and a preparation method thereof. The invention also discloses the anticancer use of the novel antibacterial peptide.
Description
Technical field
The invention belongs to pharmaceutical field, invention describes class novel anti-bacterial peptide and preparation method thereof, the present invention also takes off
Show described antibacterial peptide purposes in terms of prophylactic treatment cancer.
Background technology
Antibacterial peptide (Peptaibols) is a class natural chain oligopeptide, has three kinds of exemplary chemical structures features, including: contain
There are a high proportion of nonprotein amino acid residue or fat propylhomoserin, are especially enriched in α-aminoacid (Aib)；There is alkylating N
End and hydroxylated C-terminal；Having linear alpha-helix, molecular weight is mostly 500～2000D, and molecular length is at 5～20 residues
Between.Because it has amphipathic-helical architectural feature, therefore can optionally be incorporated into cell membrane and form cross-film duct, destroy cell
Film integrality, causes intracellular organic matter outflow to cause cell death (Chugh J K, et
al.Biochem.Sco.Trans.2001；Degenkolb T, et al.Chem.Biodivesity 2007).Antibacterial peptide has
Multiple biological activity, as the various biological such as antibacterial, antiviral, nematicide, inducing apoptosis of tumour cell, inducing plant resistance is lived
Property (Daniel J F, et al.Nat.Prod.Rep.2007), is a kind of important active skull cap components.
Most antibacterial peptides come from land microorganism, are primarily present in fungus Trichoderma.Inventor passes through
Early-stage Study, is found that the antibacterial peptide that a class formation is brand-new first from deep-sea bacterium.Lived by In Vitro Anti human tumor cell line
Property screening, find its have significant inhibiting tumour cells activity, have further medicinal study exploitation potentiality.
Summary of the invention
An object of the present invention is to provide two brand-new antibacterial peptides.
The two of the purpose of the present invention are to provide the preparation method of such antibacterial peptide.
The three of the purpose of the present invention are to disclose the anticancer purpose of above-mentioned novel antimicrobial peptide.
Compound shown in first aspect present invention offer Formulas I～Formula II:
Second aspect present invention provides the preparation method of above-claimed cpd, comprises the steps:
1) with deep-sea bacterium Microbacterium sediminis sp.nov.YLB-01TAs engineering strain, pass through
Liquid fermentation and culture, it is thus achieved that the fermented product containing above-claimed cpd；
2) fermented product extraction is concentrated, by silica gel, gel, HPLC isochromatic spectrum means isolation and purification, from strain fermentation
Formula I, Formula II compound in thing extractum；
Third aspect present invention originally have rated above-mentioned Formulas I by MTT method, the human tumor cells suppression of Formula II compound is lived
Property.Test result indicate that Bel-7402, BGC-823 and A549 three-type-person's tumor cell line is had by compound of formula I significantly to press down
Make and use；Five kinds of human tumor cell lines of HCT-8, Bel-7402, BGC-823, A549 and A2780 are had bright by Formula II compound
Aobvious inhibitory action.
Accompanying drawing explanation
The high resolution mass spectrum of Fig. 1: compound of formula I；
The Pyrolysis Mass Spectrometry of Fig. 2: compound of formula I m/z 1166 fragment；
The Pyrolysis Mass Spectrometry of Fig. 3: compound of formula I m/z 712 fragment；
Fig. 4: the high resolution mass spectrum of Formula II compound；
Fig. 5: the Pyrolysis Mass Spectrometry of Formula II compound m/z 1180 fragment；
Fig. 6: the Pyrolysis Mass Spectrometry of Formula II compound m/z 712 fragment；
Specific embodiments:
According to technology contents disclosed in this invention, those skilled in the art will be better understood when this patent essence, following
Embodiment only makees example.
Example 1: deep-sea bacterium Microbacterium sediminis sp.nov.YLB-01TLiquid fermentation and culture
1. strain background
Microbacterium sediminis sp.nov.YLB-01TBeing isolatable from the Indian Ocean, south trench, bacterial strain size is
0.4-0.7 × 0.8-1.7 μm, optimum growth temp 28 DEG C, optimum growh pH value 7.0, culture presevation is in Chinese Sea microorganism
DSMZ (MCCC:1A06153T)。
2. liquid culture matrix manufacturing:
Soluble starch 2g, KNO30.1g, MgSO40.05g, K2HPO40.05g, FeSO40.001g, Urea 0.03g, molten
In 80mL deionized water, adjust pH value 6.8.
3. strain fermentation:
By Microbacterium sediminis sp.nov.YLB-01TStrain is inoculated in the triangle containing above-mentioned culture medium
In Ping, at 28 DEG C, lucifuge constant temperature culture 12d, shaking speed 120r/min, obtain tunning.
Example 2: the preparation of compound
Fermentation liquid 40L ethyl acetate ultrasonic extraction 3 times, each 1.5h, concentrating under reduced pressure obtains crude extract (4.2g).Extractum is used
By ODS column chromatography with methanol/water for flowing phase (10%～100%, v/v) gradient elution, 90% elution fraction (1.1g) is through half
Preparative high-performance liquid chromatographic is with 84% methanol/water for flowing phase Gradient elution, and purification obtains compound of formula I (23.4mg) and formula
II compound (11.8mg).
Table 1. compound of formula I1H NMR,13C NMR data belongs to
AHMA=3-amino-2-hydroxy-3-methylbutanoic acid；
AE=aminoethanol
Table 2. Formula II compound1H NMR,13C NMR data belongs to
Formulas I, Formula II compound mass spectrometric data with reference to accompanying drawing illustrate in Fig. 1～Fig. 6.
Example 3: Formulas I, the antitumor activity of Formula II compound:
Mtt assay is used to test Formulas I and Formula II compound to HCT-8, Bel-7402, BGC-823, A549 and A2780
The cytotoxicity of five kinds of human tumor cell lines, concretely comprises the following steps:
1. inoculating cell: be made into individual cells suspension with obtaining culture fluid containing 10% tire calf serum, with every hole 1000-
10000 cells are inoculated into 96 orifice plates, every pore volume 200 μ L.
2. cultivate cell: with general condition of culture, cultivate 3-5 days (when can determine to cultivate according to test objective and requirement
Between).
3. colour generation: after cultivating 3-5 days, every hole adds MTT solution (5mg/ml PBS prepares, pH=7.4) 20 μ L. and continues to incubate
Educating 4h, termination is cultivated, and careful suction abandons culture supernatant in hole, inhales again and abandon culture supernatant in hole after needing to be centrifuged for suspension cell
Liquid.
4. every hole adds 150 μ L DMSO, and vibrate 10min, makes crystal fully melt.
5. colorimetric: select 490nm wavelength, measure each hole absorbance value on enzyme linked immunological monitor, records result, with time
Between be abscissa, light absorption value be vertical coordinate draw cell growth curve.
Table 3. Formulas I, the human tumor cell line inhibitory activity of Formula II compound
Test result indicate that: Formulas I, Formula II compound have good anti-tumor activity.
Claims (3)
1. a Formulas I and Formula II compound
2. the preparation method of the compound described in claim 1, comprises the steps:
1) with marine bacteria Microbacterium sediminis sp.nov.YLB-01TFor strain, liquid fermentation is utilized to train
Support, it is thus achieved that the fermented product containing above-claimed cpd；
2) by fermented product by silica gel, gel, high-efficient liquid phase isochromatic spectrum means isolation and purification, in same fermented product in preparation
State compound.
3. compound shown in Formulas I and Formula II, its pharmaceutically acceptable salt or the purposes of their mixture, be used for preparing medicine
Thing, described medicine is used for preventing or treating cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510057977.1A CN105837660A (en) | 2015-02-03 | 2015-02-03 | Preparation method and anticancer use of novel antibacterial peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510057977.1A CN105837660A (en) | 2015-02-03 | 2015-02-03 | Preparation method and anticancer use of novel antibacterial peptide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105837660A true CN105837660A (en) | 2016-08-10 |
Family
ID=56580213
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510057977.1A Pending CN105837660A (en) | 2015-02-03 | 2015-02-03 | Preparation method and anticancer use of novel antibacterial peptide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105837660A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110170045A (en) * | 2019-06-28 | 2019-08-27 | 福州大学 | Application of the antibacterial peptide Lchamp2-3 in anti-tumour cell proliferative drug |
-
2015
- 2015-02-03 CN CN201510057977.1A patent/CN105837660A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110170045A (en) * | 2019-06-28 | 2019-08-27 | 福州大学 | Application of the antibacterial peptide Lchamp2-3 in anti-tumour cell proliferative drug |
| CN110170045B (en) * | 2019-06-28 | 2020-10-09 | 福州大学 | Application of antibacterial peptide Lchamp2-3 in anti-tumor cell proliferation medicine |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ebrahim et al. | Metabolites from the fungal endophyte Aspergillus austroafricanus in axenic culture and in fungal–bacterial mixed cultures | |
| Eltamany et al. | New antibacterial xanthone from the marine sponge-derived Micrococcus sp. EG45 | |
| Fang et al. | A new isocoumarin from metabolites of the endophytic fungus Alternaria tenuissima (Nees & T. Nees: Fr.) Wiltshire | |
| Wu et al. | N-carboxamido-staurosporine and selina-4 (14), 7 (11)-diene-8, 9-diol, new metabolites from a marine Streptomyces sp. | |
| Khamthong et al. | An antibacterial cytochalasin derivative from the marine-derived fungus Diaporthaceae sp. PSU-SP2/4 | |
| Wu et al. | Cytotoxic metabolites from Perenniporia tephropora, an endophytic fungus from Taxus chinensis var. mairei | |
| Luo et al. | Cytochalasins from mangrove endophytic fungi Phomopsis spp. xy21 and xy22 | |
| Montenegro et al. | Cytotoxic activity of fungal strains isolated from the ascidian Eudistoma vannamei | |
| Rodrigues et al. | Bioprospection of cytotoxic compounds in fungal strains recovered from sediments of the Brazilian coast | |
| Nasfi et al. | Soil bacteria isolated from tunisian arid areas show promising antimicrobial activities against gram-negatives | |
| Buttachon et al. | Secondary metabolites from the culture of the marine sponge-associated fungi Talaromyces tratensis and Sporidesmium circinophorum | |
| Pore et al. | Production, purification, identification of prodigiosin from Serratia sp. and its antimicrobial activity | |
| Atalla et al. | Production of some biologically active secondary metabolites from marine-derived fungus Varicosporina ramulosa | |
| Liu et al. | Antimicrobial potential of endophytic fungi from Astragalus chinensis | |
| CN115595343A (en) | Active secondary metabolite in cherry tomato rhizosphere soil microorganism and preparation and application thereof | |
| Oktiansyah et al. | Antibacterial and Antioxidant Activity of Endophytic Fungi Extract Isolated from Leaves of Sungkai (Peronema canescens) | |
| CN105837660A (en) | Preparation method and anticancer use of novel antibacterial peptide | |
| CN111533777B (en) | Method for preparing steroid compound from mangrove endophytic fungi | |
| Kim et al. | Penidioxolanes A and B, 1, 3-dioxolane containing azaphilone derivatives from marine-derived Penicillium sp. KCB12C078 | |
| Zhang et al. | Antimicrobial and cytotoxic activity of endophytic fungi from lagopsis supina | |
| Huang et al. | Antimicrobial compounds from the endophytic fungus Fusarium sp. Ppf4 isolated from the medicinal plant Paris polyphylla var. yunnanensis | |
| CN103724290A (en) | Cyclopeptide compound clavatustide A as well as producing strain, preparation method and application thereof | |
| CN109576174A (en) | Bacillus subtilis Bacillus subtilis CS30 and its application | |
| CN115806881A (en) | Penicillium fungus and application thereof in preparation of antibacterial drugs | |
| Thumar et al. | Antimicrobial potential and metabolite profiling of marine actinobacteria |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160810 |