Fusion immunomodulatory proteins and methods for making same
OwnerNagaraj Govindappa, Maria Melina Soares +1inventors
InventorsNagaraj Govindappa, Maria Melina Soares, Kedernath Sastry
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Abstract
The present invention relates generally to the field of generating recombinant chimeric fusion proteins to be used in the cancer therapy, and more specifically, to fusion molecules of Anti-EGFR1-TGFβRII, Anti-EGFR1-PD1 and Anti-CTLA4-PD1 and methods of generating same, wherein the methods reduce production costs and increase homogeneity of the recombinant chimeric fusion proteins.
Claims6
A method for preparing therapeutically active fusion protein, wherein the fusion protein comprises a tumor targeting moiety and an immunomodulatory molecule wherein the immunomodulatory molecule counteracts immune tolerance of a cancer cell, wherein the tumor targeting moiety is an antibody that binds to Cytotoxic T-Lymphocyte Antigen 4 (CTLA- 4) and wherein the fusion protein is prepared by the following steps:
- preparing a codon optimized nucleotide sequence encoding the fusion protein, wherein the codon optimized nucleotide sequence has been modified to increase CG sequences, wherein the codon optimized nucleotide sequence for the antibody is lacking nucleotides for expression of a lysine at the C-terminal end of the heavy chain of the antibody, wherein the immunomodulatory molecule binds to transforming growth factor beta (TGF-β) or Programmed Death Ligand-1 (PDL1) and wherein the codon optimized nucleotide sequences consist of SEQ ID NOs:
- 5, 6, 4 and 3, or SEQ ID NOs:5, 6, 4 and 7;
- cloning the codon optimized nucleotide sequence of said fusion protein in a CHO host cell capable of transient or stable expression;
- growing the host cell in a media under suitable conditions for growing and allowing the host cell to express the fusion protein;
- and
- collecting secreted fusion proteins and optionally for further purification.
A method of treating cancer in a subject, the method comprising:
- a) preparing a therapeutically active fusion protein having anti-proliferative activity, wherein the fusion protein comprises a tumor targeting moiety and at least one immunomodulatory molecule, wherein the tumor targeting moiety is an antibody that binds to CTLA-4 and wherein the fusion protein is prepared by the following steps:
- i) preparing a codon optimized nucleotide sequence encoding the fusion protein, wherein the codon optimized nucleotide sequence has been modified to increase CG sequences, wherein the codon optimized nucleotide sequence for the antibody is lacking nucleotides for expression of a lysine at the C-terminal end of the heavy chain of the antibody, wherein the immmunomodulatory molecule binds to transforming growth factor beta (TGF-β) or PD-L1 and wherein the codon optimized nucleotide sequences consisting of SEQ ID NOs:
- 5, 6, 4 and 3, or SEQ ID NOs:5, 6, 4 and 7;
- ii) cloning the codon optimized nucleotide sequence of said fusion protein in a CHO host cell capable of transient or stable expression;
- iii) growing the host cell in a media under suitable conditions for growing and allowing the host cell to express the fusion protein;
- and
- iv) collecting secreted fusion proteins for optional purification;
- and
- b) administering a therapeutically active amount of the fusion proteins to the subject.
Dependencies are inferred from the claim text.