ANALOGUES OF PARATHYROID HORMONE (1-34) THAT FUNCTION AS AGONISTS OF THE PARATHYROID HORMONE RECEPTOR-1 AND DISPLAY MODIFIED ACTIVITY PROFILES
Described are polypeptide analogs of parathyroid hormone (PTH) that include an unnatural amino acid substitution at positions 7 or 8 from the N-terminus of the polypeptide. Also described are pharmaceutical compositions useful for treating hypoparathyroidism that contain the analogs and methods of using the analogs to treat hypoparathyroidism.
1 . An isolated, unnatural peptide analogue comprising:
PTH, a parathyroid hormone receptor (PTHR-1, PTHR-2) agonist-effective fragment of PTH, a parathyroid hormone related protein (PTHrP), a PTHR-1 or PTHR-2 agonist-effective fragment of PTHrP, M-PTH, a PTHR-1 or PTHR-2 agonist-effective fragment of M-PTH, BA058, or a PTHR-1 or PTHR-2 agonist-effective fragment of BA058, in which at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a β-amino acid residue or a (D)-α-amino acid; and
salts thereof.
2 . The peptide analogue of claim 1 , wherein the at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a β 2 -amino acid residue.
3 . The peptide analogue of claim 2 , wherein the at least one naturally occurring (L)-α-amino acid residue at 6, 7, or 8 from the N-terminus is replaced with a β 2 -amino acid residue having a side-chain identical to the (L)-α-amino acid residue it replaces.
4 . The peptide analogue of claim 1 , wherein the at least one naturally occurring (L)-α-amino acid residue at 6, 7, or 8 from the N-terminus is replaced with a β 3 -amino acid residue.
5 . The peptide analogue of claim 4 , wherein the at least one naturally occurring (L)-α-amino acid residue at 6, 7, or 8 from the N-terminus is replaced with a β 3 -amino acid residue having a side-chain identical to the (L)-α-amino acid residue it replaces.
6 . The peptide analogue of claim 1 , wherein the at least one naturally occurring (L)-α-amino acid residue at 6, 7, or 8 from the N-terminus is replaced with a (D)-α-amino acid.
7 . The peptide analogue of claim 6 , wherein the at least one naturally occurring (L)-α-amino acid residue at 6, 7, or 8 from the N-terminus is replaced with a (D)-α-amino acid residue having a side-chain identical to the (L)-α-amino acid residue it replaces.
8 . The peptide analogue of claim 1 , selected from the group consisting of
(SEQ. ID. NO: 8)
SVSEIQ L MHNLGKHLNSMERVEWLRKKLQDVHNF
(SEQ. ID. NO: 10)
SVSEIQ MHNLGKHLNSMERVEWLRKKLQDVHNF
and
(SEQ. ID. NO: 12)
SVSEIQL n L HNLGKHLNSMERVEWLRKKLQDVHNF
(SEQ. ID. NO: 14)
S VSEIQ L MHNLGKHLNSMERVEWLRKKLQDVHNF
(SEQ. ID. NO: 15)
S V SEI Q LMHNLGKHLNSMERVEWLRKKLQDVHNF
wherein L=(R*) β 2 leucine;
L=(D)-leucine; and
n L=(R*)-β 2 -norleucine.
Q=β 3 glutamine
S=β 2 serine.
9 . The peptide analogue of claim 1 , in which at least one additional naturally occurring (L)-α-amino acid residue at position 1 or 2 from the N-terminus is replaced with a β-amino acid residue or a (D)-α-amino acid.
10 . An isolated, unnatural peptide analogue consisting of:
PTH, a parathyroid hormone receptor (PTHR-1, PTHR-2) agonist-effective fragment of PTH, a parathyroid hormone related protein (PTHrP), a PTHR-1 or PTHR-2 agonist-effective fragment of PTHrP, M-PTH, a PTHR-1 or PTHR-2 agonist-effective fragment of M-PTH, BA058, or a PTHR-1 or PTHR-2 agonist-effective fragment of BA058, in which at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a β-amino acid residue or a (D)-α-amino acid; and
salts thereof.
11 . The peptide analogue of claim 10 , wherein the at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a β 2 -amino acid residue.
12 . The peptide analogue of claim 11 , wherein the at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a β 2 -amino acid residue having a side-chain identical to the (L)-α-amino acid residue it replaces.
13 . The peptide analogue of claim 10 , wherein the at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a β 3 -amino acid residue.
14 . The peptide analogue of claim 13 , wherein the at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a β 3 -amino acid residue having a side-chain identical to the (L)-α-amino acid residue it replaces.
15 . The peptide analogue of claim 10 , wherein the at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a (D)-α-amino acid.
16 . The peptide analogue of claim 15 , wherein the at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a (D)-α-amino acid residue having a side-chain identical to the (L)-α-amino acid residue it replaces.
17 . The peptide analogue of claim 10 , selected from the group consisting of
(SEQ. ID. NO: 8)
SVSEIQ L MHNLGKHLNSMERVEWLRKKLQDVHNF
(SEQ. ID. NO: 10)
SVSEIQ MHNLGKHLNSMERVEWLRKKLQDVHNF
and
(SEQ. ID. NO: 12)
SVSEIQL n L HNLGKHLNSMERVEWLRKKLQDVHNF
(SEQ. ID. NO: 14)
S VSEIQ L MHNLGKHLNSMERVEWLRKKLQDVHNF
(SEQ. ID. NO: 15)
S V SEI Q LMHNLGKHLNSMERVEWLRKKLQDVHNF
wherein L=(R*) β 2 leucine;
L=(D)-leucine; and
n L=(R*)-β 2 -norleucine.
Q=β 3 glutamine
S=β 2 serine.
18 . The peptide analogue of claim 10 , in which at least one additional naturally occurring (L)-α-amino acid residue at position 1 or 2 from the N-terminus is replaced with a β-amino acid residue or a (D)-α-amino acid.
19 . A pharmaceutical composition for treating hypoparathyroidism, the composition comprising a parathyroid hormone receptor agonist-effective amount of PTH, a parathyroid hormone receptor (PTHR-1, PTHR-2) agonist-effective fragment of PTH, a parathyroid hormone related protein (PTHrP), a PTHR-1 or PTHR-2 agonist-effective fragment of PTHrP, M-PTH, a PTHR-1 or PTHR-2 agonist-effective fragment of M-PTH, BA058, or a PTHR-1 or PTHR-2 agonist-effective fragment of BA058, in which at least one naturally occurring (L)-α-amino acid residue at position 6, 7, or 8 from the N-terminus is replaced with a β-amino acid residue or a (D)-α-amino acid; and pharmaceutically suitable salts thereof,
in combination with a pharmaceutically suitable carrier.
20 . A method of treating hypoparathyroidism in a mammalian subject, including a human subject, the method comprising administering to the subject a parathyroid hormone receptor agonist-effective amount of a pharmaceutical composition as recited in claim 19 .