CN114181279A - 抗菌多肽化合物、医疗器械、水凝胶及其应用 - Google Patents
抗菌多肽化合物、医疗器械、水凝胶及其应用 Download PDFInfo
- Publication number
- CN114181279A CN114181279A CN202110407232.9A CN202110407232A CN114181279A CN 114181279 A CN114181279 A CN 114181279A CN 202110407232 A CN202110407232 A CN 202110407232A CN 114181279 A CN114181279 A CN 114181279A
- Authority
- CN
- China
- Prior art keywords
- leu
- compound
- arg
- hydrogel
- pro
- 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
- 150000001875 compounds Chemical class 0.000 title claims abstract description 151
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 90
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 90
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 81
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 77
- 239000000017 hydrogel Substances 0.000 title claims abstract description 75
- 239000003814 drug Substances 0.000 claims abstract description 34
- 229940079593 drug Drugs 0.000 claims abstract description 25
- 239000003102 growth factor Substances 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 14
- 230000029663 wound healing Effects 0.000 claims abstract description 9
- 239000000835 fiber Substances 0.000 claims abstract description 6
- 229940124599 anti-inflammatory drug Drugs 0.000 claims abstract description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 24
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 20
- 150000001413 amino acids Chemical group 0.000 claims description 13
- 230000000845 anti-microbial effect Effects 0.000 claims description 11
- 239000011780 sodium chloride Substances 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 8
- 239000007836 KH2PO4 Substances 0.000 claims description 7
- 239000007853 buffer solution Substances 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 7
- 239000008055 phosphate buffer solution Substances 0.000 claims description 6
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 5
- 230000001737 promoting effect Effects 0.000 claims description 5
- 238000004873 anchoring Methods 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000004132 cross linking Methods 0.000 claims description 4
- 238000006116 polymerization reaction Methods 0.000 claims description 4
- GLGNXYJARSMNGJ-VKTIVEEGSA-N (1s,2s,3r,4r)-3-[[5-chloro-2-[(1-ethyl-6-methoxy-2-oxo-4,5-dihydro-3h-1-benzazepin-7-yl)amino]pyrimidin-4-yl]amino]bicyclo[2.2.1]hept-5-ene-2-carboxamide Chemical compound CCN1C(=O)CCCC2=C(OC)C(NC=3N=C(C(=CN=3)Cl)N[C@H]3[C@H]([C@@]4([H])C[C@@]3(C=C4)[H])C(N)=O)=CC=C21 GLGNXYJARSMNGJ-VKTIVEEGSA-N 0.000 claims description 3
- UNILWMWFPHPYOR-KXEYIPSPSA-M 1-[6-[2-[3-[3-[3-[2-[2-[3-[[2-[2-[[(2r)-1-[[2-[[(2r)-1-[3-[2-[2-[3-[[2-(2-amino-2-oxoethoxy)acetyl]amino]propoxy]ethoxy]ethoxy]propylamino]-3-hydroxy-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-[(2r)-2,3-di(hexadecanoyloxy)propyl]sulfanyl-1-oxopropan-2-yl Chemical compound O=C1C(SCCC(=O)NCCCOCCOCCOCCCNC(=O)COCC(=O)N[C@@H](CSC[C@@H](COC(=O)CCCCCCCCCCCCCCC)OC(=O)CCCCCCCCCCCCCCC)C(=O)NCC(=O)N[C@H](CO)C(=O)NCCCOCCOCCOCCCNC(=O)COCC(N)=O)CC(=O)N1CCNC(=O)CCCCCN\1C2=CC=C(S([O-])(=O)=O)C=C2CC/1=C/C=C/C=C/C1=[N+](CC)C2=CC=C(S([O-])(=O)=O)C=C2C1 UNILWMWFPHPYOR-KXEYIPSPSA-M 0.000 claims description 3
- 230000003110 anti-inflammatory effect Effects 0.000 claims description 3
- 229940124350 antibacterial drug Drugs 0.000 claims description 3
- 229940125758 compound 15 Drugs 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- -1 rinses Substances 0.000 claims description 3
- 239000007921 spray Substances 0.000 claims description 3
- SZUVGFMDDVSKSI-WIFOCOSTSA-N (1s,2s,3s,5r)-1-(carboxymethyl)-3,5-bis[(4-phenoxyphenyl)methyl-propylcarbamoyl]cyclopentane-1,2-dicarboxylic acid Chemical compound O=C([C@@H]1[C@@H]([C@](CC(O)=O)([C@H](C(=O)N(CCC)CC=2C=CC(OC=3C=CC=CC=3)=CC=2)C1)C(O)=O)C(O)=O)N(CCC)CC(C=C1)=CC=C1OC1=CC=CC=C1 SZUVGFMDDVSKSI-WIFOCOSTSA-N 0.000 claims description 2
- GHYOCDFICYLMRF-UTIIJYGPSA-N (2S,3R)-N-[(2S)-3-(cyclopenten-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-hydroxy-3-(4-methoxyphenyl)-2-[[(2S)-2-[(2-morpholin-4-ylacetyl)amino]propanoyl]amino]propanamide Chemical compound C1(=CCCC1)C[C@@H](C(=O)[C@@]1(OC1)C)NC([C@H]([C@@H](C1=CC=C(C=C1)OC)O)NC([C@H](C)NC(CN1CCOCC1)=O)=O)=O GHYOCDFICYLMRF-UTIIJYGPSA-N 0.000 claims description 2
- QFLWZFQWSBQYPS-AWRAUJHKSA-N (3S)-3-[[(2S)-2-[[(2S)-2-[5-[(3aS,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]-3-methylbutanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-4-[1-bis(4-chlorophenoxy)phosphorylbutylamino]-4-oxobutanoic acid Chemical compound CCCC(NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@@H](NC(=O)CCCCC1SC[C@@H]2NC(=O)N[C@H]12)C(C)C)P(=O)(Oc1ccc(Cl)cc1)Oc1ccc(Cl)cc1 QFLWZFQWSBQYPS-AWRAUJHKSA-N 0.000 claims description 2
- 230000004888 barrier function Effects 0.000 claims description 2
- 229940125904 compound 1 Drugs 0.000 claims description 2
- 229940125773 compound 10 Drugs 0.000 claims description 2
- 229940125797 compound 12 Drugs 0.000 claims description 2
- 229940126543 compound 14 Drugs 0.000 claims description 2
- 229940125782 compound 2 Drugs 0.000 claims description 2
- 229940126214 compound 3 Drugs 0.000 claims description 2
- 229940125898 compound 5 Drugs 0.000 claims description 2
- 239000008298 dragée Substances 0.000 claims description 2
- 239000006196 drop Substances 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 238000012377 drug delivery Methods 0.000 claims description 2
- 239000004744 fabric Substances 0.000 claims description 2
- 239000006260 foam Substances 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- ZLVXBBHTMQJRSX-VMGNSXQWSA-N jdtic Chemical compound C1([C@]2(C)CCN(C[C@@H]2C)C[C@H](C(C)C)NC(=O)[C@@H]2NCC3=CC(O)=CC=C3C2)=CC=CC(O)=C1 ZLVXBBHTMQJRSX-VMGNSXQWSA-N 0.000 claims description 2
- 239000004005 microsphere Substances 0.000 claims description 2
- 239000002674 ointment Substances 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 230000000379 polymerizing effect Effects 0.000 claims description 2
- 239000008174 sterile solution Substances 0.000 claims description 2
- 239000007940 sugar coated tablet Substances 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 239000003106 tissue adhesive Substances 0.000 claims description 2
- 229940075469 tissue adhesives Drugs 0.000 claims description 2
- 239000004599 antimicrobial Substances 0.000 claims 2
- 239000002260 anti-inflammatory agent Substances 0.000 claims 1
- 239000000872 buffer Substances 0.000 claims 1
- 206010052428 Wound Diseases 0.000 abstract description 13
- 208000027418 Wounds and injury Diseases 0.000 abstract description 13
- 230000002439 hemostatic effect Effects 0.000 abstract description 10
- 238000002360 preparation method Methods 0.000 abstract description 8
- 230000015572 biosynthetic process Effects 0.000 abstract description 4
- 238000011068 loading method Methods 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 3
- 102000018386 EGF Family of Proteins Human genes 0.000 abstract description 2
- 108010066486 EGF Family of Proteins Proteins 0.000 abstract description 2
- 231100000241 scar Toxicity 0.000 abstract description 2
- 230000006444 vascular growth Effects 0.000 abstract description 2
- 239000003910 polypeptide antibiotic agent Substances 0.000 description 33
- 241000699670 Mus sp. Species 0.000 description 21
- 241000588724 Escherichia coli Species 0.000 description 18
- 230000000694 effects Effects 0.000 description 18
- 239000012528 membrane Substances 0.000 description 17
- 210000004027 cell Anatomy 0.000 description 16
- 230000004083 survival effect Effects 0.000 description 15
- 108700042778 Antimicrobial Peptides Proteins 0.000 description 14
- 102000044503 Antimicrobial Peptides Human genes 0.000 description 14
- 230000001580 bacterial effect Effects 0.000 description 13
- 230000035699 permeability Effects 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- YBYIRNPNPLQARY-UHFFFAOYSA-N 1H-indene Chemical compound C1=CC=C2CC=CC2=C1 YBYIRNPNPLQARY-UHFFFAOYSA-N 0.000 description 10
- 241000894006 Bacteria Species 0.000 description 10
- 208000032843 Hemorrhage Diseases 0.000 description 10
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- 241000699666 Mus <mouse, genus> Species 0.000 description 9
- 239000013642 negative control Substances 0.000 description 9
- 230000000740 bleeding effect Effects 0.000 description 8
- 238000011160 research Methods 0.000 description 8
- 239000011347 resin Substances 0.000 description 8
- 229920005989 resin Polymers 0.000 description 8
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 7
- 238000011156 evaluation Methods 0.000 description 7
- 230000002949 hemolytic effect Effects 0.000 description 7
- 108010025306 histidylleucine Proteins 0.000 description 7
- 230000002401 inhibitory effect Effects 0.000 description 7
- 239000007788 liquid Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- XOWMDXHFSBCAKQ-SRVKXCTJSA-N Leu-Ser-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(C)C XOWMDXHFSBCAKQ-SRVKXCTJSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 6
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 6
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 230000003013 cytotoxicity Effects 0.000 description 6
- 231100000135 cytotoxicity Toxicity 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 238000001727 in vivo Methods 0.000 description 6
- 210000004185 liver Anatomy 0.000 description 6
- 239000013641 positive control Substances 0.000 description 6
- 238000012449 Kunming mouse Methods 0.000 description 5
- 230000007059 acute toxicity Effects 0.000 description 5
- 231100000403 acute toxicity Toxicity 0.000 description 5
- 125000003275 alpha amino acid group Chemical group 0.000 description 5
- 230000000843 anti-fungal effect Effects 0.000 description 5
- KUWPCJHYPSUOFW-YBXAARCKSA-N 2-nitrophenyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC=C1[N+]([O-])=O KUWPCJHYPSUOFW-YBXAARCKSA-N 0.000 description 4
- XTWYTFMLZFPYCI-KQYNXXCUSA-N 5'-adenylphosphoric acid Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XTWYTFMLZFPYCI-KQYNXXCUSA-N 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- XTWYTFMLZFPYCI-UHFFFAOYSA-N Adenosine diphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(O)=O)C(O)C1O XTWYTFMLZFPYCI-UHFFFAOYSA-N 0.000 description 4
- 241000233866 Fungi Species 0.000 description 4
- 206010018910 Haemolysis Diseases 0.000 description 4
- 108010036176 Melitten Proteins 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- 229920004890 Triton X-100 Polymers 0.000 description 4
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 4
- 210000003743 erythrocyte Anatomy 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000008588 hemolysis Effects 0.000 description 4
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 4
- VDXZNPDIRNWWCW-JFTDCZMZSA-N melittin Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(N)=O)CC1=CNC2=CC=CC=C12 VDXZNPDIRNWWCW-JFTDCZMZSA-N 0.000 description 4
- 238000010172 mouse model Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000005303 weighing Methods 0.000 description 4
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 3
- 241000415078 Anemone hepatica Species 0.000 description 3
- 241000222122 Candida albicans Species 0.000 description 3
- 239000007995 HEPES buffer Substances 0.000 description 3
- UEKYKRQIAQHOOZ-KBPBESRZSA-N Pro-Trp Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)[O-])C(=O)[C@@H]1CCC[NH2+]1 UEKYKRQIAQHOOZ-KBPBESRZSA-N 0.000 description 3
- 206010040047 Sepsis Diseases 0.000 description 3
- 210000000683 abdominal cavity Anatomy 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 229940095731 candida albicans Drugs 0.000 description 3
- 230000004663 cell proliferation Effects 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 229910000397 disodium phosphate Inorganic materials 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 230000002538 fungal effect Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 206010034674 peritonitis Diseases 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000006467 substitution reaction Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- YEDUAINPPJYDJZ-UHFFFAOYSA-N 2-hydroxybenzothiazole Chemical compound C1=CC=C2SC(O)=NC2=C1 YEDUAINPPJYDJZ-UHFFFAOYSA-N 0.000 description 2
- JDDWRLPTKIOUOF-UHFFFAOYSA-N 9h-fluoren-9-ylmethyl n-[[4-[2-[bis(4-methylphenyl)methylamino]-2-oxoethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamate Chemical compound COC1=CC(OC)=CC=C1C(C=1C=CC(OCC(=O)NC(C=2C=CC(C)=CC=2)C=2C=CC(C)=CC=2)=CC=1)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 JDDWRLPTKIOUOF-UHFFFAOYSA-N 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 2
- 229930182566 Gentamicin Natural products 0.000 description 2
- QUAAUWNLWMLERT-IHRRRGAJSA-N Leu-Arg-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(C)C)C(O)=O QUAAUWNLWMLERT-IHRRRGAJSA-N 0.000 description 2
- BGZCJDGBBUUBHA-KKUMJFAQSA-N Leu-Lys-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O BGZCJDGBBUUBHA-KKUMJFAQSA-N 0.000 description 2
- 108060003100 Magainin Proteins 0.000 description 2
- 206010034133 Pathogen resistance Diseases 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- IWIANZLCJVYEFX-RYUDHWBXSA-N Pro-Phe Chemical compound C([C@@H](C(=O)O)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 IWIANZLCJVYEFX-RYUDHWBXSA-N 0.000 description 2
- LGMBKOAPPTYKLC-JYJNAYRXSA-N Pro-Phe-Arg Chemical compound C([C@@H](C(=O)N[C@@H](CCCNC(=N)N)C(O)=O)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 LGMBKOAPPTYKLC-JYJNAYRXSA-N 0.000 description 2
- SWRNSCMUXRLHCR-ULQDDVLXSA-N Pro-Phe-Lys Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 SWRNSCMUXRLHCR-ULQDDVLXSA-N 0.000 description 2
- XNJVJEHDZPDPQL-BZSNNMDCSA-N Pro-Trp-Arg Chemical compound NC(=N)NCCC[C@H](NC(=O)[C@H](Cc1c[nH]c2ccccc12)NC(=O)[C@@H]1CCCN1)C(O)=O XNJVJEHDZPDPQL-BZSNNMDCSA-N 0.000 description 2
- 239000013504 Triton X-100 Substances 0.000 description 2
- 241000222126 [Candida] glabrata Species 0.000 description 2
- 210000001015 abdomen Anatomy 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 208000032343 candida glabrata infection Diseases 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 229960003405 ciprofloxacin Drugs 0.000 description 2
- 238000006482 condensation reaction Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 231100000517 death Toxicity 0.000 description 2
- 238000001804 debridement Methods 0.000 description 2
- 238000011033 desalting Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical group [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 229960002518 gentamicin Drugs 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000007928 intraperitoneal injection Substances 0.000 description 2
- 210000001985 kidney epithelial cell Anatomy 0.000 description 2
- 108010034529 leucyl-lysine Proteins 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000012452 mother liquor Substances 0.000 description 2
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 2
- 230000002980 postoperative effect Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000005086 pumping Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 239000012266 salt solution Substances 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- ZGYICYBLPGRURT-UHFFFAOYSA-N tri(propan-2-yl)silicon Chemical compound CC(C)[Si](C(C)C)C(C)C ZGYICYBLPGRURT-UHFFFAOYSA-N 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- ZPGDWQNBZYOZTI-SFHVURJKSA-N (2s)-1-(9h-fluoren-9-ylmethoxycarbonyl)pyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 ZPGDWQNBZYOZTI-SFHVURJKSA-N 0.000 description 1
- XXMYDXUIZKNHDT-QNGWXLTQSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(1-tritylimidazol-4-yl)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C(N=C1)=CN1C(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 XXMYDXUIZKNHDT-QNGWXLTQSA-N 0.000 description 1
- REITVGIIZHFVGU-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[(2-methylpropan-2-yl)oxy]propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](COC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 REITVGIIZHFVGU-IBGZPJMESA-N 0.000 description 1
- PKAUMAVONPSDRW-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[(2-methylpropan-2-yl)oxycarbonylamino]propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 PKAUMAVONPSDRW-IBGZPJMESA-N 0.000 description 1
- SJVFAHZPLIXNDH-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-phenylpropanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C1=CC=CC=C1 SJVFAHZPLIXNDH-QFIPXVFZSA-N 0.000 description 1
- LIWKOFAHRLBNMG-FQEVSTJZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-[(2-methylpropan-2-yl)oxycarbonylamino]butanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 LIWKOFAHRLBNMG-FQEVSTJZSA-N 0.000 description 1
- CBPJQFCAFFNICX-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-methylpentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(C)C)C(O)=O)C3=CC=CC=C3C2=C1 CBPJQFCAFFNICX-IBGZPJMESA-N 0.000 description 1
- JOOIZTMAHNLNHE-NRFANRHFSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-5-[(2-methylpropan-2-yl)oxycarbonylamino]pentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 JOOIZTMAHNLNHE-NRFANRHFSA-N 0.000 description 1
- UMRUUWFGLGNQLI-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-6-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCCNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 UMRUUWFGLGNQLI-QFIPXVFZSA-N 0.000 description 1
- HNICLNKVURBTKV-NDEPHWFRSA-N (2s)-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C HNICLNKVURBTKV-NDEPHWFRSA-N 0.000 description 1
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 1
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 1
- MSWZFWKMSRAUBD-VFUOTHLCSA-N 2-amino-2-deoxy-beta-D-galactopyranose Chemical compound N[C@H]1[C@H](O)O[C@H](CO)[C@H](O)[C@@H]1O MSWZFWKMSRAUBD-VFUOTHLCSA-N 0.000 description 1
- GOJUJUVQIVIZAV-UHFFFAOYSA-N 2-amino-4,6-dichloropyrimidine-5-carbaldehyde Chemical group NC1=NC(Cl)=C(C=O)C(Cl)=N1 GOJUJUVQIVIZAV-UHFFFAOYSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 206010000117 Abnormal behaviour Diseases 0.000 description 1
- 208000002874 Acne Vulgaris Diseases 0.000 description 1
- 206010048946 Anal abscess Diseases 0.000 description 1
- 206010002153 Anal fissure Diseases 0.000 description 1
- 208000016583 Anus disease Diseases 0.000 description 1
- 241000256844 Apis mellifera Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000222173 Candida parapsilosis Species 0.000 description 1
- 241000222178 Candida tropicalis Species 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 208000034656 Contusions Diseases 0.000 description 1
- 241001135265 Cronobacter sakazakii Species 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 208000035874 Excoriation Diseases 0.000 description 1
- 208000009531 Fissure in Ano Diseases 0.000 description 1
- XQFRJNBWHJMXHO-RRKCRQDMSA-N IDUR Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(I)=C1 XQFRJNBWHJMXHO-RRKCRQDMSA-N 0.000 description 1
- 101800002029 Jellein-1 Proteins 0.000 description 1
- 108010014144 Jelleine I Proteins 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- CSFVADKICPDRRF-KKUMJFAQSA-N Leu-His-Leu Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C([O-])=O)CC1=CN=CN1 CSFVADKICPDRRF-KKUMJFAQSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- QGMRQYFBGABWDR-UHFFFAOYSA-M Pentobarbital sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC(=O)[N-]C1=O QGMRQYFBGABWDR-UHFFFAOYSA-M 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 241000235645 Pichia kudriavzevii Species 0.000 description 1
- 108010093965 Polymyxin B Proteins 0.000 description 1
- FYXCBXDAMPEHIQ-FHWLQOOXSA-N Pro-Trp-Lys Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)N[C@@H](CCCCN)C(=O)O FYXCBXDAMPEHIQ-FHWLQOOXSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 208000004680 Rectal Fistula Diseases 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 206010040844 Skin exfoliation Diseases 0.000 description 1
- 206010041925 Staphylococcal infections Diseases 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 241000191963 Staphylococcus epidermidis Species 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 1
- 206010053615 Thermal burn Diseases 0.000 description 1
- 241000130764 Tinea Species 0.000 description 1
- 208000002474 Tinea Diseases 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 235000005811 Viola adunca Nutrition 0.000 description 1
- 240000009038 Viola odorata Species 0.000 description 1
- 235000013487 Viola odorata Nutrition 0.000 description 1
- 235000002254 Viola papilionacea Nutrition 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- 206010000496 acne Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000037354 amino acid metabolism Effects 0.000 description 1
- 206010002156 anal fistula Diseases 0.000 description 1
- 229910052786 argon Inorganic materials 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012888 bovine serum Substances 0.000 description 1
- 229940055022 candida parapsilosis Drugs 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 208000019065 cervical carcinoma Diseases 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000000701 coagulant Substances 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000009519 contusion Effects 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 230000035618 desquamation Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 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
- 239000002552 dosage form Substances 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 230000000214 effect on organisms Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 230000005182 global health Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 208000014617 hemorrhoid Diseases 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 208000015688 methicillin-resistant staphylococcus aureus infectious disease Diseases 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 239000002324 mouth wash Substances 0.000 description 1
- 229940051866 mouthwash Drugs 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- 231100000989 no adverse effect Toxicity 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229960002275 pentobarbital sodium Drugs 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920000024 polymyxin B Polymers 0.000 description 1
- 229960005266 polymyxin b Drugs 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 229940109850 royal jelly Drugs 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000036559 skin health Effects 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
- 231100000057 systemic toxicity Toxicity 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0034—Urogenital system, e.g. vagina, uterus, cervix, penis, scrotum, urethra, bladder; Personal lubricants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
- A61K9/006—Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0028—Polypeptides; Proteins; Degradation products thereof
- A61L26/0047—Specific proteins or polypeptides not covered by groups A61L26/0033 - A61L26/0042
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/0066—Medicaments; Biocides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/008—Hydrogels or hydrocolloids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/04—Macromolecular materials
- A61L31/043—Proteins; Polypeptides; Degradation products thereof
- A61L31/047—Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/08—Materials for coatings
- A61L31/10—Macromolecular materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/145—Hydrogels or hydrocolloids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/16—Biologically active materials, e.g. therapeutic substances
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P41/00—Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
- C08J3/075—Macromolecular gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/41—Anti-inflammatory agents, e.g. NSAIDs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2389/00—Characterised by the use of proteins; Derivatives thereof
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Dispersion Chemistry (AREA)
- Surgery (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Materials Engineering (AREA)
- Heart & Thoracic Surgery (AREA)
- Vascular Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Dermatology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Inorganic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Physiology (AREA)
- Nutrition Science (AREA)
- Urology & Nephrology (AREA)
- Reproductive Health (AREA)
- Gynecology & Obstetrics (AREA)
- Biomedical Technology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一种抗菌多肽化合物、医疗器械、水凝胶及其应用,该抗菌多肽化合物具有高抗菌活性,酶解稳定性高,生物利用度高,且细胞毒性低。本发明的水凝胶不粘连伤口,具抗菌活性和止血性能,可用于药物包载与缓释,如可以负载消炎药物或者表皮生长因子、血管生长因子等,加速伤口愈合,减少瘢痕组织纤维的形成。同时本发明的水凝胶的制备方法工艺步骤少,原料种类简单,操作方便。
Description
技术领域
本发明属于生物化学技术领域,涉及一种抗菌多肽化合物,具体涉及一种可用于抗菌、止血的抗菌多肽化合物、水凝胶及其应用,及该抗菌多肽化合物或水凝胶适用的医疗器械。
背景技术
随着抗生素在医疗、农业和食品工业的过量使用和滥用,导致了细菌对抗生素耐药性的惊人增长,使得现有抗生素的治疗效果大大降低,严重威胁全球公共健康和人类生命安全。当前全球每年约有70万人的死亡是耐药菌感染造成的,据估计,到2050年每年将有1000万人因此而死亡,经济损失将超数万亿美元。世卫组织已经将细菌耐药问题视为“当今全球健康、食品安全和发展面临的最大威胁之一”(生物工程学报,2018,34(8):1346-1360)。为应对细菌耐药给人类健康和生命安全带来的挑战,研发新型的抗菌药物已然成为了目前最紧迫的医学问题之一。
鉴于多肽分子与小分子药物相比具有结合面更大、作用靶向更强、更安全、副作用更小、不易引起严重免疫反应等优点,当前抗菌肽分子已成为新兴的抗菌药物研发的热门领域。然而天然抗菌肽往往由于抗菌活性低、系统毒性不明而限制了其临床应用。抗菌肽肽链中氨基酸构型、侧链的改变及化学修饰往往对其抗菌活性、毒性等产生不可预知的改变。因此,对抗菌肽化合物进行氨基酸修饰和替换,也是获得高效、低毒的抗菌肽衍生物的一个重要策略。
近年来,水凝胶被认为是一种非常有潜力的用于抗菌、止血、促伤口愈合、抗粘连等的生物材料。一般地,制备水凝胶材料主要有两大类,一类是合成高分子,一类是天然生物材料如多糖和蛋白质、多肽等。其中多肽水凝胶在体内易被蛋白酶水解为氨基酸,对机体不会产生不良影响,其低副作用的特性要优于已有的高分子水凝胶。
抗菌肽J-1(Jelleine-1)是最早从蜜蜂Apismellifera蜂王浆的一种天然抗菌肽,其具有广谱抗菌、抗真菌等活性(Peptides,2004,25:919–928)。
然而,抗菌肽J-1同其它天然抗菌肽一样存在抗菌活性低,生物利用度低等缺点,严重限制了其应用(Peptides,2019,112:56-66)。为提高其生物活性及和临床应用性,本发明基于抗菌肽J-1设计合成了系列抗菌肽J-1衍生物。我们研究发现,这些衍生物能表现出比抗菌肽J-1更好的抗菌活性、更低的细胞毒性,更好的酶解稳定性,且都能通过简单方法制备成水凝胶,非常有潜力发展成为具有抗菌、止血、促伤口愈合等功能的生物敷料。
发明内容
本发明的主要目的在于提供一种抗菌多肽化合物,该抗菌多肽化合物具有高抗菌活性,酶解稳定性高,生物利用度高,且细胞毒性低。
为达上述目的,本发明提供一种抗菌多肽化合物,所述抗菌多肽化合物具有以下氨基酸序列表示的母体肽:
Pro-Xaa2-Xaa3-Leu-Xaa5-Leu-Xaa7-Leu-NH2
其中,Xaa2=Phe,homo-Phe或Trp;
Xaa3=Lys,Aib,Orn,Dab,Dap或Arg;
Xaa5=Ser,Lys,Orn,Dab,Dap或Arg;
Xaa7=His,Lys,Orn,Dab,Dap或Arg;
且当Xaa2=Phe,Xaa3=Lys,Xaa5=Ser时,Xaa7≠His。
当Xaa2=Phe,Xaa3=Lys,Xaa5=Ser且Xaa7=His时,所示氨基酸序列为天然抗菌肽J-1的氨基酸序列,该氨基酸序列不在本发明的保护范围内。
当Xaa2为Phe,Xaa3为Lys,Xaa5为Lys,Xaa7为His时,标记抗菌多肽化合物为化合物1(涉及SEQ ID NO:1):
Pro-Phe-Lys-Leu-Lys-Leu-His-Leu-NH2
PFKLKLHL-NH2。
当Xaa2为Phe,Xaa3为Orn,Xaa5为Ser,Xaa7为Lys时,标记抗菌多肽化合物为化合物2(涉及SEQ ID NO:2):
Pro-Phe-Orn-Leu-Ser-Leu-Lys-Leu-NH2
PF-Orn-LSLKL-NH2。
当Xaa2为Phe,Xaa3为Dab,Xaa5为Lys,Xaa7为Lys时,标记抗菌多肽化合物为化合物3(涉及SEQ ID NO:3):
Pro-Phe-Dab-Leu-Lys-Leu-Lys-Leu-NH2
PF-Dab-LKLKL-NH2。
当Xaa2为Phe,Xaa3为Arg,Xaa5为Ser,Xaa7为His时,标记抗菌多肽化合物为化合物4(涉及SEQ ID NO:4):
Pro-Phe-Arg-Leu-Ser-Leu-His-Leu-NH2
PFRLSLHL-NH2。
当Xaa2为Phe,Xaa3为Arg,Xaa5为Arg,Xaa7为His时,标记抗菌多肽化合物为化合物5(涉及SEQ ID NO:5):
Pro-Phe-Arg-Leu-Arg-Leu-His-Leu-NH2
PFRLRLHL-NH2。
当Xaa2为Phe,Xaa3为Arg,Xaa5为Ser,Xaa7为Arg时,标记抗菌多肽化合物为化合物6(涉及SEQ ID NO:6):
Pro-Phe-Arg-Leu-Ser-Leu-Arg-Leu-NH2
PFRLSLRL-NH2。
当Xaa2为Phe,Xaa3为Arg,Xaa5为Arg,Xaa7为Arg时,标记抗菌多肽化合物为化合物7(涉及SEQ ID NO:7):
Pro-Phe-Arg-Leu-Arg-Leu-Arg-Leu-NH2
PFRLRLRL-NH2。
当Xaa2为Trp,Xaa3为Lys,Xaa5为Ser,Xaa7为His时,标记抗菌多肽化合物为化合物8(涉及SEQ ID NO:8):
Pro-Trp-Lys-Leu-Ser-Leu-His-Leu-NH2
PWKLSLHL-NH2。
当Xaa2为Trp,Xaa3为Orn,Xaa5为Orn,Xaa7为His时,标记抗菌多肽化合物为化合物9(涉及SEQ ID NO:9):
Pro-Trp-Orn-Leu-Orn-Leu-His-Leu-NH2
PW-Orn-L-Orn-LHL-NH2。
当Xaa2为Trp,Xaa3为Dab,Xaa5为Ser,Xaa7为Dab时,标记抗菌多肽化合物为化合物10(涉及SEQ ID NO:10):
Pro-Trp-Dab-Leu-Ser-Leu-Dab-Leu-NH2
PW-Dab-LSL-Dab-L-NH2。
当Xaa2为Trp,Xaa3为Dap,Xaa5为Dap,Xaa7为Dap时,标记抗菌多肽化合物为化合物11(涉及SEQ ID NO:11):
Pro-Trp-Dap-Leu-Dap-Leu-Dap-Leu-NH2
PW-Dap-L-Dap-L-Dap-L-NH2。
当Xaa2为Trp,Xaa3为Arg,Xaa5为Ser,Xaa7为His时,标记抗菌多肽化合物为化合物12(涉及SEQ ID NO:12):
Pro-Trp-Arg-Leu-Ser-Leu-His-Leu-NH2
PWRLSLHL-NH2。
当Xaa2为Trp,Xaa3为Arg,Xaa5为Arg,Xaa7为His时,标记抗菌多肽化合物为化合物13(涉及SEQ ID NO:13):
Pro-Trp-Arg-Leu-Arg-Leu-His-Leu-NH2
PWRLRLHL-NH2。
当Xaa2为Trp,Xaa3为Arg,Xaa5为Ser,Xaa7为Arg时,标记抗菌多肽化合物为化合物14(涉及SEQ ID NO:14):
Pro-Trp-Arg-Leu-Ser-Leu-Arg-Leu-NH2
PWRLSLHL-NH2。
当Xaa2为Trp,Xaa3为Arg,Xaa5为Arg,Xaa7为Arg时,标记抗菌多肽化合物为化合物15(涉及SEQ ID NO:15):
Pro-Trp-Arg-Leu-Arg-Leu-Arg-Leu-NH2
PWRLRLRL-NH2。
上述化合物1-15及天然抗菌肽J-1的氨基酸序列如下表1所示:
表1化合物1-15及天然抗菌肽J-1的氨基酸序列
本发明的另一个目的在于提供一种水凝胶的制备方法。
本发明的发明人经过大量的实验研究,证明该水凝胶具有抗菌、止血性能,可负载各类药物或生长因子,实现敷料的功能化治疗、实现创面治疗中抗菌、止血、为创面提供湿性环境等功能。
为达上述目的,本发明提供的水凝胶是由上述抗菌多肽化合物与缓冲液经反应聚合而成。
本发明的水凝胶具体通过以下方法制备:
步骤S1:将抗菌多肽化合物溶于二甲基亚砜,得到抗菌多肽化合物的溶解液,备用;
步骤S2:将抗菌多肽化合物的溶解液加入至缓冲液中,在超声或搅拌条件下进行离子交联聚合反应,得到水凝胶。
本发明的水凝胶中的溶剂主要为水,次要为二甲基亚砜(DMSO),其中二甲基亚砜的体积含量小于5%。
本发明的制备方法,优选地还包括以下步骤:
步骤S3:缓冲液中还可加入药物和/或生长因子,得到负载药物或生长因子的水凝胶。
本发明的药物优选为抗菌药物或消炎药物,生长因子优选为促伤口愈合生长因子。
本发明的缓冲液可以为碳酸盐溶液、亚硫酸盐溶液、以及磷酸盐缓冲液等,优选磷酸盐缓冲液;其中磷酸盐缓冲液为将Na2HPO4、KH2PO4、KCl以及NaCl按比例溶解于去离子水中制得;抗菌多肽化合物与磷酸盐缓冲液的组分及配比以摩尔比计为抗菌多肽化合物:Na2HPO4:KH2PO4:KCl:NaCl=(1-50):(1-10):(1-5):(1-5):(50-200);优选抗菌多肽化合物:Na2HPO4:KH2PO4:KCl:NaCl=1-50:10:2:2.7:137。
优选地,本发明的磷酸盐缓冲液的组分还包括二磷酸腺苷(ADP),该磷酸盐缓冲液的组分及配比以摩尔比计为Na2HPO4:KH2PO4:KCl:NaCl:ADP=(1-10):(1-5):(1-5):(50-200):1;优选ADP与Na2HPO4的摩尔比为1:10。
本发明的反应可以为物理反应或化学反应,优选离子交联聚合反应,反应温度为0-60℃,反应时间为1-120min。
本发明的再一个目的在于提供一种水凝胶在防粘连药物中的应用,该防粘连药物包含负载有药物或生长因子的水凝胶和至少一种药学上可接受的药用载体和/或辅料。
本发明的防粘连药物为片剂、糖衣片剂、粒剂、滴剂、喷雾剂、冲洗剂、漱口剂、用于皮肤表面的油膏和药贴、以及用于注射的无菌溶液中的至少一种剂型。
本发明的药物为抗菌药物或消炎药物,所述生长因子为促伤口愈合生长因子。
本发明的水凝胶可以直接对创面进行冲洗、喷涂、湿敷或覆盖,制作成方便使用的喷雾剂,直接喷敷于创面形成保护膜,能瞬间止血、保持创面湿润,创造利于上皮细胞的生长和愈合的低氧环境,加速伤口愈合;同时水凝胶中的抗菌肽,起到快速广谱持久的杀菌作用,伤口愈合后抗菌肽分解为氨基酸代谢,避免粘连和残留。
此外,本发明的水凝胶还可依据病症或创面的位置,选择合适的使用方法及制作成相应的适用剂型;例如,创伤、挫伤、擦伤、术后伤口、烧烫伤、溃疡清创后,可将本发明的水凝胶喷涂换出、或湿敷并包扎;痔疮、肛脓肿、肛瘘、肛裂、造口、造瘘、会阴侧切、包皮环切术后可将本发明的水凝胶喷涂或湿敷包扎;放疗前后,可将本发明的水凝胶对局部皮肤进行喷涂或湿敷;糖尿病足、脉管炎、老年性褥疮慢性不愈合伤口,清创后可将本发明的水凝胶喷涂患处;口腔异味、口腔术后护理可将本发明的水凝胶制作成漱口剂直接含于口腔漱口后排出;癣、疱疹、粉刺等,可将本发明的水凝胶喷涂或湿敷于创面;因刺激皮肤出现不适、痛痒、干燥、脱皮等现象,可将本发明的水凝胶直接喷涂或湿敷,以此改善皮肤健康。
本发明的水凝胶还可负载各类药物或生长因子,从而实现功能化治疗。
本发明的又一个目的在于提供一种医疗器械,该医疗器械具有上述抗菌多肽化合物或水凝胶。
本发明的抗菌多肽化合物或水凝胶可涂覆于医疗器械的至少一个表面上形成材料。
本发明的医疗器械的形式为由医用敷料、纤维、网片、粉末、微球、薄片、海绵、泡沫、缝合锚定器械、导管、支架、外科手术平头钉、板和螺丝、药物递送器械、防粘屏障和组织粘合剂组成的群组中的任一者。
本发明的纤维为织物;薄片为膜或夹片;缝合锚定器械为缝合线或U形钉。
本发明的抗菌多肽化合物具有高抗菌活性,酶解稳定性高,生物利用度高,且细胞毒性低,且在特定条件下能形成水凝胶。
本发明的水凝胶不粘连伤口,具抗菌活性和止血性能,可用于药物包载与缓释,如可以负载消炎药物或者表皮生长因子、血管生长因子等,加速伤口愈合,减少瘢痕组织纤维的形成。同时本发明的水凝胶的制备方法工艺步骤少,原料种类简单,操作方便。本发明的医疗器械具有该抗菌多肽化合物或水凝胶,从而实现更加方便、高效的治疗作用,能广泛应用于临床中。
附图说明
以下,结合附图来详细说明本发明的实施方案,其中:
图1:为本发明的化合物2、8、9、13、15在腹膜炎脓毒症小鼠模型中对小鼠存活率的影响。
图2:为本发明的溶血率评价结果柱状图。
图3:为本发明的化合物2、8、9、13、15与天然抗菌肽J-1对所测试细胞增殖的影响图。
图4:为本发明的化合物2、8、9、13、15与天然抗菌肽J-1对大肠杆菌内、外膜通透性的影响。
图5:为小鼠腹腔分别注射化合物2、8、9、13、15(100mg/kg、300mg/kg、500mg/kg、700mg/kg、1000mg/kg)后,各组小鼠的存活率曲线图。
图6:本发明的抗菌多肽化合物的水凝胶图片及电镜图片。
图7:为本发明的水凝胶对小鼠肝脏出血模型中的止血性能图。
具体实施方式
下面将结合实施例对本发明的实施方案进行详细描述,但是本领域技术人员将会理解,下列实施例仅用于说明本发明,而不应视为限定本发明的范围。本发明中所用仪器包括:高效液相色谱仪为美国waters公司的Delta 600型,所用制备柱为反相C18柱(XBridgeBEH 130Prep),19mm*250mm;分析柱为反相C8柱,4.6mm*250mm;质谱仪为BrukerMaxis 4G质谱仪;所用试剂包括:树脂为天津南开和成公司的Rink-amide-MBHA-resin(取代值为0.43mol/g,200~400目);N-A-Fmoc保护的氨基酸、N-羟基苯并三氮唑(HOBt)、氧-苯并三氮唑-N,N,N’,N’-四甲基脲-六氟磷酸盐(HBTU)、二异丙基乙胺(DIEA)和三异丙基硅烷(TIS)均购自上海吉尔生化有限公司;二氯甲烷、N,N-二甲基甲酰胺(DMF)、六氢吡啶(哌啶)、甲醇、三氟乙酸(TFA)、苯酚和吡啶均购自天津第二试剂厂;茚三酮购自上海试剂三厂。其它所用仪器或试剂未注明生产厂商者,均为可以通过市购获得的常规产品。实施例中未注明具体条件者,按照常规条件或制造商建议条件进行。
一、抗菌多肽化合物的制备
为方便阐述,以化合物1-15为例,本发明的抗菌多肽化合物的制备方法包括但不限于以下步骤:
(1)树脂溶胀及检测:一定量的Rink-amide-MBHA树脂(取代值为0.45mmol/g)置于合成仪中,加入适量重蒸的二氯甲烷,浸泡搅拌30min后使其完全溶胀,随后用重蒸的DMF清洗四次(每次两分钟),然后茚检,检测树脂是否干净(茚检试剂为吡啶:茚三酮:苯酚=2:1:1)。
(2)脱除F-moc保护基团:用20%重蒸的六氢吡啶+80%重蒸的DMF(v/v)清洗四遍(每次三分钟)脱去F-moc保护基,再用重蒸DMF清洗四遍(每次两分钟),然后茚检,检测保护基是否脱去。
(3)氨基酸缩合反应:称取3倍过量的缩合剂HOBT、HBTU和氨基酸(N-alpha-Fmoc保护)用重蒸DMF溶解后,加入六倍过量的启动剂DIEA混匀后,加入到步骤2中脱去Fmoc的树脂中,开始缩合反应,整个反应过程用氩气保护,搅拌反应一小时。抽干溶剂后用重蒸DMF清洗四遍(每次两分钟),然后茚检,检测氨基酸是否缩合完全。每合成一个氨基酸时重复上述脱F-moc保护和缩合反应过程,直到化合物1-15的各序列完成缩合。所用到的氨基酸有Fmoc-Pro-OH、Fmoc-Phe-OH、Fmoc-Lys(Boc)-OH、Fmoc-Orn(Boc)-OH、Fmoc-Dab(Boc)-OH、Fmoc-Dap(Boc)-OH、Fmoc-Arg(pbf)-OH、Fmoc-His(Trt)-OH、Fmoc-Leu-OH、Fmoc-Ser(tBu)-OH。
(4)肽链切割:当最后一个氨基酸合成完毕后用20%重蒸的六氢吡啶脱去F-moc保护,茚检,然后用重蒸的二氯甲烷和重蒸甲醇交替清洗两遍(每次三分钟),将合成仪密封用水泵抽三小时,待树脂完全抽干,然后加入切割试剂(TFA:Tris:H2O=95:2.5:2.5),反应三小时,反应期间每隔二十分钟搅拌一分钟,三小时后用旋转蒸发仪将其旋干,放入-20℃冰箱保存。
(5)萃取:将产物从-20℃冰箱取出,加入冰乙醚,剧烈摇晃后静置十分钟,待完全分层后,用双蒸水先萃取上清层,然后再萃取下层,将萃取后的液体分装至50ml烧杯放入-80℃过夜,然后冷冻干燥得到粗肽粉末。
(6)多肽纯化:①脱盐,用G-25葡聚糖凝胶柱对粗肽进行脱盐以除去小分子杂质。称取一定量粗肽(40mg左右)用5%的冰醋酸/水溶液(1ml)溶解,然后上样,用5%的冰醋酸/水溶液洗脱,观察紫外检测仪220nm处吸收值,收集主峰,然后将收集的液体分装至50ml烧杯放入-80℃过夜,然后真空抽干得到脱盐后的粗肽粉末。②HPLC制备,称取一定量脱盐后的粗肽(40mg左右)用20%乙腈/水溶液(5ml)溶解,用滤器过滤后上样,该纯化过程所用的柱子为μ-bondapaktm C18反向柱(19mm×300mm),用20%-80%乙腈/水溶液梯度洗脱,观察紫外检测仪220nm吸收值,收集主峰,将收集的液体分装置50ml烧杯放入-80℃过夜,然后真空抽干得纯肽粉末。所合成的化合物1-15以及天然抗菌肽J-1的氨基酸序列和质谱表征如下表2所示。
表2化合物1-15的氨基酸序列及质谱表征
二、抗菌多肽化合物最小抑菌浓度的测定
抗菌多肽化合物的抗菌活性和抗真菌活性的测定所用细菌菌株有:大肠杆菌(ATCC 25922)、大肠杆菌(ATCC 43837)、铜绿假单胞菌(ATCC 27853),肺炎克雷伯菌(ATCC700603)、坂崎肠杆菌(ATCC 29544)、金黄色葡萄球菌(ATCC 29213)、枯草芽孢杆菌(ATCC23857)和表皮葡萄球菌(ATCC 12228);所用真菌菌株为白色念珠菌(ATCC 14053)、光滑念珠菌(ATCC 2001)、近平滑念珠菌(ATCC 22019)、热带念珠菌(ATCC 750)和克柔念珠菌(ATCC 6258),以上菌株均为标准菌株,均来自于美国菌种保藏中心。此外,耐药菌株S-1a,S-2a,S-3a,S-4a,E-1b,E-2b,E-3b,E-4b,E-5b,P-1c,P-2c,P-3c,P-4c,P-5c,及临床分离真菌菌株光滑念珠菌2-1、白色念珠菌14-1、14-2、14-3均为临床分离菌株。
化合物对细菌和真菌的最小抑菌浓度(MIC)值都是采用美国临床和实验室标准协会(NCCLS)推荐的标准二倍稀释法进行测定的。简单地,取适量细菌或真菌冻存液至新鲜MH/SD培养基中,将菌液置于37℃摇床上,180rpm培养过夜。将上述菌液进行二次转接并在摇床继续培养4-5小时,得到对数生长期细菌。然后取1×105CFU/ml菌液加入96孔板内,每孔100μL,之后按二倍稀释的方式每孔100μL两倍于终浓度的不同浓度的多肽(1-256μM)。阴性对照为新鲜培养液,每个浓度做三个平行。加药后将96孔板放入37℃恒温恒湿培养箱内培养12小时即可观测结果,以肉眼可见显浑浊孔后第一个透亮孔的化合物浓度记为该化合物最小抑菌浓度,即其对该菌的MIC值。化合物1-15对标准细菌菌株的最小抑菌浓度结果详见表3,化合物1-15对真菌的抑菌活性结果详见表4。以抗菌肽J-1、环丙沙星(ciprofloxacin)、庆大霉素(gentamicin)作为对照组,化合物2、8、9、13、15对耐药菌株的抑菌活性结果详见表5。
由表3和表5的结果可以看出,化合物1-15对所测试的细菌及真菌均表现出了很好的抗菌、抗真菌活性。其中,化合物7和化合物15的抗菌、抗真菌活性较天然抗菌肽J-1提高了8-16倍,本发明的其他化合物的抗菌、抗真菌活性较抗菌肽J-1提高了1-8倍。值得一提的是,化合物1-15均对临床分离的耐药菌株表现出良好的抗菌活性。
表3化合物1-15对标准细菌菌株的最小抑菌浓度
表4化合物1-15对真菌的抑菌活性
表5化合物2、8、9、13、15对耐药菌株的抑菌活性
a为临床分离的耐药株MRSA;b为临床分离的耐药株ESBL;c为临床分离的耐药株PA。
三、抗菌多肽化合物体内抗菌评价
抗菌多肽化合物体内抗菌活性的测定是通过检测细菌感染小鼠的存活率反映其体内抗菌活性。
本发明以化合物2、8、9、13、15为例,研究抗菌多肽化合物体内抗菌效果。本实验所用小鼠为昆明系小鼠,这些小鼠被安置在一个恒温环境中(22±1℃)。简单地,将36只昆明小鼠(体重18~22克)随机分为6组,第1天腹腔注射0.2ml浓度为3×108CFU/mL大肠杆菌(ATCC 25922),建立腹膜炎脓毒症小鼠模型。接种细菌1h后腹腔内给药一次,然后20mg/kg体重的剂量腹腔注射化合物2、8、9、13、15,选取多粘菌素B(20mg/kg)作为阳性对照药物,阴性对照组采用腹腔注射无菌生理盐水,随后观察7天,计算各给药组和对照组小鼠存活率并绘制生存曲线。
图1为本发明的化合物2、8、9、13、15在腹膜炎脓毒症小鼠模型中对小鼠存活率的影响。从图1的结果可以看出,本发明的化合物13和阳性对照组小鼠存活率最高,为100%;化合物9和15的存活率相当,在90%以上;化合物2和8的存活率在70%以上;其次是天然抗菌肽J-I存活率在50%左右;阴性对照组小鼠的存活率最差,在第3天时,小鼠全部死亡。相较于阴性对照组和天然抗菌肽J-I,本发明的化合物2、8、9、13、15在20mg/kg的浓度下均可以有效提高小鼠的生存率。由此可以证明,本发明的抗菌多肽化合物体内抗菌活性高。
四、抗菌多肽化合物溶血毒性评价
溶血活性是通过测定抗菌多肽化合物对小鼠血红细胞的溶血浓度来测定的。
本发明以化合物1-15为例,研究抗菌多肽化合物溶血毒性。将小鼠红细胞浓度稀释至8%,将红细胞悬液加入到96孔板中(每孔100μL),然后每孔加入化合物100μL。孵育1h后,取每孔中的上清置于新的96孔板中,然后使用酶标仪于490nm处测定样品上清液的吸光值。本发明以PBS和2%Triton-X100分别作为阴性对照和阳性对照。
图2为本发明的溶血率评价结果柱状图.其中Triton X-100用作阳性对照,其溶血率定为100%;PBS用作阴性对照,其溶血率为0。由图2的结果可以看出,本发明的化合物1-15对小鼠红细胞几乎不表现溶血活性,溶血率均<5%。由此可以证明,本发明的抗菌多肽化合物溶血活性低。
五、抗菌多肽化合物的细胞毒性评价
细胞毒性是通过MTT方法测定各化合物对人宫颈癌细胞Hela、人胚肾上皮细胞HEK293细胞和小鼠单核巨噬细胞RAW264.7的增殖影响而测定的。
本发明以化合物2、8、9、13、15为例,研究抗菌多肽化合物的细胞毒性。本发明所用的三种细胞株为人宫颈癌细胞Hela、人胚肾上皮细胞HEK293细胞和小鼠单核巨噬细胞RAW264.7。Hela细胞培养在含有10%新生牛血清的RPMI 1640培养基,HEK293细胞和RAW264.7细胞培养在含有10%胎牛血清的DMEM培养基中。简单地,将细胞(1×104细胞/孔)接种于96孔板中,放入37℃5%CO2培养箱中孵育,24h后加入不同浓度的化合物后放入5%CO2培养箱中孵育,24h后每孔加入20μL浓度为5mg/mL的MTT溶液孵育,4h后吸出培养基加入150μL DMSO用于溶解蓝紫色结晶甲瓒。用振荡器震荡5min后用酶标仪检测吸光值(570nm),然后计算存活率=(OD加药组/OD对照组)×100%(n=3)。
图3为本发明的化合物2、8、9、13和15与天然抗菌肽J-1对所测试细胞增殖的影响图。其中左图为化合物2、8、9、13和15与天然抗菌肽J-1对Hela细胞增殖的影响图;中间图为化合物2、8、9、13和15对RAW 264.7细胞增殖的影响图;右图为化合物2、8、9、13和15对HEK293细胞增殖的影响图。由图3的结果可以看出,化合物2、8、9、13和15对所测试的三种细胞均不表现抑制活性。由此可以证明,本发明的抗菌多肽化合物不具有细胞毒性。
六、抗菌多肽化合物大肠杆菌外膜(OM)渗透性评价
细菌细胞外膜的破坏作用是通过使用疏水性荧光探针NPN(1-N-phenylnaphthylamine)测定化合物对大肠杆菌(ATCC 25922)外膜渗透性的影响而测定的。
本发明以化合物2、8、9、13和15为例,研究抗菌多肽化合物大肠杆菌外膜(OM)渗透性。将大肠杆菌细胞用PBS溶液清洗两遍后用5mM HEPES(PH 7.2)重悬,然后将其稀释到浓度为5*104CFU/mL。化合物浓度为1×MIC-8×MIC,实验组:100μL菌液+50μL Jelleine-I+50μL NPN(40μM);对照组:100μL菌液+50μL HEPES(5mM)+50μL NPN(40μM)。激发和发射波长分别设置为350nm和420nm,用多功能酶标仪每分钟检测一次,连续检测十五分钟。
图4为本发明的化合物2、8、9、13和15与天然抗菌肽J-1对大肠杆菌内、外膜通透性的影响。其中,图4(A、C、E、G、I、K)分别为不同剂量的天然抗菌肽J-1与化合物2、8、9、13和15对大肠杆菌外膜通透性的影响,其中HEPES缓冲液用作阴性对照;由图4(A、C、E、G、I、K)的结果可以看出,化合物2、8、9、13和15与抗菌肽J-1一样均能剂量依赖的破坏大肠杆菌外膜完整性。
七、抗菌多肽化合物大肠杆菌内膜(IM)渗透性评价
对细菌内膜的完整性的影响是通过使用ONPG(邻硝基苯-β-D-半乳糖苷)测定大肠杆菌ML-35(ATCC 43837)胞内β-半乳糖甘酶的释放量从而检测化合物对大肠杆菌内膜渗透性影响来测定的。
本发明以化合物2、8、9、13和15为例,研究抗菌多肽化合物大肠杆菌内膜(IM)渗透性。将大肠杆菌ML-35细胞用PBS溶液清洗两遍然后用0.9%氯化钠溶液重悬。化合物浓度为1×MIC-8×MIC,实验组:100μL菌液+90μLJelleine-I+10μL ONPG(30mM);阳性对照组:100μL菌液+90μL 1%Triton-X100+10μLONPG(30mM);阴性对照组:100μL菌液+90μL 0.5%氯化钠溶液+10μL ONPG(30mM)。用多功能酶标仪检测420nm处吸光度,每五分钟检测一次,连续检测九十分钟。
图4为本发明的化合物2、8、9、13和15与天然抗菌肽J-1对大肠杆菌内、外膜通透性的影响。图4(B、D、F、H、J、L)分别为不同剂量的抗菌肽J-1和化合物2、8、9、13和15对大肠杆菌内膜通透性的影响,其中triton X-100用作阳性对照,生理盐水用作阴性对照;由图4(B、D、F、H、J、L)的结果可以看出,化合物2、8、9、13和15与天然抗菌肽J-1相似,能剂量依赖的破坏大肠杆菌细胞内膜的完整性。
八、抗菌多肽化合物急性毒性评价
化合物的急性毒性是通过测定各化合物对小鼠的急性毒性而测定的。
本发明以化合物2、8、9、13和15为例,研究抗菌多肽化合物大肠杆菌内膜(IM)渗透性。本实验所用小鼠为昆明系小鼠,这些小鼠被安置在一个恒温环境中(22±1℃),可以自由的获取水和食物。简单地,132只健康昆明小鼠随机分为11组,每组12只(雌雄各半),其中5组腹腔注射0.2ml不同剂量的化合物(100-1000mg/kg),对照组腹腔注射生理盐水,本实验中以蜂毒肽作为对照药物,另外5组腹腔注射不同剂量的蜂毒肽(10-32mg/kg),给药2h后观察记录小鼠状态,之后一天观察两次,连续观察14天,记录小鼠存活率。
图5为小鼠腹腔分别注射化合物2、8、9、13和15(100mg/kg、300mg/kg、500mg/kg、700mg/kg、1000mg/kg)后,各组小鼠的存活率曲线图。由图5的结果可以看出,化合物2、8、9、13和15在最高剂量1000mg/kg处理组有1-3只小鼠死亡,其它均无明显的异常行为。根据寇氏法计算蜂毒肽Magainin的LD50为20mg/kg,化合物2、8、9、13和15的LD50大于1000mg/kg,其急性毒性比蜂毒肽Magainin至少低50倍。由此可以证明,本发明的抗菌多肽化合物的急性毒性低。
九、水凝胶的制备
本发明的水凝剂的原料及配比按照摩尔比计为:抗菌多肽化合物1-50mM;Na2HPO4:1-10mM;KH2PO4:1-5mM;KCl:1-5mM;NaCl:50-200mM;所述水凝胶溶剂主要为水,次要为二甲基亚砜(DMSO),其比例低于5%(v/v)。
本发明的水凝胶的制备方法,具体包括以下步骤:
步骤一:将化合物溶于DMSO,配成多肽化合物溶解液储液;将Na2HPO4;KH2PO4;KCl;NaCl;按上述浓度溶解于去离子水中备用;
步骤二:将DMSO溶解的抗菌肽J-1溶液用加入到上述盐溶液中,化合物终浓度为1.5-40mM,搅拌均匀即可得水凝胶;
步骤三:在步骤二制备水凝胶的过程中,提前在盐溶液中加入药物或生长因子,可制得负载药物或生长因子的水凝胶缓释敷料。
本发明以化合物8为例,首先将化合物8溶解于二甲基亚砜中配成化合物高浓度母液,然后将化合物高浓度母液加入到磷酸缓冲溶液(PBS,pH6.0~9.0)中,终浓度稀释到10mM,搅拌放置适当时间即可成水凝胶。
图6:本发明的抗菌多肽化合物的水凝胶图片及电镜图片。其中图6中A为液体状态的抗菌体;图6中B为本发明的抗菌多肽化合物的水凝胶图片;图6中C为本发明的抗菌多肽化合物溶解于去离子水中,室温干燥后所呈现的电镜图片;图6中D为本发明的水凝胶在室温经干燥后所呈现的电镜图片。
十、水凝胶在小鼠肝脏出血模型中止血性能测定
选用上述过程制备的水凝胶对止血性能进行测定。测定水凝胶止血性能所用小鼠为雄性昆明系小鼠,体重18-22g,这些小鼠在温度22~24℃,相对温度45%~55%环境下饲养,术前12h对实验小鼠禁食。
肝脏出血模型的建立:实验共分两组,分别为对照组、化合物8水凝胶组,每组8只小鼠。用40mg/kg体重的戊巴比妥钠麻醉小鼠,然后将小鼠固定在手术台上,腹部备皮,使用碘伏对手术区域进行消毒;然后在腹部取一个直径为1.5cm左右的纵向切口,逐层分离,充分暴露肝脏右页,随后将事先称好重量的滤纸片垫在肝脏右页下方,用21G的针头将肝脏右页正中间刺穿,随后立即敷200ul水凝胶在伤口处(对照组不做任何处理),拍照记录肝脏出血过程。
图7:为本发明的水凝胶对小鼠肝脏出血模型中的止血性能图;图7中A为生理盐水对照组;图7中B为本发明的水凝胶处理组。由图7的结果可以看出,与对照组相比,本发明的水凝胶能明显抑制肝脏出血。由此可以证明,本发明的水凝胶具有良好的止血性能。
虽然本发明已以实施例揭露如上然其并非用以限定本发明,任何所属技术领域中具有公知常识者,在不脱离本发明的精神和范围内,当可作些许的更动与润饰,故本发明的保护范围当视所附的权利要求书所界定的范围为准。
序列表
<110> 广州图微科创生物科技有限公司
<120> 抗菌多肽化合物、医疗器械、水凝胶及其应用
<130> GD2387-20P125134
<150> 202011613292.8
<151> 2020-12-30
<150> 202110240501.7
<151> 2021-03-04
<160> 16
<170> PatentIn version 3.5
<210> 1
<211> 8
<212> PRT
<213> 人工序列
<400> 1
Pro Phe Lys Leu Lys Leu His Leu
1 5
<210> 2
<211> 8
<212> PRT
<213> 人工序列
<220>
<221> MISC_FEATURE
<222> (3)..(3)
<223> X=Orn
<400> 2
Pro Phe Xaa Leu Ser Leu Lys Leu
1 5
<210> 3
<211> 8
<212> PRT
<213> 人工序列
<220>
<221> MISC_FEATURE
<222> (3)..(3)
<223> X=Dab
<400> 3
Pro Phe Xaa Leu Lys Leu Lys Leu
1 5
<210> 4
<211> 8
<212> PRT
<213> 人工序列
<400> 4
Pro Phe Arg Leu Ser Leu His Leu
1 5
<210> 5
<211> 8
<212> PRT
<213> 人工序列
<400> 5
Pro Phe Arg Leu Arg Leu His Leu
1 5
<210> 6
<211> 8
<212> PRT
<213> 人工序列
<400> 6
Pro Phe Arg Leu Ser Leu Arg Leu
1 5
<210> 7
<211> 8
<212> PRT
<213> 人工序列
<400> 7
Pro Phe Arg Leu Arg Leu Arg Leu
1 5
<210> 8
<211> 8
<212> PRT
<213> 人工序列
<400> 8
Pro Trp Lys Leu Ser Leu His Leu
1 5
<210> 9
<211> 8
<212> PRT
<213> 人工序列
<220>
<221> MISC_FEATURE
<222> (3)..(3)
<223> X=Orn
<220>
<221> MISC_FEATURE
<222> (5)..(5)
<223> X=Orn
<400> 9
Pro Trp Xaa Leu Xaa Leu His Leu
1 5
<210> 10
<211> 8
<212> PRT
<213> 人工序列
<220>
<221> MISC_FEATURE
<222> (3)..(3)
<223> X=Dab
<220>
<221> MISC_FEATURE
<222> (7)..(7)
<223> X=Dab
<400> 10
Pro Trp Xaa Leu Ser Leu Xaa Leu
1 5
<210> 11
<211> 8
<212> PRT
<213> 人工序列
<220>
<221> MISC_FEATURE
<222> (3)..(3)
<223> X=Dap
<220>
<221> MISC_FEATURE
<222> (5)..(5)
<223> X=Dap
<220>
<221> MISC_FEATURE
<222> (7)..(7)
<223> X=Dap
<400> 11
Pro Trp Xaa Leu Xaa Leu Xaa Leu
1 5
<210> 12
<211> 8
<212> PRT
<213> 人工序列
<400> 12
Pro Trp Arg Leu Ser Leu His Leu
1 5
<210> 13
<211> 8
<212> PRT
<213> 人工序列
<400> 13
Pro Trp Arg Leu Arg Leu His Leu
1 5
<210> 14
<211> 8
<212> PRT
<213> 人工序列
<400> 14
Pro Trp Arg Leu Ser Leu Arg Leu
1 5
<210> 15
<211> 8
<212> PRT
<213> 人工序列
<400> 15
Pro Trp Arg Leu Arg Leu Arg Leu
1 5
<210> 16
<211> 8
<212> PRT
<213> 人工序列
<400> 16
Pro Phe Lys Leu Ser Leu His Leu
1 5
Claims (17)
1.一种抗菌多肽化合物,其特征在于,所述抗菌多肽化合物具有以下氨基酸序列表示的母体肽:
Pro-Xaa2-Xaa3-Leu-Xaa5-Leu-Xaa7-Leu-NH2
其中,Xaa2=Phe,homo-Phe或Trp;
Xaa3=Lys,Aib,Orn,Dab,Dap或Arg;
Xaa5=Ser,Lys,Orn,Dab,Dap或Arg;
Xaa7=His,Lys,Orn,Dab,Dap或Arg;
且当Xaa2=Phe,Xaa3=Lys,Xaa5=Ser时,Xaa7≠His。
2.根据权利要求1所述的抗菌多肽化合物,其特征在于,所述抗菌多肽化合物的氨基酸如下所示:
化合物1:
Pro-Phe-Lys-Leu-Lys-Leu-His-Leu-NH2
化合物2:
Pro-Phe-Orn-Leu-Ser-Leu-Lys-Leu-NH2
化合物3:
Pro-Phe-Dab-Leu-Lys-Leu-Lys-Leu-NH2
化合物4:
Pro-Phe-Arg-Leu-Ser-Leu-His-Leu-NH2
化合物5:
Pro-Phe-Arg-Leu-Arg-Leu-His-Leu-NH2
化合物6:
Pro-Phe-Arg-Leu-Ser-Leu-Arg-Leu-NH2
化合物7:
Pro-Phe-Arg-Leu-Arg-Leu-Arg-Leu-NH2
化合物8:
Pro-Trp-Lys-Leu-Ser-Leu-His-Leu-NH2
化合物9:
Pro-Trp-Orn-Leu-Orn-Leu-His-Leu-NH2
化合物10:
Pro-Trp-Dab-Leu-Ser-Leu-Dab-Leu-NH2
化合物11:
Pro-Trp-Dap-Leu-Dap-Leu-Dap-Leu-NH2
化合物12:
Pro-Trp-Arg-Leu-Ser-Leu-His-Leu-NH2
化合物13:
Pro-Trp-Arg-Leu-Arg-Leu-His-Leu-NH2
化合物14:
Pro-Trp-Arg-Leu-Ser-Leu-Arg-Leu-NH2
化合物15:
Pro-Trp-Arg-Leu-Arg-Leu-Arg-Leu-NH2。
3.一种水凝胶,其特征在于,所述水凝胶由权利要求1-2任一项所述的抗菌多肽化合物与缓冲液经反应聚合而成。
4.根据权利要求3所述的水凝胶,其特征在于,所述水凝胶具体通过以下方法制备:
步骤S1:将抗菌多肽化合物溶于二甲基亚砜,得到抗菌多肽化合物的溶解液,备用;
步骤S2:将抗菌多肽化合物的溶解液加入至缓冲液中,在超声或搅拌条件下进行离子交联聚合反应,得到水凝胶。
5.根据权利要求4所述的水凝胶,其特征在于,还包括以下步骤:
步骤S3:缓冲液中还加入药物和/或生长因子,得到负载药物或生长因子的水凝胶。
6.根据权利要求5所述的水凝胶,其特征在于,所述药物为抗菌药物或消炎药物,所述生长因子为促伤口愈合生长因子。
7.根据权利要求4所述的水凝胶,其特征在于,所述二甲基亚砜的体积含量小于5%。
8.根据权利要求3所述的水凝胶,其特征在于,所述缓冲液为磷酸盐缓冲液;所述抗菌多肽化合物与磷酸盐缓冲液的组分及配比以摩尔比计为抗菌多肽化合物:Na2HPO4:KH2PO4:KCl:NaCl=(1-50):(1-10):(1-5):(1-5):(50-200)。
9.根据权利要求8所述的水凝胶,其特征在于,所述磷酸盐缓冲液的组分还包括ADP,以摩尔比计所述ADP与Na2HPO4的比例为1:(1-50)。
10.根据权利要求3所述的水凝胶,其特征在于,所述反应为离子交联聚合反应,反应温度为0-60℃,反应时间为1-120min。
11.一种根据权利要求3-10任一项所述的水凝胶在防粘连药物中的应用,其特征在于,所述防粘连药物包含负载有药物和/或生长因子的所述水凝胶和至少一种药学上可接受的药用载体和/或辅料。
12.根据权利要求11所述的应用,其特征在于,所述防粘连药物为片剂、糖衣片剂、粒剂、滴剂、喷雾剂、冲洗剂、漱口剂、用于皮肤表面的油膏和药贴、以及用于注射的无菌溶液中的至少一种剂型。
13.根据权利要求11所述的应用,其特征在于,所述药物为抗菌药物或消炎药物,所述生长因子为促伤口愈合生长因子。
14.一种医疗器械,其特征在于,具有权利要求1-2任一项所述的抗菌多肽化合物或权利要求3-13任一项所述的水凝胶。
15.根据权利要求14所述的医疗器械,其特征在于,所述抗菌多肽化合物或所述水凝胶涂覆于所述医疗器械的至少一个表面上形成材料。
16.根据权利要求14所述的医疗器械,其特征在于,所述医疗器械的形式为由医用敷料、纤维、网片、粉末、微球、薄片、海绵、泡沫、缝合锚定器械、导管、支架、外科手术平头钉、板和螺丝、药物递送器械、防粘屏障和组织粘合剂组成的群组中的任一者。
17.根据权利要求16所述的医疗器械,其特征在于,所述纤维为织物;所述薄片为膜或夹片;所述缝合锚定器械为缝合线或U形钉。
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/CN2021/139094 WO2022143219A1 (zh) | 2020-12-30 | 2021-12-17 | 抗菌多肽化合物、医疗器械、水凝胶及其应用 |
AU2021412202A AU2021412202A1 (en) | 2020-12-30 | 2021-12-17 | Antibacterial polypeptide compound, medical instrument, hydrogel, and application thereof |
KR1020237025869A KR20230127299A (ko) | 2020-12-30 | 2021-12-17 | 항균 폴리펩타이드 화합물, 의료 기기, 하이드로겔및 그의 응용 |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011613292 | 2020-12-30 | ||
CN2020116132928 | 2020-12-30 | ||
CN2021102405017 | 2021-03-04 | ||
CN202110240501 | 2021-03-04 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114181279A true CN114181279A (zh) | 2022-03-15 |
CN114181279B CN114181279B (zh) | 2024-04-16 |
Family
ID=80601309
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110407232.9A Active CN114181279B (zh) | 2020-12-30 | 2021-04-15 | 抗菌多肽化合物、医疗器械、水凝胶及其应用 |
Country Status (4)
Country | Link |
---|---|
KR (1) | KR20230127299A (zh) |
CN (1) | CN114181279B (zh) |
AU (1) | AU2021412202A1 (zh) |
WO (1) | WO2022143219A1 (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116763720B (zh) * | 2023-06-06 | 2023-12-12 | 广州图微科创生物科技有限公司 | 一种多肽自组装水凝胶及其制备方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20180086792A1 (en) * | 2015-04-10 | 2018-03-29 | Ustav Organicke Chemie A Biochemie Av Cr, V.V.I. | Antimicrobial peptides and their use for the treatment of topical infections |
CN109456389A (zh) * | 2018-12-25 | 2019-03-12 | 苏州大学 | 一种抗菌肽、抗菌肽水凝胶及其制备方法 |
CN112121226A (zh) * | 2020-09-17 | 2020-12-25 | 常州大学 | 一种促伤口愈合的复合水凝胶及其制备方法和应用 |
-
2021
- 2021-04-15 CN CN202110407232.9A patent/CN114181279B/zh active Active
- 2021-12-17 KR KR1020237025869A patent/KR20230127299A/ko unknown
- 2021-12-17 AU AU2021412202A patent/AU2021412202A1/en active Pending
- 2021-12-17 WO PCT/CN2021/139094 patent/WO2022143219A1/zh active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20180086792A1 (en) * | 2015-04-10 | 2018-03-29 | Ustav Organicke Chemie A Biochemie Av Cr, V.V.I. | Antimicrobial peptides and their use for the treatment of topical infections |
CN109456389A (zh) * | 2018-12-25 | 2019-03-12 | 苏州大学 | 一种抗菌肽、抗菌肽水凝胶及其制备方法 |
CN112121226A (zh) * | 2020-09-17 | 2020-12-25 | 常州大学 | 一种促伤口愈合的复合水凝胶及其制备方法和应用 |
Non-Patent Citations (3)
Title |
---|
DANUBIA BATISTA MARTINS等: "Comparing activity, toxicity and model membrane interactions of Jelleine‑I and Trp/Arg analogs: analysis of peptide aggregation" * |
FENGJING JIA等: "The effect of halogenation on the antimicrobial activity, antibiofilm activity, cytotoxicity and proteolytic stability of the antimicrobial peptide Jelleine-I" * |
MARCIA PEREZ DOS SANTOS CABRERA等: "Combining experimental evidence and molecular dynamic simulations to understand the mechanism of action of the antimicrobial octapeptide jelleine-I" * |
Also Published As
Publication number | Publication date |
---|---|
WO2022143219A1 (zh) | 2022-07-07 |
KR20230127299A (ko) | 2023-08-31 |
AU2021412202A9 (en) | 2023-08-24 |
CN114181279B (zh) | 2024-04-16 |
AU2021412202A1 (en) | 2023-07-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109232719B (zh) | 一种pH响应的抗菌肽及其制备方法和应用 | |
EP2078529B1 (en) | Antimicrobial peptides | |
JP6106737B2 (ja) | ポリペプチド及びその抗菌または消毒用途 | |
CN111484568B (zh) | 一种壳聚糖-抗菌性多肽接枝聚合物及其制备方法和应用 | |
JP5753077B2 (ja) | 抗菌ペプチド多量体 | |
JP2000217579A (ja) | 新規抗菌性ペプチド | |
CN114181279B (zh) | 抗菌多肽化合物、医疗器械、水凝胶及其应用 | |
US6358921B1 (en) | Antimicrobial peptide compositions and method | |
CN112778401B (zh) | 一种辛酸酰化修饰抗菌肽及其应用 | |
CN102766196B (zh) | 一组阳离子抗菌肽及其制备方法和应用 | |
CN115869459A (zh) | 促伤口愈合的多肽水凝胶及其制备方法与应用 | |
CN110054664B (zh) | 含d型氨基酸的侧链脂肪酸修饰抗菌肽类似物及其合成和应用 | |
CN112625106B (zh) | 一种抗菌多肽化合物、合成方法及其应用 | |
CN112625092B (zh) | 一种基于polybia-MPI的抗菌多肽化合物及其合成与应用 | |
CN111423493A (zh) | 一种棕榈酸化抗酶解抗菌肽及其制备方法和应用 | |
JP4817335B2 (ja) | 新規抗菌性ペプチド | |
WO2019085926A1 (zh) | 多黏菌素类似物及其制备方法 | |
CN110028557B (zh) | 一种Ce6标记的双链抗菌肽及其合成方法和应用 | |
CN107337713B (zh) | 一组抑菌肽及其制备方法 | |
CN116785401A (zh) | 新型两亲性脂肽纳米制剂及其应用 | |
CN101974076B (zh) | 抑制由碱性成纤维细胞生长因子诱导的细胞增殖及血管增生的环肽 | |
CN116178520B (zh) | 一种阳离子肽及其应用 | |
CN113896767B (zh) | 一种自组装抗菌肽纳米粒子及其应用 | |
US20240254175A1 (en) | Novel selective antimicrobial fusion peptides | |
JP2006272002A (ja) | 医療用手当材 |
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 |