CN115025073B - 一种苯烯烃硝基类化合物作为群体感应抑制剂的非经典抗菌活性用途 - Google Patents
一种苯烯烃硝基类化合物作为群体感应抑制剂的非经典抗菌活性用途 Download PDFInfo
- Publication number
- CN115025073B CN115025073B CN202210716740.XA CN202210716740A CN115025073B CN 115025073 B CN115025073 B CN 115025073B CN 202210716740 A CN202210716740 A CN 202210716740A CN 115025073 B CN115025073 B CN 115025073B
- Authority
- CN
- China
- Prior art keywords
- methylstyrene
- nitro
- serratia marcescens
- quorum sensing
- nitro compound
- 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
- -1 benzol nitro-compound Chemical class 0.000 title claims abstract description 47
- 229940123361 Quorum sensing inhibitor Drugs 0.000 title abstract description 15
- 230000000844 anti-bacterial effect Effects 0.000 title abstract description 9
- 241000607715 Serratia marcescens Species 0.000 claims abstract description 60
- 230000018612 quorum sensing Effects 0.000 claims abstract description 41
- 230000000694 effects Effects 0.000 claims description 48
- 150000001875 compounds Chemical class 0.000 claims description 38
- 230000002401 inhibitory effect Effects 0.000 claims description 20
- 238000004519 manufacturing process Methods 0.000 claims description 16
- HCOLPNRPCMFHOH-UHFFFAOYSA-N Prodigiosin Natural products CCCCCC1C=C(C=C/2N=C(C=C2OC)c3ccc[nH]3)N=C1C HCOLPNRPCMFHOH-UHFFFAOYSA-N 0.000 claims description 9
- TWFGRJUTAULJPZ-USZBIXTISA-N prodigiosin Chemical compound N1=C(C)C(CCCCC)=C\C1=C/C1=NC(C=2[N]C=CC=2)=C[C]1OC TWFGRJUTAULJPZ-USZBIXTISA-N 0.000 claims description 9
- 239000003814 drug Substances 0.000 claims description 8
- 108091005804 Peptidases Proteins 0.000 claims description 6
- 239000004365 Protease Substances 0.000 claims description 6
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 5
- 239000004367 Lipase Substances 0.000 claims description 4
- 102000004882 Lipase Human genes 0.000 claims description 4
- 108090001060 Lipase Proteins 0.000 claims description 4
- 235000019421 lipase Nutrition 0.000 claims description 4
- 241001465754 Metazoa Species 0.000 claims description 3
- 229920002683 Glycosaminoglycan Polymers 0.000 claims description 2
- 230000000254 damaging effect Effects 0.000 claims description 2
- 230000005764 inhibitory process Effects 0.000 abstract description 40
- 241000589517 Pseudomonas aeruginosa Species 0.000 abstract description 25
- 230000006378 damage Effects 0.000 abstract description 12
- 241000193830 Bacillus <bacterium> Species 0.000 abstract description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 29
- UHOVQNZJYSORNB-UHFFFAOYSA-N monobenzene Natural products C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 18
- 241000894006 Bacteria Species 0.000 description 17
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 12
- 239000002609 medium Substances 0.000 description 11
- 239000013641 positive control Substances 0.000 description 11
- 108010025955 Pyocins Proteins 0.000 description 10
- 230000001580 bacterial effect Effects 0.000 description 10
- 239000001963 growth medium Substances 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 8
- YQUVCSBJEUQKSH-UHFFFAOYSA-N protochatechuic acid Natural products OC(=O)C1=CC=C(O)C(O)=C1 YQUVCSBJEUQKSH-UHFFFAOYSA-N 0.000 description 8
- 238000012216 screening Methods 0.000 description 8
- WKOLLVMJNQIZCI-UHFFFAOYSA-N vanillic acid Chemical compound COC1=CC(C(O)=O)=CC=C1O WKOLLVMJNQIZCI-UHFFFAOYSA-N 0.000 description 8
- TUUBOHWZSQXCSW-UHFFFAOYSA-N vanillic acid Natural products COC1=CC(O)=CC(C(O)=O)=C1 TUUBOHWZSQXCSW-UHFFFAOYSA-N 0.000 description 8
- 230000007923 virulence factor Effects 0.000 description 8
- 239000000304 virulence factor Substances 0.000 description 8
- 239000011248 coating agent Substances 0.000 description 7
- 238000000576 coating method Methods 0.000 description 7
- 239000013642 negative control Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- 230000001105 regulatory effect Effects 0.000 description 6
- 241000894007 species Species 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- CEIUIHOQDSVZJQ-UHFFFAOYSA-N 2-heptyl-3-hydroxy-4-quinolone Chemical compound C1=CC=C2C(=O)C(O)=C(CCCCCCC)NC2=C1 CEIUIHOQDSVZJQ-UHFFFAOYSA-N 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 description 4
- PHSRRHGYXQCRPU-AWEZNQCLSA-N N-(3-oxododecanoyl)-L-homoserine lactone Chemical compound CCCCCCCCCC(=O)CC(=O)N[C@H]1CCOC1=O PHSRRHGYXQCRPU-AWEZNQCLSA-N 0.000 description 4
- 241000607720 Serratia Species 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 230000007918 pathogenicity Effects 0.000 description 4
- 239000000049 pigment Substances 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 229920002444 Exopolysaccharide Polymers 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 230000028654 Type IV pili-dependent aggregation Effects 0.000 description 3
- 230000002776 aggregation Effects 0.000 description 3
- 238000004220 aggregation Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000013078 crystal Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 108020003175 receptors Proteins 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 230000009182 swimming Effects 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 240000005979 Hordeum vulgare Species 0.000 description 2
- 235000007340 Hordeum vulgare Nutrition 0.000 description 2
- 239000006137 Luria-Bertani broth Substances 0.000 description 2
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 2
- 241001240958 Pseudomonas aeruginosa PAO1 Species 0.000 description 2
- 241000206572 Rhodophyta Species 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- 241000607598 Vibrio Species 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 101150002827 lasI gene Proteins 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000012452 mother liquor Substances 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- WLJVXDMOQOGPHL-UHFFFAOYSA-N phenylacetic acid Chemical compound OC(=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-UHFFFAOYSA-N 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- WTFXTQVDAKGDEY-UHFFFAOYSA-N (-)-chorismic acid Natural products OC1C=CC(C(O)=O)=CC1OC(=C)C(O)=O WTFXTQVDAKGDEY-UHFFFAOYSA-N 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 108020004256 Beta-lactamase Proteins 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 241000244203 Caenorhabditis elegans Species 0.000 description 1
- 241000122827 Delisea pulchra Species 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 108091006149 Electron carriers Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010061217 Infestation Diseases 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 229920000715 Mucilage Polymers 0.000 description 1
- ZJFKKPDLNLCPNP-QMMMGPOBSA-N N-[(3s)-2-Oxotetrahydrofuran-3-Yl]hexanamide Chemical compound CCCCCC(=O)N[C@H]1CCOC1=O ZJFKKPDLNLCPNP-QMMMGPOBSA-N 0.000 description 1
- ZJFKKPDLNLCPNP-UHFFFAOYSA-N N-hexanoyl-L-homoserine lactone Natural products CCCCCC(=O)NC1CCOC1=O ZJFKKPDLNLCPNP-UHFFFAOYSA-N 0.000 description 1
- 101100202644 Parasynechococcus marenigrum (strain WH8102) bsmB gene Proteins 0.000 description 1
- 108700020474 Penicillin-Binding Proteins Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 102000045595 Phosphoprotein Phosphatases Human genes 0.000 description 1
- 108700019535 Phosphoprotein Phosphatases Proteins 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 101100408257 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) phzM gene Proteins 0.000 description 1
- 101100408260 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) phzS gene Proteins 0.000 description 1
- 101100524675 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) rhlI gene Proteins 0.000 description 1
- 108700014559 Serratia liquefaciens SwrI Proteins 0.000 description 1
- 101100258337 Serratia liquefaciens swrI gene Proteins 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 244000028419 Styrax benzoin Species 0.000 description 1
- 235000000126 Styrax benzoin Nutrition 0.000 description 1
- 235000008411 Sumatra benzointree Nutrition 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 101150025572 ahyI gene Proteins 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 125000002723 alicyclic group Chemical group 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 230000000078 anti-malarial effect Effects 0.000 description 1
- 230000000842 anti-protozoal effect Effects 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 239000003430 antimalarial agent Substances 0.000 description 1
- 239000003904 antiprotozoal agent Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 229960002130 benzoin Drugs 0.000 description 1
- 102000006635 beta-lactamase Human genes 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000032770 biofilm formation Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000001851 biosynthetic effect Effects 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 101150090954 bsmA gene Proteins 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- WTFXTQVDAKGDEY-HTQZYQBOSA-N chorismic acid Chemical group O[C@@H]1C=CC(C(O)=O)=C[C@H]1OC(=C)C(O)=O WTFXTQVDAKGDEY-HTQZYQBOSA-N 0.000 description 1
- 239000003636 conditioned culture medium Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- ISAOCJYIOMOJEB-UHFFFAOYSA-N desyl alcohol Natural products C=1C=CC=CC=1C(O)C(=O)C1=CC=CC=C1 ISAOCJYIOMOJEB-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 235000019382 gum benzoic Nutrition 0.000 description 1
- 125000003187 heptyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000004051 hexyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 231100001231 less toxic Toxicity 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- VLZLOWPYUQHHCG-UHFFFAOYSA-N nitromethylbenzene Chemical compound [O-][N+](=O)CC1=CC=CC=C1 VLZLOWPYUQHHCG-UHFFFAOYSA-N 0.000 description 1
- 125000001400 nonyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 125000001147 pentyl group Chemical group C(CCCC)* 0.000 description 1
- 125000001791 phenazinyl group Chemical class C1(=CC=CC2=NC3=CC=CC=C3N=C12)* 0.000 description 1
- 229960003424 phenylacetic acid Drugs 0.000 description 1
- 239000003279 phenylacetic acid Substances 0.000 description 1
- 101150106861 pqsH gene Proteins 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000004076 pyridyl group Chemical group 0.000 description 1
- 125000000168 pyrrolyl group Chemical group 0.000 description 1
- 239000001054 red pigment Substances 0.000 description 1
- 238000009877 rendering Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000008054 signal transmission Effects 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000820 toxicity test Toxicity 0.000 description 1
- 125000002023 trifluoromethyl group Chemical group FC(F)(F)* 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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
- A61K31/04—Nitro compounds
-
- 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
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/05—Phenols
-
- 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
- A61K31/075—Ethers or acetals
- A61K31/085—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon
-
- 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
- A61K31/13—Amines
- A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
- A61K31/136—Amines having aromatic rings, e.g. ketamine, nortriptyline having the amino group directly attached to the aromatic ring, e.g. benzeneamine
-
- 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
- A61K31/404—Indoles, e.g. pindolol
-
- 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4418—Non condensed pyridines; Hydrogenated derivatives thereof having a carbocyclic group directly attached to the heterocyclic ring, e.g. cyproheptadine
-
- 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)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
本发明公开了一种苯烯烃硝基类化合物作为群体感应抑制剂的非经典抗菌活性用途,所述苯烯烃硝基类化合物可应用作为紫色杆菌、铜绿假单胞菌和粘质沙雷氏菌的群体感应抑制剂,其中对甲基苯乙烯硝基化合物对粘质沙雷氏菌的被膜抑制率最高可达80%,进一步对粘质沙雷氏菌的被膜破坏率高达85%,对铜绿假单胞菌的被膜破坏高达60%,具有很好的群体感应非经典抗菌的潜在应用价值。
Description
技术领域
本发明涉及一类化合物的群体感应活性用途,尤其涉及一种苯烯烃硝基类化合物作为群体感应抑制剂的非经典抗菌活性用途。
背景技术
由于群体感应能调节病原菌毒力因子的产生,与病原菌的感染和致病性密切相关,因此,群体感应已成为抗感染药物的新靶点。抑制细菌的群体感应,即在不影响细菌正常生长的条件下,通过抑制群体感应系统中相关酶的活性,或者竞争性抑制信号分子的传递,而达到抑制群体感应,减少毒力因子的产生,降低致病性的效果。细菌群体感应抑制剂(quorum sensing inhibitor,QSI)大致分为两种:小分子化合物和降解酶类。
铜绿假单胞菌的群体感应系统由las、rhl和pqs组成,las系统由LasI和LasR组成,rhl系统由RhlI和RhlR组成。三个系统中,las起主导作用,las系统能调控rhl和pqs。lasI基因能编码调控3-oxododecanoyl-homoserine lactone(3-oxo-C12-HSL)信号分子的合成,rhlI编码合成信号分子butryl-homoserine lactone(C4-HSL),pqs编码合成信号分子Pseudomonas quinolone signal(PQS),PQS的合成受到基因pqsABCDE和pqsH的调控,合成后的PQS与受体蛋白PqsR结合,激活一系列致病基因的表达。
铜绿假单胞菌能分泌多种胞外和胞内的毒力因子,包括蛋白酶、绿脓菌素、磷脂酶、溶血素、鼠李糖脂、β-内酰胺酶、青霉素结合蛋白、海藻酸盐等,这些毒力因子对于铜绿假单胞菌的粘附和侵染具有重要的作用。绿脓菌素是铜绿假单胞菌最重要的毒力因子之一,属于吩嗪类化合物,该化合物具有亲水和亲脂两重性质,因此很容易穿透细胞膜进入细胞内部。绿脓菌素在铜绿假单胞菌侵染宿主细胞过程中发挥着重要作用,一方面该化合物能够抑制其他细菌的生长,减少种间竞争,从而有利于自身的生长。另一方面,绿脓菌素很容易透过细胞膜进入宿主细胞,从而对宿主细胞产生毒性。此外,该化合物具有氧化还原两重性质,在细胞内部能作为电子传递体,提高宿主细胞的氧化压力,导致宿主细胞中毒死亡。研究表明,绿脓菌素的合成受群体感应的调控,其生物合成前体物质为分支酸,通过2个phzABCDEFG操纵子编码以及phzH、phzM和phzS等基因的调控将前体物质转化成三环化合物。
粘质沙雷氏菌是一种广泛存在于土壤和水环境中的兼性厌氧菌,是沙雷氏菌属中发现的第一个模式种。历史上对沙雷氏细菌的分类经历了很长一段时间的混乱:1923年第一版《伯杰细菌鉴定手册》将沙雷氏菌属定义为包含23种;1974年第八版《伯杰细菌鉴定手册》将其合并为1种,即Serrtia marcescens;2004年第九版《伯杰细菌鉴定手册》重新规定为11种,加上后来分离鉴定的新种,目前沙雷氏菌属共有15个种。粘质沙雷氏菌体呈球状或者短杆状,是最小的细菌之一。粘质沙雷氏菌分为产色素菌株和不产色素菌株。产色素菌株菌落为红色圆形凸起形态,味道微臭;不产色素菌株则呈白色或淡黄色。关于其致病性,已证实粘质沙雷氏菌能够感染人类,但其仅对免疫力低下的人群有感染能力,大多数具有耐药性的致病性粘质沙雷氏菌通常不能产生灵菌红素。
灵菌红素是具有抗癌、抗疟、抗细菌、抗真菌和抗原虫等多种生物学活性的三吡咯环结构的天然红色素家族。灵菌红素主要由粘质沙雷氏菌、假单胞菌、放线菌的一些种和某些海洋细菌产生。灵菌红素呈暗红色,为脂溶性色素,几乎不溶于水,溶于甲醇,对pH敏感,酸性条件下呈现红色,碱性条件下呈现黄色。
目前的研究表明,群体感应能影响生物被膜的形成。在细菌的初始附着阶段,金黄色葡萄球菌的群体感应系统agr能抑制黏附素的分泌,黏附素对于细菌在物体表面的初始粘附起到重要的作用,且与野生型菌株相比,agr突变菌株能分泌更多的黏附素,有利于在物体表面的附着。在液化沙雷氏菌中,swrI调控信号分子的合成,该基因突变后,所形成的被膜量减少,厚度变薄。其调控被膜形成的两个基因bsmA和bsmB也受群体感应系统所调控。被膜的外面包裹着一层厚厚的聚合物,该聚合物由多糖、糖蛋白、脂蛋白等组成,而这些聚合物的合成过程受群体感应的调控。嗜水蒸气单胞菌的生物被膜在成熟过程中也需要群体感应系统的调控,ahyI基因突变菌株形成的被膜生物量显著低于野生型菌株,当外源添加信号分子butyryl-HSL后,其表型能得到部分恢复。早在1998年,发表在《Science》上的一篇文章就已经介绍了铜绿假单胞菌las系统与被膜之间的关系。lasI基因突变后,菌株不能合成3-oxo-C12-HSL信号分子,此时形成的被膜表面平坦、松散、形状较为均一,而野生型菌株形成的被膜结构不均匀、蘑菇状突起、紧密厚实。以上的研究表明,群体感应系统与生物被膜之间确实存在着一定的联系。
植物来源的QSI最早发现于海洋红藻(Delisea pulchra),从红藻中提取分离的化合物溴代呋喃酮(halogenated furanones)为AHL信号分子的结构类似物,该化合物对大肠杆菌、费氏弧菌、铜绿假单胞菌等多种细菌的群体感应系统均具有较好的抑制活性。
从自然界中获取群体感应抑制剂存在着提取率低、产率少、容易造成资源浪费等缺陷,而且有些植物和动物资源是濒危资源,不能随意利用。因此,研究人员通过化学合成的方法来合成群体感应抑制剂,提高了获取阳性化合物的概率。通过对信号分子比如C4-HSL、3-oxo-C12-HSL进行结构修饰,能够获得一系列的信号分子结构类似物,这些结构类似物能够竞争性的与受体蛋白结合,阻断信号分子与受体蛋白的结合,从而阻断群体感应系统,降低细菌的致病性。通过改变酰基侧链的长度、饱和度、官能团,以及内脂环的构象,Geske合成了一系列的信号分子结构类似物,其研究结果表面,具有LasR抑制活性的结构类似物的特点为侧链原子数目最多8个,侧链为直链或支链为芳香环;侧链末端是苯乙酸基且第三个碳原子上含亲脂性基团。这些化合物对根癌农杆菌、铜绿假单胞菌和费氏弧菌的群体感应系统有不同程度的抑制活性。O’Loughlin等对侧链和内脂环同时进行了结构修饰,合成了一系列的信号分子结构类似物,其中mBTL的活性最好,该化合物能显著抑制铜绿假单胞菌绿脓菌素和生物被膜的生成,其作用靶点为LasR和RhlR,体内和体外的实验表明,该化合物能保护秀丽隐杆线虫和人源肺腺上皮细胞免受铜绿假单胞菌的侵染。
综上所述,抗生素的出现为感染的病人带来了福音,但是随着抗生素的广泛使用,越来越多的细菌产生了耐药性。生物被膜是细菌产生耐药性的一个重要机制,而已有的研究表明,细菌生物被膜与群体感应密切相关。因此,寻找新型群体感应抑制剂,阻断细菌群体感应,抑制生物被膜的产生是治疗耐药菌感染的一项重要途径。
发明内容
发明目的:本发明旨在提供一种苯烯烃硝基类化合物作为粘质沙雷氏菌、铜绿假单胞菌和紫色杆菌群体感应抑制剂和非经典抗菌应用。
技术方案:本发明所述的一种结构式如下的苯烯烃硝基类化合物作为群体感应抑制剂的非经典抗菌活性用应用,
其中取代基R为甲基,乙基,丙基,异丙基,丁基,戊基,己基,庚基,辛基,壬基,苯基,吡啶基,吡咯基,N,N-二苯基,-F,-Cl,-Br,-I,甲氧基,三氟甲基,硝基,羟基,氨基中的至少一种,R为所在苯环任意位置取代基。
进一步地,所述苯烯烃硝基类化合物作为粘质沙雷氏菌、铜绿假单胞菌和紫色杆菌的群体感应抑制剂。
进一步地,所述粘质沙雷氏菌来源为动物源性或人源性。
进一步地,所述苯烯烃硝基类化合物对粘质沙雷菌有被膜抑制和破坏作用。
进一步地,所述苯烯烃硝基类化合物能有效减少粘质沙雷氏菌灵菌红素的产生。
进一步地,所述苯烯烃硝基类化合物能有效减少粘质沙雷胞外多糖的产生。
进一步地,所述苯烯烃硝基类化合物能减少粘质沙雷脂肪酶的产生。
进一步地,所述苯烯烃硝基类化合物能有效减少粘质沙雷蛋白酶的产生。
进一步地,所述苯烯烃硝基类化合物能有效抑制铜绿假单胞菌的被膜抑制和破坏作用。
进一步地,所述苯烯烃硝基类化合物能有效抑制铜绿假单胞菌的毒力因子作用,进而产生群体感应作用。
有益效果:与现有技术相比,本发明具有如下显著优点:合成了一系列苯烯烃硝基类化合物;所述苯烯烃硝基类化合物作为紫色杆菌、粘质沙雷氏菌和铜绿假单胞菌新型的群体感应抑制剂和非经典抗菌应用,其中个别化合物对粘质沙雷氏菌的被膜抑制率最高可达80%,进一步对粘质沙雷氏菌的被膜破坏率高达85%,对铜绿假单胞菌的被膜破坏高达60%。说明具有很好的群体感应非经典抗菌的潜在应用价值。
附图说明
图1为苯烯烃硝基类化合物对粘质沙雷氏菌群体感应活性初筛;
图2为活性优质化合物对粘质沙雷氏菌群体感应活性的平板放大图;
图3为对甲基苯乙烯硝基化合物的核磁表征数据图;
图4为对甲基苯乙烯硝基对粘质沙雷菌的生长曲线图;
图5为对甲基苯乙烯硝基对粘质沙雷菌的被膜抑制作用图(OD);
图6为对甲基苯乙烯硝基对粘质沙雷菌的被膜抑制作用扫描电镜图(SEM);
图7为对甲基苯乙烯硝基对粘质沙雷菌的被膜抑制作用激光共聚焦图(CLSM);
图8为对甲基苯乙烯硝基对粘质沙雷菌的被膜破坏作用图(OD);
图9为对甲基苯乙烯硝基对粘质沙雷菌的被膜破坏作用扫描电镜图(SEM);
图10为对甲基苯乙烯硝基对粘质沙雷菌的被膜破坏作用激光共聚焦图(CLSM);
图11为对甲基苯乙烯硝基对粘质沙雷菌的灵菌红素的影响图;
图12为对甲基苯乙烯硝基对粘质沙雷菌的胞外多糖的影响图;
图13为对甲基苯乙烯硝基对粘质沙雷菌的脂肪的影响图;
图14为对甲基苯乙烯硝基对粘质沙雷菌的蛋白酶的影响图;
图15为对甲基苯乙烯硝基对粘质沙雷菌的表面疏水性的影响图;
图16为对甲基苯乙烯硝基处理能减低粘质沙雷菌的自动聚集率图;
图17为对甲基苯乙烯硝基处理能减低粘质沙雷菌的泳动实验;
图18为群体感应相关基因表达图;
图19为面包虫实验说明化合物的毒性实验;
图20对甲基苯乙烯硝基处理对铜绿假单胞菌的生长曲线图;
图21对甲基苯乙烯硝基处理对铜绿假单胞菌被膜抑制(OD);
图22为对甲基苯乙烯硝基处理能减低铜绿假单胞菌的泳动实验(swarming和swimming)。
具体实施方式
下面结合附图对本发明的技术方案作进一步说明。
(一)紫色杆菌CV026的活性筛选(报告指示菌株)
挑取CV026单菌落到LB液体培养基中,28℃、180rpm培养17h,调菌液OD620=0.1。将100mL LB固体培养基均匀的倒入培养皿,作为底层平板。取培养液1mL以1%的接种量接种到100mL LB固体培养基中(冷却到50℃),随后加入卡那霉素(20μg/mL)和信号分子C6-HSL(5μM),混匀后均匀倒在底层平板上面,作为上层平板。待培养基凝固后,取5μL化合物,点在上层平板上,甲醇作为阴性对照,28℃培养24h,观察培养基颜色的变化。每组实验3次重复。
(二)铜绿假单胞菌群体感应抑制活性初筛
通过在LB肉汤中接种0.1%过夜的铜绿假单胞菌培养物来定量测定32个苯烯烃硝基类化合物(化合物结构式如表1所示)对绿脓菌素的抑制效果,大麦芽碱作阳性对照,二甲基亚砜(DMSO)作阴性对照。在37℃下培养24小时后,观察绿脓菌素颜色变化。
最小抑菌浓度(MIC)的测定
苯烯烃硝基类化合物对铜绿假单胞菌的MIC测定参照临床实验标准委员会制定的方法进行(CLSI,2015)。挑取铜绿假单胞菌单菌落接种到5mL Luria-Bertani肉汤(SangonBiotech,Shanghai,China)中活化17h,作为母液。取适量母液加入LB培养基中,调OD620=0.05,此时培养基中的细菌浓度约为1.5*10^5CFU/mL。加入苯烯烃硝基类化合物,采用二倍稀释法梯度稀释,DMSO为阴性对照。每个浓度取200μL加入到96孔板中,37℃,150rpm培养24h,酶标仪测定OD620。在实验中进行了三个重复。
绿脓菌素的测定
挑取铜绿假单胞菌PAO1单菌落至5mL LB培养基中活化17h,作为种子液。种子液以0.1%的接种量接种到新的LB培养基中,加入不同浓度的苯烯烃硝基类化合物,DMSO作为阴性对照,大麦芽碱作为阳性对照。37℃、180rpm培养17h。培养液用0.22μm无菌过滤器过滤,得无菌滤液备用。绿脓菌素含量的测定:培养后的上清液加入三氯甲烷进行萃取(5/3,v/v),静置分层后,取下层有机相,加入1mL 0.2M的HCl,混合均匀,4℃、10000rpm离心10min,取200μL上层溶液酶标仪测定OD520。
被膜抑制
参照Damiano等的方法并稍作修改。挑取铜绿假单胞菌单菌落至5mL LB培养基中活化17h,作为种子液。取种子液50μL以1:1000的比例加入到50mL TSB(含1%甘油)培养基中,加入不同浓度的苯烯烃硝基类化合物,DMSO作为阴性对照。取混合后的培养液200μL加入到96孔板中,37℃静置培养24h。吸去上层培养液,用PBS水洗3遍,洗去浮游的细菌,置于60℃烘箱中烘干水分,加入200μL甲醇固定15min。吸去甲醇,60℃烘箱中干燥。加入100μL0.05%的结晶紫染色15min。吸去结晶紫,PBS水洗3遍,去掉多余的未附着的结晶紫染料,放入60℃烘箱中干燥。干燥后加入200μL 95%乙醇,摇床中37℃、180rpm脱色15min。取150μL脱色液,酶标仪测定OD570。测试结果如表2所示。
(三)粘质沙雷氏菌群体感应活性初筛
挑取NJ01单菌落到LB液体培养基中,28℃、180rpm培养17h,调菌液OD620=0.1。将100mL LB固体培养基均匀的倒入培养皿,作为底层平板。取培养液1mL以1%的接种量接种到100mL LB固体培养基中(冷却到50℃),混匀后均匀倒在底层平板上面,作为上层平板。待培养基凝固后,取5μL苯烯烃硝基类化合物生物,点在上层平板上,DMSO作为阴性对照,28℃培养24h,观察培养基颜色的变化。每组实验3次重复。
最小抑菌浓度(MIC)的测定
参照上面
灵菌红素的测定
接种S.marcescens NJ01单菌落到新鲜LB培养基中,28℃,180rpm,过夜,菌液调OD600=0.4,以1%比例加入到2mL LB培养基,向其中加入50μg/mL的苯烯烃硝基类化合物,空白对照;DMSO为阴性对照,50μg/mL的苯烯烃硝基类化合物做阳性对照。28℃,180rpm,培养16-24h,吸取1mL各组菌液至离心管,离心,10000rpm,10min,收集上清备用(或用0.22μm无菌滤头过滤,得无菌滤液),向离心管加1mL酸化乙醇(4%,1M HCl in ethanol)离心,10000rpm,10min将上清转移至96孔板,测OD534。
被膜抑制和被膜破坏
参照上面。
(四)结果与统计分析
数据表示为平均值±标准偏差。在分析之前,使用Levene的方差均匀性检验来评估所有变量的等效性。统计差异由Levene检验,方差分析(ANOVA)和Tukey-Kramer检验确定。使用Graphpad Prism进行统计分析。p≤0.05,具有统计学意义。
表1所有苯烯烃硝基类化合物结构式
表2化合物对铜绿假单胞菌PAO1的被膜抑制率结果
备注:化合物浓度统一为:50μg/mL。
下面结合附图做出以下说明和解释:
图1是苯烯烃硝基类化合物对粘质沙雷氏菌群体的感应活性初筛,大部分化合物均具有一定的群体感应活性,其中有部分化合物活性更为优质。
如图2所示,苯烯烃硝基类化合物初筛粘质沙雷氏菌群体感应活性时,表现较好的活性化合物,局部放大的平板图,进一步说明该化合物具有很好的群体感应活性。图中a)代表NO2-32化合物的群体感应活性初筛结果,b)代表NO2-6,NO2-9,NO2-25,DMSO的初筛结果图,c)代表NO2-9化合物(对甲基苯乙烯硝基化合物)的群体感应活性初筛结果。
图3对甲基苯乙烯硝基化合物的核磁表征数据1H NMR(400MHz,Chloroform-d)δ7.95(d,J=13.7Hz,1H),7.54(d,J=13.6Hz,1H),7.47-7.36(m,2H),7.24(d,J=8.0Hz,2H),2.39(s,3H).13C NMR(101MHz,Chloroform-d)δ143.2,139.2,136.3,130.2,129.3,127.3,21.7.
如图4所示为对甲基苯乙烯硝基处理对粘质沙雷菌的生长曲线图。在测定MIC为100μg/mL之后,再次进行实验。当给药浓度为12.5μg/mL时,当给药浓度为25μg/mL时,当的给药浓度为50μg/mL时,均不影响细菌的正常生长。群体感应活性即在不影响细菌生长条件下进行。
如图5所示,对甲基苯乙烯硝基对粘质沙雷菌有被膜抑制作用。当给药浓度为12.5μg/mL时,被膜抑制率为49.6%;当给药浓度为25μg/mL时,被膜抑制率为66.4%;当的给药浓度为50μg/mL时,对甲基苯乙烯硝基对的被膜抑制率为80.0%。可见,该化合物对基对粘质沙雷菌有被膜抑制作用明显,并具有很好的梯度效果。
如图6所示,对甲基苯乙烯硝基处理对粘质沙雷菌有被膜抑制作用。进一步使用SEM进行深入的评价。当给药浓度为12.5μg/mL时,当给药浓度为25μg/mL时,当的给药浓度为50μg/mL时,对甲基苯乙烯硝基对的被膜抑制情况分别如图所示,直接明了,清晰可见。
如图7所示,对甲基苯乙烯硝基处理对粘质沙雷菌有被膜抑制作用。进一步使用CLSM进行深入的评价。当给药浓度为12.5μg/mL时,当给药浓度为25μg/mL时,当的给药浓度为50μg/mL时,对甲基苯乙烯硝基对的被膜抑制情况分别如图所示,清晰直接,进一步反复确定化合物对被膜的抑制作用强,进而说明群体感应活性强。
如图8所示,对甲基苯乙烯硝基处理对粘质沙雷菌有很好的被膜破坏作用。当给药浓度为12.5μg/mL时,被膜破坏率为25.7%;当给药浓度为25μg/mL时,被膜破坏率为68.2%;当的给药浓度为50μg/mL时,对甲基苯乙烯硝基对的被膜破坏率为85.0%。由此可知,该化合物对基对粘质沙雷菌有被膜破坏作用明显,并具有很好的梯度效果。具有很好的被膜破坏应用价值。
如图9所示,对甲基苯乙烯硝基处理对粘质沙雷菌有被膜破坏作用。进一步使用SEM进行深入的评价。当给药浓度为12.5μg/mL时,当给药浓度为25μg/mL时,当的给药浓度为50μg/mL时,对甲基苯乙烯硝基对的被膜破坏情况分别如图所示,清晰明确。
如图10所示,对甲基苯乙烯硝基处理对粘质沙雷菌有被膜破坏作用。进一步使用CLSM进行深入的评价。当给药浓度为12.5μg/mL时,当给药浓度为25μg/mL时,当的给药浓度为50μg/mL时,对甲基苯乙烯硝基对的被膜破坏情况分别如图所示,准确清晰。进一步反复确定化合物对被膜破坏作用强,进而说明群体感应活性强。
如图11所示,对甲基苯乙烯硝基能有效减少粘质沙雷灵菌红素的产生,并呈剂量效应。(灵菌红素为主要毒力因子,抑制其减少说明化合物群体感应活性的强。)当给药浓度为12.5μg/mL时,抑制率为42.2%;当给药浓度为25μg/mL时,抑制率为48.4%;当的给药浓度为50μg/mL时,对甲基苯乙烯硝基对的抑制率为63.1%,均低于阳性对照香草酸的22.7%。因此,与空白对照DMSO组比较,对甲基苯乙烯硝基能显著减少粘质沙雷灵菌红素的产生。
如图12所示,对甲基苯乙烯硝基能有效减少粘质沙雷胞外多糖的产生,并呈剂量效应。当给药浓度为12.5μg/mL时,抑制率为33.9%;当给药浓度为25μg/mL时,抑制率为45.9%;当给药浓度为50μg/mL时,对甲基苯乙烯硝基对的抑制率为53.2%;阳性对照香草酸的抑制率为39.0%。因此,与空白对照DMSO组比较,对甲基苯乙烯硝基能显著减少粘质沙雷胞外多糖的产生。
如图13所示,对甲基苯乙烯硝基能减少粘质沙雷脂肪酶的产生。抑制率约为33.5%;阳性对照香草酸的抑制率为17.4%。因此,与空白对照DMSO组比较,对甲基苯乙烯硝基能减少粘质沙雷脂肪酶的产生。
如图14所示,对甲基苯乙烯硝基能有效减少粘质沙雷蛋白酶的产生。当给药浓度为12.5μg/mL时,抑制率为17.9;当给药浓度为25μg/mL时,抑制率为20.7%;当给药浓度为50μg/mL时,对甲基苯乙烯硝基对的抑制率为35.8%;阳性对照香草酸的35.1%。因此,与空白对照DMSO组比较,对甲基苯乙烯硝基能减少粘质沙雷蛋白酶的产生。
如图15所示,对甲基苯乙烯硝基处理能显著提高粘质沙雷菌的表面疏水性。当给药浓度为12.5μg/mL时,表面疏水率为17.0%;当给药浓度为25μg/mL时,表面疏水性率为25.9%;当给药浓度为50μg/mL时,对甲基苯乙烯硝基对的表面疏水率为36.7%,均高于阳性对照香草酸的15.7%和空白对照DMSO的9.2%。因此,与空白对照DMSO组比较,对甲基苯乙烯硝基能显著提高的表面疏水性。
如图16所示,对甲基苯乙烯硝基处理能减低粘质沙雷菌的自动聚集率。当给药浓度为12.5μg/mL时,自动聚集率为20.5%;当给药浓度为25μg/mL时,自动聚集率为18.6%;当给药浓度为50μg/mL时,对甲基苯乙烯硝基对的自动聚集率为16.6%;阳性对照香草酸的18.6%均低于空白对照DMSO的25.0%。因此,与空白对照DMSO组比较,对甲基苯乙烯硝基能减低的自动聚集率。
如图17所示,对甲基苯乙烯硝基处理能减低粘质沙雷菌的泳动实验。当给药浓度为12.5μg/mL,25μg/mL,50μg/mL,阳性对照香草酸时,均低于空白对照DMSO,梯度效果,趋势非常明显。因此,对甲基苯乙烯硝基处理能减低粘质沙雷菌的的泳动实验。趋势良好,说明化合物对毒力因子有强烈抑制作用,进而证明其群体感应活性良好。
图18群体感应相关基因下调,证明了化合物抑制其群体感应活性的本质原因。证明该化合物是群体感应抑制剂,进一步具有很强烈的群体感应活性。
图19的面包虫实验说明化合物的毒性问题,说明化合物毒性整体较小。
图20为对甲基苯乙烯硝基处理对铜绿假单胞菌的生长曲线图。在测定MIC为100μg/mL之后,再次进行实验。当给药浓度为12.5μg/mL时,当给药浓度为25μg/mL时,当给药浓度为50μg/mL时,均不影响细菌的正常生长。群体感应活性即在不影响细菌生长条件下进行。
如图21所示,对甲基苯乙烯硝基处理对铜绿假单胞菌有被膜抑制作用。当给药浓度为12.5μg/mL时,被膜抑制率为23%;当给药浓度为25μg/mL时,被膜抑制率为38%;当的给药浓度为50μg/mL时,对甲基苯乙烯硝基对的被膜抑制率为60%。
图22为对甲基苯乙烯硝基处理能减低铜绿假单胞菌的泳动实验(swarming和swimming)。当给药浓度为12.5μg/mL,25μg/mL,50μg/mL,阳性对照香草酸时,均低于空白对照DMSO,两组实验的结果趋势非常明显。因此,对甲基苯乙烯硝基处理能减低铜绿假单胞菌的的泳动实验趋势良好,说明化合物对毒力因子有强烈抑制作用,进而证明其群体感应活性良好。
Claims (6)
1.一种对甲基苯乙烯硝基化合物用于制备抑制粘质沙雷氏菌药物的应用,其特征在于,所述化合物的结构式如下:
该化合物对粘质沙雷菌有群体感应活性及相关的被膜抑制和破坏作用。
2.根据权利要1所述的对甲基苯乙烯硝基化合物用于制备抑制粘质沙雷氏菌药物的应用,其特征在于,所述粘质沙雷氏菌来源为动物源性。
3.根据权利要求1所述的对甲基苯乙烯硝基化合物用于制备抑制粘质沙雷氏菌药物的应用,其特征在于,所述对甲基苯乙烯硝基化合物能有效减少粘质沙雷氏菌灵菌红素的产生。
4.根据权利要求1所述的对甲基苯乙烯硝基化合物用于制备抑制粘质沙雷氏菌药物的应用,其特征在于,所述对甲基苯乙烯硝基化合物能有效减少粘质沙雷胞外多糖的产生。
5.根据权利要求1所述的对甲基苯乙烯硝基化合物用于制备抑制粘质沙雷氏菌药物的应用,其特征在于,所述对甲基苯乙烯硝基化合物能减少粘质沙雷脂肪酶的产生。
6.根据权利要求1所述的对甲基苯乙烯硝基化合物用于制备抑制粘质沙雷氏菌药物的应用,其特征在于,所述对甲基苯乙烯硝基化合物能有效减少粘质沙雷蛋白酶的产生。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210716740.XA CN115025073B (zh) | 2022-06-23 | 2022-06-23 | 一种苯烯烃硝基类化合物作为群体感应抑制剂的非经典抗菌活性用途 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210716740.XA CN115025073B (zh) | 2022-06-23 | 2022-06-23 | 一种苯烯烃硝基类化合物作为群体感应抑制剂的非经典抗菌活性用途 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115025073A CN115025073A (zh) | 2022-09-09 |
CN115025073B true CN115025073B (zh) | 2024-05-07 |
Family
ID=83126972
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210716740.XA Active CN115025073B (zh) | 2022-06-23 | 2022-06-23 | 一种苯烯烃硝基类化合物作为群体感应抑制剂的非经典抗菌活性用途 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN115025073B (zh) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3652772A (en) * | 1969-01-06 | 1972-03-28 | American Cyanamid Co | 2-hydroxy-3-methoxy-beta-nitrostyrene and 2-hydroxy - 5 - beta-nitrostyrene as antifungal and antibacterial agents |
RU2145215C1 (ru) * | 1997-04-18 | 2000-02-10 | Денисенко Петр Прокофьевич | Вещества, проявляющие антимикробную, антигрибковую, антипротозойную активности |
CN1863788A (zh) * | 2001-06-18 | 2006-11-15 | 拜欧戴姆有限公司 | 抗菌和防辐射化合物 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010127307A1 (en) * | 2009-04-30 | 2010-11-04 | Rutgers, The State University Of New Jersey | Antimicrobial agents |
US10292390B2 (en) * | 2011-11-04 | 2019-05-21 | Wisconsin Alumni Research Foundation | Inhibition and dispersion of bacterial biofilms with 2-aminobenzimidazole derivatives |
-
2022
- 2022-06-23 CN CN202210716740.XA patent/CN115025073B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3652772A (en) * | 1969-01-06 | 1972-03-28 | American Cyanamid Co | 2-hydroxy-3-methoxy-beta-nitrostyrene and 2-hydroxy - 5 - beta-nitrostyrene as antifungal and antibacterial agents |
RU2145215C1 (ru) * | 1997-04-18 | 2000-02-10 | Денисенко Петр Прокофьевич | Вещества, проявляющие антимикробную, антигрибковую, антипротозойную активности |
CN1863788A (zh) * | 2001-06-18 | 2006-11-15 | 拜欧戴姆有限公司 | 抗菌和防辐射化合物 |
Non-Patent Citations (1)
Title |
---|
Nuno Milhazes et al.."β-Nitrostyrene derivatives as potential antibacterial agents:A structur-property-activity relationship study".Bioorganic&Medicinal Chemistry.2006,第14卷(第12期),摘要、第4079页左栏第3段、第4080页右栏第2段至第4081页左栏第1段、第4084页右栏第2段以及表1-2. * |
Also Published As
Publication number | Publication date |
---|---|
CN115025073A (zh) | 2022-09-09 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Durán et al. | Chromobacterium violaceum: a review of pharmacological and industiral perspectives | |
US10577404B2 (en) | Polypeptide derivatives from grass carp interferon and application thereof | |
US20210340179A1 (en) | Antimicrobial peptide as-hepc3(48-56) of acanthopagrus schlegelii and method thereof | |
CN103665071A (zh) | 洋橄榄叶素衍生物及其在抗耐药菌和耐药结核分枝杆菌感染中的应用 | |
CN115025073B (zh) | 一种苯烯烃硝基类化合物作为群体感应抑制剂的非经典抗菌活性用途 | |
CN105663244B (zh) | 一种具有细菌群体感应淬灭活性的无花果叶提取物的用途 | |
CN107929742B (zh) | 大麦芽碱或其联合抗生素的抗感染制药用途 | |
Jalil et al. | Time-kill study and morphological changes of Proteus mirabilis cells exposed to ethyl acetate crude extract of Lasiodiplodia pseudotheobromae IBRL OS-64. | |
CN115057807B (zh) | 一种3-(2-异氰基苄基)吲哚衍生物群体感应抑制剂及其制备方法和应用 | |
Iswarya et al. | Promising antimicrobial compounds from an endophytic fungus Penicillium daleae EF 4 isolated from the seaweed Enteromorpha Flexuosa Linn | |
CN101982462A (zh) | 一种由海洋镰刀霉菌属分离提取的化合物及其应用 | |
CN113209085B (zh) | 水鬼蕉新碱a在制备抗菌药物中的应用 | |
Milanda et al. | Antibacterial activity of red yeast rice extract against Propionibacterium acnes ATCC 11827 and methicilin-resistant Staphylococcus aureus ATCC BAA-1683 | |
KR101756123B1 (ko) | 아스퍼질러스 속 균 유래의 조효소를 이용하는 안토시아닌 올리고머의 제조 방법 | |
RU2493247C1 (ru) | Ингибитор возбудителя бактериального ожога плодовых культур (erwinia amylovora) | |
DE2040141A1 (de) | Neues Antibioticum und Verfahren zu seiner Herstellung | |
Nag et al. | Evaluation of antagonistic activities of microbes from Vallapattanam and Pappinishery mangrove ecosystems of Kannur district in Kerala, India. | |
CN113832064B (zh) | 一种微生物群体感应淬灭复合剂及其应用 | |
Garba et al. | Isolation and Evaluation of In vitro Antibacterial Potential of Endophytic Fungi from the leaves of Psidium guajava (Guava) | |
NL2033150B1 (en) | Application of limonium wrightii naphthoquinone in preparing anti- haemophilus parasuis drug | |
Abbas et al. | Isolation and Screening of Actinomycetes producing Antibacterial compounds from different river sediments | |
CN117338760A (zh) | 佳味酚在制备抑制微生物感染的药物中的应用 | |
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 | |
CN107739740A (zh) | 一种海洋真菌来源的Lasiodiplodins化合物的制备方法及应用 | |
US20080090900A1 (en) | Antibiotic bushrin |
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 |