CN116675779A - Short peptide targeting Src kinase and application thereof in systemic fungal infection - Google Patents
Short peptide targeting Src kinase and application thereof in systemic fungal infection Download PDFInfo
- Publication number
- CN116675779A CN116675779A CN202310619462.0A CN202310619462A CN116675779A CN 116675779 A CN116675779 A CN 116675779A CN 202310619462 A CN202310619462 A CN 202310619462A CN 116675779 A CN116675779 A CN 116675779A
- Authority
- CN
- China
- Prior art keywords
- src
- sting
- polypeptide
- short peptide
- application
- 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.)
- Granted
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 95
- 230000009885 systemic effect Effects 0.000 title claims abstract description 27
- 208000031888 Mycoses Diseases 0.000 title claims abstract description 24
- 206010017533 Fungal infection Diseases 0.000 title claims abstract description 22
- 108010087686 src-Family Kinases Proteins 0.000 title claims description 9
- 102000001332 SRC Human genes 0.000 title claims 4
- 230000008685 targeting Effects 0.000 title description 2
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 63
- 229920001184 polypeptide Polymers 0.000 claims abstract description 60
- 102000020313 Cell-Penetrating Peptides Human genes 0.000 claims abstract description 19
- 108010051109 Cell-Penetrating Peptides Proteins 0.000 claims abstract description 19
- 230000002757 inflammatory effect Effects 0.000 claims abstract description 17
- 230000000843 anti-fungal effect Effects 0.000 claims abstract description 16
- 229940121375 antifungal agent Drugs 0.000 claims abstract description 15
- 239000003814 drug Substances 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 13
- 108090001005 Interleukin-6 Proteins 0.000 claims abstract description 11
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 8
- 125000000539 amino acid group Chemical group 0.000 claims abstract description 7
- 238000011160 research Methods 0.000 claims abstract description 5
- 101710196623 Stimulator of interferon genes protein Proteins 0.000 claims abstract 5
- 241000222122 Candida albicans Species 0.000 claims description 21
- 229940095731 candida albicans Drugs 0.000 claims description 21
- 102000004127 Cytokines Human genes 0.000 claims description 14
- 108090000695 Cytokines Proteins 0.000 claims description 14
- 230000001737 promoting effect Effects 0.000 claims description 12
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical group N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 10
- 230000009918 complex formation Effects 0.000 claims description 9
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims description 6
- 102000019034 Chemokines Human genes 0.000 claims description 6
- 108010012236 Chemokines Proteins 0.000 claims description 6
- 238000011282 treatment Methods 0.000 claims description 6
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 claims description 5
- 229960002685 biotin Drugs 0.000 claims description 5
- 235000020958 biotin Nutrition 0.000 claims description 5
- 239000011616 biotin Substances 0.000 claims description 5
- 241000233866 Fungi Species 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 244000052616 bacterial pathogen Species 0.000 claims description 3
- 230000008030 elimination Effects 0.000 claims description 3
- 238000003379 elimination reaction Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 239000007790 solid phase Substances 0.000 claims description 3
- 238000001308 synthesis method Methods 0.000 claims description 3
- 230000002194 synthesizing effect Effects 0.000 claims description 3
- 241000228212 Aspergillus Species 0.000 claims description 2
- 241001337994 Cryptococcus <scale insect> Species 0.000 claims description 2
- 241000228143 Penicillium Species 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 9
- 230000028993 immune response Effects 0.000 abstract description 9
- 244000053095 fungal pathogen Species 0.000 abstract description 7
- 108060008682 Tumor Necrosis Factor Proteins 0.000 abstract description 6
- 108020004999 messenger RNA Proteins 0.000 abstract description 6
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 abstract description 5
- 238000013518 transcription Methods 0.000 abstract description 5
- 230000035897 transcription Effects 0.000 abstract description 5
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 230000005764 inhibitory process Effects 0.000 abstract description 3
- 230000002829 reductive effect Effects 0.000 abstract description 3
- 230000003993 interaction Effects 0.000 abstract description 2
- 101000643024 Homo sapiens Stimulator of interferon genes protein Proteins 0.000 description 35
- 102100035533 Stimulator of interferon genes protein Human genes 0.000 description 35
- 241000699670 Mus sp. Species 0.000 description 34
- 210000004027 cell Anatomy 0.000 description 23
- 208000015181 infectious disease Diseases 0.000 description 14
- 238000010186 staining Methods 0.000 description 13
- 230000004913 activation Effects 0.000 description 11
- 210000003734 kidney Anatomy 0.000 description 11
- 230000002538 fungal effect Effects 0.000 description 10
- 229920002307 Dextran Polymers 0.000 description 9
- 102000004889 Interleukin-6 Human genes 0.000 description 9
- 229940100601 interleukin-6 Drugs 0.000 description 9
- 230000019491 signal transduction Effects 0.000 description 9
- 230000000638 stimulation Effects 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 241001529936 Murinae Species 0.000 description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- 239000003937 drug carrier Substances 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000007924 injection Substances 0.000 description 5
- 238000002347 injection Methods 0.000 description 5
- 102000009076 src-Family Kinases Human genes 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 241000725303 Human immunodeficiency virus Species 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 229920000392 Zymosan Polymers 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 210000004443 dendritic cell Anatomy 0.000 description 4
- 230000002708 enhancing effect Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 229920002521 macromolecule Polymers 0.000 description 4
- 210000000440 neutrophil Anatomy 0.000 description 4
- 230000007115 recruitment Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 108010050904 Interferons Proteins 0.000 description 3
- 102000000551 Syk Kinase Human genes 0.000 description 3
- 108010016672 Syk Kinase Proteins 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000008595 infiltration Effects 0.000 description 3
- 238000001764 infiltration Methods 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 210000000952 spleen Anatomy 0.000 description 3
- 108700026239 src Genes Proteins 0.000 description 3
- 230000004936 stimulating effect Effects 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- RAVVEEJGALCVIN-AGVBWZICSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-5-amino-2-[[(2s)-2-[[(2s)-2-[[(2s)-6-amino-2-[[(2s)-6-amino-2-[[(2s)-2-[[2-[[(2s)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]acetyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]hexanoyl]amino]hexanoyl]amino]-5-(diamino Chemical group NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 RAVVEEJGALCVIN-AGVBWZICSA-N 0.000 description 2
- 102000003930 C-Type Lectins Human genes 0.000 description 2
- 108090000342 C-Type Lectins Proteins 0.000 description 2
- 102000016950 Chemokine CXCL1 Human genes 0.000 description 2
- 108010014419 Chemokine CXCL1 Proteins 0.000 description 2
- 102100031256 Cyclic GMP-AMP synthase Human genes 0.000 description 2
- 108030002637 Cyclic GMP-AMP synthases Proteins 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 108700000788 Human immunodeficiency virus 1 tat peptide (47-57) Proteins 0.000 description 2
- 102000014150 Interferons Human genes 0.000 description 2
- 208000037026 Invasive Fungal Infections Diseases 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 108091000080 Phosphotransferase Proteins 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 108091023040 Transcription factor Proteins 0.000 description 2
- 102000040945 Transcription factor Human genes 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 210000001185 bone marrow Anatomy 0.000 description 2
- 210000004979 bone marrow derived macrophage Anatomy 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 230000005745 host immune response Effects 0.000 description 2
- 238000001114 immunoprecipitation Methods 0.000 description 2
- 229940079322 interferon Drugs 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 239000002523 lectin Substances 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 230000004807 localization Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 102000020233 phosphotransferase Human genes 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000000770 proinflammatory effect Effects 0.000 description 2
- 230000000069 prophylactic effect Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 210000005084 renal tissue Anatomy 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 108091023037 Aptamer Proteins 0.000 description 1
- 239000000592 Artificial Cell Substances 0.000 description 1
- 208000031648 Body Weight Changes Diseases 0.000 description 1
- 238000011740 C57BL/6 mouse Methods 0.000 description 1
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 1
- AUJXLBOHYWTPFV-BLWRDSOESA-N CS[C@H]1SC[C@H]2N(C)C(=O)[C@@H](C)NC(=O)[C@H](COC(=O)[C@@H](C(C)C)N(C)C(=O)[C@@H]1N(C)C(=O)[C@@H](C)NC(=O)[C@H](COC(=O)[C@@H](C(C)C)N(C)C2=O)NC(=O)c1cnc2ccccc2n1)NC(=O)c1cnc2ccccc2n1 Chemical compound CS[C@H]1SC[C@H]2N(C)C(=O)[C@@H](C)NC(=O)[C@H](COC(=O)[C@@H](C(C)C)N(C)C(=O)[C@@H]1N(C)C(=O)[C@@H](C)NC(=O)[C@H](COC(=O)[C@@H](C(C)C)N(C)C2=O)NC(=O)c1cnc2ccccc2n1)NC(=O)c1cnc2ccccc2n1 AUJXLBOHYWTPFV-BLWRDSOESA-N 0.000 description 1
- 206010007134 Candida infections Diseases 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 208000030453 Drug-Related Side Effects and Adverse reaction Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 108010009858 Echinomycin Proteins 0.000 description 1
- 102100030013 Endoribonuclease Human genes 0.000 description 1
- 101710199605 Endoribonuclease Proteins 0.000 description 1
- 206010063560 Excessive granulation tissue Diseases 0.000 description 1
- 108091029865 Exogenous DNA Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 102000002227 Interferon Type I Human genes 0.000 description 1
- 108010014726 Interferon Type I Proteins 0.000 description 1
- 241000581650 Ivesia Species 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- 108090000744 Mitogen-Activated Protein Kinase Kinases Proteins 0.000 description 1
- 102000004232 Mitogen-Activated Protein Kinase Kinases Human genes 0.000 description 1
- 125000001429 N-terminal alpha-amino-acid group Chemical group 0.000 description 1
- 108010014632 NF-kappa B kinase Proteins 0.000 description 1
- 206010028851 Necrosis Diseases 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 1
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 1
- 101710113029 Serine/threonine-protein kinase Proteins 0.000 description 1
- 206010070863 Toxicity to various agents Diseases 0.000 description 1
- 108010003533 Viral Envelope Proteins Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 239000002518 antifoaming agent Substances 0.000 description 1
- -1 antiseptic Substances 0.000 description 1
- 150000003851 azoles Chemical class 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229920000249 biocompatible polymer Polymers 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000004579 body weight change Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000000701 coagulant Substances 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000013601 cosmid vector Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000002716 delivery method Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 210000002288 golgi apparatus Anatomy 0.000 description 1
- 210000001126 granulation tissue Anatomy 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 229920006008 lipopolysaccharide Polymers 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 210000004779 membrane envelope Anatomy 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 229920005615 natural polymer Polymers 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000027405 negative regulation of phosphorylation Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 210000000680 phagosome Anatomy 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 230000006461 physiological response Effects 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 150000004291 polyenes Chemical class 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- AUJXLBOHYWTPFV-UHFFFAOYSA-N quinomycin A Natural products CN1C(=O)C(C)NC(=O)C(NC(=O)C=2N=C3C=CC=CC3=NC=2)COC(=O)C(C(C)C)N(C)C(=O)C2N(C)C(=O)C(C)NC(=O)C(NC(=O)C=3N=C4C=CC=CC4=NC=3)COC(=O)C(C(C)C)N(C)C(=O)C1CSC2SC AUJXLBOHYWTPFV-UHFFFAOYSA-N 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
- 238000012385 systemic delivery Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- MPLHNVLQVRSVEE-UHFFFAOYSA-N texas red Chemical compound [O-]S(=O)(=O)C1=CC(S(Cl)(=O)=O)=CC=C1C(C1=CC=2CCCN3CCCC(C=23)=C1O1)=C2C1=C(CCC1)C3=[N+]1CCCC3=C2 MPLHNVLQVRSVEE-UHFFFAOYSA-N 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000012384 transportation and delivery Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 102000003390 tumor necrosis factor Human genes 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The application belongs to the technical fields of biological medicine and molecular biology, and particularly relates to a Src-targeted short peptide and application thereof in systemic fungal infection. According to the application, research shows that the polypeptide can promote host antifungal immune response by inhibiting STING-Src interaction. The polypeptide is derived from amino acid residues 1-18 of STING protein, and TAT cell penetrating peptide is fused at the N end of the polypeptide. After the polypeptide enters cells, the polypeptide competitively binds with Src to inhibit the formation of Src and STING complexes, so that the inhibition effect of STING on CLR channels is reduced; the mRNA transcription level of inflammatory cytokines TNF-alpha, IL-6 and the like is obviously improved, and the clearance capacity to pathogenic fungi is promoted, so that the method has good practical application value.
Description
Technical Field
The application belongs to the technical fields of biological medicine and molecular biology, and particularly relates to a Src-targeted short peptide and application thereof in systemic fungal infection.
Background
The information disclosed in the background of the application is only for enhancement of understanding of the general background of the application and is not necessarily to be taken as an admission or any form of suggestion that this information forms the prior art already known to a person of ordinary skill in the art.
Candida is the most common cause of fungal infection in humans, resulting in about 400000 systemic invasive infections annually, with a mortality rate of about 40%. Among them, candida albicans is the most common pathogenic species. Systemic infection of candida still has high mortality due to problems of drug toxicity (polyene) and drug resistance (azoles and echinomycin). There is therefore an urgent need to develop new antifungal therapies.
Interferon gene stimulatory factor STING (Stimulator of interferon genes) is an aptamer on the endoplasmic reticulum that specifically recognizes a natural cyclic dinucleotide ligand synthesized by cyclic GMP-AMP synthase (cGAS), thereby activating STING. STING, which binds to the ligand, is transported to the golgi apparatus and initiates a cascade of downstream signals, including recruitment and activation of serine/threonine protein kinase (TBK 1) and interferon regulated transcription factors (IRF 3) and nuclear factors (NF- κb), thereby producing type I interferons and pro-inflammatory cytokines, including interleukin 6 (IL-6), tumor necrosis factor (TNF- α), and the like. However, there is still little research on STING against fungal signal pathways, and further research is still needed.
Disclosure of Invention
Aiming at the defects in the prior art, the application aims to provide a Src-targeting short peptide and application thereof in systemic fungal infection. Specifically, it promotes host antifungal immune responses by inhibiting STING-Src interactions. The polypeptide is derived from amino acid residues 1-18 of STING protein, and TAT cell penetrating peptide is fused at the N end of the polypeptide. After the polypeptide enters cells, the polypeptide competitively binds with Src to inhibit the formation of Src and STING complexes, so that the inhibition effect of STING on CLR channels is reduced; significantly improves the mRNA transcription level of inflammatory cytokines TNF-alpha, IL-6 and the like, and promotes the clearance ability to pathogenic fungi. Based on the above results, the present application has been completed.
In order to achieve the technical purpose, the technical scheme provided by the application is as follows:
in a first aspect of the application there is provided a Src kinase-targeting short peptide comprising at least a peptide derived from
Amino acid residues 1-18 (including human hN18 and murine mN 18) at the N-terminal of STING protein are fused with cell penetrating peptide to enhance cell penetrating ability. Experiments prove that the N18 polypeptide cell penetrating peptide specifically binds to Src kinase through a PXXP structural domain.
In particular, cell penetrating peptides are a class of short peptides that carry macromolecular substances into cells, including naturally occurring cell penetrating peptides as well as synthetic cell penetrating peptides, and in one embodiment of the application, the cell penetrating peptides used are derived from amino acid residues 47-57 (YGRKRRRRR) of Human Immunodeficiency Virus (HIV) -1, which have been widely used for delivering exogenous macromolecules into cells.
Further, to facilitate tracking of the localization of the polypeptide after it has entered the cell, the N-terminus of the polypeptide is labeled with Biotin (Biotin). Thus, the polypeptide sequence of the application is any one of the following:
1.hN18:Biotin-YGRKKRRQRRR-MPHSSLHPSIPCPRGHGA(SEQ ID NO.1)；
2.mN18:Biotin-YGRKKRRQRRR-MPYSNLHPAIPRPRGHRS(SEQ ID NO.2)。
specifically, the N18 polypeptide provided by the application has been shown to inhibit STING-Src binding at the cellular level, promoting recruitment and activation of downstream Syk kinase, thereby enhancing activation of the C-type lectin receptor pathway; remarkably enhances the mRNA transcription level of inflammatory cytokines TNF-alpha, IL-6 and the like, plays a role in enhancing host antifungal immune response, and is helpful for eliminating pathogenic bacteria.
In a second aspect of the present application, there is provided a process for producing the above-mentioned short peptide, which comprises synthesizing the above-mentioned polypeptide by a chemical method, specifically by a solid-phase polypeptide synthesis method.
In a third aspect of the application, there is provided the use of the above-described short peptide in the preparation of a systemic fungal infection product.
The function of the product is at least one of the following (a 1) - (a 4):
(a1) Specifically binds Src, thereby inhibiting Src binding to STING;
(a2) Promoting Src-Syk complex formation;
(a3) Promoting expression of antifungal inflammatory cytokines and chemokines;
(a4) Promoting the elimination of pathogenic fungi in invasive fungal infections.
In a fourth aspect of the application there is provided a medicament for the treatment of systemic fungal infections comprising at least the above-described short peptide.
In a fifth aspect of the application, there is provided a method of treating a systemic fungal infection, the method comprising administering to a subject the above-described short peptide or the above-described medicament.
The subject of the present application is an animal, preferably a mammal, most preferably a human, who has been the subject of treatment, observation or experiment.
The beneficial technical effects of one or more of the technical schemes are as follows:
(1) The N18 polypeptide in the technical scheme specifically binds to Src kinase and inhibits the binding of Src and STING, so that the inhibiting effect of STING on a CLR signal path is relieved; enhanced expression (mRNA transcription level and protein secretion level) of inflammatory cytokines (TNF-alpha, IL-6); the immune response of the host against fungal infection is enhanced at both the cellular and animal level.
(2) Compared with the full-length protein, the N18 polypeptide in the technical scheme has small molecular weight, is easy to process and synthesize, has weak immunogenicity and relatively weak side effect; and the cell penetrating peptide is better in absorption, and the polypeptide mark is easy to trace and observe, so that a reference is provided for clinically treating the fungal disease.
Drawings
The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this specification, illustrate embodiments of the application and together with the description serve to explain the application.
Fig. 1: sting KO Systemic infection of mice with low-dose Candida albicans (1×10) 5 After/only) its survival rate was significantly increased over 21 days, with a slower decrease in body weight than the wild type.
Fig. 2: sting KO Systemic infection of mice with high-dose Candida albicans (2×10) 5 After/only) its survival rate was significantly increased over 21 days, with a slower decrease in body weight than the wild type.
Fig. 3: sting KO And wild-type mice systematically infected with Candida albicans (1X 10) 5 After/only), the fungal load in kidney, liver and spleen was significantly lower than in wild type mice.
Fig. 4: sting KO And wild-type mice systematically infected with Candida albicans (1X 10) 5 After/only), the kidneys were histopathologically stained. H&E staining visible Sting KO Is significantly lower than the wild type; PAS dyeing shows Sting KO Only a small number of hyphae were visible in the kidneys of mice compared to the wild type; ly6G staining showed less neutrophil infiltration.
Fig. 5: in vitro culture Sting KO And bone marrow derived dendritic cells (BMDC) from wild-type mice, and stimulated with dextran Zymosan, heat-killed candida albicans Yeast (Yeast), and hyphal (Hyphae), respectively. And an exogenous DNA (interferon stimulatory DNA, ISD) activating the STING signal pathway was added as a positive control. qPCR detection found Sting KO After fungal stimulation of BMDC secretion, the ability to secrete inflammatory cytokines is enhanced.
Fig. 6: in vitro culture Sting KO And bone of wild type miceMedullary derived dendritic cells (BMDC) stimulated with dextran Zymosan, heat-killed Candida albicans yeast (HKCA-Y) and mycelium (HKCA-H), respectively, ELISA detection found Sting KO The ability of BMDC to secrete inflammatory cytokines after stimulation by fungi is enhanced.
Fig. 7: in vitro culture Sting KO And bone marrow derived dendritic cells (BMDC) from wild type mice, stimulated with dextran Zymd for 15, 30, 60 minutes, respectively, and Western blot detection found Sting KO After fungal stimulation, activation of CLR signaling pathways is enhanced.
Fig. 8: immunofluorescence of distribution of Biotin-mN18 polypeptide in BMDC cells;
fig. 9: BMDC was pretreated with mN18 polypeptide (5. Mu.M, 10. Mu.M) and mN18 mutant polypeptide (10. Mu.M) for 2 hours, and dextran particles Zymosan were added thereto, and heated to stimulate the lethal candida albicans Yeast state (Yeast) and Hyphae state (Hyphae) for two hours. qPCR measures the transcript levels of the inflammatory cytokines TNF- α, IL-6 and chemokine CXCL1 and finds that polypeptide-treated BMDCs have an increased ability to secrete these factors upon fungal stimulation.
Fig. 10: BMDCs were pretreated with mN18 polypeptide (10. Mu.M) and mN18 mutant polypeptide (10. Mu.M) for 2 hours and then stimulated with dextran pellet Zymd for 15 and 30 minutes. Syk-Src complex formation was detected by immunoprecipitation techniques. It was found that complex formation was enhanced upon fungal stimulation of the polypeptide-treated BMDCs.
Fig. 11: c57BL/6 mice build a model of Candida albicans systemic infection. mN18 polypeptide (5 mg/kg) was injected intraperitoneally three days before the mN18 group, mN18 mutant polypeptide (5 mg/kg) was injected intraperitoneally three days before the mutant group mice, and PBS solution was injected daily three days before the control group mice. The mice were measured daily for 21 days for their body weight change patterns and survival patterns.
Fig. 12: c57BL/6 mice build a model of Candida albicans systemic infection. mN18 polypeptide (5 mg/kg) was injected intraperitoneally three days before the mN18 group, mN18 mutant polypeptide (5 mg/kg) was injected intraperitoneally three days before the mutant group mice, and PBS solution was injected daily three days before the control group mice. The fifth day kidney, liver and spleen were ground and the fungal load in the viscera was measured.
Fig. 13: c57BL/6 mice build a model of Candida albicans systemic infection. mN18 polypeptide (5 mg/kg) was injected intraperitoneally three days before the mN18 group, mN18 mutant polypeptide (5 mg/kg) was injected intraperitoneally three days before the mutant group mice, and PBS solution was injected daily three days before the control group mice. Kidneys were taken on day five for H & E staining, PAS staining and Ly6G staining.
Detailed Description
It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the application. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present application. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.
As previously mentioned, there are still few studies on antifungal signaling pathways for STING, and further studies are still needed.
In view of this, the present application has found that STING exerts the effect of negatively regulating the host immune response in mice with systemic infection patterns by the pathogenic fungus candida albicans. The action mechanism mainly forms a complex with tyrosine kinase Src through 18 amino acid residues at the N end of the self (humanized STING:1-MPHSSLHPSIPCPRGHGA-18; murine STING: 1-MPYSNLHPAIPRPRGHRS-18), so that the downstream kinase Syk cannot be recruited and activated by the Src. Syk plays a pivotal role in the key signaling pathway of antifungal infection, the C-lectin (CLR) signaling pathway, inhibition of which activity results in failure of the host to normally exert an antifungal immune response. With respect to STING having the property of blocking Src-Syk complex formation, researchers synthesized amino acids (hN 18 (human) and mN18 (murine)) at positions 1-18 of STING protein and fused TAT transmembrane peptide (YGRKKRRQRRR) at the N-terminus. Cell penetrating peptide enhances the ability of the short peptide to enter cells, improving the absorbability of the short peptide. After the short peptide enters cells, the short peptide competitively binds with Src to inhibit the formation of Src and STING complexes, so that the inhibiting effect of STING on antifungal signal paths is lightened, and the short peptide has important significance for promoting host antifungal immune response and controlling infection progress.
Specifically, in one exemplary embodiment of the present application, a Src kinase-targeted short peptide is provided, which includes at least residues 1-18 (including human hN18 and murine mN 18) from the N-terminal amino acid of STING protein, and is fused to a cell-penetrating peptide to enhance cell penetration. Experiments prove that the N18 polypeptide cell penetrating peptide specifically binds to Src kinase through a PXXP structural domain.
Among them, cell penetrating peptides are a class of short peptides capable of carrying macromolecular substances into cells, including naturally occurring cell penetrating peptides as well as artificially synthesized cell penetrating peptides, and in one embodiment of the present application, the cell penetrating peptides used are derived from amino acid residues 47-57 (YGRKKRRQRRR) of Human Immunodeficiency Virus (HIV) -1, which have been widely used for delivering exogenous macromolecules into cells.
Further, to facilitate tracking of the localization of the polypeptide after it has entered the cell, the N-terminus of the polypeptide is labeled with Biotin (Biotin). Therefore, the short peptide sequence of the application is any one of the following:
1.hN18:Biotin-YGRKKRRQRRR-MPHSSLHPSIPCPRGHGA(SEQ ID NO.1)；
2.mN18:Biotin-YGRKKRRQRRR-MPYSNLHPAIPRPRGHRS(SEQ ID NO.2)。
specifically, the N18 polypeptide provided by the application has been shown to inhibit STING-Src binding at the cellular level, promoting recruitment and activation of downstream Syk kinase, thereby enhancing activation of the C-type lectin receptor pathway; remarkably enhances the mRNA transcription level of inflammatory cytokines TNF-alpha, IL-6 and the like, plays a role in enhancing host antifungal immune response, and is helpful for eliminating pathogenic bacteria.
It should be noted that although the present application uses STING proteins of human and murine origin as examples, it is apparent that other sources of STING proteins are also within the scope of the present application.
In still another embodiment of the present application, there is provided a method for producing the above-mentioned short peptide, which comprises synthesizing the above-mentioned polypeptide by a chemical method, specifically, by a solid-phase polypeptide synthesis method.
In yet another embodiment of the present application, there is provided the use of the above-described short peptides in the preparation of a systemic fungal infection product.
The function of the product is at least one of the following (a 1) - (a 4):
(a1) Specifically binds Src, thereby inhibiting Src binding to STING;
(a2) Promoting Src-Syk complex formation;
(a3) Promoting expression of antifungal inflammatory cytokines and chemokines;
(a4) Promoting the elimination of pathogenic fungi in invasive fungal infections.
In the present application, the antifungal inflammatory cytokines include, but are not limited to, TNF- α, IL-6, and the chemokines include, but are not limited to, CXCL-1.
Such fungi include, but are not limited to, fungi (i.e., pathogenic fungi) pathogenic to animals (including humans) such as candida, aspergillus, cryptococcus, and penicillium, and further candida such as candida albicans.
The product may be a pharmaceutical or experimental agent that may be used for basic research, such as for related studies on the mechanism of fungal infection.
In yet another embodiment of the present application, there is provided a medicament for treating systemic fungal infections comprising at least the above-described short peptide.
In particular, the medicament may further comprise a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier can be buffer, emulsifying agent, suspending agent, stabilizer, antiseptic, excipient, filler, coagulating agent and concocting agent
And a neutralizing agent, a surfactant, a dispersing agent, or a defoaming agent.
The medicament may also include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be a virus, a microcapsule, a liposome, a nanoparticle, or a polymer, and any combination thereof. The delivery vehicle for the pharmaceutically acceptable carrier may be a liposome, biocompatible polymer (including natural and synthetic polymers), lipoprotein, polysaccharide, lipopolysaccharide, artificial viral envelope, inorganic (including metallic) particles, as well as bacterial or viral (e.g., baculovirus, adenovirus, and retrovirus), phage, cosmid, or plasmid vector.
In the present application, the medicament may also be used in combination with other medicaments for the prevention and/or treatment of fungal infections, and other prophylactic and/or therapeutic compounds may be administered simultaneously with the main active ingredient, even in the same composition.
The medicament may also be administered alone in separate compositions or in a dosage form different from the primary active ingredient, with other prophylactic and/or therapeutic compounds. A partial dose of the principal component may be administered simultaneously with other therapeutic compounds, while other doses may be administered separately. The dosage of the medicament of the application may be adjusted during the course of treatment according to the severity of the symptoms, the frequency of recurrence and the physiological response of the treatment regimen.
The medicament of the application may be administered to the body in a known manner. For example, by intravenous systemic delivery or local injection into the tissue of interest. Alternatively via intravenous, transdermal, intranasal, mucosal or other delivery methods. Such administration may be via single or multiple doses. It will be appreciated by those skilled in the art that the actual dosage to be administered in the present application may vary greatly depending on a variety of factors, such as the target cell, the type of organism or tissue thereof, the general condition of the subject to be treated, the route of administration, the mode of administration, and the like.
In yet another embodiment of the present application, there is provided a method of treating a systemic fungal infection, the method comprising administering to a subject the above-described short peptide or the above-described drug.
The subject of the present application is an animal, preferably a mammal, most preferably a human, who has been the subject of treatment, observation or experiment.
The application will now be further illustrated with reference to specific examples, which are given for the purpose of illustration only and are not intended to be limiting in any way. If experimental details are not specified in the examples, it is usually the case that the conditions are conventional or recommended by the reagent company; reagents, consumables, etc. used in the examples described below are commercially available unless otherwise specified.
The application is further illustrated by the following examples, which are not to be construed as limiting the application. It is to be understood that these examples are illustrative of the present application and are not intended to limit the scope of the present application. The following examples are test methods in which specific conditions are noted, and are generally conducted under conventional conditions.
Example 1: STING inhibits host anti-fungal immune responses
By constructing a model of mice systematically infected with candida albicans, STING is found to negatively regulate the immune response of hosts against fungal infection at animal level and cell level.
1.1 STING plays a role in down-regulation in a model of systemic fungal infection
C57BL/6j mice with Sting whole body knockdown (Sting) KO ) A systemic infection model was constructed by tail vein injection of Candida albicans. Sting compared to wild-type mice KO Mice were significantly improved in survival over 21 days after low/high dose candida albicans infection, respectively (figures 1, 2); fungal loads were significantly lower in kidney, liver and spleen than in wild type mice (fig. 3). H for kidney after infection&E staining, see Sting KO Is significantly lower than the wild type; PAS dyeing shows Sting KO Only a small number of hyphae were visible in the kidneys of mice compared to the wild type; ly6G staining showed less neutrophil infiltration (fig. 4). The above results suggest that STING is involved in and negatively regulates the host immune response against fungal infection.
1.2 STING inhibits activation of the C-lectin receptor (CLR) signaling pathway
Sting was found by in vitro culturing Bone Marrow Derived Macrophages (BMDM) and dendritic cells (BMDC) and stimulating with dextran, heat-killed Candida albicans, etc., respectively KO Is to be immunized with (a)The cells have an enhanced ability to secrete inflammatory cytokines (FIGS. 5, 6). The inhibition of phosphorylation of important kinases in CLR pathways by STING, including Syk, PLC- γ, was demonstrated by Western blot experiments, which in turn inhibited activation of transcription factors NF- κb and MAPK kinase (fig. 7). These results suggest that STING negatively regulates activation of CLR signaling pathway.
Example 2: STING protein binds Src kinase through 18 amino acids at the N-terminus
STING relies primarily on its N-terminal 18 amino acids to interact with Src, blocking Src recruitment and activation of downstream Syk kinase, and inhibiting activation of downstream NF- κb signaling pathways. Therefore, the N-terminal 18 amino acid polypeptide of STING protein is designed, and the polypeptide with cell permeability is synthesized, and the sequence is as follows. Critical for binding Src 8 -PXXPXP- 13 The domain mutation was AXXAXA (underlined). After the design is completed, the Nanjing source peptide limited company is entrusted to synthesize and purify the polypeptide based on the sequences.
1.hN18:Biotin-YGRKKRRQRRR-MPHSSLHPSIPCPRGHGA(SEQ ID NO.1)；
2.hN18 mutant：Biotin-YGRKKRRQRRR-MPHSSLHASIACARGHGA(SEQ ID NO.3)
3.mN18:Biotin-YGRKKRRQRRR-MPYSNLHPAIPRPRGHRS(SEQ ID NO.2)
4.mN18 mutant:Biotin-YGRKKRRQRRR-MPYSNLHAAIARARGHRS(SEQ ID NO.4)
Mouse BMDC cells were taken, incubated with Biotin-mN18 polypeptide (5. Mu.M) for 2 hours, and the fluorescence intensity of the polypeptide and the distribution in the cells were examined by a laser confocal microscope, and the results showed that: 2 hours after the addition of the polypeptide, biotin-mN18 enters the cells and is uniformly distributed in the cells; after 20 minutes of stimulation with red fluorescent-labeled dextran particles (Texas red-labeled Zymosan), the polypeptide translocates to the phagosome membrane and co-localizes with Src (fig. 8).
Example 3: verification of the Effect of Polypeptides on inflammatory cytokine expression
Biotin-mN18 polypeptide (5. Mu.M, 10. Mu.M) was added to mice BMDC for pretreatment for 2 hours, and then dextran, heat-killed Candida albicans, etc. were added to stimulate the cells. mRNA transcript levels of the inflammatory cytokines TNF- α, IL-6 and chemokine CXCL-1 were detected by RT-PCR after 2 hours. FIG. 9 shows that mN18 polypeptide pretreatment significantly promotes secretion of pro-inflammatory cytokines and chemokines.
Example 4: verification of Effect of Polypeptides on Syk-Src Complex formation
Biotin-mN18 polypeptide (5. Mu.M) was added to mouse BMDC for pretreatment for 2 hours, dextran was added for 15 and 30 minutes, and the effect of Syk-Src complex formation was examined by immunoprecipitation. FIG. 10 shows that pretreatment with mN18 polypeptide promotes complex formation.
Example 5: verifying the role of the polypeptide in the model of Candida albicans systemic infection
5.1 construction of a model of systemic fungal infection in mice
SPF grade, 7-8 week old C57BL/6 male mice, weighing 20-22g, purchased from Beijing Vittolihua, fed to SPF animal center at Shandong university, divided into 3 groups: mN18 (15) and mN18 mutant group (11) and control group (15), respectively. Three groups were each injected by mouse tail vein with candida albicans (2 x 10 per mouse injection) 5 Individual fungal cells). mN18 polypeptide (5 mg/kg) was injected intraperitoneally three days before the mN18 group, mN18 mutant polypeptide (5 mg/kg) was injected intraperitoneally three days before the mutant group mice, and PBS solution was injected daily three days before the control group mice. The weight change and survival curves of each group of mice were recorded daily for a total of 21 days, starting with the model of systemic fungal infection in the mice.
5.2 histopathological observations and assessment
Candida albicans was injected intravenously via the tail of mice (2 x 10 per mouse injection) 5 Individual fungal cells) to construct a systemic infection model. mN18 polypeptide (5 mg/kg) was intraperitoneally injected daily three days before the mN18 group, and mN18 mutant polypeptide (5 mg/kg) was intraperitoneally injected daily three days before the mutant group mice. Taking kidney tissue, fixing in 4% paraformaldehyde, and delivering to hematoxylin-eosin (H) of Wohai Weir Biotech company&E) Staining, glycogen staining (schiff periodic acid shiff, PAS) and neutrophil (Ly 6G) staining. Pathological conditions of kidney tissue were observed using a panoramic slice scanner and basic pathological changes such as congestion, blood stasis, and the like in the sections were evaluated,Bleeding, edema, degeneration, necrosis, hyperplasia, fibrosis, granulation tissue, inflammatory changes, etc. Each parameter is rated by severity on a scale of 0-4, the higher the scale the more severe the pathological damage to the colon tissue.
The results show that compared with the intraperitoneal injection of the mutant polypeptide group, the injection of the mN18 polypeptide group significantly reduces the weight of the mice (figure 11), the clearance capacity of kidneys to pathogenic fungi is significantly improved (figure 12), and the H & E staining results show that the kidney damage of the mN18 polypeptide group is significantly reduced compared with the control group for H & E staining; PAS staining showed that only a small number of hyphae were visible in the kidneys of the mN18 polypeptide group; ly6G staining suggested less renal neutrophil infiltration of the mN18 polypeptide group (FIG. 13).
The application is not a matter of the known technology.
The above embodiments are provided to illustrate the technical concept and features of the present application and are intended to enable those skilled in the art to understand the content of the present application and implement the same, and are not intended to limit the scope of the present application. All equivalent changes or modifications made in accordance with the spirit of the present application should be construed to be included in the scope of the present application.
Claims (10)
1. A Src kinase-targeted short peptide, which at least comprises amino acid residues 1-18 at the N-terminal of STING protein, and is further modified with a cell-penetrating peptide.
2. A Src kinase targeted short peptide according to claim 1 wherein the cell penetrating peptide is TAT cell penetrating peptide.
3. A Src kinase targeted short peptide according to claim 1, wherein said short peptide is further modified with biotin.
4. A Src kinase targeted short peptide according to any one of claims 1 to 3, wherein said short peptide is as shown in any one of SEQ ID No.1 to 2.
5. The method for producing a short peptide according to any one of claims 1 to 4, wherein the method comprises synthesizing by a chemical method; further, the chemical method includes a solid-phase polypeptide synthesis method.
6. Use of the short peptide according to any one of claims 1 to 4 for the preparation of a product for systemic fungal infection.
7. The use according to claim 6, wherein the function of the product is at least one of the following (a 1) - (a 4):
(a1) Specifically binds Src, thereby inhibiting Src binding to STING;
(a2) Promoting Src-Syk complex formation;
(a3) Promoting expression of antifungal inflammatory cytokines and chemokines;
(a4) Promoting the elimination of pathogenic bacteria in invasive fungus infection models.
8. The use of claim 7, wherein the antifungal inflammatory cytokine comprises TNF- α, IL-6 and the chemokine comprises CXCL-1;
the fungi include candida, aspergillus, cryptococcus and penicillium, and further candida including candida albicans.
9. The use according to any one of claims 6 to 8, wherein the product is a pharmaceutical or experimental agent for use in basic research.
10. A medicament for the treatment of systemic fungal infections, characterized in that it contains at least a short peptide according to any one of claims 1-4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310619462.0A CN116675779B (en) | 2023-05-26 | Short peptide targeting Src kinase and application thereof in systemic fungal infection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310619462.0A CN116675779B (en) | 2023-05-26 | Short peptide targeting Src kinase and application thereof in systemic fungal infection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116675779A true CN116675779A (en) | 2023-09-01 |
| CN116675779B CN116675779B (en) | 2024-07-09 |
Family
ID=
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020045191A1 (en) * | 2000-09-15 | 2002-04-18 | Schneider Robert J. | Inhibition of the SRC kinase family pathway as a method of treating HBV infection and hepatocellular carcinoma |
| CN108137641A (en) * | 2015-08-13 | 2018-06-08 | 默沙东公司 | Cyclic annular dinucleotide compound as sting agonists |
| CN111944032A (en) * | 2020-07-29 | 2020-11-17 | 扬州大学 | Polypeptide capable of inducing anti-chicken STING protein antibody and application thereof |
| CN111978408A (en) * | 2020-07-14 | 2020-11-24 | 山东大学 | IKK beta-targeting short peptides and application thereof in inflammatory diseases |
| CN116003524A (en) * | 2022-09-20 | 2023-04-25 | 山东大学齐鲁医院 | Short peptide targeting cGAS, fusion polypeptide and application |
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020045191A1 (en) * | 2000-09-15 | 2002-04-18 | Schneider Robert J. | Inhibition of the SRC kinase family pathway as a method of treating HBV infection and hepatocellular carcinoma |
| CN108137641A (en) * | 2015-08-13 | 2018-06-08 | 默沙东公司 | Cyclic annular dinucleotide compound as sting agonists |
| CN111978408A (en) * | 2020-07-14 | 2020-11-24 | 山东大学 | IKK beta-targeting short peptides and application thereof in inflammatory diseases |
| CN111944032A (en) * | 2020-07-29 | 2020-11-17 | 扬州大学 | Polypeptide capable of inducing anti-chicken STING protein antibody and application thereof |
| CN116003524A (en) * | 2022-09-20 | 2023-04-25 | 山东大学齐鲁医院 | Short peptide targeting cGAS, fusion polypeptide and application |
Non-Patent Citations (2)
| Title |
|---|
| ISHIKAWA, H等: "STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling", 《NATURE》, vol. 455, no. 7213, 2 October 2008 (2008-10-02), pages 674 - 678, XP055021546, DOI: 10.1038/nature07317 * |
| 赵若楠等: "STING受体在布鲁氏菌感染固有免疫中的作用机制研究进展", 《中国人兽共患病学报》, vol. 39, no. 01, 31 January 2023 (2023-01-31), pages 67 - 72 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3038636B1 (en) | Administration of kynurenine depleting enzymes for tumor therapy | |
| JP6680760B2 (en) | Antagonist IC PD-1 aptamer and its application for use in cancer therapy | |
| WO2021027704A1 (en) | Application of polypeptide or derivative thereof | |
| US20230270824A1 (en) | Interferon-based cancer treatment method and pharmaceutical composition | |
| JP2020519230A (en) | Envelope virus resistant to complement inactivation for the treatment of cancer | |
| KR20180056695A (en) | Treatment of neurodegenerative diseases | |
| CN110393791B (en) | Anti-infection effect of hnRNPA2B1 and application thereof | |
| CN113769063B (en) | Application of polypeptide PTPR in preparation of tumor immunotherapy medicament | |
| US20070281041A1 (en) | Compositions and Methods Involving MDA-7 for the Treatment of Cancer | |
| KR20200143455A (en) | Human kinureninase enzyme and usage thereof | |
| JP2015091826A (en) | Combination products for treating cancer | |
| CN116675779B (en) | Short peptide targeting Src kinase and application thereof in systemic fungal infection | |
| US11510999B2 (en) | Treatment of neuropathy with DNA constructs expressing IGF-1 isoforms | |
| JP7362156B2 (en) | Pharmaceutical compositions, kits and methods for treating tumors | |
| CN116675779A (en) | Short peptide targeting Src kinase and application thereof in systemic fungal infection | |
| US10533179B2 (en) | Antagonistic CTLA-4 aptamers and applications thereof in enhancing immune activity | |
| EP2510091B1 (en) | Method and composition to increase radiation-induced tumor therapeutic effects | |
| WO2013126872A1 (en) | Anti-cancer therapeutic strategy to overcome cancer resistance and to enable tailoring treatment to patients | |
| US20090221675A1 (en) | Use of iex-1 for the treatment of glioma tumors | |
| AU2020343018A1 (en) | Method for increasing lymphocyte count by using IL-7 fusion protein in tumors | |
| EP1854474A1 (en) | Immunopotentiating agent | |
| KR20150054543A (en) | Liver-targeted nanoparticles, a preparation method thereof, and a pharmaceutical composition comprising the nanoparticles | |
| WO2024065651A1 (en) | Method for loading dimer cd24 into hek293 cell exosome with adam10 gene knocked out | |
| WO2007127951A2 (en) | Compositions and methods involving mda-7 for the treatment of cancer | |
| WO2019146805A1 (en) | Therapeutic agent for frontotemporal lobar degeneration, method for screening therapeutic agent for frontotemporal lobar degeneration and method for treating frontotemporal lobar degeneration |
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 |