PD-L1 analog fusion proteins for antigen specific immunotherapy and methods of use
The present disclosure provides recombinantly manufactured fusion proteins comprising a Programmed Death-Ligand 1 (PD-L1) protein fragment or an analog thereof linked to a canine Fc fragment. Embodiments include the administration of the fusion proteins to patients as a treatment for cancers, tumors or other diseases associated with expression of the PD-L1 protein in dogs. Exemplary Fc fusion proteins and pharmaceutical formulations of exemplary Fc fusion proteins are provided, in addition to methods of use and preparation.
1 . A fusion protein comprising a PD-L1 analog and an Fc fragment, wherein the PD-L1 analog and the Fc fragment are connected by a peptide linker, wherein the PD-L1 analog consists of the sequence:
(SEQ ID NO: 25)
FTITVSKDLYVVEYGGNVTMECKFPVEKQLNLFALIVYWEMEDKKIIQFV
NGKEDLKVQHSSYSQRAQLLKDQLFLGKAALQITDVRLQDAGVYRCLIGY
GGADYKRITLKVHAPYRNISQRISVDPVTSEHELMCQAEGYPEAEVIWTS
SDHRVLSGKTTITNSNREEKLFNVTSTLNINATANEIFYCTFQRSGPEEN
NTAELVIPERLPVPASERTHFMILGPF,
and
wherein the Fc fragment comprises the sequence:
(SEQ ID NO: 1)
DCPKCPAPEMLGGPSVFIFPPKPKDTLLIARTPEVTCVVVDLDPEDPEVQ
ISWFVDGKQMQTAKTQPREEQFNGTYRVVSVLPIGHQDWLKGKQFTCKVN
NKALPSPIERTISKARGQAHQPSVYVLPPSREELSKNTVSLTCLIKDFFP
PDIDVEWQSNGQQEPESKYRTTPPQLDEDGSYFLYSKLSVDKSRWQRGDT
FICAVMHEALHNHYTQESLSHSPG.
2 . The fusion protein of claim 1 , wherein the PD-L1 analog and the Fc fragment are connected by the peptide linker comprising the sequence:
(SEQ ID NO: 7)
QGGGSGGQ.
3 . A fusion protein comprising a PD-L1 analog consisting of SEQ ID NO: 25 and an Fc fragment, wherein the PD-L1 analog and the Fc fragment are connected by a peptide linker, wherein the fusion protein comprises the sequence:
(SEQ ID NO: 26)
FTITVSKDLYVVEYGGNVTMECKFPVEKQLNLFALIVYWEMEDKKIIQFV
NGKEDLKVQHSSYSQRAQLLKDQLFLGKAALQITDVRLQDAGVYRCLIGY
GGADYKRITLKVHAPYRNISQRISVDPVTSEHELMCQAEGYPEAEVIWTS
SDHRVLSGKTTITNSNREEKLFNVTSTLNINATANEIFYCTFQRSGPEEN
NTAELVIPERLPVPASERTHFMILGPFQGGGSGGQDCPKCPAPEMLGGPS
VFIFPPKPKDTLLIARTPEVTCVVVDLDPEDPEVQISWFVDGKQMQTAKT
QPREEQFNGTYRVVSVLPIGHQDWLKGKQFTCKVNNKALPSPIERTISKA
RGQAHQPSVYVLPPSREELSKNTVSLTCLIKDFFPPDIDVEWQSNGQQEP
ESKYRTTPPQLDEDGSYFLYSKLSVDKSRWQRGDTFICAVMHEALHNHYT
QESLSHSPG.
4 . The fusion protein of claim 1 , wherein the fusion protein is a homodimer comprising two identical monomers bound together via one or more disulfide bonds.
5 . The fusion protein of claim 1 , wherein the Fc fragment is glycosylated.
6 . An immunogenic composition comprising a fusion protein according to claim 1 and a pharmaceutically acceptable carrier.
7 . The immunogenic composition of claim 6 , further comprising an adjuvant.
8 . A fusion protein comprising the sequence of SEQ ID NO: 26 or pharmaceutical composition thereof for use in treatment for cancers or tumors in dogs.
9 . A method of treating cancers or tumors in a dog, the method comprising administering a fusion protein according to claim 1 or pharmaceutical composition thereof to a dog in need thereof.
10 . The method of claim 9 , wherein said fusion protein or pharmaceutical composition thereof is administered via injection.
11 . The method of claim 10 , wherein said fusion protein or pharmaceutical composition thereof is administered subcutaneously or intramuscularly.
12 . The method of claim 9 , wherein said fusion protein or pharmaceutical composition thereof is administered as a priming vaccine, the method further comprising administering a booster of said fusion protein or pharmaceutical composition thereof 7 to 14 days after administering said priming vaccine.
13 . The method of claim 9 , wherein said dog is antibody-positive to PD-L1 before said administering step, wherein said fusion protein or pharmaceutical composition thereof is administered as a booster vaccine to said dog.
14 . A method of treating cancers or tumors in a dog, the method comprising administering a fusion protein according to claim 3 or pharmaceutical composition thereof to a dog in need thereof.
15 . The method of claim 14 , wherein said fusion protein or pharmaceutical composition thereof is administered via injection.
16 . The method of claim 15 , wherein said fusion protein or pharmaceutical composition thereof is administered subcutaneously or intramuscularly.
17 . The method of claim 14 , wherein said fusion protein or pharmaceutical composition thereof is administered as a priming vaccine, the method further comprising administering a booster of said fusion protein or pharmaceutical composition thereof 7 to 14 days after administering said priming vaccine.
18 . The method of claim 14 , wherein said dog is antibody-positive to PD-L1 before said administering step, wherein said fusion protein or pharmaceutical composition thereof is administered as a booster vaccine to said dog.