LONG LASTING NATRIURETIC PEPTIDE DERIVATIVES

1 . A natriuretic peptide derivative comprising a NP peptide and a reactive entity coupled to the NP peptide, the reactive entity being capable of covalently bonding with a functionality on a blood component; wherein the NP peptide has a sequence of formula:

wherein

X 1 is Thr or absent;

X 2 is Ser, Thr, Ala or absent;

X 3 is Pro, Hpr, Val, or absent;

X 4 is Lys, D-Lys, Arg, D-Arg, Asn, Gln or absent;

X 5 is Met, Leu, Ile, an oxidatively stable Met-replacement amino acid, Ser, Thr or absent;

X 6 is Val, Ile, Leu, Met, Phe, Ala, D-Ala, Nle or absent;

X 7 is Gln, Asn, Arg, D-Arg, Asp, Lys, D-Lys or absent;

X 8 is Gly, Pro, Ala, D-Ala, Arg, D-Arg, Asp, Lys, D-Lys, Gln, Asn or absent;

X 9 is Ser, Thr or absent;

X 10 is Gly, Pro, Ala, D-Ala, Ser, Thr or absent;

X 12 is Phe, Tyr, Leu, Val, IIe, Ala, D-Ala, Phe with an isosteric replacement of its amide bond selected from the group consisting of N-α-methyl, methyl amino, hydroxylethyl, hydrazino, ethylene, sulfonamide and N-alkyl-α-aminopropionic acid, or a Phe-replacement amino acid conferring on said analog resistance to NEP enzyme;

X 13 is Gly, Ala, D-Ala or Pro;

X 14 is Arg, Lys, D-Lys, Asp, Gly, Ala, D-Ala or Pro;

X 15 is Lys, D-Lys, Arg, D-Arg, Asn, Gln or Asp;

X 16 is Met, Leu, IIe or an oxidatively stable Met-replacement amino acid;

X 20 is Ser, Gly, Ala, D-Ala or Pro;

X 21 is Ser, Gly, Ala, D-Ala, Pro, Val, Leu, or Ile;

X 22 is Ser, Gly, Ala, D-Ala, Pro, Gln or Asn;

X 24 is Gly, Ala, D-Ala or Pro;

X 26 is Gly, Ala, D-Ala or Pro;

X 28 is Lys, D-Lys, Arg, D-Arg, Asn, Gln, H is or absent;

X 29 is Val, Ile, Leu, Met, Phe, Ala, D-Ala, Nle, Ser, Thr or absent;

X 30 is Leu, Nle, IIe, Val, Met, Ala, D-Ala, Phe, Tyr or absent;

X 31 is Arg, D-Arg, Asp, Lys, D-Lys or absent;

X 32 is Arg, D-Arg, Asp, Lys, D-Lys, Tyr, Phe, Trp, Thr, Ser or absent;

X 33 is H is, Asn, Gln, Lys, D-Lys, Arg, D-Arg or absent;

R 1 is NH 2 or a N-terminal blocking group;

R 2 is COOH, CONH 2 or a C-terminal blocking group;

where a peptidic bond links Argl 8 and Ile 19 and the line between CyslI and Cys 27 represents a direct disulfide bridge.

2 . The derivative defined in claim 1 wherein:

X 1 is Thr or absent;

X 2 is Ala or absent;

X 3 is Pro or absent;

X 4 is Arg or absent;

X 5 is Ser, Thr or absent;

X 6 is Leu, IIe, Nle, Met, Val, Ala, Phe or absent;

X 7 is Arg, D-Arg, Asp, Lys, D-Lys, Gln, Asn or absent;

X 8 is Arg, D-Arg, Asp, Lys, D-Lys, Gln, Asn or absent;

X 9 is Ser, Thr or absent;

X 10 is Ser, Thr or absent;

X 12 is Phe, Tyr, Leu, Val, Ile, Ala, D-Ala, Phe with an isosteric replacement of its amide bond selected from the group consisting of N-α-methyl, methyl amino, hydroxylethyl, hydrazino, ethylene, sulfonamide and N-alkyl-α-aminopropionic acid, or a Phe-replacement amino acid conferring on said analog resistance to NEP enzyme;

X 13 is Gly, Ala, D-Ala or Pro;

X 14 is Gly, Ala, D-Ala or Pro;

X 15 is Arg, Lys, D-Lys, or Asp;

X 16 is Met, Leu, IIe or an oxidatively stable Met-replacement amino acid;

X 20 is Gly, Ala, D-Ala or Pro;

X 21 is Ala, D-Ala, Val, Leu, or Ile;

X 22 is Gln or Asn;

X 24 is Gly, Ala, D-Ala or Pro;

X 26 is Gly, Ala, D-Ala or Pro;

X 28 is Asn, Gln, H is, Lys, D-Lys, Arg, D-Arg or absent;

X 29 is Ser, Thr or absent;

X 30 is Phe, Tyr, Leu, Val, Ile, Ala or absent;

X 31 is Arg, D-Arg, Asp, Lys, D-Lys or absent;

X 32 is Tyr, Phe, Trp, Thr, Ser or absent;

X 33 is absent;

R 1 is NH 2 or a N-terminnal blocking group;

R 2 is COOH, CONH 2 or a C-terminal blocking group.

3 . The derivative of claim 2 wherein

X 1 is Thr or absent;

X 2 is Ala or absent;

X 3 is Pro or absent;

X 4 is Arg or absent;

X 5 is Ser or absent;

X 6 is Leu or absent;

X 7 is Arg, Asp or absent;

X 8 is Arg, Asp or absent;

X 9 is Ser or absent;

X 10 is Ser or absent;

X 12 is Phe or Phe with an isosteric replacement of its amide bond selected from the group consisting of N-α-methyl, methyl amino, hydroxylethyl, hydrazino, ethylene, sulfonamide and N-alkyl-β-aminopropionic acid;

X 13 is Gly;

X 14 is Gly;

X 15 is Arg or Asp;

X 16 is Met or Ile;

X 20 is Gly;

X 21 is Ala;

X 22 is Gln;

X 24 is Gly;

X 26 is Gly;

X 28 is Asn or absent;

X 29 is Ser or absent;

X 30 is Phe or absent;

X 31 is Arg, Asp or absent;

X 32 is Tyr or absent;

X 33 is absent;

R 1 is NH 2 or a N-terminal blocking group;

R 2 is COOH, CONH 2 or a C-terminal blocking group.

4 . The derivative of claim 3 , wherein the NP peptide is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 8, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17 and SEQ ID NO: 19.

5 . The derivative of claim 1 , selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO:18 and SEQ ID NO: 20.

6 . The derivative defined in claim 1 , wherein:

X 1 is absent;

X 2 is Ser, Thr, or absent;

X 3 is Pro, Hpr, Val, or absent;

X 4 is Lys, D-Lys, Arg, D-Arg, Asn, Gln or absent;

X 5 is Met, Leu, Ile, an oxidatively stable Met-replacement amino acid, or absent;

X 6 is Val, Ile, Leu, Met, Phe, Ala, D-Ala, Nle or absent;

X 7 is Gln, Asn, or absent;

X 8 is Gly, Pro, Ala, D-Ala, or absent;

X 9 is Ser, Thr or absent;

X 10 is Gly, Pro, Ala, D-Ala, or absent;

X 12 is Phe, Tyr, Leu, Val, Ile, Ala, D-Ala, Phe with an isosteric replacement of its amide bond selected from the group consisting of N-α-methyl, methyl amino, hydroxylethyl, hydrazino, ethylene, sulfonamide and N-alkyl-α-aminopropiomc acid, or a Phe-replacement amino acid conferring on said analog resistance to NEP enzyme;

X 13 is Gly, Ala, D-Ala or Pro;

X 14 is Arg, Lys, D-Lys, or Asp;

X 15 is Lys, D-Lys, Arg, D-Arg, Asn, or Gln;

X 16 is Met, Leu, IIe or an oxidatively stable Met-replacement amino acid;

X 20 is Ser, Gly, Ala, D-Ala or Pro;

X 21 is Ser, Gly, Ala, D-Ala, or Pro;

X 22 is Ser, Gly, Ala, D-Ala, or Pro;

X 24 is Gly, Ala, D-Ala or Pro;

X 26 is Gly, Ala, D-Ala or Pro;

X 28 is Lys, D-Lys, Arg, D-Arg, Asn, Gln, or absent;

X 29 is Val, Ile, Leu, Met, Phe, Ala, D-Ala, Nle, or absent;

X 30 is Leu, Nle, Ile, Val, Met, Ala, D-Ala, Phe, or absent;

X 31 is Arg, D-Arg, Asp, Lys, D-Lys or absent;

X 32 is Arg, D-Arg, Asp, Lys, D-Lys, or absent;

X 33 is H is, Asn, Gln, Lys, D-Lys, Arg, D-Arg or absent;

R 1 is NH 2 or a N-terminal blocking group;

R 2 is COOH, CONH 2 or a C-terminal blocking group.

7 . The derivative of claim 6 wherein:

X 1 is absent;

X 2 is Ser or absent;

X 3 is Pro or absent;

X 4 is Lys or absent;

X 5 is Met, ile or absent;

X 6 is Val or absent;

X 7 is Gln or absent;

X 8 is Gly or absent;

X 9 is Ser or absent;

X 10 is Gly or absent;

X 12 is Phe or Phe with an isosteric replacement of its amide bond selected from the group consisting of N-α-methyl, methyl amino, hydroxylethyl, hydrazino, ethylene, sulfonamide and N-alkyl-β-aminopropionic acid;

X 13 is Gly;

X 14 is Arg or Asp;

X 15 is Lys or Arg;

X 16 is Met or IIe;

X 20 is Ser;

X 21 is Ser;

X 22 is Ser;

X 24 is Gly;

X 26 is Gly;

X 28 is Lys, Arg or absent;

X 29 is Val or absent;

X 30 is Leu or absent;

X 31 is Arg, Asp or absent;

X 32 is Arg, Asp or absent;

X 33 is H is or absent.

8 . The derivative of 7 wherein the NP peptide is selected from the group consisting of SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ IDI NO: 25, SEQ ID NO: 28, SEQ ID NO: 31, SEQ ID NO: 34, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 42, SEQ ID NO: 45, SEQ ID NO: 48 and SEQ ID NO: 51.

9 . The derivative of claim 1 selected from the group consisting of

SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56 and SEQ ID NO: 57.

10 . The derivative of claim 1 , being capable of selectively covalently bonding with a single functionality on the blood component with a degree of selectivity of 80% or more.

11 . The derivative defined in any one of claim 10 , wherein the derivative bonds the blood component in a ratio 1:1 derivative:blood component.

12 . The derivative of claim 1 , wherein the reactive entity is a maleimide or a maleimido-containing group.

13 . The derivative of claim 13 , wherein the reactive entity is MPA.

14 . A pharmaceutical composition comprising the derivative of claim 1 in combination with a pharmaceutically acceptable carrier.

15 . The composition of claim 14 for the treatment of congestive heart failure.

16 . The composition of claim 14 for the treatment of hypertension.

17 . A method for the treatment of congestive heart failure in a subject comprising 13 , alone or in combination with a pharmaceutically acceptable carrier.

18 . A conjugate comprising the derivative of claim 1 covalently bonded to a blood component, where the covalent bond is performed in vivo or ex vivo.

19 . The conjugate of claim 18 , wherein the reactive entity is a maleimide or a maleimidocontaining group and the blood component is a blood protein.

20 . The conjugate of claim 19 , wherein the blood protein is serum albumin.

21 . A method for the treatment of congestive heart failure in a subject comprising administering to a subject an effective amount of the conjugate of claim 18 alone or in combination with a pharmaceutically acceptable carrier.

22 . A method for extending the in vivo half-life of a NP peptide claim 1 , the method comprising coupling to the NP peptide a reactive group which is capable of forming a covalent bond with a blood component, and covalently bonding in vivo or ex vivo the NP peptide to a blood component.

23 . The method as claimed in claim 22 , wherein the blood component is serum albumin.

24 . A method for the treatment of renal disorder in a subject comprising administering to a subject an effective amount of the derivative of claim 1 , alone or in combination with a pharmaceutical carrier.

25 . A method for the treatment of hypertension in a subject comprising administering to a subject an effective amount of the derivative of claim 1 , alone or in combination with a pharmaceutical carrier.

26 . A method for the treatment of asthma in a subject comprising administering to a subject an effective amount of the derivative of claim 1 alone or in combination with a pharmaceutical carrier.

27 . A method for the treatment of renal disorder in a subject comprising administering to a subject an effective amount of the conjugate of claims 18 , alone or in combination with a pharmaceutical carrier.

28 . A method for the treatment of hypertension in a subject comprising administering to a subject an effective amount of the conjugate of claims 18 , alone or in combination with a pharmaceutical carrier

29 . A method for the treatment of asthma in a subject comprising administering to a subject an effective amount of the conjugate of claims 18 , alone or in combination with a pharmaceutical carrier