Pharmaceutical compositions of polypeptide conjugates and methods of uses thereof

A pharmaceutical composition, comprising a polypeptide conjugate and an absorption enhancing excipient, wherein:

• the polypeptide conjugate comprises a polypeptide portion and a conjugate portion,
• the polypeptide portion comprises a single biologically active peptide and a peptide linker, wherein the biologically active peptide is attached to N-terminus of the peptide linker and comprises GLP-1;
• and
• the conjugate portion comprises a first clearance-reducing moiety (CRM) conjugated to a first CRM residue in the peptide linker, and optionally a second CRM conjugated to a second CRM residue in the polypeptide portion, wherein the first CRM residue and the second CRM residue are both lysine residues,
• wherein:
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 52, and is conjugated with the first CRM at 76K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 55, and is conjugated with the first CRM at 68K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 56, and is conjugated with the first CRM at 60K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 60, and is conjugated with the first CRM at 76K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 61, and is conjugated with the first CRM at 68K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 62, and is conjugated with the first CRM at 60K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 51, and is conjugated with the first CRM and the second CRM respectively at 26K and 76K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 53, and is conjugated with the first CRM and the second CRM respectively at 26K and 84K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 54, and is conjugated with the first CRM and the second CRM respectively at 26K and 68K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 57, and is conjugated with the first CRM and the second CRM respectively at 26K and 76K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 58, and is conjugated with the first CRM and the second CRM respectively at 26K and 84K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 59, and is conjugated with the first CRM and the second CRM respectively at 26K and 68K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 85, and is conjugated with the first CRM and the second CRM respectively at 26K and 96K;
• the polypeptide portion comprises the amino acid sequence of SEQ ID NO:
• 86, and is conjugated with the first CRM and the second CRM respectively at 26K and 60K;
• wherein the first clearance-reducing moiety (CRM) and the second CRM both have the structure of below formula: