Natriuretic compounds, conjugates, and uses thereof
Modified natriuretic compounds and conjugates thereof are disclosed in the present invention. In particular, conjugated forms of hBNP are provided that include at least one modifying moiety attached thereto. The modified natriuretic compound conjugates retain activity for stimulating cGMP production, binding to NPR-A receptor, and in some embodiments an improved half-life in circulation as compared to unmodified counterpart natriuretic compounds. Oral, parenteral, enteral, subcutaneous, pulmonary, and intravenous forms of the compounds and conjugates may be prepared as treatments and/or therapies for heart conditions particularly congestive heart failure. Modifying moieties comprising oligomeric structures having a variety of lengths and configurations are also disclosed. Analogs of the natriuretic compound are also disclosed, having an amino acid sequence that is other than the native sequence.
1 . A natriuretic compound conjugate comprising:
a biologically active natriuretic compound comprising:
a natriuretic molecule NPR-A binding site; and
at least one modifying moiety conjugation site; and
at least one modifying moiety attached to said modifying moiety conjugation site;
wherein said natriuretic compound conjugate exhibits one or more advantages selected from the group consisting of increased resistance to enzymatic degradation relative to a corresponding unconjugated natriuretic compound, increased circulating half life, increased bioavailability, and prolonged duration of effect.
2 . The natriuretic compound conjugate of claim 1 wherein the natriuretic compound comprises a peptide or a biologically active peptide segment of brain natriuretic peptide, atrial natriuretic peptide, C-type natriuretic peptide, or dendroaspis natriuretic peptide.
3 . The natriuretic compound conjugate of claim 1 wherein the natriuretic compound comprises a native hBNP sequence having one or more mutations selected from the group consisting of Lys3Arg, Lys14Arg, Arg30Lys, Lys27Arg, and Arg31Lys.
4 . The natriuretic compound conjugate of claim 1 wherein the natriuretic compound comprises a native hBNP sequence, having one or more insertions, deletions or extensions.
5 . The natriuretic compound conjugate of claim 1 wherein the modifying moiety has a formula:
wherein
each C is independently selected and is an alkyl moiety having m carbons and m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; and
each PAG is independently selected and is a polyalkylene glycol moiety having n subunits and n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25; and
each X is independently selected and is a linking moiety.
6 . The natriuretic compound conjugate of claim 5 , wherein the modifying moiety has a formula:
7 . The natriuretic compound conjugate of claim 5 , wherein the modifying moiety has a formula:
8 . A method of treating a condition characterized by an excessive level of extracellular fluid, the method comprising administering to a subject in need thereof a pharmaceutically acceptable amount of a natriuretic compound conjugate of claim 5 .
9 . The natriuretic compound conjugate of claim 1 , wherein the modifying moiety has a formula:
10 . The natriuretic compound conjugate of claim 5 , wherein the modifying moiety has a formula:
11 . The natriuretic compound conjugate of claim 1 , wherein the modifying moiety has a formula:
12 . The natriuretic compound conjugate of claim 5 , wherein the modifying moiety has a formula:
13 . The natriuretic compound conjugate of claim 5 , wherein the modifying moiety has a formula:
and the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.
14 . The natriuretic compound conjugate of claim 9 wherein the natriuretic compound is hBNP.
15 . The natriuretic compound conjugate of claim 10 wherein the natriuretic compound is hBNP.
16 . The natriuretic compound conjugate of claim 11 wherein the natriuretic compound is hBNP.
17 . The natriuretic compound conjugate of claim 12 wherein the natriuretic compound is hBNP.
18 . The natriuretic compound conjugate of claim 9 wherein the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.
19 . The natriuretic compound conjugate of claim 11 wherein the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.
20 . The natriuretic compound conjugate of claim 12 wherein the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.
21 . The natriuretic compound conjugate of claim 9 wherein the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.
22 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 9 and a pharmaceutically acceptable carrier.
23 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 10 and a pharmaceutically acceptable carrier.
24 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 11 and a pharmaceutically acceptable carrier.
25 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 12 and a pharmaceutically acceptable carrier.
26 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 14 and a pharmaceutically acceptable carrier.
27 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 15 and a pharmaceutically acceptable carrier.
28 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 16 and a pharmaceutically acceptable carrier.
29 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 17 and a pharmaceutically acceptable carrier.
30 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 18 and a pharmaceutically acceptable carrier.
31 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 19 and a pharmaceutically acceptable carrier.
32 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 20 and a pharmaceutically acceptable carrier.
33 . A pharmaceutical composition comprising the natriuretic compound conjugate of claim 21 and a pharmaceutically acceptable carrier.
34 . A method of making the natriuretic compound conjugate of claim 9 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
35 . A method of making the natriuretic compound conjugate of claim 10 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
36 . A method of making the natriuretic compound conjugate of claim 11 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
37 . A method of making the natriuretic compound conjugate of claim 12 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
38 . A method of making the natriuretic compound conjugate of claim 14 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
39 . A method of making the natriuretic compound conjugate of claim 15 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
40 . A method of making the natriuretic compound conjugate of claim 16 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
41 . A method of making the natriuretic compound conjugate of claim 17 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
42 . A method of making the natriuretic compound conjugate of claim 18 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
43 . A method of making the natriuretic compound conjugate of claim 19 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
44 . A method of making the natriuretic compound conjugate of claim 20 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
45 . A method of making the natriuretic compound conjugate of claim 21 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.
46 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 9 .
47 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 10 .
48 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 11 .
49 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 12 .
50 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 14 .
51 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 15 .
52 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 16 .
53 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 17 .
54 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 18 .
55 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 19 .
56 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 20 .
57 . A pharmaceutically pure composition of the natriuretic compound conjugate of claim 21 .
58 . The natriuretic compound conjugate of claim 5 wherein m is 1, X is —O—, n is 4, and C is —CH 3 .