CN113024643A - 一种人工拟肽及其制备方法和应用 - Google Patents
一种人工拟肽及其制备方法和应用 Download PDFInfo
- Publication number
- CN113024643A CN113024643A CN202110481513.9A CN202110481513A CN113024643A CN 113024643 A CN113024643 A CN 113024643A CN 202110481513 A CN202110481513 A CN 202110481513A CN 113024643 A CN113024643 A CN 113024643A
- Authority
- CN
- China
- Prior art keywords
- peptidomimetic
- artificial
- solid phase
- cyprkr
- dgrc
- 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
- 239000000816 peptidomimetic Substances 0.000 title claims abstract description 91
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 58
- 239000003814 drug Substances 0.000 claims abstract description 25
- 230000003278 mimic effect Effects 0.000 claims abstract description 18
- 108090001007 Interleukin-8 Proteins 0.000 claims abstract description 12
- 102000003777 Interleukin-1 beta Human genes 0.000 claims abstract description 11
- 108090000193 Interleukin-1 beta Proteins 0.000 claims abstract description 11
- 230000037182 bone density Effects 0.000 claims abstract description 11
- 206010061218 Inflammation Diseases 0.000 claims abstract description 9
- 230000004054 inflammatory process Effects 0.000 claims abstract description 9
- 230000008961 swelling Effects 0.000 claims abstract description 7
- 206010051728 Bone erosion Diseases 0.000 claims abstract description 5
- 239000007790 solid phase Substances 0.000 claims description 26
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 25
- 239000004475 Arginine Substances 0.000 claims description 16
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 16
- 239000002243 precursor Substances 0.000 claims description 16
- 150000001413 amino acids Chemical class 0.000 claims description 15
- 235000001014 amino acid Nutrition 0.000 claims description 14
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 13
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 12
- -1 9-Fluorenylmethyloxycarbonyl (FMOC) Chemical class 0.000 claims description 10
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 10
- 235000018417 cysteine Nutrition 0.000 claims description 10
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 10
- 239000011347 resin Substances 0.000 claims description 10
- 229920005989 resin Polymers 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- AENSJDFBDBQUMU-UHFFFAOYSA-N COC1=C(C=CC(=C1)OC)C(C1=CC=C(OCC(=O)NN(C(C2=CC=CC=C2)C2=CC=CC=C2)C)C=C1)(N)C(=O)OCC1=CC=CC=2C3=CC=CC=C3CC12 Chemical compound COC1=C(C=CC(=C1)OC)C(C1=CC=C(OCC(=O)NN(C(C2=CC=CC=C2)C2=CC=CC=C2)C)C=C1)(N)C(=O)OCC1=CC=CC=2C3=CC=CC=C3CC12 AENSJDFBDBQUMU-UHFFFAOYSA-N 0.000 claims description 8
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 8
- 125000006239 protecting group Chemical group 0.000 claims description 8
- 239000000825 pharmaceutical preparation Substances 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
- 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 claims description 5
- 239000004471 Glycine Substances 0.000 claims description 4
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims description 4
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 4
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims description 4
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 4
- 239000004472 Lysine Substances 0.000 claims description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 4
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 4
- 235000003704 aspartic acid Nutrition 0.000 claims description 4
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 4
- 150000001875 compounds Chemical class 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 239000003826 tablet Substances 0.000 claims description 4
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 4
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
- 150000003839 salts Chemical class 0.000 claims description 3
- 239000012453 solvate Substances 0.000 claims description 3
- 239000008215 water for injection Substances 0.000 claims description 3
- CMWTZPSULFXXJA-UHFFFAOYSA-N Naproxen Natural products C1=C(C(C)C(O)=O)C=CC2=CC(OC)=CC=C21 CMWTZPSULFXXJA-UHFFFAOYSA-N 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000006071 cream Substances 0.000 claims description 2
- 229960003957 dexamethasone Drugs 0.000 claims description 2
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 claims description 2
- 229960001259 diclofenac Drugs 0.000 claims description 2
- DCOPUUMXTXDBNB-UHFFFAOYSA-N diclofenac Chemical compound OC(=O)CC1=CC=CC=C1NC1=C(Cl)C=CC=C1Cl DCOPUUMXTXDBNB-UHFFFAOYSA-N 0.000 claims description 2
- 239000000945 filler Substances 0.000 claims description 2
- 239000000499 gel Substances 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 239000008176 lyophilized powder Substances 0.000 claims description 2
- 229960002009 naproxen Drugs 0.000 claims description 2
- CMWTZPSULFXXJA-VIFPVBQESA-N naproxen Chemical compound C1=C([C@H](C)C(O)=O)C=CC2=CC(OC)=CC=C21 CMWTZPSULFXXJA-VIFPVBQESA-N 0.000 claims description 2
- 239000002674 ointment Substances 0.000 claims description 2
- 230000003204 osmotic effect Effects 0.000 claims description 2
- 229960005205 prednisolone Drugs 0.000 claims description 2
- OIGNJSKKLXVSLS-VWUMJDOOSA-N prednisolone Chemical compound O=C1C=C[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 OIGNJSKKLXVSLS-VWUMJDOOSA-N 0.000 claims description 2
- 239000003381 stabilizer Substances 0.000 claims description 2
- 229960002117 triamcinolone acetonide Drugs 0.000 claims description 2
- YNDXUCZADRHECN-JNQJZLCISA-N triamcinolone acetonide Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@H]3OC(C)(C)O[C@@]3(C(=O)CO)[C@@]1(C)C[C@@H]2O YNDXUCZADRHECN-JNQJZLCISA-N 0.000 claims description 2
- 230000003247 decreasing effect Effects 0.000 claims 1
- 230000005764 inhibitory process Effects 0.000 claims 1
- 230000000069 prophylactic effect Effects 0.000 claims 1
- 230000001225 therapeutic effect Effects 0.000 claims 1
- 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 abstract description 5
- 108060008682 Tumor Necrosis Factor Proteins 0.000 abstract description 5
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 abstract description 5
- 230000002401 inhibitory effect Effects 0.000 abstract description 5
- 239000002994 raw material Substances 0.000 abstract description 5
- 230000009467 reduction Effects 0.000 abstract description 4
- 210000002966 serum Anatomy 0.000 description 20
- 241000699666 Mus <mouse, genus> Species 0.000 description 18
- 241000699670 Mus sp. Species 0.000 description 18
- 238000011282 treatment Methods 0.000 description 18
- 229940079593 drug Drugs 0.000 description 16
- 239000000203 mixture Substances 0.000 description 15
- 102000008186 Collagen Human genes 0.000 description 12
- 108010035532 Collagen Proteins 0.000 description 12
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 12
- 229920001436 collagen Polymers 0.000 description 12
- 102000004890 Interleukin-8 Human genes 0.000 description 10
- 230000008859 change Effects 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- 201000010099 disease Diseases 0.000 description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 9
- 210000001503 joint Anatomy 0.000 description 9
- 210000001258 synovial membrane Anatomy 0.000 description 9
- 102000004127 Cytokines Human genes 0.000 description 8
- 108090000695 Cytokines Proteins 0.000 description 8
- 206010003246 arthritis Diseases 0.000 description 8
- 241000287828 Gallus gallus Species 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 239000003995 emulsifying agent Substances 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- 230000003628 erosive effect Effects 0.000 description 6
- 238000011156 evaluation Methods 0.000 description 6
- 238000004128 high performance liquid chromatography Methods 0.000 description 6
- 239000013642 negative control Substances 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- 229920001184 polypeptide Polymers 0.000 description 6
- 102000004196 processed proteins & peptides Human genes 0.000 description 6
- 238000004007 reversed phase HPLC Methods 0.000 description 6
- 210000001188 articular cartilage Anatomy 0.000 description 5
- 238000001819 mass spectrum Methods 0.000 description 5
- 238000001356 surgical procedure Methods 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 4
- 241000233866 Fungi Species 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000002271 resection Methods 0.000 description 4
- 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 4
- 238000002965 ELISA Methods 0.000 description 3
- 206010028813 Nausea Diseases 0.000 description 3
- 208000002193 Pain Diseases 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 239000002250 absorbent Substances 0.000 description 3
- 230000002745 absorbent Effects 0.000 description 3
- 230000002411 adverse Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 230000002538 fungal effect Effects 0.000 description 3
- 230000002757 inflammatory effect Effects 0.000 description 3
- 238000010172 mouse model Methods 0.000 description 3
- 230000008693 nausea Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 230000002441 reversible effect Effects 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical group C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 2
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 208000009386 Experimental Arthritis Diseases 0.000 description 2
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 2
- 102000003929 Transaminases Human genes 0.000 description 2
- 108090000340 Transaminases Proteins 0.000 description 2
- 206010047700 Vomiting Diseases 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 230000010100 anticoagulation Effects 0.000 description 2
- 239000003435 antirheumatic agent Substances 0.000 description 2
- 239000003124 biologic agent Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000007664 blowing Methods 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- 210000000845 cartilage Anatomy 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 244000053095 fungal pathogen Species 0.000 description 2
- 239000003862 glucocorticoid Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 238000010603 microCT Methods 0.000 description 2
- 229940021182 non-steroidal anti-inflammatory drug Drugs 0.000 description 2
- 238000010606 normalization Methods 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 239000002023 wood Substances 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 206010060968 Arthritis infective Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 208000008964 Chemical and Drug Induced Liver Injury Diseases 0.000 description 1
- 206010008479 Chest Pain Diseases 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 102000000541 Defensins Human genes 0.000 description 1
- 108010002069 Defensins Proteins 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 206010072268 Drug-induced liver injury Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 206010015150 Erythema Diseases 0.000 description 1
- 208000010201 Exanthema Diseases 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 101000974353 Mus musculus Nuclear receptor coactivator 5 Proteins 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 206010030302 Oliguria Diseases 0.000 description 1
- 208000007117 Oral Ulcer Diseases 0.000 description 1
- 241001197443 Paecilomyces purpureus Species 0.000 description 1
- 241001111421 Pannus Species 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 208000009921 Rheumatoid Nodule Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 208000018359 Systemic autoimmune disease Diseases 0.000 description 1
- 206010047249 Venous thrombosis Diseases 0.000 description 1
- 208000019790 abdominal distention Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000009098 adjuvant therapy Methods 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 208000022531 anorexia Diseases 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 208000002399 aphthous stomatitis Diseases 0.000 description 1
- 230000006793 arrhythmia Effects 0.000 description 1
- 206010003119 arrhythmia Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 229940049638 carbomer homopolymer type c Drugs 0.000 description 1
- 229940043234 carbomer-940 Drugs 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000007211 cardiovascular event Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000009693 chronic damage Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 201000005884 exanthem Diseases 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003414 extremity Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- KGPPDNUWZNWPSI-UHFFFAOYSA-N flurotyl Chemical compound FC(F)(F)COCC(F)(F)F KGPPDNUWZNWPSI-UHFFFAOYSA-N 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 229960000598 infliximab Drugs 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000008407 joint function Effects 0.000 description 1
- 206010023332 keratitis Diseases 0.000 description 1
- 238000013150 knee replacement Methods 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 201000002364 leukopenia Diseases 0.000 description 1
- 231100001022 leukopenia Toxicity 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 208000008494 pericarditis Diseases 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 206010037844 rash Diseases 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000008085 renal dysfunction Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000009528 severe injury Effects 0.000 description 1
- 208000018320 severe joint pain Diseases 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000012089 stop solution Substances 0.000 description 1
- 210000004304 subcutaneous tissue Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 229940037128 systemic glucocorticoids Drugs 0.000 description 1
- 239000007916 tablet composition Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 238000011541 total hip replacement Methods 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 230000009278 visceral effect Effects 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- 210000002517 zygapophyseal joint Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/10—Tetrapeptides
- C07K5/1021—Tetrapeptides with the first amino acid being acidic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Rheumatology (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- General Chemical & Material Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Physical Education & Sports Medicine (AREA)
- Mycology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Immunology (AREA)
- Pain & Pain Management (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Description
技术领域
本发明涉及生物医药领域,尤其是指一种人工拟肽及其制备方法和应用。
背景技术
类风湿关节炎(rheumatoid arthritis,RA)是一种慢性全身性自身免疫性疾病,主要引起关节的压痛,肿胀和破坏,从而导致残疾。该病好发于女性,且年龄主要集中在30-50岁之间。世界范围内,约1%的人口患有该疾病。RA的病灶部位多为小关节,具有对称分布的特性,早期的临床症状主要为红肿、疼痛、活动范围受限,随着疾病的进展逐步发展为关节活动能力下降,部分患者关节完全失去活动能力。如病情未能很好的控制,RA还可能导致除关节之外的器官受累,诸如:角膜炎、类风湿性结节、心膜炎等疾病。
尽管RA的发病机制尚不完全清楚,但是目前已经明确RA的发生是由遗传基因素与环境因素之间相互作用而发生发展的,除此之外,该病亦与激素的分泌、病毒与微生物感染等有关。在上述一种或几种因素的作用及影响下,人体会产生自主的免疫,引发滑膜产生炎症,最终致使局部关节受到破坏。病理上的特征主要在于炎性因子全身性的异常变化,使关节局部的血管发生异常的增生,从而在关节滑膜的形成新的血管翳,导致关节滑膜的炎症、增生及软骨的慢性损伤破坏。
当前治疗RA的主要方法为手术治疗和药物治疗:手术治疗是利用手术将相应的关节部分或全部切除或完全置换从而使患者恢复行动能力,是治疗该类疾病的重要方式,多用于中、晚期药物治疗效果不佳的软骨严重损伤缺失的患者,手术的方式主要包括滑膜切除和关节置换。滑膜切除是将含有病灶的关节打开,对于炎性浸润的滑膜进行切除,主要可通过放射滑膜切除和关节镜滑膜切除,其中关节镜滑膜切除术创伤较小,术后恢复较快,且能缩短住院的天数,减轻患者的痛苦。临床上常常在术后使用抗风湿药物,可进一步提高患者的关节功能和生活质量。RA进展到末期时,采用药物和滑膜切除术无法改善病情或预后不佳时,则需人开展工关节置换术,其中最常见的是全髋关节和全膝关节置换术。关节置换术后也有一定的风险(诸如人工关节感染、静脉血栓和心血管事件等不良事件)。因此,关节置换术对于因晚期RA而导致严重关节疼痛的患者更为合适,尽管置换关节可以使患者减轻痛苦,但RA具有全身性、多关节受累的特点,同样在术后需要使用药物进行辅助治疗。
大多数患者采用药物来治疗RA,该疗法也从单一的药物逐步发展为多种药物联合治疗。目前认为RA是无法治愈的终身疾病,患者需要终生使用抗炎或/和免疫调节药,目前临床上使用较多的治疗RA的药物主要有:非甾体抗炎药、抗风湿药、糖皮质激素、生物制剂、植物药制剂,详见表1。
表1目前临床治疗类风湿性关节炎的代表药物
依照患者疾病的具体情况综合选择合理的药物,并采用适当的药物治疗策略,可以一定程度控制RA病程的进展。但由于RA患者需要长期用药,且药物都有一定的副作用,比如长期使用非甾体抗炎药可诱发口腔溃疡、药物性肝损伤(肝脏酶异常升高)、腹痛、腹泻、恶心等症状;RA患者长时间使用糖皮质激素可增加其心血管疾病、糖尿病、感染等的风险,对骨骼人体发育均有影响;抗风湿药,如甲氨蝶呤有胃肠道反应(主要表现为恶心、呕吐、腹泻等)、骨髓抑制(白细胞降低、皮肤或内脏出血)、肾脏损害等不良反应;生物制剂如英夫利昔单抗在临床试验中曾发生皮疹、头痛、恶心、呼吸道感染、血压异常、转氨酶升高等不良事件,同时患者使用生物制剂的医疗费用是该疾病的平均治疗费用的3-5倍,经济负担较重;植物药制剂,如雷公藤多苷片也存在呕吐、乏力、食欲不振、腹胀、转氨酶升高、白细胞、血小板下降、少尿或多尿、水肿、肾功能异常、心悸、胸闷、心律失常、头昏、头晕等不良反应。
综上所述,鉴于手术治疗和药物治疗中存在的问题和该病的发病率居高不下,为了解决上述治疗过程中需要的问题和瓶颈,发现新型的具有良好的治疗类风湿性关节炎潜力的活性候选药物分子是亟待解决的重要科学问题,亦具有重要的社会意义和经济价值。
发明内容
本发明旨在至少在一定程度上解决现有技术中存在的技术问题之一,在本发明的第一方面,本发明提供了一种人工拟肽,所述人工拟肽的结构式如式1所示:
本发明所述拟肽的模板分子来源于条件致病真菌淡紫色拟青霉Purpureocilliumlilacinum中的多肽Prulisin(Gene Bank ID:PWI75532.1),完整的氨基酸序列(N端→C端)如下:
[参考文献:Shen B,Cao Z,Wu Y,Yi W,Zhu Z,Lv Z,Zhu C,Yu Y*.Purlisin,atoxin-like defensin derived from clinical pathogenic fungusPurpureocilliumlilacinum with both antimicrobial and potassium channelinhibitory activities.FASEB J.2020,(11):15093-15107.]
将天然多肽截短,利用该天然多肽中含有多聚碱性氨基酸的肽段CYPRKR(方框中的部分),将N端半胱氨酸残基中游离的氨基(-NH2)甲酰化,制备天然多肽衍生物,即拟肽中间体HCONH-CYPRKR。将上述来源于真菌多肽Purlisin截短拟肽中间体HCONH-CYPRKR与前导拟肽DGRC-COOC2H5(即天然多肽DGRC的C端半胱氨酸残基中的羧基乙酯化的衍生物)两者通过半胱氨酸中的巯基相偶联,连接成完整的全新结构的分子,其具体结构为:
本发明的第一方面所述的人工拟肽的制备方法的基本思路为:首先通过固相合成技术制备前导拟肽DGRC-COOC2H5和真菌来源的天然截短拟肽HCONH-CYPRKR,再通过半胱氨酸侧链的巯基形成分子间的二硫键使两个片段相偶联。反向高效液相色谱(Reversed-phase High Performance Liquid Chromatography)纯化后,经电喷雾质谱法(Electrospray ionisation tandem mass spectrometry,ESI-MS)测定分子量。
由此,本发明的第二方面,在于提供了一种在本发明的第一方面所述的人工拟肽的制备方法,其由前导拟肽中间体DGRC-COOC2H5和天然截短拟肽中间体HCONH-CYPRKR制备得到,所述前导拟肽中间体DGRC-COOC2H5的结构式如式2所示:
所述天然截短拟肽中间体HCONH-CYPRKR的结构式如式3所示:
在本发明的一个或多个实施例中,本发明的第一方面所述的人工拟肽的制备方法,包括如下步骤:
步骤1):以9-芴甲氧羰基(FMOC)作为氨基端保护基团,以4-(2′,4′-二甲氧基苯基-芴甲氧羰基-氨甲基)-苯氧基乙酰氨基-甲基二苯甲胺树脂为起始氨基酸固相载体,依次接连天冬氨酸、甘氨酸、精氨酸,得到氨基酸固相载体,最后将羧基乙酰化的半胱氨酸的氨基与所述氨基酸固相载体的精氨酸R末端的羧基形成酰胺键,得到键合于固相载体上的DGRC-COOC2H5,去掉固相载体,纯化,即得到前导拟肽中间体DGRC-COOC2H5;
步骤2):以9-芴甲氧羰基(FMOC)作为氨基端保护基团,以4-(2′,4′-二甲氧基苯基-芴甲氧羰基-氨甲基)-苯氧基乙酰氨基-甲基二苯甲胺树脂(Rink amide MBHA resin)为起始氨基酸固相载体,依次接连以氨基甲酰化的半胱氨酸、酪氨酸、脯氨酸、精氨酸、赖氨酸和精氨酸,得到键合于固相载体上的HCONH-CYPRKR,去掉固相载体,纯化,即得到天然截短拟肽中间体HCONH-CYPRKR;
步骤3):将步骤1)得到的前导拟肽中间体DGRC-COOC2H5和步骤2)得到的天然截短拟肽中间体HCONH-CYPRKR在26℃~28℃,60~80rpm下孵育36~48小时,即得所述人工拟肽。
另外,发明人通过实验证实,本发明所涉及的人工拟肽,具有抑制类风湿性关节炎模型小鼠血清中炎症相关细胞因子IL-1β、IL-8和TNF-α的作用。在动物水平上,够能显著缓解实验小鼠病灶关节处的红肿或/和肿胀或/和骨侵蚀或/和关节变形或/和骨密度下降等症状,具有作为候选药物进一步研究和开发的价值。
由此,本发明的第三方面,在于提供了一种本发明的第一方面所述的人工拟肽在制备类风湿性关节炎药物中的应用。
本发明还提供了一种本发明的第一方面所述的人工拟肽在制备预防或治疗关节处的红肿或/和关节处的肿胀或/和关节处的骨侵蚀或/和关节变形或/和关节骨密度下降的药物中的应用。
本发明的第四方面,在于本发明提供了一种本发明的第一方面所述的人工拟肽在制备预防或治疗抑制炎症相关细胞因子IL-1β或/和IL-8或/和TNF-α的药物中的应用。
本发明的第五方面,在于本发明提供一种药物制剂,所述药物制剂包含本发明的第一方面所述的人工拟肽或所述人工拟肽在药学上可接受的盐或所述人工拟肽的水合物或所述人工拟肽的溶剂化物。
本发明的一个或多个实施例中,所述药物制剂还包括药学上可接受的辅料。
本发明的一个或多个实施例中,所述药学上可接受的辅料选自填充剂、pH调节剂、稳定剂、注射用水和渗透压调节剂中的一种或多种。
本发明的一个或多个实施例中,所述药物制剂为注射剂、片剂、粉剂、颗粒剂、胶囊剂、口服液、膏剂、霜剂、凝胶剂、冻干粉中的一种。
本发明的第六方面,在于本发明提供了一种药物组合物,包括地塞米松、泼尼松龙、曲安奈德、萘普生、双氯酚酸中的一种或多种与本发明的第一方面所述的人工拟肽的组合。
在本发明的第七方面,本发明提供一种制备本发明的第一方面所述的人工拟肽的前导拟肽中间体DGRC-COOC2H5化合物,其特征在于,所述前导拟肽中间体DGRC-COOC2H5的结构式如式2所示:
本发明的一个或多个实施例中,上述前导拟肽中间体DGRC-COOC2H5的制备方法包括如下步骤:以9-芴甲氧羰基(FMOC)作为氨基端保护基团,以4-(2′,4′-二甲氧基苯基-芴甲氧羰基-氨甲基)-苯氧基乙酰氨基-甲基二苯甲胺树脂为起始氨基酸固相载体,依次接连天冬氨酸、甘氨酸、精氨酸,得到氨基酸固相载体,最后将羧基乙酰化的半胱氨酸的氨基与所述氨基酸固相载体的精氨酸R末端的羧基形成酰胺键,得到键合于固相载体上的DGRC-COOC2H5,去掉固相载体,纯化,即得到前导拟肽中间体DGRC-COOC2H5;
本发明还提供一种制备本发明的第一方面所述的人工拟肽的天然截短拟肽中间体HCONH-CYPRKR化合物,其特征在于,所述天然截短拟肽中间体HCONH-CYPRKR的结构式如式3所示:
本发明的一个或多个实施例中,上述天然截短拟肽中间体HCONH-CYPRKR的制备方法包括如下步骤:以9-芴甲氧羰基(FMOC)作为氨基端保护基团,以4-(2′,4′-二甲氧基苯基-芴甲氧羰基-氨甲基)-苯氧基乙酰氨基-甲基二苯甲胺树脂(Rink amide MBHA resin)为起始氨基酸固相载体,依次接连以氨基甲酰化的半胱氨酸、酪氨酸、脯氨酸、精氨酸、赖氨酸和精氨酸,得到键合于固相载体上的HCONH-CYPRKR,去掉固相载体,纯化,即得到天然截短拟肽中间体HCONH-CYPRKR。
与现有技术相比,本发明具有以下优点和有益效果:
1、本发明提供一种人工拟肽,其具有药用价值。
2、本发明提供上述人工拟肽的制备方法,其原料选择合理,易得,制备工艺简单,易于产业化。
3、本发明提供一种上述人工拟肽在制备预防或治疗关节处的红肿或/和关节处的肿胀或/和关节处的骨侵蚀或/和关节变形或/和关节骨密度下降的药物中的应用。
4、本发明提供了一种上述的人工拟肽在制备预防或治疗抑制炎症相关细胞因子IL-1β或/和IL-8或/和TNF-α的药物中的应用。
5、本发明还提供一种包含上述人工拟肽或所述人工拟肽在药学上可接受的盐或所述人工拟肽的水合物或所述人工拟肽的溶剂化物的药物制剂。
附图说明
图1为本发明实施例1制备的前导拟肽中间体DGRC-COOC2H5的HPLC图;
图2为本发明实施例1制备的前导拟肽中间体DGRC-COOC2H5的质谱图;
图3为本发明实施例2制备的天然截短拟肽中间体HCONH-CYPRKR的HPLC图;
图4为本发明实施例2制备的天然截短拟肽中间体HCONH-CYPRKR的质谱图;
图5为本发明实施例3制备的人工拟肽完整分子的HPLC图;
图6为本发明实施例3制备的人工拟肽完整分子的质谱图;
图7为正常组、模型组及人工拟肽干预组关节炎指数得分图;图7说明:*P与模型组小鼠相比(P<0.05);
图8为正常组、模型组及人工拟肽干预组的小鼠关节软骨侵蚀程度及骨密度评估得分图;图8说明:*P与正常组相比(P<0.05);
图9为经本发明实施例3制备的人工拟肽治疗后模型小鼠血清细胞因子IL-1β的变化;图9说明:*P与正常组相比(P<0.05),#P与模型组相比(P<0.05);
图10为经本发明实施例3制备的人工拟肽治疗后模型小鼠血清细胞因子IL-8的变化;图10说明:*P与正常组相比(P<0.05),#P与模型组相比(P<0.05);
图11为经本发明实施例3制备的人工拟肽治疗后模型小鼠血清细胞因子TNF-α的变化;图11说明:*P与正常组相比(P<0.05),#P与模型组相比(P<0.05)。
具体实施方式
下面将结合实施例和对比例对本发明的方案进行解释。本领域技术人员将会理解,下面的实施例和对比例仅用于说明本发明,而不应视为限定本发明的范围。实施例中未注明具体技术或条件的,按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。使用的方法如无特别说明,均为本领域公知的常规方法,使用的耗材和试剂如无特别说明,均为市场购得。除非另有说明,本文中所用的专业与科学术语与本领域熟练人员所熟悉的意义相同。此外,任何与所记载内容相似或均等的方法或材料也可应用于本发明中。
实施例1:前导拟肽中间体DGRC-COOC2H5的制备
前导拟肽中间体DGRC-COOC2H5的结构式如下式2所示:
具体制备步骤如下所示:
在前导拟肽中间体制备中,以9-芴甲氧羰基(FMOC)作为氨基端保护基团,以4-(2′,4′-二甲氧基苯基-芴甲氧羰基-氨甲基)-苯氧基乙酰氨基-甲基二苯甲胺树脂(Rinkamide MBHA resin)为固相载体,缩合剂为1-羟基苯并三唑/二环己基碳二亚胺(HOBt/DCC),由羧基端向氨基端方向延长肽链,依次接连天然氨基酸天冬氨酸、甘氨酸、精氨酸。最后用羧基乙酰化的非天然的半胱氨酸作原料,以其氨基与精氨酸R末端的羧基形成酰胺键。用质量百分比(m/m)为94%三氟乙酸(TFA)、3%的水和3%的三异丙甲硅烷(TIA)的混合液,将该前导拟肽中间体分子从MBHA树脂上裂解。经过乙醚反复沉淀后,用制备型RP-HPLC进行纯化。使用C18反向半制备柱(300mm×30mm,10μm);流动相为1.5‰的三氟乙酸,体积百分比(v/v)为0~55%的乙腈为流动相,2mL/min的流速进行梯度洗脱。再通过分析型RP-HPLC进行检测,保留时间(retention time,RT)为9.64min,经色谱峰面积归一化法计算,其纯度为96.54%(制备的前导拟肽中间体DGRC-COOC2H5的HPLC图如图1所示),冻干后置于-20℃冰箱存放备用。经ESI/MS测定,其分子量为m/z 477.84[M+H]+,与拟制备的前导拟肽中间体理论分子量一致(制备的前导拟肽中间体DGRC-COOC2H5的质谱图如图2所示)。
实施例2:天然截短拟肽中间体HCONH-CYPRKR的制备
天然截短拟肽中间体HCONH-CYPRKR的结构式如下式3所示:
具体制备步骤如下所示:
在天然截短拟肽中间体的制备中,以9-芴甲氧羰基(FMOC)作为氨基端保护基团,以4-(2′,4′-二甲氧基苯基-芴甲氧羰基-氨甲基)-苯氧基乙酰氨基-甲基二苯甲胺树脂(Rink amide MBHA resin)为固相载体,缩合剂为1-羟基苯并三唑/二环己基碳二亚胺(HOBt/DCC),由羧基端向氨基端方向延长肽链,首先以氨基甲酰化的半胱氨酸为原料,再依次接连酪氨酸、脯氨酸、精氨酸、赖氨酸和精氨酸。用质量百分比(m/m)为96%三氟乙酸(TFA)、2%的水和2%的三异丙甲硅烷(TIA)的混合液,将该前导拟肽中间体分子从MBHA树脂上裂解。经过乙醚反复沉淀后,用制备型RP-HPLC进行纯化。使用C18反向半制备柱(300mm×30mm,10μm);流动相为2.0‰的三氟乙酸,体积百分比(v/v)为0~65%的乙腈为流动相,2mL/min的流速进行梯度洗脱。再经分析型RP-HPLC进行检测,保留时间(retention time,RT)为14.36min,经色谱峰面积归一化法计算,其纯度为97.21%。(制备的天然截短拟肽中间体HCONH-CYPRKR的HPLC图如图3所示),冻干后置于-20℃冰箱存放备用。经ESI/MS测定,其分子量为m/z 851.58[M+H]+,与拟制备的天然截短拟肽中间体理论分子量一致(制备的天然截短拟肽中间体HCONH-CYPRKR的质谱图如图4所示)。
实施例3:人工拟肽的合成
人工拟肽的结构式如下式1所示:
具体合成步骤如下所示:以上述实施例1和实施例2中制备的前导拟肽中间体DGRC-COOC2H5以及天然截短拟肽中间体HCONH-CYPRKR为原料。分别取10mg前导拟肽中间体和18mg天然截短拟肽中间体于50mL棕色蓝盖试剂瓶中(两者的摩尔质量比约为1:1),加入25mL浓度为0.1M pH=7.1的Tris-HCl缓冲液。拧紧瓶盖后用Pharfim封口膜密封,确认无漏液后颠倒试剂瓶以加速多肽溶解。将盛有前导拟肽中间体和天然截短拟肽中间体溶液的试剂瓶置于恒温摇床中于26℃~28℃,60~80rpm孵育36~48小时,促进分子间二硫键的形成。于4℃10000rpm高速离心5min,取上清液。使用C18反向制备柱(300mm×30mm,10μm),经制备型RP-HPLC纯化。流动相为1.5%三氟乙酸,体积百分比0%~75%(v/v)的乙腈为流动相,以流速3.0mL/min的流速进行梯度洗脱,收集主峰洗脱液,冻干后备用。经分析型RP-HPLC检测,完整的人工拟肽分子保留时间RT为16.56min,其纯度为95.29%(制备的人工拟肽完整分子的HPLC图如图5所示)。经ESI/MS测定,其分子量为m/z 1326.04[M+H]+,663.91[M+2H]2+,与真菌来源的人工拟肽分子的理论分子量一致(制备的人工拟肽完整分子的质谱图如图6所示)。
实施例4:类风湿性关节炎模型小鼠的构建及人工拟肽的干预
使用6周龄的SPF级雄性DBA/1共18只,体重约(15±3)g,饲养过程中保持恒温恒湿且通风达标的环境,所有实验小鼠饮水进食均自由。上述小鼠饲养1周后,随机分成空白对照组和模型组。空白对照组小鼠不作任何干预与处理,模型组利用鸡II型胶原免疫两次以诱导类风湿性关节炎的模型,具体方式如下:
1.免疫胶原乳化剂的制备
首次免疫胶原乳化剂的制备:将冷冻保存的50mg鸡胶原蛋白取出后溶于浓度为0.1M的20mL醋酸中,置于4℃层析柜中溶解过夜后在冰浴中按体积比1:1~1.5滴加完全弗氏佐剂(Complete Freunds Adjuvant,CFA)形成乳状混合物,接着用移液枪反复吹吸混合物使其混匀,使其成为W/O的乳剂,即为初次免疫乳化剂。
二次免疫胶原乳化剂的制备:将冷冻保存的50mg鸡胶原蛋白取出后溶于浓度为0.1M的20mL醋酸中,置于4℃层析柜中溶解过夜后在冰浴中按体积比1:1~1.5滴加不完全弗氏佐剂(Freund incomplete adjuvant,FIA)形成乳状混合物,接着用移液枪反复吹吸混合物使其混匀,使其成为W/O的乳剂,即为二次免疫乳化剂。
2.胶原诱导性关节炎(collagen induced arthritis,CIA)小鼠模型的建立及拟肽干预
将12只待造模的小鼠用固定器固定好后,于尾部皮下注射首次免疫胶原乳化剂,每只小鼠的注射0.25mL,在首次注射免疫后三周,进行第二次免疫。运用同样的方法将事先制备好的二次免疫胶原乳化剂注射到小鼠为根部皮下组织中。经过二次免疫后的小鼠一周后可用本发明实施例3制备得到的人工拟肽进行干预治疗。
3.人工拟肽进行干预治疗
将12只模型小鼠分为阴性对照组(模型组)和干预治疗组。模型组不作任何干预,每日灌喂适量生理盐水,治疗组用本发明实施例3制备得到人工拟肽进行关节腔微量注射(浓度150μg/mL,注射量5μL),每周注射2次,连续注射给药2周。
实施例5:小鼠CIA模型及治疗效果评估
于造模前后以及本发明所述人工拟肽干预前后,对小鼠实施观察,记录各组小鼠关节炎指标评分,以如下表2中Wood关节炎量表中各个项目进行评价(按分值分为0~4五级),最后根据计算出四肢的累计评分为小鼠关节炎指数的得分(正常组、模型组及人工拟肽干预组关节炎指数得分图如图7所示)。
表2 Wood关节炎量表
另一方面,利用Micro-CT对正常组、模型组及人工拟肽干预组的小鼠关节(后肢关节)进行扫描,并对扫描的样品三维重构。以如下表3中关节软骨的侵蚀程度及骨密度评分标准及对应分值表中的评分标准及对应分值表对关节骨损伤的评价标准根据关节软骨的侵蚀程度及骨密度进行分析。记录并统计各组小鼠的评分结果(正常组、模型组及人工拟肽干预组的小鼠关节软骨侵蚀程度及骨密度评估得分图如图8所示)。
表3关节软骨的侵蚀程度及骨密度评分标准及对应分值表
可以看出,按照实施例4中的方法造模成功,免疫后小鼠的关节炎指数以及Micro-CT侵蚀程度及骨密度评分的得分明显增高,且差异具有统计学意义(P<0.05);经本发明所述的真菌来源的人工拟肽治疗后小鼠关节炎指数明显降低,且具有时间依赖性和统计学意义(P<0.05)(图7、图8)。
实施例6:人工拟肽治疗前后模型小鼠血清细胞因子IL-1β、IL-8及TNF-α水平变化
炎症因子在类风湿性关节炎的发生和进展过程中起着十分关键的作用,利用酶联免疫吸附测定法(enzyme linked immunosorbent assay,ELISA),测定小鼠血清中细胞因子IL-1β、IL-8及TNF-α的变化。
阴性对照组、模型组、实施例3制备得到人工拟肽干预组的小鼠,利用眼眶静脉采血的方法将各组小鼠血液样本收集与抗凝管中,颠倒数次,防止血液凝固。将抗凝管在室温在放置30min后于离心机中,6000rpm离心8min,使红细胞沉淀于管底,收集上清于干净无菌的离心管中,置于-20℃冰箱中保存。
将ELISA检测试剂盒从冰箱取出后室温放置20min,按需拆封板条,每孔加入250μL洗涤液清洗板孔,静置45s后将板中液体弃去,并在吸水滤纸上拍干。在板孔中加入适量的不同浓度的标准品,样品板孔中同样加入等体积的待测样品(如样品浓度过大,超过标曲范围,则应将样品稀释后再测定),封板膜封口后室温下静置1小时。将板孔中液体弃去,并在吸水滤纸上倒扣拍干孔中的液体,每孔加300μL洗涤液,静置45s后弃去洗涤液,重复洗板3次后将板孔中的剩余液体拍干,每孔加入辣根过氧化物酶(Horseradish Peroxidase,HRP)标记的抗体100μL,室温下静置30min。将板孔中的液体弃去,并在吸水滤纸上倒扣拍干孔中的液体,每孔加300μL洗涤液,静置45s后弃去,重复3次后将板孔中的液体拍干。在每孔中加入100μL3,3',5,5'-四甲基联苯胺(Tetramethylbenzidine,TMB),室温下避光孵育10min后于每孔中加入100μL终止液,用全波长酶标仪测定各孔在波长为450nm处的OD值。绘制标准曲线,以标准品浓度为X轴,OD450值为Y轴,按所得的方程计算出各孔中的浓度。
图9是经本发明实施例3制备得到的人工拟肽治疗后模型小鼠血清细胞因子IL-1β的变化(即阴性对照组、模型组、实施例3制备得到人工拟肽干预组模型小鼠血清细胞因子IL-1β的变化)。图10是经本发明实施例3制备得到的人工拟肽治疗后模型小鼠血清细胞因子IL-8的变化(即阴性对照组、模型组、实施例3制备得到人工拟肽干预组模型小鼠血清细胞因子IL-8的变化)。图11是经发明的人工拟肽治疗后模型小鼠血清细胞因子TNF-α的变化(即阴性对照组、模型组、实施例3制备得到人工拟肽干预组模型小鼠血清细胞因子TNF-α的变化)。由实验结果可知,正常小鼠(阴性对照组)血清中炎症相关细胞因子IL-1β、IL-8及TNF-α均处于较低水平,分别为(68.68±9.21)pg/mL、(54.25±8.06)pg/mL、(51.20±6.34)pg/mL。而模型组小鼠血清中,上述三种细胞因子水平均呈现出较大幅度的升高,分别为(302.11±39.45)pg/mL、(281.23±30.15)pg/mL、(354.26±16.94)pg/mL,且差异具有统计学意义(P<0.05)。经本发明所述实施例3制备得到人工拟肽治疗后两周后,以上三种细胞因子在血清中的水平分别为(140.65±19.44)pg/mL、(124.23±14.43)pg/mL、(180.01±12.28)pg/mL,较之于模型组,血清细胞因子水平均有显著的降低,且差异具有统计学意义(P<0.05)。由此可见,本发明实施例3制备得到的人工拟肽具有明显抑制血清炎症相关细胞因子的作用。
实施例7:片剂
取本发明实施例3制备得到的真菌来源的人工拟肽0.25g、淀粉3.0g、糊精4.0g充分混合均匀,加入质量浓度为45%(m/m)的药用级聚乙烯吡咯烷酮(PVP)作为粘合剂,制粒、整粒、压片,既得片剂。
实施例8:凝胶剂
取10mL蒸馏水,将药用级卡波姆940均匀撒于液面上,搅拌使之充分溶胀。加入丙二醇混合均匀,接着边搅拌边滴入三乙胺制得凝胶基质。将本发明实施例3制备得到的真菌来源的具有治疗类风湿性关节炎作用的人工拟肽0.15g溶于5mL PBS溶液中(0.15M,pH=7.4),在不断搅拌下将其缓慢加入凝胶基质中。最后补加灭菌蒸馏水,即得约20g本发明所述人工拟肽凝胶。
实施例9:注射用冻干粉
取本发明实施例3制备得到的真菌来源的具有治疗类风湿性关节炎作用的人工拟肽2g、甘露醇20g,置于容器中,加适量PBS缓冲液(0.15M,pH 7.4)溶解,加注射用水至300mL,充分混匀,加8~12g针用活性炭,室温搅拌50分钟,粗滤后用0.22μm灭菌滤膜过滤除菌,分装,每瓶2.0mL。以速冻的方式,每分钟降温10~20℃,降温至-50~-55℃,维持3小时,抽真空,在真空状态下缓慢升温,升温速度为每小时3~5℃,温度缓慢升高至(25±2)℃时停止升温,待温度接近室温后将样品取出,加盖密封,即可得到冻干粉针剂。
尽管上面已经示出和描述了本发明的实施例和对比例,可以理解的是,上述实施例和对比例是示例性的,不能理解为对本发明的限制,本领域的普通技术人员在本发明的范围内可以对上述实施例进行变化、修改、替换和变型。
Claims (10)
5.根据权利要求4所述的人工拟肽的制备方法,其特征在于,包括如下步骤:
步骤1):以9-芴甲氧羰基(FMOC)作为氨基端保护基团,以4-(2′,4′-二甲氧基苯基-芴甲氧羰基-氨甲基)-苯氧基乙酰氨基-甲基二苯甲胺树脂为起始氨基酸固相载体,依次接连天冬氨酸、甘氨酸、精氨酸,得到氨基酸固相载体,最后将羧基乙酰化的半胱氨酸的氨基与所述氨基酸固相载体的精氨酸R末端的羧基形成酰胺键,得到键合于固相载体上的DGRC-COOC2H5,去掉固相载体,纯化,即得到前导拟肽中间体DGRC-COOC2H5;
步骤2):以9-芴甲氧羰基(FMOC)作为氨基端保护基团,以4-(2′,4′-二甲氧基苯基-芴甲氧羰基-氨甲基)-苯氧基乙酰氨基-甲基二苯甲胺树脂(Rink amide MBHA resin)为起始氨基酸固相载体,依次接连以氨基甲酰化的半胱氨酸、酪氨酸、脯氨酸、精氨酸、赖氨酸和精氨酸,得到键合于固相载体上的HCONH-CYPRKR,去掉固相载体,纯化,即得到天然截短拟肽中间体HCONH-CYPRKR;
步骤3):将步骤1)得到的前导拟肽中间体DGRC-COOC2H5和步骤2)得到的天然截短拟肽中间体HCONH-CYPRKR在26℃~28℃,60~80rpm下孵育36~48小时,即得所述人工拟肽。
6.一种权利要求1所述的人工拟肽在制备类风湿性关节炎药物中的应用。
7.一种权利要求1所述的人工拟肽在制备预防或治疗关节处的红肿或/和关节处的肿胀或/和关节处的骨侵蚀或/和关节变形或/和关节骨密度下降的药物中的应用。
8.一种权利要求1所述的人工拟肽在制备预防或治疗抑制炎症相关细胞因子IL-1β或/和IL-8或/和TNF-α的药物中的应用。
9.一种药物制剂,其特征在于,包含权利要求1所述的人工拟肽或所述人工拟肽在药学上可接受的盐或所述人工拟肽的水合物或所述人工拟肽的溶剂化物;
优选地,所述药物制剂还包括药学上可接受的辅料,所述药学上可接受的辅料选自填充剂、pH调节剂、稳定剂、注射用水和渗透压调节剂中的一种或多种;
优选地,所述药物制剂为注射剂、片剂、粉剂、颗粒剂、胶囊剂、口服液、膏剂、霜剂、凝胶剂、冻干粉中的一种。
10.一种药物组合物,其特征在于,包括地塞米松、泼尼松龙、曲安奈德、萘普生、双氯酚酸中的一种或多种与权利要求1所述的人工拟肽的组合。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110481513.9A CN113024643B (zh) | 2021-04-30 | 2021-04-30 | 一种人工拟肽及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110481513.9A CN113024643B (zh) | 2021-04-30 | 2021-04-30 | 一种人工拟肽及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113024643A true CN113024643A (zh) | 2021-06-25 |
CN113024643B CN113024643B (zh) | 2022-05-13 |
Family
ID=76454805
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110481513.9A Active CN113024643B (zh) | 2021-04-30 | 2021-04-30 | 一种人工拟肽及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113024643B (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113583098A (zh) * | 2021-07-09 | 2021-11-02 | 武汉大学 | 一种来源于真菌的环状拟肽及其制备方法和应用 |
CN116003541A (zh) * | 2022-07-27 | 2023-04-25 | 武汉大学 | 多功能真菌防御素修饰肽、其制备方法及应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102449481A (zh) * | 2009-05-29 | 2012-05-09 | 德克萨斯大学系统董事会 | 用于分离和处理自身免疫性t细胞的拟肽配体 |
CN102796173A (zh) * | 2012-06-27 | 2012-11-28 | 中国人民解放军军事医学科学院基础医学研究所 | 一种类风湿关节炎的抗原表位及其应用 |
EP2853537A4 (en) * | 2011-12-19 | 2016-05-25 | Shanghai 1St Peoples Hospital | SMALL MOLECULAR POLYPEPTIDE FOR THE PREVENTION AND INHIBITION OF INFLAMMATION AND APPLICATION THEREOF |
US20170037086A1 (en) * | 2014-04-09 | 2017-02-09 | Aileron Therapeutics, Inc. | Peptidomimetic macrocycles with pth activity |
-
2021
- 2021-04-30 CN CN202110481513.9A patent/CN113024643B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102449481A (zh) * | 2009-05-29 | 2012-05-09 | 德克萨斯大学系统董事会 | 用于分离和处理自身免疫性t细胞的拟肽配体 |
EP2853537A4 (en) * | 2011-12-19 | 2016-05-25 | Shanghai 1St Peoples Hospital | SMALL MOLECULAR POLYPEPTIDE FOR THE PREVENTION AND INHIBITION OF INFLAMMATION AND APPLICATION THEREOF |
CN102796173A (zh) * | 2012-06-27 | 2012-11-28 | 中国人民解放军军事医学科学院基础医学研究所 | 一种类风湿关节炎的抗原表位及其应用 |
US20170037086A1 (en) * | 2014-04-09 | 2017-02-09 | Aileron Therapeutics, Inc. | Peptidomimetic macrocycles with pth activity |
Non-Patent Citations (2)
Title |
---|
杨麟等: "IL-6受体对类风湿关节炎成纤维样滑膜细胞致病信号及功能的抑制作用", 《中国现代应用药学》 * |
王洁等: "肿瘤坏死因子及其受体在类风湿性关节炎中的研究进展", 《药学实践杂志》 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113583098A (zh) * | 2021-07-09 | 2021-11-02 | 武汉大学 | 一种来源于真菌的环状拟肽及其制备方法和应用 |
CN113583098B (zh) * | 2021-07-09 | 2023-03-14 | 武汉大学 | 一种来源于真菌的环状拟肽及其制备方法和应用 |
CN116003541A (zh) * | 2022-07-27 | 2023-04-25 | 武汉大学 | 多功能真菌防御素修饰肽、其制备方法及应用 |
CN116003541B (zh) * | 2022-07-27 | 2024-05-28 | 武汉大学 | 多功能真菌防御素修饰肽、其制备方法及应用 |
Also Published As
Publication number | Publication date |
---|---|
CN113024643B (zh) | 2022-05-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN113024643B (zh) | 一种人工拟肽及其制备方法和应用 | |
EP2426139B1 (en) | CXCR4 antagonist and use thereof | |
KR100234520B1 (ko) | 종양괴사인자 매개병 치료용 의약품 조성물 | |
AU2011272137B2 (en) | Novel peptide and use thereof | |
EP0443404A1 (en) | Peptide fragments and analogs of thrombospondin | |
JP3818322B2 (ja) | コラーゲン分解促進剤 | |
EP2987804A1 (en) | Solid phase synthesis of h[gly2]glp-2 | |
EP2894163A1 (en) | A glp-1 analogue, its preparation methods and use thereof | |
US20200188471A1 (en) | Application of selective tnfr1 antagonist peptide sn10 in preparation of drugs for preventing and treating rheumatoid arthritis | |
CN105906709A (zh) | 狭鳕鱼皮活性寡肽及其合成方法和应用 | |
JPH02174798A (ja) | 細胞接着活性コア配列の繰り返し構造からなるポリペプチド | |
EP0524834A2 (en) | Immunosuppressive drugs containing a cysteine protease | |
EP0792886B1 (en) | Novel peptide and therapeutic agent | |
US7659246B2 (en) | Treatment of osteoarthritis | |
US4981950A (en) | Vasoconstrictor peptide | |
EP2464656A1 (en) | Novel peptide and use thereof | |
CN101696234A (zh) | 抗类风湿性关节炎的多肽及其在制药中的应用 | |
JPH08509960A (ja) | 骨原性成長オリゴペプチドおよびそれを含む医薬組成物 | |
WO2005086578A2 (en) | Anti-inflammatory peptides and methods of use thereof | |
CN112125952B (zh) | 一种猪源ace抑制活性多肽与药物组合物或食品及应用 | |
CN110669105B (zh) | 长效多肽构建及其抗急性肾损伤和糖尿病并发肾病的应用 | |
WO2011119008A2 (en) | Peptides for promoting angiogenesis and an use thereof | |
JP2000191700A (ja) | 持続型消化管運動ペプチド | |
CN113583098B (zh) | 一种来源于真菌的环状拟肽及其制备方法和应用 | |
US20230149510A1 (en) | Method for alleviating arthritis using epidermal growth factor |
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 | ||
TR01 | Transfer of patent right |
Effective date of registration: 20231031 Address after: 430022 room 35, floor 3, building 1 and 2, Vanke golden home phase II, east of Qianjin 1st Road, Jianghan District, Wuhan City, Hubei Province Patentee after: Wuhan Inverse Time Medical Health Investment Co.,Ltd. Address before: 430072 Hubei Province, Wuhan city Wuchang District of Wuhan University Luojiashan Patentee before: WUHAN University |
|
TR01 | Transfer of patent right |