Parathyroid hormone polypeptide conjugates and methods of their use

1. A conjugate, or a pharmaceutically acceptable salt thereof, comprising a polypeptide and a fatty acid acyl covalently linked to the polypeptide, wherein the polypeptide comprises a sequence of formula (I):

(I)

(SEQ ID NO: 9)

X 01 -X 02 -X 03 -X 04 -X 05 -X 06 -Leu-X 08 -His-X10-X 11 -X 12 -X 13 -X 14 -

X 15 -X 16 -X 17 -X 18 -X 19 -Arg-X 21 -X 22 -X 23 -Leu-X 25 -X 26 -X 27 -X 28 -

X 29 -X 30 -X 31 -His-X 33 -X 34 -X 35 -X 36 -X 37 ,

wherein

X 01 is absent;

X 02 is absent;

X 03 is absent;

X 04 is absent;

X 05 is absent;

X 06 is absent;

X 08 is Nle;

X 10 is Asn, Ala, Val, Asp, Glu, or Gln;

X 11 is Leu, Ala, Val, Met, Lys, Ile, Arg, Har, or Trp;

X 12 is dTrp;

X 13 is Lys, Ala, Leu, Gln, Arg, His, or Trp;

X 14 is His, Leu, Arg, Phe, Trp, or Ser;

X 15 is Leu or Ile;

X 16 is Asn or Gln;

X 17 is Ser or Asp;

X 18 is Met, Ala, Leu, Glu, Ser, or Phe;

X 19 is Glu or Arg;

X 21 is Val or Arg;

X 22 is Glu, Ala, Phe, Ser, Leu, Asn, Trp, or Lys;

X 23 is Trp, Phe, or Leu;

X 25 is Arg, His, Leu, Glu, Trp, or Lys;

X 26 is Lys, His, Ala, Ser, Asn, or Arg;

X 27 is Lys or Leu;

X 28 is Leu or Ile;

X 29 is Gln, Ala, or Aib;

X 30 is Asp, Glu, or Lys;

X 31 is Val, Leu, or Ile;

X 33 is Asn or Thr;

X 34 is Phe or Ala;

X 35 is Glu;

X 36 is Ile; and

X 37 is absent,

wherein the fatty acid acyl is covalently linked to the C-terminus of the polypeptide through a linker of the structure:

wherein

n is an integer from 0 to 10;

each of m and q is an integer from 2 to 5;

each AA 1 and each AA 2 is independently a proteinogenic amino acid; and

R is a bond to the carbonyl carbon of the fatty acid acyl.

2. The conjugate of claim 1 , further comprising a dye.

3. The conjugate of claim 1 , wherein the fatty acid acyl is a very long chain fatty acid or a long chain fatty acid acyl.

4. The conjugate of claim 3 , wherein the long chain fatty acid acyl is palmitoyl.

5. The conjugate of claim 1 , wherein X 23 is Phe; or X 23 is Trp; or X 11 is Leu and X 23 is Trp; or X 11 is Leu, X 13 is Lys, and X 23 is Trp.

6. The conjugate of claim 1 , wherein the polypeptide has the amino acid sequence Leu-Nle-His-Gln-Leu-dTrp-Lys-Trp-Ile-Gln-Asp-Ala-Arg-Arg-Arg-Ala-Trp-Leu-His-Lys-Leu-Ile-Ala-Glu-Ile-His-Thr-Ala-Glu-Ile (SEQ ID NO: 5); or Leu-Nle-His-Gln-Leu-dTrp-Lys-Trp-Ile-Gln-Asp-Ala-Arg-Arg-Arg-Ala-Trp-Leu-Leu-Lys-Leu-Ile-Ala-Glu-Ile-His-Thr-Ala-Glu-Ile (SEQ ID NO: 7).

7. A pharmaceutical composition comprising the conjugate of claim 1 and one or more pharmaceutically acceptable carriers or excipients.