Erythropoietin Analogs For Veterinary Use
AssigneeKindred Biosciences, Inc.
InventorsHangjun Zhan, Lam Nguyen, Qingyi Chu, Shyr Jiann Li
Life of this patent
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Abstract
Provided are various embodiments relating to erythropoietin (EPO) polypeptide analogs with one or more additional glycosylation sites and/or additional cysteine residues and methods of producing and using the same to treat anemia in companion animals. Also provided are various embodiments relating to polypeptides comprising an extracellular domain of EPO receptor and methods of using the same for treating polycythemia in mammals.
Claims178
An erythropoietin (EPO) polypeptide comprising the amino acid sequence of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4, SEQ ID NO:
- 7, or SEQ ID NO:
- 8, except for the presence of at least one N-linked glycosylation site not present in SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4, SEQ ID NO:
- 7, or SEQ ID NO:
- 8, wherein the N-linked glycosylation site comprises the sequence asparagine-xaa-serine or asparagine-xaa-threonine, wherein xaa is any amino acid except proline, and wherein one N-linked glycosylation site does not overlap with another N-linked glycosylation site.
An EPO polypeptide comprising the amino acid sequence of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4, SEQ ID NO:
- 7, or SEQ ID NO:
- 8 except for the presence of at least one cysteine not present in SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4, SEQ ID NO:
- 7, or SEQ ID NO:
- 8.
An EPO polypeptide comprising the amino acid sequence of SEQ ID NO:
- 7 or SEQ ID NO:
- 8 except for the presence of an amino acid other than a cysteine at position 165 of SEQ ID NO:
- 7 or at position 139 of SEQ ID NO:
- 8.
The contiguous polypeptide of any one of the claims 60 to 80, wherein the variant IgG Fc polypeptide comprises:
- a) a proline at position 5, a glycine at position 38, an arginine at position 39, an isoleucine at position 97, and/or a glycine at position 98 of SEQ ID NO:
- 54;
- or
- b) a proline at position 5, a glycine at position 38, an arginine at position 39, an isoleucine at position 97, and/or a glycine at position 98 of SEQ ID NO:
- 56.
The contiguous polypeptide of any one of claims 60 to 81, wherein the variant IgG Fc polypeptide comprises the amino acid sequence of SEQ ID NO:
- 53, SEQ ID NO:
- 54, SEQ ID NO:
- 55, SEQ ID NO:
- 56, SEQ ID NO:
- 57, SEQ ID NO:
- 58, SEQ ID NO:
- 59, SEQ ID NO:
- 60, SEQ ID NO:
- 61, SEQ ID NO:
- 62, SEQ ID NO:
- 63, SEQ ID NO:
- 64, SEQ ID NO:
- 65, SEQ ID NO:
- 66, SEQ ID NO:
- 67, SEQ ID NO:
- 68, SEQ ID NO:
- 69, SEQ ID NO:
- 70, SEQ ID NO:
- 71, SEQ ID NO:
- 72, SEQ ID NO:
- 73, SEQ ID NO:
- 74, SEQ ID NO:
- 75, SEQ ID NO:
- 76, SEQ ID NO:
- 77, SEQ ID NO:
- 78, SEQ ID NO:
- 79, SEQ ID NO:
- 80, SEQ ID NO:
- 81, SEQ ID NO:
- 82, SEQ ID NO:
- 83, SEQ ID NO:
- 84, SEQ ID NO:
- 85, SEQ ID NO:
- 86, SEQ ID NO:
- 87, SEQ ID NO:
- 88, SEQ ID NO:
- 89, SEQ ID NO:
- 90, SEQ ID NO:
- 91, SEQ ID NO:
- 92, SEQ ID NO:
- 93, SEQ ID NO:
- 94, SEQ ID NO:
- 95, SEQ ID NO:
- 96, SEQ ID NO:
- 97, SEQ ID NO:
- 98, SEQ ID NO:
- 99, SEQ ID NO:
- 100, SEQ ID NO:
- 101, SEQ ID NO:
- 102, SEQ ID NO:
- 103, SEQ ID NO:
- 104, SEQ ID NO:
- 105, SEQ ID NO:
- 106, SEQ ID NO:
- 107, SEQ ID NO:
- 108, SEQ ID NO:
- 109, SEQ ID NO:
- 110, or SEQ ID NO:
- 111.
An isolated nucleic acid encoding an EPO polypeptide comprising the amino acid sequence of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, or SEQ ID NO:
- 4;
- and a heterologous regulatory sequence, wherein the heterologous regulatory sequence is not a constitutive promoter.
An expression system comprising a first vector comprising a nucleic acid encoding an EPO polypeptide comprising the amino acid sequence of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, or SEQ ID NO:
- 4;
- and a second vector comprising a regulatory sequence.
A method of expressing an EPO polypeptide in a target cell, comprising
- a) transferring a nucleic acid, a vector, or an expression system into the target cell, wherein the nucleic acid, vector, or expression system comprises:
- i) a nucleic acid encoding an EPO polypeptide having the amino acid sequence of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, or SEQ ID NO:
- 4, and
- ii) a regulatory sequence, wherein the regulatory sequence is not a constitutive promoter;
- and
- b) culturing the cell under conditions supportive for expression of the EPO polypeptide.
A polypeptide comprising an extracellular domain of a canine, equine, or feline erythropoietin receptor (EPOR) polypeptide, wherein the canine, equine, or feline EPOR polypeptide comprises the amino acid sequence of SEQ ID NO:
- 33, SEQ ID NO:
- 37, SEQ ID NO:
- 41, SEQ ID NO:
- 44, SEQ ID NO:
- 47, or SEQ ID NO:
- 50;
- and a heterologous polypeptide sequence.
A polypeptide comprising the amino acid sequence of SEQ ID NO:
- 34, SEQ ID NO:
- 35, SEQ ID NO:
- 38, SEQ ID NO:
- 39, SEQ ID NO:
- 42, SEQ ID NO:
- 43, SEQ ID NO:
- 45, SEQ ID NO:
- 46, SEQ ID NO:
- 48, SEQ ID NO:
- 49, SEQ ID NO:
- 51, or SEQ ID NO:
- 52;
- and a heterologous polypeptide sequence.
Dependencies are inferred from the claim text.