CN113956346B - 一种重组白介素-15变体 - Google Patents
一种重组白介素-15变体 Download PDFInfo
- Publication number
- CN113956346B CN113956346B CN202111246297.6A CN202111246297A CN113956346B CN 113956346 B CN113956346 B CN 113956346B CN 202111246297 A CN202111246297 A CN 202111246297A CN 113956346 B CN113956346 B CN 113956346B
- Authority
- CN
- China
- Prior art keywords
- ile
- leu
- glu
- interleukin
- ser
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 102000003812 Interleukin-15 Human genes 0.000 title claims abstract description 115
- 108090000172 Interleukin-15 Proteins 0.000 title claims abstract description 115
- 241000588724 Escherichia coli Species 0.000 claims abstract description 9
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 24
- 239000013604 expression vector Substances 0.000 claims description 22
- 230000014509 gene expression Effects 0.000 claims description 21
- 101001055157 Homo sapiens Interleukin-15 Proteins 0.000 claims description 17
- 102000056003 human IL15 Human genes 0.000 claims description 17
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 17
- 229920001184 polypeptide Polymers 0.000 claims description 15
- 208000015181 infectious disease Diseases 0.000 claims description 14
- 239000008194 pharmaceutical composition Substances 0.000 claims description 13
- 206010028980 Neoplasm Diseases 0.000 claims description 12
- 208000035473 Communicable disease Diseases 0.000 claims description 9
- 108091033319 polynucleotide Proteins 0.000 claims description 9
- 102000040430 polynucleotide Human genes 0.000 claims description 9
- 239000002157 polynucleotide Substances 0.000 claims description 9
- 208000023275 Autoimmune disease Diseases 0.000 claims description 8
- 201000011510 cancer Diseases 0.000 claims description 8
- 208000027866 inflammatory disease Diseases 0.000 claims description 8
- 102000004169 proteins and genes Human genes 0.000 claims description 7
- 108090000623 proteins and genes Proteins 0.000 claims description 7
- 206010006187 Breast cancer Diseases 0.000 claims description 6
- 239000003085 diluting agent Substances 0.000 claims description 6
- 208000014951 hematologic disease Diseases 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 230000002265 prevention Effects 0.000 claims description 6
- 208000026310 Breast neoplasm Diseases 0.000 claims description 5
- 206010009944 Colon cancer Diseases 0.000 claims description 5
- 201000007270 liver cancer Diseases 0.000 claims description 5
- 208000014018 liver neoplasm Diseases 0.000 claims description 5
- 201000001441 melanoma Diseases 0.000 claims description 5
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 5
- 208000035143 Bacterial infection Diseases 0.000 claims description 4
- 208000015943 Coeliac disease Diseases 0.000 claims description 4
- 206010009900 Colitis ulcerative Diseases 0.000 claims description 4
- 208000001333 Colorectal Neoplasms Diseases 0.000 claims description 4
- 208000011231 Crohn disease Diseases 0.000 claims description 4
- 201000004624 Dermatitis Diseases 0.000 claims description 4
- 206010017533 Fungal infection Diseases 0.000 claims description 4
- 208000007882 Gastritis Diseases 0.000 claims description 4
- 208000031886 HIV Infections Diseases 0.000 claims description 4
- 208000037357 HIV infectious disease Diseases 0.000 claims description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 4
- 206010025323 Lymphomas Diseases 0.000 claims description 4
- 241001465754 Metazoa Species 0.000 claims description 4
- 206010028116 Mucosal inflammation Diseases 0.000 claims description 4
- 201000010927 Mucositis Diseases 0.000 claims description 4
- 208000031888 Mycoses Diseases 0.000 claims description 4
- 201000003793 Myelodysplastic syndrome Diseases 0.000 claims description 4
- 201000004681 Psoriasis Diseases 0.000 claims description 4
- 208000006265 Renal cell carcinoma Diseases 0.000 claims description 4
- 208000000453 Skin Neoplasms Diseases 0.000 claims description 4
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 4
- 201000006704 Ulcerative Colitis Diseases 0.000 claims description 4
- 206010046851 Uveitis Diseases 0.000 claims description 4
- 241000700647 Variola virus Species 0.000 claims description 4
- 208000007502 anemia Diseases 0.000 claims description 4
- 208000022362 bacterial infectious disease Diseases 0.000 claims description 4
- 208000003167 cholangitis Diseases 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 206010017758 gastric cancer Diseases 0.000 claims description 4
- 208000033519 human immunodeficiency virus infectious disease Diseases 0.000 claims description 4
- 208000032839 leukemia Diseases 0.000 claims description 4
- 201000005202 lung cancer Diseases 0.000 claims description 4
- 208000020816 lung neoplasm Diseases 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 201000006417 multiple sclerosis Diseases 0.000 claims description 4
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 4
- 201000000306 sarcoidosis Diseases 0.000 claims description 4
- 201000000849 skin cancer Diseases 0.000 claims description 4
- 201000011549 stomach cancer Diseases 0.000 claims description 4
- 238000003259 recombinant expression Methods 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- 239000001963 growth medium Substances 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims 1
- 239000003937 drug carrier Substances 0.000 claims 1
- 238000003306 harvesting Methods 0.000 claims 1
- 210000004899 c-terminal region Anatomy 0.000 abstract description 6
- 125000000539 amino acid group Chemical group 0.000 abstract description 5
- 238000000746 purification Methods 0.000 abstract description 3
- 125000003275 alpha amino acid group Chemical group 0.000 abstract 1
- 238000002955 isolation Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 26
- 150000001413 amino acids Chemical class 0.000 description 20
- 235000001014 amino acid Nutrition 0.000 description 14
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 12
- 108010057083 glutamyl-aspartyl-leucine Proteins 0.000 description 12
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 12
- 239000007788 liquid Substances 0.000 description 10
- 230000001580 bacterial effect Effects 0.000 description 9
- 241000282414 Homo sapiens Species 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- LBJYAILUMSUTAM-ZLUOBGJFSA-N Ala-Asn-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O LBJYAILUMSUTAM-ZLUOBGJFSA-N 0.000 description 6
- GKAZXNDATBWNBI-DCAQKATOSA-N Ala-Met-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)O)N GKAZXNDATBWNBI-DCAQKATOSA-N 0.000 description 6
- DYXOFPBJBAHWFY-JBDRJPRFSA-N Ala-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C)N DYXOFPBJBAHWFY-JBDRJPRFSA-N 0.000 description 6
- IOFVWPYSRSCWHI-JXUBOQSCSA-N Ala-Thr-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](C)N IOFVWPYSRSCWHI-JXUBOQSCSA-N 0.000 description 6
- KXFCBAHYSLJCCY-ZLUOBGJFSA-N Asn-Asn-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O KXFCBAHYSLJCCY-ZLUOBGJFSA-N 0.000 description 6
- BZWRLDPIWKOVKB-ZPFDUUQYSA-N Asn-Leu-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BZWRLDPIWKOVKB-ZPFDUUQYSA-N 0.000 description 6
- AMGQTNHANMRPOE-LKXGYXEUSA-N Asn-Thr-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O AMGQTNHANMRPOE-LKXGYXEUSA-N 0.000 description 6
- YQPSDMUGFKJZHR-QRTARXTBSA-N Asn-Trp-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CC(=O)N)N YQPSDMUGFKJZHR-QRTARXTBSA-N 0.000 description 6
- JNCRAQVYJZGIOW-QSFUFRPTSA-N Asn-Val-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JNCRAQVYJZGIOW-QSFUFRPTSA-N 0.000 description 6
- KVMPVNGOKHTUHZ-GCJQMDKQSA-N Asp-Ala-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KVMPVNGOKHTUHZ-GCJQMDKQSA-N 0.000 description 6
- ITGFVUYOLWBPQW-KKHAAJSZSA-N Asp-Thr-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O ITGFVUYOLWBPQW-KKHAAJSZSA-N 0.000 description 6
- ZXCAQANTQWBICD-DCAQKATOSA-N Cys-Lys-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CS)N ZXCAQANTQWBICD-DCAQKATOSA-N 0.000 description 6
- RESAHOSBQHMOKH-KKUMJFAQSA-N Cys-Phe-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CS)N RESAHOSBQHMOKH-KKUMJFAQSA-N 0.000 description 6
- DFRYZTUPVZNRLG-KKUMJFAQSA-N Gln-Met-Phe Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CCC(=O)N)N DFRYZTUPVZNRLG-KKUMJFAQSA-N 0.000 description 6
- OKQLXOYFUPVEHI-CIUDSAMLSA-N Gln-Ser-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N OKQLXOYFUPVEHI-CIUDSAMLSA-N 0.000 description 6
- SDSMVVSHLAAOJL-UKJIMTQDSA-N Gln-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCC(=O)N)N SDSMVVSHLAAOJL-UKJIMTQDSA-N 0.000 description 6
- ZOXBSICWUDAOHX-GUBZILKMSA-N Glu-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCC(O)=O ZOXBSICWUDAOHX-GUBZILKMSA-N 0.000 description 6
- JVSBYEDSSRZQGV-GUBZILKMSA-N Glu-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCC(O)=O JVSBYEDSSRZQGV-GUBZILKMSA-N 0.000 description 6
- PKYAVRMYTBBRLS-FXQIFTODSA-N Glu-Cys-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(O)=O PKYAVRMYTBBRLS-FXQIFTODSA-N 0.000 description 6
- MUSGDMDGNGXULI-DCAQKATOSA-N Glu-Glu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CCC(O)=O MUSGDMDGNGXULI-DCAQKATOSA-N 0.000 description 6
- LGYZYFFDELZWRS-DCAQKATOSA-N Glu-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CCC(O)=O LGYZYFFDELZWRS-DCAQKATOSA-N 0.000 description 6
- IRXNJYPKBVERCW-DCAQKATOSA-N Glu-Leu-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IRXNJYPKBVERCW-DCAQKATOSA-N 0.000 description 6
- SWRVAQHFBRZVNX-GUBZILKMSA-N Glu-Lys-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O SWRVAQHFBRZVNX-GUBZILKMSA-N 0.000 description 6
- MRWYPDWDZSLWJM-ACZMJKKPSA-N Glu-Ser-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O MRWYPDWDZSLWJM-ACZMJKKPSA-N 0.000 description 6
- KQDMENMTYNBWMR-WHFBIAKZSA-N Gly-Asp-Ala Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O KQDMENMTYNBWMR-WHFBIAKZSA-N 0.000 description 6
- LGQZOQRDEUIZJY-YUMQZZPRSA-N Gly-Cys-Lys Chemical compound NCCCC[C@H](NC(=O)[C@H](CS)NC(=O)CN)C(O)=O LGQZOQRDEUIZJY-YUMQZZPRSA-N 0.000 description 6
- YOSQCYUFZGPIPC-PBCZWWQYSA-N His-Asp-Thr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YOSQCYUFZGPIPC-PBCZWWQYSA-N 0.000 description 6
- VTZYMXGGXOFBMX-DJFWLOJKSA-N His-Ile-Asp Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O VTZYMXGGXOFBMX-DJFWLOJKSA-N 0.000 description 6
- IWXMHXYOACDSIA-PYJNHQTQSA-N His-Ile-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O IWXMHXYOACDSIA-PYJNHQTQSA-N 0.000 description 6
- YEKYGQZUBCRNGH-DCAQKATOSA-N His-Pro-Ser Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CN=CN2)N)C(=O)N[C@@H](CO)C(=O)O YEKYGQZUBCRNGH-DCAQKATOSA-N 0.000 description 6
- YBJWJQQBWRARLT-KBIXCLLPSA-N Ile-Gln-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O YBJWJQQBWRARLT-KBIXCLLPSA-N 0.000 description 6
- KIMHKBDJQQYLHU-PEFMBERDSA-N Ile-Glu-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N KIMHKBDJQQYLHU-PEFMBERDSA-N 0.000 description 6
- TWPSALMCEHCIOY-YTFOTSKYSA-N Ile-Ile-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(=O)O)N TWPSALMCEHCIOY-YTFOTSKYSA-N 0.000 description 6
- KLBVGHCGHUNHEA-BJDJZHNGSA-N Ile-Leu-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)O)N KLBVGHCGHUNHEA-BJDJZHNGSA-N 0.000 description 6
- ADDYYRVQQZFIMW-MNXVOIDGSA-N Ile-Lys-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ADDYYRVQQZFIMW-MNXVOIDGSA-N 0.000 description 6
- JZNVOBUNTWNZPW-GHCJXIJMSA-N Ile-Ser-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N JZNVOBUNTWNZPW-GHCJXIJMSA-N 0.000 description 6
- AUIYHFRUOOKTGX-UKJIMTQDSA-N Ile-Val-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N AUIYHFRUOOKTGX-UKJIMTQDSA-N 0.000 description 6
- GPICTNQYKHHHTH-GUBZILKMSA-N Leu-Gln-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O GPICTNQYKHHHTH-GUBZILKMSA-N 0.000 description 6
- QVFGXCVIXXBFHO-AVGNSLFASA-N Leu-Glu-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O QVFGXCVIXXBFHO-AVGNSLFASA-N 0.000 description 6
- WQWSMEOYXJTFRU-GUBZILKMSA-N Leu-Glu-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O WQWSMEOYXJTFRU-GUBZILKMSA-N 0.000 description 6
- QNBVTHNJGCOVFA-AVGNSLFASA-N Leu-Leu-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCC(O)=O QNBVTHNJGCOVFA-AVGNSLFASA-N 0.000 description 6
- KPYAOIVPJKPIOU-KKUMJFAQSA-N Leu-Lys-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O KPYAOIVPJKPIOU-KKUMJFAQSA-N 0.000 description 6
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 6
- BGGTYDNTOYRTTR-MEYUZBJRSA-N Leu-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CC(C)C)N)O BGGTYDNTOYRTTR-MEYUZBJRSA-N 0.000 description 6
- AIPHUKOBUXJNKM-KKUMJFAQSA-N Lys-Cys-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O AIPHUKOBUXJNKM-KKUMJFAQSA-N 0.000 description 6
- CRNNMTHBMRFQNG-GUBZILKMSA-N Lys-Glu-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CS)C(=O)O)N CRNNMTHBMRFQNG-GUBZILKMSA-N 0.000 description 6
- DUTMKEAPLLUGNO-JYJNAYRXSA-N Lys-Glu-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O DUTMKEAPLLUGNO-JYJNAYRXSA-N 0.000 description 6
- GAHJXEMYXKLZRQ-AJNGGQMLSA-N Lys-Lys-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GAHJXEMYXKLZRQ-AJNGGQMLSA-N 0.000 description 6
- RIPJMCFGQHGHNP-RHYQMDGZSA-N Lys-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCCCN)N)O RIPJMCFGQHGHNP-RHYQMDGZSA-N 0.000 description 6
- QWTGQXGNNMIUCW-BPUTZDHNSA-N Met-Asn-Trp Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O QWTGQXGNNMIUCW-BPUTZDHNSA-N 0.000 description 6
- NHDMNXBBSGVYGP-PYJNHQTQSA-N Met-His-Ile Chemical compound CSCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(O)=O)CC1=CN=CN1 NHDMNXBBSGVYGP-PYJNHQTQSA-N 0.000 description 6
- RSOMVHWMIAZNLE-HJWJTTGWSA-N Met-Phe-Ile Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O RSOMVHWMIAZNLE-HJWJTTGWSA-N 0.000 description 6
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 6
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 6
- RSPUIENXSJYZQO-JYJNAYRXSA-N Phe-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 RSPUIENXSJYZQO-JYJNAYRXSA-N 0.000 description 6
- FXEKNHAJIMHRFJ-ULQDDVLXSA-N Phe-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N FXEKNHAJIMHRFJ-ULQDDVLXSA-N 0.000 description 6
- CZCCVJUUWBMISW-FXQIFTODSA-N Pro-Ser-Cys Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O CZCCVJUUWBMISW-FXQIFTODSA-N 0.000 description 6
- BGOWRLSWJCVYAQ-CIUDSAMLSA-N Ser-Asp-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O BGOWRLSWJCVYAQ-CIUDSAMLSA-N 0.000 description 6
- SWSRFJZZMNLMLY-ZKWXMUAHSA-N Ser-Asp-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O SWSRFJZZMNLMLY-ZKWXMUAHSA-N 0.000 description 6
- MUARUIBTKQJKFY-WHFBIAKZSA-N Ser-Gly-Asp Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O MUARUIBTKQJKFY-WHFBIAKZSA-N 0.000 description 6
- SVWQEIRZHHNBIO-WHFBIAKZSA-N Ser-Gly-Cys Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CS)C(O)=O SVWQEIRZHHNBIO-WHFBIAKZSA-N 0.000 description 6
- BEAFYHFQTOTVFS-VGDYDELISA-N Ser-Ile-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CO)N BEAFYHFQTOTVFS-VGDYDELISA-N 0.000 description 6
- ZIFYDQAFEMIZII-GUBZILKMSA-N Ser-Leu-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O ZIFYDQAFEMIZII-GUBZILKMSA-N 0.000 description 6
- YUJLIIRMIAGMCQ-CIUDSAMLSA-N Ser-Leu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YUJLIIRMIAGMCQ-CIUDSAMLSA-N 0.000 description 6
- QMCDMHWAKMUGJE-IHRRRGAJSA-N Ser-Phe-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O QMCDMHWAKMUGJE-IHRRRGAJSA-N 0.000 description 6
- GFDUZZACIWNMPE-KZVJFYERSA-N Thr-Ala-Met Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O GFDUZZACIWNMPE-KZVJFYERSA-N 0.000 description 6
- ONNSECRQFSTMCC-XKBZYTNZSA-N Thr-Glu-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O ONNSECRQFSTMCC-XKBZYTNZSA-N 0.000 description 6
- LVFZXRQQQDTBQH-IRIUXVKKSA-N Tyr-Thr-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O LVFZXRQQQDTBQH-IRIUXVKKSA-N 0.000 description 6
- DBOXBUDEAJVKRE-LSJOCFKGSA-N Val-Asn-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N DBOXBUDEAJVKRE-LSJOCFKGSA-N 0.000 description 6
- OVBMCNDKCWAXMZ-NAKRPEOUSA-N Val-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](C(C)C)N OVBMCNDKCWAXMZ-NAKRPEOUSA-N 0.000 description 6
- UVHFONIHVHLDDQ-IFFSRLJSSA-N Val-Thr-Glu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N)O UVHFONIHVHLDDQ-IFFSRLJSSA-N 0.000 description 6
- 108010034529 leucyl-lysine Proteins 0.000 description 6
- 108010009298 lysylglutamic acid Proteins 0.000 description 6
- 108010073101 phenylalanylleucine Proteins 0.000 description 6
- 108010071207 serylmethionine Proteins 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- SWLOHUMCUDRTCL-ZLUOBGJFSA-N Asn-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N SWLOHUMCUDRTCL-ZLUOBGJFSA-N 0.000 description 5
- XVAUJOAYHWWNQF-ZLUOBGJFSA-N Ser-Asn-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O XVAUJOAYHWWNQF-ZLUOBGJFSA-N 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 230000004071 biological effect Effects 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 108010054155 lysyllysine Proteins 0.000 description 4
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 4
- 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 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 102000004556 Interleukin-15 Receptors Human genes 0.000 description 3
- 108010017535 Interleukin-15 Receptors Proteins 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- BCYUHPXBHCUYBA-CUJWVEQBSA-N Thr-Ser-His Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O BCYUHPXBHCUYBA-CUJWVEQBSA-N 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 239000003086 colorant Substances 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 210000003000 inclusion body Anatomy 0.000 description 3
- 229930027917 kanamycin Natural products 0.000 description 3
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 3
- 229960000318 kanamycin Drugs 0.000 description 3
- 229930182823 kanamycin A Natural products 0.000 description 3
- 210000003292 kidney cell Anatomy 0.000 description 3
- 239000002773 nucleotide Substances 0.000 description 3
- 125000003729 nucleotide group Chemical group 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 108010048818 seryl-histidine Proteins 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 241000282693 Cercopithecidae Species 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- JPXNYFOHTHSREU-UWVGGRQHSA-N Gly-Arg-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CN JPXNYFOHTHSREU-UWVGGRQHSA-N 0.000 description 2
- VEPBEGNDJYANCF-QWRGUYRKSA-N Gly-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCCN VEPBEGNDJYANCF-QWRGUYRKSA-N 0.000 description 2
- VCBWXASUBZIFLQ-IHRRRGAJSA-N His-Pro-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O VCBWXASUBZIFLQ-IHRRRGAJSA-N 0.000 description 2
- PLCAEMGSYOYIPP-GUBZILKMSA-N His-Ser-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CN=CN1 PLCAEMGSYOYIPP-GUBZILKMSA-N 0.000 description 2
- SYPULFZAGBBIOM-GVXVVHGQSA-N His-Val-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N SYPULFZAGBBIOM-GVXVVHGQSA-N 0.000 description 2
- ZFNLIDNJUWNIJL-WDCWCFNPSA-N Leu-Glu-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZFNLIDNJUWNIJL-WDCWCFNPSA-N 0.000 description 2
- HRTRLSRYZZKPCO-BJDJZHNGSA-N Leu-Ile-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HRTRLSRYZZKPCO-BJDJZHNGSA-N 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 2
- AHERARIZBPOMNU-KATARQTJSA-N Thr-Ser-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O AHERARIZBPOMNU-KATARQTJSA-N 0.000 description 2
- 238000007792 addition Methods 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 210000003719 b-lymphocyte Anatomy 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 210000003527 eukaryotic cell Anatomy 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 210000005229 liver cell Anatomy 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 210000004962 mammalian cell 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
- 238000000329 molecular dynamics simulation Methods 0.000 description 2
- 210000000822 natural killer cell Anatomy 0.000 description 2
- 239000002687 nonaqueous vehicle Substances 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000004088 simulation Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 239000000375 suspending agent Substances 0.000 description 2
- 108010061238 threonyl-glycine Proteins 0.000 description 2
- 239000011800 void material Substances 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- ZORQXIQZAOLNGE-UHFFFAOYSA-N 1,1-difluorocyclohexane Chemical compound FC1(F)CCCCC1 ZORQXIQZAOLNGE-UHFFFAOYSA-N 0.000 description 1
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-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
- WBSCNDJQPKSPII-UHFFFAOYSA-N 6-amino-2-[[6-amino-2-(2,6-diaminohexanoylamino)hexanoyl]amino]hexanoic acid Chemical compound NCCCCC(N)C(=O)NC(CCCCN)C(=O)NC(CCCCN)C(O)=O WBSCNDJQPKSPII-UHFFFAOYSA-N 0.000 description 1
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 description 1
- MMGCRPZQZWTZTA-IHRRRGAJSA-N Arg-His-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N MMGCRPZQZWTZTA-IHRRRGAJSA-N 0.000 description 1
- DXVMJJNAOVECBA-WHFBIAKZSA-N Asn-Gly-Asn Chemical compound NC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O DXVMJJNAOVECBA-WHFBIAKZSA-N 0.000 description 1
- 210000001266 CD8-positive T-lymphocyte Anatomy 0.000 description 1
- 241000282465 Canis Species 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- GPSHCSTUYOQPAI-JHEQGTHGSA-N Glu-Thr-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O GPSHCSTUYOQPAI-JHEQGTHGSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 description 1
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- AKAPKBNIVNPIPO-KKUMJFAQSA-N His-His-Lys Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@@H](N)CC=1NC=NC=1)C1=CN=CN1 AKAPKBNIVNPIPO-KKUMJFAQSA-N 0.000 description 1
- 239000004354 Hydroxyethyl cellulose Substances 0.000 description 1
- 229920000663 Hydroxyethyl cellulose Polymers 0.000 description 1
- JDAWAWXGAUZPNJ-ZPFDUUQYSA-N Ile-Glu-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N JDAWAWXGAUZPNJ-ZPFDUUQYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 108010002586 Interleukin-7 Proteins 0.000 description 1
- 108010002335 Interleukin-9 Proteins 0.000 description 1
- 241000235058 Komagataella pastoris Species 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- HGFGEMSVBMCFKK-MNXVOIDGSA-N Leu-Ile-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O HGFGEMSVBMCFKK-MNXVOIDGSA-N 0.000 description 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- 240000007472 Leucaena leucocephala Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 241001195348 Nusa Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- VTFXTWDFPTWNJY-RHYQMDGZSA-N Pro-Leu-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VTFXTWDFPTWNJY-RHYQMDGZSA-N 0.000 description 1
- 102000001708 Protein Isoforms Human genes 0.000 description 1
- 108010029485 Protein Isoforms Proteins 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- VAUMZJHYZQXZBQ-WHFBIAKZSA-N Ser-Asn-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O VAUMZJHYZQXZBQ-WHFBIAKZSA-N 0.000 description 1
- OJPHFSOMBZKQKQ-GUBZILKMSA-N Ser-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CO OJPHFSOMBZKQKQ-GUBZILKMSA-N 0.000 description 1
- WSTIOCFMWXNOCX-YUMQZZPRSA-N Ser-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CO)N WSTIOCFMWXNOCX-YUMQZZPRSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- GXDLGHLJTHMDII-WISUUJSJSA-N Thr-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(O)=O GXDLGHLJTHMDII-WISUUJSJSA-N 0.000 description 1
- MYNYCUXMIIWUNW-IEGACIPQSA-N Thr-Trp-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MYNYCUXMIIWUNW-IEGACIPQSA-N 0.000 description 1
- UUIYFDAWNBSWPG-IHPCNDPISA-N Trp-Lys-Lys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)O)N UUIYFDAWNBSWPG-IHPCNDPISA-N 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- OQWNEUXPKHIEJO-NRPADANISA-N Val-Glu-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N OQWNEUXPKHIEJO-NRPADANISA-N 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000010933 acylation Effects 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- CEGOLXSVJUTHNZ-UHFFFAOYSA-K aluminium tristearate Chemical compound [Al+3].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CEGOLXSVJUTHNZ-UHFFFAOYSA-K 0.000 description 1
- 229940063655 aluminum stearate Drugs 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 239000008135 aqueous vehicle Substances 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 238000006664 bond formation reaction Methods 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 238000001311 chemical methods and process Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 229920005565 cyclic polymer Polymers 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 230000017858 demethylation Effects 0.000 description 1
- 238000010520 demethylation reaction Methods 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 229940029030 dendritic cell vaccine Drugs 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000005755 formation reaction Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000022244 formylation Effects 0.000 description 1
- 238000006170 formylation reaction Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 235000019447 hydroxyethyl cellulose Nutrition 0.000 description 1
- 230000033444 hydroxylation Effects 0.000 description 1
- 238000005805 hydroxylation reaction Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000001571 immunoadjuvant effect Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 239000000568 immunological adjuvant Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000007972 injectable composition Substances 0.000 description 1
- 229940102223 injectable solution Drugs 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 201000004514 liver lymphoma Diseases 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 201000005249 lung adenocarcinoma Diseases 0.000 description 1
- 210000005265 lung cell Anatomy 0.000 description 1
- 239000012931 lyophilized formulation Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 210000003071 memory t lymphocyte Anatomy 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 229960002216 methylparaben Drugs 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 230000007498 myristoylation Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 230000006320 pegylation Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000004481 post-translational protein modification Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 230000009465 prokaryotic expression Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 1
- 238000007363 ring formation reaction Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 239000001593 sorbitan monooleate Substances 0.000 description 1
- 235000011069 sorbitan monooleate Nutrition 0.000 description 1
- 229940035049 sorbitan monooleate Drugs 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000019635 sulfation Effects 0.000 description 1
- 238000005670 sulfation reaction Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 238000010798 ubiquitination Methods 0.000 description 1
- 239000012646 vaccine adjuvant Substances 0.000 description 1
- 229940124931 vaccine adjuvant Drugs 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5443—IL-15
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/06—Antipsoriatics
-
- 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
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
-
- 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]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/06—Antianaemics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- General Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Virology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Rheumatology (AREA)
- Hematology (AREA)
- Dermatology (AREA)
- Neurosurgery (AREA)
- Diabetes (AREA)
- Physical Education & Sports Medicine (AREA)
- Orthopedic Medicine & Surgery (AREA)
- AIDS & HIV (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Pain & Pain Management (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
Abstract
本公开涉及一种重组白介素‑15变体,其在野生型白介素‑15氨基酸序列的末端添加、删除、取代一个或多个氨基酸残基,从而使得白介素‑15变体的羧基端较野生型白介素‑15有更加稳定的结构,使白介素‑15变体可以在大肠杆菌中高表达,更容易分离和纯化。本公开的白介素‑15变体的表达量为野生型白介素‑15的10至20倍。
Description
技术领域
本公开属于分子生物学领域,具体涉及一种重组白介素-15变体。
背景技术
白细胞介素-15(IL-15)是一种约为12-14kD的细胞因子。与IL-2、IL-4、IL-7、IL-9、粒细胞集落刺激因子、粒细胞-巨噬细胞集落刺激因子同属于4a螺旋细胞因子家族。
天然成熟人IL-15成熟肽含有114个氨基酸,包括4个半胱氨酸残基(Cys35与Cys85、Cys42与Cys88),半胱氨酸残基形成的两对分子内二硫键对于保持IL-15的空间构象和生物学活性起重要作用。
IL-15受体由3个亚基组成(α、β和γ亚基)。IL-15受体的α亚基与IL-15有很高的亲和力,生理状态下IL-15与α亚基形成复合物(IL-15-Rα),并增强IL-15对受体的β链和γ链亚基的亲和力,激活T细胞和NK细胞。因此,一些制药公司将IL-15与IL-15受体α亚基(或其部分)形成复合物,并在动物试验中显示了良好的生物效价及稳定性。
IL-15可在机体正常的免疫应答中发挥作用,如促进T细胞、B细胞、自然杀伤(NK)细胞的发育等。炎症与肿瘤的发生密切相连,由于IL-15可促进NK细胞、B淋巴细胞及T淋巴细胞的增殖并可维持这些免疫细胞功能,因此IL-15对于治疗一些恶性肿瘤存在一定疗效。在多种实验动物肿瘤模型(肺腺癌、黑素瘤、结肠癌、肝癌、淋巴瘤)中,利用IL-15治疗均可以促进肿瘤的消退,减少肿瘤的转移,提高存活率。
在感染性疾病模型的临床研究中发现IL-15是一种有效的疫苗佐剂。Steel等人开发了一种靶向性治疗neu+乳腺癌的DC疫苗(可组成型表达IL-15)。研究人员将IL-15作为一种刺激体内免疫细胞活性的免疫佐剂进行评估,现阶段已有的临床结果表明培养基中添加IL-15培养出的NK、CD8+T细胞、CD8+记忆T细胞、树突状细胞转移至动物体内后,生物活性显著提高。
随着对IL-15作用机制及临床应用研究的不断深入,IL-15作为药物及免疫佐剂其需求量也大大增加。然而,天然野生型的IL-15存在明显的药物开发瓶颈,包括原核生物和真核生物表达的表达量低、纯化困难、半衰期短等。市场上主要利用原核表达***对其进行表达,大肠杆菌的表达产物通常以包涵体形式存在,需通过变性-复性的复杂操作才能得到活性形式的IL-15,此方法极大的限制了活性蛋白的产率。要得到高纯度的蛋白往往需要经过多步纯化操作,导致蛋白的产率下降,生物活性也会随之降低。所以,本领域有必要提供一种更加稳定的IL-15。
发明内容
IL-15变体
本文以下所用的氨基酸单字母代码时本领域公知的,如JBC,243,p3558,1968中所述。
本公开的一方面提供了一种IL-15变体,和野生型人IL-15相比,在野生型人IL-15的羧基端添加一段多肽,所述多肽由1至30个氨基酸残基组成。
在一些实施方案中,不限于具体的理论,在野生型人IL-15的羧基端添加不超过30个氨基酸长度的多肽时,有助于形成alpha螺旋;通过提高IL-15羧基端结构的稳定性来增加其在宿主(尤其是原核宿主)中的表达量。在一个示例中,表达量约为野生型IL-15的10至20倍,甚至更高。
在一些实施方案中,提供了一种IL-15变体,和野生型人IL-15相比,在野生型人IL-15的羧基端添加至少一个碱性亲水性氨基酸残基。
在一些实施方案中,所述碱性亲水性氨基酸选自以下任一个:H、R和K。
根据一些实施方案,提供了一种IL-15变体,其包含式I或式II所示的肽结构,从左至右按氨基端至羧基端方向:
X1-J-X2-X3(式I)
X1-X2-J-X3(式II)
其中:
X1表示野生型人IL-15;
-表示共价键,优选酰胺键;
X2表示随机序列或空缺,所述随机序列的长度是4至8个氨基酸长;所述随机序列不含亲水性肽残基;
X3表示碱性亲水性肽残基,其选自以下任一个或其组合:KK、HH、RH、KKK、WKK、HHK;
J表示接头或空缺。
接头和随机序列的顺序可互换,而不影响IL-15变体的改进的性能。
术语“接头(Linker)”在本公开中用于连接野生型IL-15和添加肽,以保证蛋白的正确折叠和稳定性。如果接头的序列太短,可能影响肽链的折叠,从而相互干扰;如果接头的序列太长,又涉及免疫原性的问题等。
在一些实施方案中,所述接头包含选自G和S的至少一个氨基酸残基。
在一些实施方案中,所述接头选自以下任一个或其组合:(G)n、(GGGGS)n、(GS)n、(GGS)n,其中n为1至10的整数(1、2、3、4、5、6、7、8、9、10)。
在一些具体的实施方案中,提供了一种IL-15变体,其包含选自以下的任一个结构(或如以下结构所示):
X1-HPLT(SEQ ID NO:13)-WKK、
X1-LRLISG(SEQ ID NO:14)-RH、
X1-LIER(SEQ ID NO:15)-HHK、
X1-HVESG(SEQ ID NO:16)-KKK、
X1-HSQLETG(SEQ ID NO:17)-KK;
其中,X1表示野生型人IL-15。
在本申请中,变体是指在母体多肽的基础上引入氨基酸残基的取代、添加、缺失或修饰后所产生的多肽。相较于母体多肽的物理、化学或生物活性,变体具有改进的活性。
在一些实施方案中,野生型人IL-15选自以下的任一项:天然存在的人IL-15、天然存在的人IL-15剪接变体。
在一些具体的实施方案中,所述野生型人IL-15是野生型人IL-15的成熟形式。在一些具体的实施方案中,所述野生型人IL-15是SEQ ID NO:1所示(首位甲硫氨酸是任选的,SEQ ID NO:2)。
在本申请中,术语“白介素-15”、“白细胞介素-15”、“IL-15”指白介素-15蛋白或多肽,尤其是人白介素-15蛋白或多肽。未成熟形式的IL-15包含162个氨基酸,其中第1-29位氨基酸构成信号肽,第30-48位氨基酸构成前肽;IL-15的成熟形式对应于氨基酸49至162。未成熟形式的IL-15可在UniProtKB登录号P40933获得。人成熟IL-15的氨基酸序列对应于SEQ ID NO:1或去除首位M的SEQ ID NO:2。术语“IL-15”还包括由细胞天然表达的IL-15的任何变体或同种型。值得注意的是,已经报道了IL-15的若干种剪接的转录变体。
在本申请中,“多肽”理解为由至少两个氨基酸通过肽键连接而形成的线性或环状聚合物。在一些情况下,多肽包含天然的20种氨基酸;在另一些情况下,多肽可以包含除遗传密码定义的20种氨基酸以外的氨基酸;在又一些情况下,多肽可包含修饰的氨基酸(如翻译后修饰、或通过化学过程)。这些修饰可以出现在多肽的任何位置:在肽骨架中、在侧链中或在羧基或氨基末端;例如但不限于酰化、核糖基化、二硫键形成、脱甲基化、甲酰化、甲基化、肉豆蔻酰化、氧化、羟基化、磷酸化、糖基化、聚乙二醇化、GPI锚形成、硫酸化、泛素化、共价或非共价交联、环化、固定至脂质或脂质衍生物。
在一些实施方案中,本公开的IL-15变体包含选自以下任一项所示的多肽:SEQ IDNO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12。
在一些实施方案中,本公开的IL-15变体与SEQ ID NO:3至12中任一个具有至少85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%的同一性。
多核苷酸
根据在一些实施方案中,本公开提供了一种多核苷酸,其编码本公开的IL-15变体。
术语“多核苷酸”或“核酸分子”是指包含核苷酸的聚合物。多核苷酸的示例是,例如但不限于DNA、RNA、LNA、cDNA。
技术人员理解,编码相同氨基酸序列的核苷酸序列可以不同,由于密码子的兼并性和不同宿主的密码子偏好性,不同的核苷酸序列能够编码相同的氨基酸序列,这些序列均属于本公开的范畴内。
表达载体
根据在一些实施方案中,本公开提供了一种重组表达载体,其包含本公开的多核苷酸。
表达载体可以是原核或真核表达载体,也可以是穿梭载体。
表达载体是本领域熟知的,通常可以市售获得。在用于表达本申请的活性分子时,技术人员将编码序列***表达载体的适当位点。
表达载体的示例,例如但不限于:PET表达载体,例如pET Dsb 39b、pETExpression System 33b、pET GST Fusion Systems 41、pET NusA Fusion Systems 43.1;PGEX表达载体,例如pGEX-2T、pGEX-2TK、pGEX-3X、pGEX-4T-1、pGEX-4T-2、pGEX-4T-3、pGEX-5X-1、pGEX-5X-2、pGEX-5X-3、pGEX-6P-1、pGEX-6P-2、pGEX-6P-3;PTYB表达载体,例如PTYB1、PTYB2、PTYB11、PTYB12;真核表达载体,例如pCDNA3.1(-)、pCDNA3.1(+)、pPICZalpha A、pGAPZαA、PYES2.0、pBI121、pEGFP-N1、pEGFP-C1、pPIC9K、pPIC3.5K。
本申请对表达载体没有特别的限制,技术人员根据公知常识有能力根据后续应用、宿主类型、生产规模等因素,而选择适当的表达载体。
宿主细胞
本公开提供一种宿主细胞,其包含根据本公开的表达载体、或表达根据本公开的IL-15变体。
一些实施方案中,宿主细胞包含(如已被转化或转染有)表达载体,所述表达载体包含根据本公开的多核苷酸。
一些实施方案中,所述宿主细胞是原核细胞。
一些实施方案中,所述宿主细胞是真核细胞。
一些具体实施方案中,宿主细胞为细菌、酵母菌或哺乳动物细胞,具体可以为巴斯德毕赤酵母或酿酒酵母。
一些具体实施方案中,宿主细胞是原核微生物(如大肠杆菌)。
一些具体实施方案中,宿主细胞真核生物细胞。
一些实施方案中,使用表达糖基化多肽的宿主细胞,例如植物和昆虫细胞。脊椎动物细胞也可以用作宿主细胞,例如,悬浮生长的哺乳动物细胞系、猴肾CV1系(COS-7)、人胚胎肾系(293或293T细胞)、幼仑鼠肾细胞(BHK)、小鼠塞托利(sertoli)细胞(TM4细胞)、猴肾细胞(CV1)、VERO-76、人***细胞(HELA)、犬肾细胞(MDCK)、buffalo大鼠肝细胞(BRL3A)、人肺细胞(W138)、人肝细胞(Hep G2)、小鼠***肿瘤细胞(MMT060562)、MRC5细胞、FS4细胞、CHO细胞、骨髓瘤细胞系(如YO、NS0、P3X63和Sp2/0)。
本申请中,宿主细胞不能发育成动物个体或植物个体。
生产方法
本公开提供一种生产IL-15变体的方法。
一些实施方案中,将编码IL-15变体的多核苷酸与表达载体相连,转入表达宿主中,通过诱导而表达IL-15变体。
一些实施方案中,IL-15变体以包涵体的方式表达,对包涵体进行复性。
一些实施方案中,包涵体溶解于尿素溶液(例如,8M)中,通过离子交换和/或反相色谱法纯化IL-15变体。
一些具体的实施方案中,表达载体为原核pET41a,表达宿主为大肠杆菌BL21(DE3)或C41(DE3)。
药物组合物
本公开提供一种药物组合物,其含有根据本公开的IL-15变体;任选地还包含药学上可接受的稀释剂、载体或赋形剂。
术语“药学上可接受的”是指不具有不期望的体内或体外作用。
术语“药学上可接受的稀释剂、载体或赋形剂”是指存在于药物制剂(或药物组合物)中而不是活性成分的任何组分。因此,可以提及稀释剂、粘合剂、润滑剂、崩解剂、填充剂、着色剂、润湿剂、乳化剂、pH缓冲剂、防腐剂等。药学上可接受的稀释剂、载体或赋形剂在教科书Remington's The Science and Practice of Pharmacy中有详细描述。
在示例性组合物中,可包含一种或多种药学上可接受的成分,例如稳定剂、抗微生物剂、缓冲剂、着色剂、调味剂、佐剂等。
本公开的活性分子(IL-15变体)与常规使用的载体(或稀释剂、或赋形剂)一起制成药物组合物及其单位剂量的形式。
组合物可以是固体形式,例如片剂或填充胶囊、冷冻干燥形式。
本公开的组合物是固体制剂时,在使用前用合适的媒介物重构。
组合物可以是液体形式(例如注射剂、溶液、悬浮液、乳液)。示例性组合物用于口服、或胃肠外(包括皮下)施用的无菌注射溶液。本公开的组合物是液体制剂时,包括但不限于水性或油性悬浮液、溶液、乳液、糖浆。适于口服给药的液体形式可包括含有缓冲剂、悬浮剂和分散剂、着色剂、调味剂等水性或非水性媒介物。悬浮剂包括但不限于山梨糖醇浆、甲基纤维素、葡萄糖、明胶、羟乙基纤维素、羧甲基纤维素、硬脂酸铝凝胶。乳化剂包括但不限于卵磷脂、脱水山梨糖醇单油酸酯、***胶。非水性媒介物包括但不限于食用油、丙二醇、乙醇。防腐剂包括但不限于对羟基苯甲酸甲酯、对羟基苯甲酸丙酯、山梨酸。
在一些具体实施方案中,所述药物组合物可以为冻干制剂或可注射溶液。
在一些具体实施方案中,可注射的组合物通常基于可注射的无菌盐水或磷酸盐缓冲盐水。
在一些具体实施方案中,药物组合物单位计量中可含有按质量计1%至10%的IL-15变体;具体而言1%±10%、2%±10%、3%±10%、4%±10%、5%±10%、6%±10%、7%±10%、8%±10%、9%±10%、10%±10%。
在一些具体实施方案中,药物组合物单位剂量中含有的IL-15变体的量为0.25μg/kg至100μg/kg;例如但不限于1μg/kg、5μg/kg、10μg/kg、20μg/kg、25μg/kg、30μg/kg、35μg/kg、40μg/kg、45μg/kg、50μg/kg、55μg/kg、60μg/kg、65μg/kg、70μg/kg、75μg/kg、80μg/kg、85μg/kg、90μg/kg、95μg/kg、100μg/kg±10%。
在一些实施方案中,本公开的IL-15变体或药物组合物分多次施用。在一些实施方案中,第一周期(28天/周期)的剂量为0.25μg/kg,然后连续周期的剂量水平分别为0.5μg/kg、1μg/kg、2μg/kg、4μg/kg和8μg/kg。
在一些实施方案中,第一周期的剂量为1μg/kg,第二周期中的剂量为2μg/kg,第三周期中的剂量为4μg/kg,第四周期中的剂量为8μg/kg。例如,受试者可以持续两周中每周三次接受相同的剂量,然后在每个治疗周期之间,间隔两周作为休息。在另一些实施方案中,可以皮下s.c.施用本公开的IL-15变体或药物组合物。
用途、治疗方法
本公开提供一种IL-15变体,其用于预防或治疗感染性疾病。在一些实施方案中,所述感染性疾病选自:天花病毒感染、HIV感染、细菌感染、真菌感染、HBV感染。
本公开提供一种IL-15变体,其用于预防或治疗癌症。在一些实施方案中,所述癌症选自:黑色素瘤、结直肠癌、皮肤癌、淋巴瘤、肾细胞癌、肝癌、肺癌、胃癌、乳腺癌。
本公开提供一种IL-15变体,其用于预防或治疗血液病。在一些实施方案中,所述血液病选自:贫血、白血病、骨髓增生异常综合征。
本公开提供一种IL-15变体,其用于预防或治疗炎性疾病。在一些实施方案中,所述炎性疾病选自:自身免疫性疾病、乳糜泻、结节病、溃疡性结肠炎、克罗恩病、胆管炎、葡萄膜炎、皮炎;在一些实施方案中,所述自身免疫性疾病选自:多发性硬化症、银屑病、风湿性关节炎、胃炎、黏膜炎。
本公开提供前述IL-15变体在制备药物中的用途,所述药物用于预防或治疗选自以下的疾病:感染性疾病、癌症、血液病、炎性疾病。在一些实施方案中,所述感染性疾病选自:天花病毒感染、HIV感染、细菌感染、真菌感染、HBV感染。在一些实施方案中,所述癌症选自:黑色素瘤、结直肠癌、皮肤癌、淋巴瘤、肾细胞癌、肝癌、肺癌、胃癌、乳腺癌。在一些实施方案中,所述血液病选自:贫血、白血病、骨髓增生异常综合征。在一些实施方案中,所述炎性疾病选自:自身免疫性疾病、乳糜泻、结节病、溃疡性结肠炎、克罗恩病、胆管炎、葡萄膜炎、皮炎;在一些实施方案中,所述自身免疫性疾病选自:多发性硬化症、银屑病、风湿性关节炎、胃炎、黏膜炎。
本公开提供一种预防或治疗以下疾病的方法:感染性疾病、癌症、血液病、炎性疾病。所述方法包括步骤:向受试者施用预防或治疗有效量的本公开的IL-15变体。在一些实施方案中,所述感染性疾病选自:天花病毒感染、HIV感染、细菌感染、真菌感染、HBV感染。在一些实施方案中,所述癌症选自:黑色素瘤、结直肠癌、皮肤癌、淋巴瘤、肾细胞癌、肝癌、肺癌、胃癌、乳腺癌。在一些实施方案中,所述血液病选自:贫血、白血病、骨髓增生异常综合征。在一些实施方案中,所述炎性疾病选自:自身免疫性疾病、乳糜泻、结节病、溃疡性结肠炎、克罗恩病、胆管炎、葡萄膜炎、皮炎;在一些实施方案中,所述自身免疫性疾病选自:多发性硬化症、银屑病、风湿性关节炎、胃炎、黏膜炎。
一些实施方案中,需要治疗的受试者(如患者或个体)通常为哺乳动物,例如人。
一些实施方案中,每日至少2次、每日至少1次、每48小时至少1次、每72小时至少一次、每周至少一次、每2周至少一次、每个月至少一次、每2个月至少一次、或每3个月至少一次向受试者施用本公开的IL-15变体、药物组合物。
一些实施方案中,通过任何途径施用本公开的IL-15变体、药物组合物。例如,通过肠胃外注射(例如,皮下、静脉注射)施用。
附图说明
图1:野生型IL-15在不同大肠杆菌单克隆中诱导表达的SDS-PAGE电泳图。
图2A至图2E:本公开的IL-15变体在大肠杆菌单克隆中表达的SDS-PAGE电泳图。图2A至图2E分别表示变体1至变体5。
图3:HPLC定量结果。
图4:本公开的IL-15变体的立体结构图。在水溶液的极性环境中,IL-15变体能稳定保持链内氢键,降低水分子造成的解螺旋效应。
具体实施方式
以下将结合实施例对本公开作进一步地说明,应理解这些实施例仅作为例证的目的,不用于限制本公开的保护范围。
下列实施例中未注明具体条件的实验方法,通常按照常规条件,例如Sambrook等分子克隆:实验室手册,Cold Spring Harbor Laboratory Press,1989中所述的条件,或按照制造厂商所建议的条件。所采用的试剂,若无特殊说明,均为市售或公开渠道可以获得的试剂。
实施例
实施例1.野生型IL-15及IL-15变体在大肠杆菌中的表达
1.表达载体的构建
野生型IL-15及IL-15变体的核苷酸序列由天津卡梅德生物技术有限公司合成。通过在反向引物中引入需要添加的序列来修饰野生型IL-15的羧基端。当技术人员知道所需添加的氨基酸序列时,可以根据公知的引物设计原则,使用任何可用的引物设计软件来设计并合成引物,这属于本领域技术人员能力范围内。
野生型IL-15、构建的IL-15变体的氨基酸序列如表1所示。
表1.野生型IL-15、IL-15变体的氨基酸序列
2.按常规方法进行PCR扩增。
3.对表达载体进行酶切:反应体系为30μL,和限制性酶混匀后37℃过夜。
4.回收PCR产物和酶切载体。
5.载体与片段的摩尔比为2:1,37℃反应30min,立即置于冰上冷却,转化入DH5α中。
6.在1.5mL的管中加入0.5mL含Amp抗性的LB培养基,向其中接种单菌落,共5管;在37℃摇床中进行振荡培养。培养3h以后,取1μL做模板,进行PCR。PCR后,用琼脂糖凝胶电泳进行检测,选择阳性克隆进行测序。
7.阳性菌液保存及质粒的提取:将测序正确的阳性克隆接种于5mL Amp抗性的LB液体培养基中,接种量为10μL。取500μL的菌液加入1.5mL的管中,加入500μL 40%甘油放入-80℃保存。离心菌液,收集菌体,用于质粒提取。
8.野生型IL-15和IL-15变体的表达
a)取10μl质粒加入100μl BL21(DE3)感受态细胞中,立即混匀,冰上放置30min;
b)42℃热激90s,迅速冰浴5min;涂布LB(含50μg/ml卡那霉素)平板,37℃培养14h后挑取单克隆进行诱导培养。
9.小规模表达
a)从LB平板中挑取单克隆,加入2ml LB液体培养基(含50μg/ml卡那霉素)活化;
b)将过夜培养的菌液按1:50比例转接到20ml含Amp的LB培养基中,37℃,220rpm,培养至至菌体OD600为0.6-0.8;转接到5ml LB液体培养基(含50μg/ml卡那霉素)培养至菌体OD600为0.6-0.8;加入浓度为1mM的IPTG进行37℃诱导表达4h。
10.SDS-PAGE鉴定表达量
a)测定培养液的OD600,取10OD菌液,10000rpm,2min离心,去掉上清;
b)用1mL裂解液(10mM Tris-HCl,pH 8.0)重悬菌体,置于冰上进行超声裂解细胞,超声条件:130W、4min、on 3s、off 3s;
c)超声结束后,取80μl样品加入20μl 5×上样缓冲液,95℃加热5min,每个样品取12.5μl(0.1OD)进行SDS-PAGE电泳。
IL-15野生型的表达如图1所示:无论是在裂解液、上清或沉淀中,都检测不到野生型IL-15的表达。图1的1、2、3、4代表四种不同的表达载体。相比于野生型白介素-15,IL-15的变体1至变体17的表达水平在大肠杆菌大中有统计学上显著的改善(p<0.05),约提高10至20倍(图2A至图2E所示)。
11.HPLC鉴定表达量
a)表达后收取10OD细胞,用1ml 10mM Tris-HCl,pH8.0的缓冲液重悬;
b)超声破碎;
c)破碎后的样品,经12000rpm离心10min,弃上清;
d)沉淀用1ml新鲜配制的8M尿素/10mM Tris-HCl,pH8.0,10mM DTT室温振荡溶解约1h;
e)用0.2μm过滤器过滤,上样进行高效液相色谱分析。
利用C4分析柱,流动相A为含0.1%TFA的去离子水,流动相B为含0.1%TFA的乙腈;用15分钟从20%B到60%B的梯度来进行分析。HPLC定量结果如图3所示,IL-15变体1至5的表达量要高于野生型IL-15的表达量20倍左右。
实施例2.IL-15变体在水溶液中的分子动力学
对于IL-15变体1至变体5,利用SIWSS-model进行同源建模。使用同源建模得到的结构作为初始结构,使用GROMACS软件进行200ns的分子动力学模拟。立场选用OPLS-AA,水分子模型选用TIP4P模型。
经过长达200ns的长程动力学模拟,得到最终的IL-15变体的平均结构。使用pymol软件分析蛋白质结构,可以看到在蛋白质羧基端添加的HHH特征序列在长时间水溶液环境下的动力学模拟中保持了alpha螺旋的结构。这说明本公开中设计的添加肽在水溶液的极性环境中能稳定保持链内氢键结构,使得水分子造成的解螺旋效应降低(图4所示变体4,其他变体结果一致)。
序列表
<110> 西安龙腾景云生物科技有限公司
<120> 一种重组白介素-15变体
<130> 310206CG
<160> 17
<170> SIPOSequenceListing 1.0
<210> 1
<211> 115
<212> PRT
<213> Homo sapiens
<400> 1
Met Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
1 5 10 15
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
20 25 30
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
35 40 45
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
50 55 60
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
65 70 75 80
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
85 90 95
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
100 105 110
Asn Thr Ser
115
<210> 2
<211> 114
<212> PRT
<213> Homo sapiens
<400> 2
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 3
<211> 122
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> VARIANT
<222>
<223> IL-15变体
<400> 3
Met Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
1 5 10 15
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
20 25 30
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
35 40 45
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
50 55 60
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn
65 70 75 80
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
85 90 95
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
100 105 110
Asn Thr Ser His Pro Leu Thr Trp Lys Lys
115 120
<210> 4
<211> 121
<212> PRT
<213> 人工序列(artificial sequence)
<400> 4
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser His Pro Leu Thr Trp Lys Lys
115 120
<210> 5
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> VARIANT
<222>
<223> IL-15变体
<400> 5
Met Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
1 5 10 15
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
20 25 30
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
35 40 45
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
50 55 60
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn
65 70 75 80
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
85 90 95
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
100 105 110
Asn Thr Ser Leu Arg Leu Ile Ser Gly Arg His
115 120
<210> 6
<211> 122
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> VARIANT
<222>
<223> IL-15变体
<400> 6
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser Leu Arg Leu Ile Ser Gly Arg His
115 120
<210> 7
<211> 124
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> VARIANT
<222>
<223> IL-15变体
<400> 7
Met Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
1 5 10 15
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
20 25 30
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
35 40 45
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
50 55 60
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn
65 70 75 80
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
85 90 95
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
100 105 110
Asn Thr Ser His Ser Gln Leu Glu Thr Gly Lys Lys
115 120
<210> 8
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> VARIANT
<222>
<223> IL-15变体
<400> 8
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser His Ser Gln Leu Glu Thr Gly Lys Lys
115 120
<210> 9
<211> 122
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> VARIANT
<222>
<223> IL-15变体
<400> 9
Met Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
1 5 10 15
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
20 25 30
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
35 40 45
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
50 55 60
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn
65 70 75 80
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
85 90 95
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
100 105 110
Asn Thr Ser Leu Ile Glu Arg His His Lys
115 120
<210> 10
<211> 121
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> VARIANT
<222>
<223> IL-15变体
<400> 10
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser Leu Ile Glu Arg His His Lys
115 120
<210> 11
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> VARIANT
<222>
<223> IL-15变体
<400> 11
Met Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
1 5 10 15
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
20 25 30
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
35 40 45
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
50 55 60
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn
65 70 75 80
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
85 90 95
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
100 105 110
Asn Thr Ser His Val Glu Ser Gly Lys Lys Lys
115 120
<210> 12
<211> 122
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> VARIANT
<222>
<223> IL-15变体
<400> 12
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Ala Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser His Val Glu Ser Gly Lys Lys Lys
115 120
<210> 13
<211> 4
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> UNSURE
<222>
<223> 随机序列
<400> 13
His Pro Leu Thr
1
<210> 14
<211> 6
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> UNSURE
<222>
<223> 随机序列
<400> 14
Leu Arg Leu Ile Ser Gly
1 5
<210> 15
<211> 4
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> UNSURE
<222>
<223> 随机序列
<400> 15
Leu Ile Glu Arg
1
<210> 16
<211> 5
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> UNSURE
<222>
<223> 随机序列
<400> 16
His Val Glu Ser Gly
1 5
<210> 17
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<220>
<221> UNSURE
<222>
<223> 随机序列
<400> 17
His Ser Gln Leu Glu Thr Gly
1 5
Claims (9)
1. 一种白介素-15变体,其是选自以下的任一个结构所示,从左至右按氨基端至羧基端方向:
X1-HPLT-WKK、
X1-LIER-HHK、
X1-HVESG-KKK、
X1-LRLISG-RH、
X1-HSQLETG-KK;
X1表示野生型人白介素-15;
所述野生型人白介素-15是SEQ ID NO:1或2所示。
2.根据权利要求1所述的白介素-15变体,其是选自以下任一项所示的多肽:SEQ IDNO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12。
3.一种多核苷酸,其编码权利要求1至2中任一项所述的白介素-15变体。
4.一种重组表达载体,其包含权利要求3所述的多核苷酸。
5.一种宿主细胞,其:
- 包含权利要求4所述的重组表达载体;或
- 表达权利要求1至2中任一项所述的白介素-15变体;
所述宿主细胞不能发育成动物个体或植物个体。
6.一种药物组合物,其包含:
权利要求1至2中任一项所述的白介素-15变体;
任选地,可药用载体或赋形剂或稀释剂。
7.一种蛋白复合物,其包含权利要求1至2中任一项所述的白介素-15变体。
8.权利要求1至2中任一项所述的白介素-15变体在制备药物中的用途;所述药物用于预防或治疗选自以下的疾病:感染性疾病、癌症、血液病、炎性疾病;
所述感染性疾病选自:天花病毒感染、HIV感染、细菌感染、真菌感染、HBV感染;
所述癌症选自:黑色素瘤、结直肠癌、皮肤癌、淋巴瘤、肾细胞癌、肝癌、肺癌、胃癌、乳腺癌;
所述血液病选自:贫血、白血病、骨髓增生异常综合征;
所述炎性疾病选自:自身免疫性疾病、乳糜泻、结节病、溃疡性结肠炎、克罗恩病、胆管炎、葡萄膜炎、皮炎;所述自身免疫性疾病选自:多发性硬化症、银屑病、风湿性关节炎、胃炎、黏膜炎。
9.一种提高白介素-15在宿主细胞中表达量的方法,包括步骤:
- 在培养介质中,培养权利要求5所述的宿主细胞;
- 收获权利要求1至2中任一项所述的白介素-15变体;
所述宿主细胞是大肠杆菌。
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111246297.6A CN113956346B (zh) | 2021-10-26 | 2021-10-26 | 一种重组白介素-15变体 |
PCT/CN2022/111669 WO2023071390A1 (zh) | 2021-10-26 | 2022-08-11 | 一种重组白介素-15变体 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111246297.6A CN113956346B (zh) | 2021-10-26 | 2021-10-26 | 一种重组白介素-15变体 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113956346A CN113956346A (zh) | 2022-01-21 |
CN113956346B true CN113956346B (zh) | 2024-03-01 |
Family
ID=79466919
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202111246297.6A Active CN113956346B (zh) | 2021-10-26 | 2021-10-26 | 一种重组白介素-15变体 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN113956346B (zh) |
WO (1) | WO2023071390A1 (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113956346B (zh) * | 2021-10-26 | 2024-03-01 | 西安龙腾景云生物科技有限公司 | 一种重组白介素-15变体 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107636015A (zh) * | 2015-03-06 | 2018-01-26 | 德国癌症研究中心 | 包含结合蛋白和对IL‑15Rα具有降低亲和力的白介素‑15多肽的融合蛋白及其治疗用途 |
CN112513070A (zh) * | 2018-02-28 | 2021-03-16 | 辉瑞公司 | Il-15变体及其用途 |
CN112552391A (zh) * | 2019-09-25 | 2021-03-26 | 北京志道生物科技有限公司 | 一种重组白介素-15类似物 |
CN113185600A (zh) * | 2021-05-28 | 2021-07-30 | 苏州复融生物技术有限公司 | 一种新型白介素15突变体多肽的开发及其应用 |
CN114573707A (zh) * | 2020-11-28 | 2022-06-03 | 北京志道生物科技有限公司 | 一种用于抗肿瘤的新型il-15融合蛋白 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2006017853A2 (en) * | 2004-08-11 | 2006-02-16 | Beth Israel Deaconess Medical Center, Inc. | Mutant interleukin-15-containing compositions and suppression of an immune response |
CN104212808B (zh) * | 2014-09-05 | 2017-03-08 | 中国科学院广州生物医药与健康研究院 | 重组人白介素15长肽段及其生产方法 |
US11230699B2 (en) * | 2020-01-28 | 2022-01-25 | Immunitybio, Inc. | Chimeric antigen receptor-modified NK-92 cells targeting EGFR super-family receptors |
CN113403339B (zh) * | 2021-06-16 | 2022-04-01 | 武汉大学 | 一种c端带有表位标签m的表达载体及其构建方法与应用 |
CN113956346B (zh) * | 2021-10-26 | 2024-03-01 | 西安龙腾景云生物科技有限公司 | 一种重组白介素-15变体 |
-
2021
- 2021-10-26 CN CN202111246297.6A patent/CN113956346B/zh active Active
-
2022
- 2022-08-11 WO PCT/CN2022/111669 patent/WO2023071390A1/zh unknown
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107636015A (zh) * | 2015-03-06 | 2018-01-26 | 德国癌症研究中心 | 包含结合蛋白和对IL‑15Rα具有降低亲和力的白介素‑15多肽的融合蛋白及其治疗用途 |
CN112513070A (zh) * | 2018-02-28 | 2021-03-16 | 辉瑞公司 | Il-15变体及其用途 |
CN112552391A (zh) * | 2019-09-25 | 2021-03-26 | 北京志道生物科技有限公司 | 一种重组白介素-15类似物 |
WO2021057826A1 (zh) * | 2019-09-25 | 2021-04-01 | 北京志道生物科技有限公司 | 一种重组白介素-15类似物 |
CN114573707A (zh) * | 2020-11-28 | 2022-06-03 | 北京志道生物科技有限公司 | 一种用于抗肿瘤的新型il-15融合蛋白 |
CN113185600A (zh) * | 2021-05-28 | 2021-07-30 | 苏州复融生物技术有限公司 | 一种新型白介素15突变体多肽的开发及其应用 |
Also Published As
Publication number | Publication date |
---|---|
CN113956346A (zh) | 2022-01-21 |
WO2023071390A1 (zh) | 2023-05-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2004101619A1 (ja) | 機能的糖ペプチドの合理的設計および合成 | |
JPH01502397A (ja) | M―csfの生産方法 | |
JPS60115528A (ja) | ヒトインタ―ロイキン―2蛋白質を含有する抗腫瘍用または免疫機能低下疾患治療用組成物 | |
EP4015526A1 (en) | Recombinant interleukin-15 analog | |
CN113956346B (zh) | 一种重组白介素-15变体 | |
CN107056929A (zh) | 经过修饰的猪促生长素多肽和其用途 | |
CN107674121A (zh) | 经过修饰的牛促生长素多肽和其用途 | |
Gomes et al. | Expression of recombinant human mutant granulocyte colony stimulating factor (Nartograstim) in Escherichia coli | |
JPH04164098A (ja) | 化学修飾顆粒球コロニー刺激因子誘導体 | |
CN114369153A (zh) | 一种白介素-2突变体 | |
JPH0463595A (ja) | ヒトインターロイキン3誘導体 | |
CN105884876B (zh) | 一种蚯蚓多肽、其编码序列及其应用 | |
CN113501862B (zh) | 一种多肽及其在制备免疫调节药物中的应用 | |
US7655437B2 (en) | Synthetic gene coding for human granulocyte-colony stimulating factor for the expression in E. coli | |
CN108690123A (zh) | 短肽在制备免疫调节药物中的应用 | |
US8623348B2 (en) | Interferon-α (IFN-α) fused proteins comprising IFN-α and a cytoplasmic transduction peptide (CTP) | |
HUT52154A (en) | Recombinant interleukin-2 hybrid proteins. | |
KR20170077252A (ko) | Hnp-1 또는 hnp-2 또는 hnp-3을 인코딩하는 플라스미드 dna, 박테리아 제조체, 진통제 (변이체) | |
US11732016B2 (en) | Polypeptide exhibiting granulocyte-colony stimulating factor activity | |
CN108250277B (zh) | 微小rna31前体编码多肽的截短体在制备免疫调节药物中的应用 | |
Su et al. | Clinical grade production and characterization of a fusion protein comprised of the chemokine CCL2-ligand genetically fused to a mutated and truncated form of the Shiga A1 subunit | |
EP3500284A1 (en) | Bovine fibroblast growth factor 21 and ketosis in dairy cattle | |
WO2024046280A1 (zh) | 一种聚乙二醇修饰的il-21衍生物及其应用 | |
US20230322882A1 (en) | Fusion Polypeptide and Polypeptide Dimer, and Use Thereof | |
EP3381935B1 (en) | Anti-gamma mutant protein against endogenous human interferon- gamma |
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 |