PARATHYROID HORMONE RECEPTOR 1 ANTAGONIST AND INVERSE AGONIST POLYPEPTIDES AND METHODS OF THEIR USE

1 . A polypeptide or a pharmaceutically acceptable salt thereof comprising an N-terminally truncated PTH/PTHrP hybrid peptide or a fragment thereof, wherein said polypeptide is a PTHR1 antagonist or a PTHR1 inverse agonist.

2 . The polypeptide of claim 1 , wherein the N-terminally truncated PTH/PTHrP hybrid peptide is of formula (I):

(I)

Ile-Gln-Leu-X 01 -His-X 02 -X 03 -X 04 -X 05 -X 06 -X 07 -X 08 -X 09 -

X 10 -Arg-Arg-Arg-X 11 -X 12 -Leu-X 13 -X 14 -Leu-Ile-Ala-Glu-

Ile-His-Thr-Ala-Glu-X 15 -Cys,

wherein

X 01 is Met, Leu, or Nle;

X 02 is Asn, Ala, Val, Asp, Glu, or Gln;

X 03 is Leu, Ala, Val, Met, Lys, Ile, Arg, Har, or Trp;

X 04 is Gly, Ala, His, Arg, or dTrp;

X 05 is Lys, Ala, Leu, Gln, Arg, His, or Trp;

X 06 is His, Leu, Arg, Phe, Trp, or Ala;

X 07 is Ile or Leu;

X 08 is Gln or Asn;

X 00 is Asp or Ser;

X 10 is Ala, Leu, Met, Glu, Ser, or Phe;

X 11 is Ala, Phe, Glu, Ser, Leu, Asn, Trp, or Lys;

X 12 is Phe or Trp;

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

X 14 is Lys, His, Ala, Ser, Asn, or Arg; and

X 15 is Ile, Cys, or Tyr;

or a fragment thereof comprising from 24 to 32 contiguous amino acid residues of formula (I).

3 . The polypeptide of claim 2 , wherein the polypeptide is a fragment of the N-terminally truncated PTH/PTHrP hybrid peptide comprising amino acid residues 1-32 of formula (I).

4 . The polypeptide of claim 2 , wherein the polypeptide is a fragment of the N-terminally truncated PTH/PTHrP hybrid peptide comprising amino acid residues 3-32 of formula (I).

5 . The polypeptide of claim 2 , wherein the polypeptide is a fragment of the N-terminally truncated PTH/PTHrP hybrid peptide comprising amino acid residues 3-33 of formula (I).

6 . The polypeptide of any one of claims 2 to 5 , wherein X 01 is Met, X 04 is Ala, X 12 is Phe, and X 15 is Ile.

7 . The polypeptide of any one of claims 2 to 5 , wherein X 01 is Met, X 04 is dTrp, X 12 is Trp, and X 14 is Ile.

8 . The polypeptide of any one of claims 2 to 5 , wherein X 01 is Nle, X 04 is dTrp, X 12 is Trp, and X 15 is Tyr.

9 . The polypeptide of any one of claims 2 to 5 , wherein X 01 is Nle, X 04 is dTrp, X 12 is Trp, and X 15 is Cys.

10 . The polypeptide of any one of claims 2 to 5 , wherein X 01 is Nle, X 04 is dTrp, X 12 is Trp, and X 15 is Ile.

11 . The polypeptide of any one of claims 2 to 5 , wherein X 01 is Met, X 04 is dTrp, X 12 is Trp, and X 15 is Ile.

12 . The polypeptide of claim 2 , wherein the N-terminally truncated PTH/PTHrP hybrid peptide has the amino acid sequence

(SEQ ID NO: 1)

Ile-Gln-Leu-Met-His-Gln-Arg-Ala-Lys-Trp-Ile-Gln-

Asp-Ala-Arg-Arg-Arg-Ala-Phe-Leu-His-Lys-Leu-Ile-

Ala-Glu-Ile-His-Thr-Ala-Glu-Ile;

(SEQ ID NO: 2)

Ile-Gln-Leu-Met-His-Gln-Arg-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: 3)

Ile-Gln-Leu-Nle-His-Gln-Arg-dTrp-Lys-Trp-Ile-Gln-

Asp-Ala-Arg-Arg-Arg-Ala-Trp-Leu-His-Lys-Leu-Ile-

Ala-Glu-Ile-His-Thr-Ala-Glu-Tyr;

(SEQ ID NO: 4)

Ile-Gln-Leu-Nle-His-Gln-Arg-dTrp-Lys-Trp-Ile-Gln-

Asp-Ala-Arg-Arg-Arg-Ala-Trp-Leu-His-Lys-Leu-Ile-

Ala-Glu-Ile-His-Thr-Ala-Glu-Cys;

(SEQ ID NO: 5)

Ile-Gln-Leu-Nle-His-Gln-Arg-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-Cys;

(SEQ ID NO: 6)

Ile-Gln-Leu-Met-His-Gln-Arg-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-Cys;

(SEQ ID NO: 14)

Leu-Met-His-Gln-Arg-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: 15)

Leu-Nle-His-Gln-Arg-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: 16)

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: 17)

Ile-Gln-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: 18)

Leu-Met-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;

or

(SEQ ID NO: 19)

Ile-Gln-Leu-Met-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;

or a 3-32 or 3-33 fragment thereof.

13 . The polypeptide of claim 1 or 2 , further comprising a radionuclide, a polyethylene glycol, or a dye.

14 . A pharmaceutical composition comprising the polypeptide of any one of claims 1 to 13 and a pharmaceutically acceptable carrier.

15 . A method of antagonizing or inversely agonizing the activity of parathyroid hormone receptor 1 (PTHR1) in a cell, the method comprising contacting the cell with the polypeptide of any one of claims 1 to 13 .

16 . The method of claim 15 , wherein the cell is a human cell.

17 . A method of treating a disease or condition associated with PTHR1 signaling overactivity, the method comprising administering to the subject an effective amount of the polypeptide of any one of claims 1 to 13 or the pharmaceutical composition of claim 14 .

18 . The method of claim 17 , wherein the disease or condition is hypercalcemia, hypophosphatemia, hyperparathyroidism, or Jansen's chondrodysplasia.

19 . The method of claim 17 or 18 , wherein the administering comprises subcutaneous, intravenous, intranasal, transpulmonary, transdermal, transmucosal, or oral administration of the polypeptide or the pharmaceutical composition to the subject.