Compositions comprising alkaline phosphatase and/or natriuretic peptide and methods of use thereof

Abstract

The present invention provides methods, compositions, and kits for the treatment of neurocutaneous syndromes, such as neurofibromatosis type I; disorders associated with overactivation of FGFR3, such as achondroplasia; bone or cartilage disorders; or vascular smooth muscle disorders; or for the elongation of bone. In some embodiments, the present invention provides polypeptides having an alkaline phosphatase peptide fused to an Fc domain of an immunoglobulin or a natriuretic peptide fused to an Fc domain of an immunoglobulin. Such polypeptides can be administered to subjects, e.g., subcutaneously, to treat a neurocutaneous syndrome, a disorder associated with overactivation of FGFR3, a bone or cartilage disorder, or a vascular smooth muscle disorder, or to elongate bone. The invention also features nucleic acid molecules encoding such polypeptides and the use of the nucleic acid molecules for treating neurocutaneous syndromes, disorders associated with overactivation of FGFR3, bone or cartilage disorders, or vascular smooth muscle disorders, or for elongating bone.

What is claimed is: 
     
         1 . A method of treating a neurocutaneous syndrome in a subject, said method comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition comprising:
 (a) a polypeptide comprising the structure A-sALP-B; and   (b) a pharmaceutically acceptable excipient,   wherein sALP is the extracellular domain of an alkaline phosphatase,   A is absent or is an amino acid sequence of at least one amino acid, and   B is absent or is an amino acid sequence of at least one amino acid,   thereby treating said syndrome in said subject.   
     
     
         2 . The method of  claim 1 , wherein the amino acid sequence of said polypeptide comprises the amino acid sequence of SEQ ID NOs: 1204 or 1221. 
     
     
         3 . The method of  claim 1 , wherein
 the amino acid sequence of said sALP comprises amino acid residues 23-508 of SEQ ID NO: 1215, amino acid residues 18-498 of SEQ ID NO: 1216, amino acid residues 23-508 of SEQ ID NO: 1218, or amino acid residues 18-498 of SEQ ID NO: 1219, or   the amino acid sequence of said sALP consists of amino acid residues 23-512 of SEQ ID NO: 1215, amino acid residues 18-502 of SEQ ID NO: 1216, amino acid residues 23-512 of SEQ ID NO: 1218, or amino acid residues 18-502 of SEQ ID NO: 1219.   
     
     
         4 . The method of  claim 1  or  3 , wherein the amino acid sequence of said sALP comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 1205, or at least 95% sequence identity to SEQ ID NO: 1205, or at least 99% sequence identity to SEQ ID NO: 1205. 
     
     
         5 . The method of  claim 4 , wherein the amino acid sequence of said sALP comprises or consists of the amino acid sequence of SEQ ID NO: 1205. 
     
     
         6 . The method of any one of  claims 1 - 5 , wherein A and/or B are absent. 
     
     
         7 . The method of any one of  claims 1 - 6 , wherein A or B comprises a fragment crystallizable region (Fc). 
     
     
         8 . The method of any one of  claims 1 - 7 , wherein said Fc comprises a C H2  domain, a C H3  domain, and a hinge region, or wherein said Fc is a constant domain of an immunoglobulin selected from the group consisting of IgG-1, IgG-2, IgG-3, and IgG-4. 
     
     
         9 . The method of  claim 8 , wherein the amino acid sequence of said Fc comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 401, or at least 95% sequence identity to SEQ ID NO: 401, or at least 99% sequence identity to SEQ ID NO: 401. 
     
     
         10 . The method of  claim 9 , wherein the amino acid sequence of said Fc comprises or consists of the amino acid sequence of SEQ ID NO: 401. 
     
     
         11 . The method of any one of  claims 1 - 10 , wherein A or B comprises I n , and wherein I represents an aspartic acid or a glutamic acid and n=10 to 16. 
     
     
         12 . The method of any one of  claims 1 - 10 , wherein said polypeptide does not comprise a polyaspartic acid or polyglutamic acid region longer than three consecutive aspartic acid or glutamic acid residues, or wherein said polypeptide does not comprise a polyaspartic acid or polyglutamic acid region longer than two consecutive aspartic acid or glutamic acid residues. 
     
     
         13 . The method of any one of  claims 1 - 10 , wherein said polypeptide does not comprise a bone-targeting moiety. 
     
     
         14 . The method of any one of  claims 1 - 10 , wherein said polypeptide comprises the structure C-sALP-D-Fc-E or the structure C-Fc-D-sALP-E,
 C is absent or is an amino acid sequence of at least one amino acid,   D is absent or is an amino acid sequence of at least one amino acid, and   E is absent or is an amino acid sequence of at least one amino acid.   
     
     
         15 . The method of  claim 14 , wherein C and/or E are absent. 
     
     
         16 . The method of any one of  claims 1 - 10 , wherein said polypeptide comprises the structure C-sALP-D-Fc-G-I n -H or the structure C-Fc-D-sALP-G-I n -H,
 C is absent or is an amino acid sequence of at least one amino acid,   D is absent or is an amino acid sequence of at least one amino acid,   G is absent or is an amino acid sequence of at least one amino acid,   H is absent or is an amino acid sequence of at least one amino acid,   I represents an aspartic acid or a glutamic acid, and n=10 to 16.   
     
     
         17 . The method of  claim 16 , wherein C and/or H are absent. 
     
     
         18 . The method of  claim 16  or  17 , wherein G is two amino acid residues. 
     
     
         19 . The method of  claim 18 , wherein G is aspartic acid-isoleucine. 
     
     
         20 . The method of any one of  claims 16 - 19 , wherein I is aspartic acid and n=10. 
     
     
         21 . The method of any one of  claims 14 - 20 , wherein D is two amino acid residues. 
     
     
         22 . The method of  claim 21 , wherein D is leucine-lysine. 
     
     
         23 . The method of any one of  claims 1 - 22 , wherein the amino acid sequence of said polypeptide consists of the amino acid sequence of SEQ ID NOs: 1201, 1204, 1220, or 1221. 
     
     
         24 . The method of  claim 23 , wherein the amino acid sequence of said polypeptide consists of the amino acid sequence of SEQ ID NO: 1204. 
     
     
         25 . A method of treating a neurocutaneous syndrome in a subject, said method comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition comprising:
 (a) a polypeptide comprising the structure V-NP-W; and   (b) a pharmaceutically acceptable excipient,   wherein NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B),   V is absent or is an amino acid sequence of at least one amino acid, and   W is absent or is an amino acid sequence of at least one amino acid,   thereby treating said syndrome in said subject.   
     
     
         26 . The method of  claim 25 , wherein the amino acid sequence of said polypeptide comprises the amino acid sequence of SEQ ID NOs: 504, 512, 530, or 572. 
     
     
         27 . The method of  claim 25 , wherein said NP comprises the structure:
 [N-terminal extension]-[short segment]-[ring domain]-[C-terminal extension],   wherein said ring domain comprises the amino acid sequence of SEQ ID NO: 6, amino acid residues 11-27 of SEQ ID NO: 30, or SEQ ID NO: 95, and each of said N-terminal extension, short segment, and C-terminal extension is, independently, absent or is an amino acid sequence of at least one amino acid.   
     
     
         28 . The method of  claim 27 , wherein said ring domain comprises amino acid residues 6-22 of SEQ ID NO: 126. 
     
     
         29 . The method of  claim 28 , wherein the amino acid at position 17 of SEQ ID NO: 126 is Phe, Leu, Ile, Thr, Val, Ala, Ser, Glu, Arg, Tyr, Cys, Pro, or Asp. 
     
     
         30 . The method of  claim 27 , wherein said ring domain comprises the amino acid sequence of SEQ ID NO: 12. 
     
     
         31 . The method of any one of  claims 27 - 30 , wherein said short segment and said ring domain together comprise the amino acid sequence of any one of SEQ ID NOs: 4, 13-30, 119-122, 126, or 156-161. 
     
     
         32 . The method of any one of  claims 27 - 31 , wherein the amino acid sequence of said short segment consists of amino acid residues 1-5, 2-5, 3-5, 4-5, or 5 of SEQ ID NO: 4, amino acid residues 1-10 of SEQ ID NO: 17, amino acid residues 1-5 of SEQ ID NO: 19, amino acid residues 1-3 of SEQ ID NO: 20, amino acid residues 1-5 of SEQ ID NO: 21, or amino acid residues 1-6 of SEQ ID NO: 29. 
     
     
         33 . The method of any one of  claims 27 - 32 , wherein the amino acid sequence of said N-terminal extension comprises amino acid residues 1-31 or 17-31 of SEQ ID NO: 11. 
     
     
         34 . The method of any one of  claims 27 - 32 , wherein the amino acid sequence of said N-terminal extension comprises KGANKK (SEQ ID NO: 314) or KGANQK (SEQ ID NO: 315). 
     
     
         35 . The method of  claim 27 , wherein said N-terminal extension, short segment, and ring domain together comprise the amino acid sequence of SEQ ID NO: 11. 
     
     
         36 . The method of any one of  claims 27 - 35 , wherein said C-terminal extension comprises the amino acid sequence of SEQ ID NOs: 117 or 118 or comprises amino acid residues 23-37 selected from any one of SEQ ID NOs: 101-116. 
     
     
         37 . The method of  claim 27 , wherein the amino acid sequence of said NP consists of SEQ ID NOs: 4 or 11, or the amino acid sequence of any one of SEQ ID NOs: 31-94, or a fragment thereof comprising at least a ring domain, or the amino acid sequence of any one of SEQ ID NOs: 13-29, 100-116, 119-125, 127-233, or 1001-1155. 
     
     
         38 . The method of any one of  claims 27 - 37 , wherein V and/or W are absent. 
     
     
         39 . The method of any one of  claims 27 - 38 , wherein V or W comprises a fragment crystallizable region (Fc). 
     
     
         40 . The method of  claim 39 , wherein said Fc comprises a C H2  domain, a C H3  domain, and a hinge region, or wherein said Fc is a constant domain of an immunoglobulin selected from the group consisting of IgG-1, IgG-2, IgG-3, and IgG-4. 
     
     
         41 . The method of  claim 40 , wherein the amino acid sequence of said Fc comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 401, or at least 95% sequence identity to SEQ ID NO: 401, or at least 99% sequence identity to SEQ ID NO: 401. 
     
     
         42 . The method of  claim 41 , wherein the amino acid sequence of said Fc comprises or consists of the amino acid sequence of SEQ ID NO: 401. 
     
     
         43 . The method of any one of  claims 27 - 42 , wherein V or W comprises a glycine-rich region. 
     
     
         44 . The method of  claim 43 , wherein the amino acid sequence of V or W consists of one or more glycines and one or more serines. 
     
     
         45 . The method of  claim 43  or  44 , wherein the amino acid sequence of V or W comprises [(Gly) m (Ser)] n (Gly) p  or (Gly) p [(Ser)(Gly) m ] n , and wherein each of m, n, and p is, independently, between 0 and 20. 
     
     
         46 . The method of  claim 45 , wherein m is between 1 and 6; n is between 1 and 10; and p is between 0 and 4. 
     
     
         47 . The method of  claim 46 , wherein m is 4 and n is 1-6. 
     
     
         48 . The method of  claim 46 , wherein combinations of m, n, and p are selected from a single row of Table 2, or wherein the amino acid sequence of V or W comprises the amino acid sequence of any one of SEQ ID NOs: 301-391. 
     
     
         49 . The method of any one of  claims 27 - 48 , wherein V or W does not comprise a bone-targeting moiety. 
     
     
         50 . The method of any one of  claims 27 - 48 , wherein V or W comprises a bone-targeting moiety. 
     
     
         51 . The method of  claim 50 , wherein said bone-targeting moiety comprises six consecutive acidic residues. 
     
     
         52 . The method of  claim 51 , wherein said bone-targeting moiety comprises ten consecutive acidic residues. 
     
     
         53 . The method of  claim 51  or  52 , wherein said acidic residues are aspartic acid or glutamic acid. 
     
     
         54 . The method of  claim 53 , wherein said bone-targeting moiety comprises E 6 , E 10 , D 6 , or D 10 . 
     
     
         55 . The method of any one of  claims 27 - 54 , wherein V or W comprises a cathepsin cleavage sequence. 
     
     
         56 . The method of  claim 55 , wherein said cathepsin cleavage sequence comprises a cathepsin K cleavage sequence. 
     
     
         57 . The method of  claim 55  or  56 , wherein said cathepsin cleavage sequence is HGPQG (SEQ ID NO: 374) or HKLRG (SEQ ID NO: 375). 
     
     
         58 . The method of any one of  claims 27 - 57 , wherein said polypeptide comprises the structure V-NP-W,
 NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B),   each of V and W is, independently, absent or is an amino acid sequence of at least one amino acid, and   said NP comprises the amino acid sequence of any one of SEQ ID NOs: 17-29, 31-40, 42-94, 101-116, 119-122, 128-161, or 163-233, or V or W comprises the amino acid sequence of any one of SEQ ID NOs: 304-313, 322-333, or 337-391.   
     
     
         59 . The method of any one of  claims 27 - 57 , wherein said polypeptide comprises the structure V-NP or NP-W,
 NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B), and   each of V and W comprises, independently, the amino acid sequence of any one of SEQ ID NOs: 304-313, 322-333, or 337-391.   
     
     
         60 . The method of any one of  claims 27 - 57 , wherein said polypeptide comprises the structure X-Fc-Y-NP-Z or the structure X-NP-Y-Fc-Z,
 NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B), and   each of X, Y, and Z is, independently, absent or is an amino acid sequence of at least one amino acid.   
     
     
         61 . The method of  claim 60 , wherein Y comprises a glycine-rich region. 
     
     
         62 . The method of  claim 61 , wherein the amino acid sequence of Y consists of one or more glycines and one or more serines. 
     
     
         63 . The method of  claim 62 , wherein the amino acid sequence of Y comprises [(Gly) m (Ser)] n (Gly) p  or (Gly) p [(Ser)(Gly) m ] n , and wherein each of m, n, and p is, independently, between 0 and 20. 
     
     
         64 . The method of any one of  claims 60 - 63 , wherein X is absent, Z is absent, or X and Z are both absent. 
     
     
         65 . The method of any one of  claims 60 - 64 , wherein X, Y, or Z comprises a bone-targeting moiety. 
     
     
         66 . The method of  claim 65 , wherein said bone-targeting moiety comprises six or ten consecutive acidic residues. 
     
     
         67 . The method of  claim 66 , wherein said acidic residues are aspartic acid or glutamic acid. 
     
     
         68 . The method of any one of  claims 65 - 67 , wherein said bone-targeting moiety comprises E 6 , E 10 , D 6 , or D 10 . 
     
     
         69 . The method of any one of  claims 60 - 68 , wherein X, Y, or Z comprises a cathepsin cleavage sequence. 
     
     
         70 . The method of  claim 69 , wherein said cathepsin cleavage sequence comprises a cathepsin K cleavage sequence. 
     
     
         71 . The method of any one of  claims 25 - 70 , wherein the amino acid sequence of said polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 501-608. 
     
     
         72 . The method of  claim 71 , wherein the amino acid sequence of said polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 502, 504, 506, 512, 514, 516, 530, 560, 562, 564, 572, 574, 576, 584, 586, 588, 596, 598, 600, or 608. 
     
     
         73 . The method of  claim 72 , wherein the amino acid sequence of said polypeptide comprises or consists of the amino acid sequence of SEQ ID NOs: 504, 512, 530, or 572. 
     
     
         74 . The method of any one of  claims 1 - 73 , wherein said polypeptide is in dimeric form. 
     
     
         75 . The method of any one of  claims 1 - 74 , wherein said polypeptide is glycosylated or pegylated. 
     
     
         76 . The method of any one of  claims 1 - 75 , wherein said pharmaceutical composition is administered in a dosage between about 0.2 mg/kg to about 20 mg/kg of said polypeptide. 
     
     
         77 . The method of any one of  claims 1 - 75 , wherein said pharmaceutical composition is administered in a dosage between about 0.5 mg/kg to about 500 mg/kg of said polypeptide. 
     
     
         78 . The method of any one of  claims 1 - 75 , wherein said pharmaceutical composition is administered in a dosage between about 10 μg/kg to about 1,000 μg/kg of said polypeptide. 
     
     
         79 . The method of any one of  claims 1 - 78 , wherein said pharmaceutical composition is administered subcutaneously. 
     
     
         80 . The method of any one of  claims 1 - 79 , wherein said pharmaceutical composition is administered one time, two times, or three times per week. 
     
     
         81 . The method of any one of  claims 1 - 80 , wherein said subject is human. 
     
     
         82 . The method of any one of  claims 1 - 81 , wherein said neurocutaneous syndrome is neurofibromatosis type I. 
     
     
         83 . A composition comprising a first polypeptide and a second polypeptide, wherein
 a) said first polypeptide comprises the structure A-sALP-B, wherein
 i) sALP is the extracellular domain of an alkaline phosphatase, 
 ii) A is absent or is an amino acid sequence of at least one amino acid, and 
 iii) B is absent or is an amino acid sequence of at least one amino acid; and 
   b) said second polypeptide comprises the structure V-NP-W, wherein
 i) NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B), 
 ii) V is absent or is an amino acid sequence of at least one amino acid, and 
 iii) W is absent or is an amino acid sequence of at least one amino acid. 
   
     
     
         84 . The composition of  claim 83 , wherein the amino acid sequence of said first polypeptide comprises the amino acid sequence of SEQ ID NOs: 1204 or 1221 and the amino acid sequence of said second polypeptide comprises the amino acid sequence of SEQ ID NOs: 504, 512, 530, or 572. 
     
     
         85 . The composition of  claim 83 , wherein the amino acid sequence of said sALP of said first polypeptide comprises amino acid residues 23-508 of SEQ ID NO: 1215, amino acid residues 18-498 of SEQ ID NO: 1216, amino acid residues 23-508 of SEQ ID NO: 1218, or amino acid residues 18-498 of SEQ ID NO: 1219, or
 the amino acid sequence of said sALP of said first polypeptide consists of amino acid residues 23-512 of SEQ ID NO: 1215, amino acid residues 18-502 of SEQ ID NO: 1216, amino acid residues 23-512 of SEQ ID NO: 1218, or amino acid residues 18-502 of SEQ ID NO: 1219, or   the amino acid sequence of said sALP of said first polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 1205, or at least 95% sequence identity to SEQ ID NO: 1205, or at least 99% sequence identity to SEQ ID NO: 1205.   
     
     
         86 . The composition of  claim 83  or  85 , wherein A and/or B of said first polypeptide are absent. 
     
     
         87 . The composition of any one of  claims 83 - 86 , wherein A or B of said first polypeptide comprises a fragment crystallizable region (Fc). 
     
     
         88 . The composition of any one of  claims 83 - 87 , wherein A or B of said first polypeptide comprises I n , and wherein I represents an aspartic acid or a glutamic acid and n=10 to 16. 
     
     
         89 . The composition of any one of  claims 83 - 88 , wherein said first polypeptide comprises the structure C-sALP-D-Fc-G-I n -H,
 C is absent or is an amino acid sequence of at least one amino acid,   D is absent or is an amino acid sequence of at least one amino acid,   G is absent or is an amino acid sequence of at least one amino acid,   H is absent or is an amino acid sequence of at least one amino acid,   I represents an aspartic acid or a glutamic acid, and n=10 to 16.   
     
     
         90 . The composition of any one of  claims 83 - 89 , wherein the amino acid sequence of said first polypeptide comprises or consists of the amino acid sequence of SEQ ID NO: 1204. 
     
     
         91 . The composition of any one of  claims 83 - 90 , wherein said NP of said second polypeptide comprises the structure:
 [N-terminal extension]-[short segment]-[ring domain]-[C-terminal extension],   wherein said ring domain comprises the amino acid sequence of SEQ ID NO: 6, amino acid residues 11-27 of SEQ ID NO: 30, or SEQ ID NO: 95, and each of said N-terminal extension, short segment, and C-terminal extension is, independently, absent or is an amino acid sequence of at least one amino acid.   
     
     
         92 . The composition of  claim 91 , wherein said ring domain comprises amino acid residues 6-22 of SEQ ID NO: 126. 
     
     
         93 . The composition of  claim 92 , wherein the amino acid at position 17 of SEQ ID NO: 126 is Phe, Leu, Ile, Thr, Val, Ala, Ser, Glu, Arg, Tyr, Cys, Pro, or Asp. 
     
     
         94 . The composition of any one of  claims 91 - 93 , wherein the amino acid sequence of said N-terminal extension comprises amino acid residues 1-31 or 17-31 of SEQ ID NO: 11, KGANKK (SEQ ID NO: 314), or KGANQK (SEQ ID NO: 315). 
     
     
         95 . The composition of any one of  claims 91 - 94 , wherein said C-terminal extension comprises the amino acid sequence of SEQ ID NOs: 117 or 118 or comprises amino acid residues 23-37 selected from any one of SEQ ID NOs: 101-116. 
     
     
         96 . The composition of  claim 91 , wherein the amino acid sequence of said NP consists of SEQ ID NOs: 4 or 11, or the amino acid sequence of any one of SEQ ID NOs: 31-94, or a fragment thereof comprising at least a ring domain, or the amino acid sequence of any one of SEQ ID NOs: 13-29, 100-116, 119-125, 127-233, or 1001-1155. 
     
     
         97 . The composition of any one of  claims 83 - 96 , wherein V and/or W of said second polypeptide are absent. 
     
     
         98 . The composition of any one of  claims 83 - 97 , wherein V or W of said second polypeptide comprises a fragment crystallizable region (Fc). 
     
     
         99 . The composition of any one of  claims 83 - 98 , wherein V or W of said second polypeptide comprises a glycine-rich region. 
     
     
         100 . The composition of any one of  claims 83 - 99 , wherein V or W of said second polypeptide comprises a bone-targeting moiety. 
     
     
         101 . The composition of any one of  claims 83 - 100 , wherein V or W of said second polypeptide comprises a cathepsin cleavage sequence. 
     
     
         102 . The composition of any one of  claims 83 - 101 , wherein said second polypeptide comprises the structure V-NP-W,
 NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B),   each of V and W is, independently, absent or is an amino acid sequence of at least one amino acid, and   said NP comprises the amino acid sequence of any one of SEQ ID NOs: 17-29, 31-40, 42-94, 101-116, 119-122, 128-161, or 163-233, or V or W comprises the amino acid sequence of any one of SEQ ID NOs: 304-313, 322-333, or 337-391.   
     
     
         103 . The composition of any one of  claims 83 - 102 , wherein said second polypeptide comprises the structure V-NP or NP-W,
 NP is said natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B), and   each of V and W comprises, independently, the amino acid sequence of any one of SEQ ID NOs: 304-313, 322-333, or 337-391.   
     
     
         104 . The composition of any one of  claims 83 - 103 , wherein said second polypeptide comprises the structure X-Fc-Y-NP-Z or the structure X-NP-Y-Fc-Z,
 NP is said natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B), and   each of X, Y, and Z is, independently, absent or is an amino acid sequence of at least one amino acid.   
     
     
         105 . The composition of  claim 104 , wherein Y comprises a glycine-rich region. 
     
     
         106 . The composition of  claim 104  or  105 , wherein X is absent, Z is absent, or X and Z are both absent. 
     
     
         107 . The composition of any one of  claims 104 - 106 , wherein X, Y, or Z comprises a bone-targeting moiety. 
     
     
         108 . The composition of any one of  claims 104 - 107 , wherein X, Y, or Z comprises a cathepsin cleavage sequence. 
     
     
         109 . The composition of any one of  claims 83 - 108 , wherein the amino acid sequence of said second polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 501-608. 
     
     
         110 . The composition of  claim 109 , wherein the amino acid sequence of said second polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 502, 504, 506, 512, 514, 516, 530, 560, 562, 564, 572, 574, 576, 584, 586, 588, 596, 598, 600, or 608. 
     
     
         111 . The composition of  claim 110 , wherein the amino acid sequence of said second polypeptide comprises or consists of the amino acid sequence of SEQ ID NO: 512. 
     
     
         112 . The composition of any one of  claims 83 - 111 , wherein said first polypeptide and/or said second polypeptide are in dimeric form. 
     
     
         113 . The composition of any one of  claims 83 - 112 , wherein said first polypeptide and/or said second polypeptide are glycosylated or pegylated. 
     
     
         114 . The composition of any one of  claims 83 - 113 , wherein the composition is a pharmaceutical composition comprising a pharmaceutically acceptable excipient. 
     
     
         115 . The composition of any one of  claims 83 - 114 , wherein said composition is lyophilized. 
     
     
         116 . The composition of any one of  claims 83 - 115 , wherein said first polypeptide is present in a dosage between about 0.2 mg/kg to about 20 mg/kg and said second polypeptide is present in a dosage between about 0.5 mg/kg to about 500 mg/kg. 
     
     
         117 . The composition of any one of  claims 83 - 116 , wherein the amino acid sequence of said first polypeptide comprises the amino acid sequence of SEQ ID NO: 1204 and wherein the amino acid sequence of said second polypeptide comprises the amino acid sequence of SEQ ID NOs: 504, 512, 530, or 572. 
     
     
         118 . A method of treating a disease or a condition in a subject, said method comprising administering to said subject a therapeutically effective amount of a first polypeptide and a second polypeptide, wherein
 a) said first polypeptide comprises the structure A-sALP-B, wherein
 i) sALP is the extracellular domain of an alkaline phosphatase, 
 ii) A is absent or is an amino acid sequence of at least one amino acid, and 
 iii) B is absent or is an amino acid sequence of at least one amino acid; and 
   b) said second polypeptide comprises the structure V-NP-W, wherein
 i) NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B), 
 ii) V is absent or is an amino acid sequence of at least one amino acid, and 
 iii) W is absent or is an amino acid sequence of at least one amino acid; and 
   said disease or condition is selected from the group consisting of a neurocutaneous syndrome, a disorder associated with overactivation of FGFR3, a bone or cartilage disorder, a vascular smooth muscle disorder, and a condition for elongation of bone,   thereby treating said disease or said condition in said subject.   
     
     
         119 . The method of  claim 118 , wherein said first polypeptide and said second polypeptide are administered within ten days, five days, or twenty-four hours of each other. 
     
     
         120 . The method of  claim 118 , wherein said first polypeptide and said second polypeptide are administered simultaneously. 
     
     
         121 . The method of any one of  claims 118 - 120 , wherein said first polypeptide and said second polypeptide are formulated together in a composition or each separately in a composition. 
     
     
         122 . The method of  claim 121 , wherein said composition is a pharmaceutical composition comprising a pharmaceutically acceptable excipient. 
     
     
         123 . The method of  claim 122 , wherein said composition is lyophilized. 
     
     
         124 . The method of any one of  claims 121 - 123 , wherein said composition is the composition of any one of  claims 83 - 117 . 
     
     
         125 . The method of any one of  claims 118 - 124 , wherein the amino acid sequence of said first polypeptide comprises the amino acid sequence of SEQ ID NO: 1204 and wherein the amino acid sequence of said second polypeptide comprises the amino acid sequence of SEQ ID NOs: 504, 512, 530, or 572. 
     
     
         126 . The method of any one of  claims 118 - 125 , wherein said first polypeptide is present in a dosage between about 0.2 mg/kg to about 20 mg/kg and said second polypeptide is present in a dosage between about 0.5 mg/kg to about 500 mg/kg. 
     
     
         127 . The method of any one of  claims 118 - 125 , wherein said first polypeptide is present in a dosage between about 0.2 mg/kg to about 20 mg/kg and said second polypeptide is present in a dosage between about 10 μg/kg to about 1,000 μg/kg. 
     
     
         128 . The method of any one of  claims 118 - 127 , wherein said first polypeptide and said second polypeptide are administered subcutaneously. 
     
     
         129 . The method of any one of  claims 118 - 128 , wherein said first polypeptide and said second polypeptide are administered one time, two times, or three times per week. 
     
     
         130 . The method of any one of  claims 118 - 129 , wherein said subject is human. 
     
     
         131 . The method of any one of  claims 118 - 130 , wherein said disease is said neurocutaneous syndrome. 
     
     
         132 . The method of  claim 131 , wherein said neurocutaneous syndrome is neurofibromatosis type I. 
     
     
         133 . A kit comprising:
 a) a first polypeptide comprising the structure A-sALP-B, wherein
 i) sALP is the extracellular domain of an alkaline phosphatase, 
 ii) A is absent or is an amino acid sequence of at least one amino acid, and 
 iii) B is absent or is an amino acid sequence of at least one amino acid; and 
   b) instructions for administering said first polypeptide to a patient diagnosed with or at risk of developing a neurocutaneous syndrome.   
     
     
         134 . The kit of  claim 133 , further comprising:
 (c) a second polypeptide comprising the structure V-NP-W, wherein
 i) NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B), 
 ii) V is absent or is an amino acid sequence of at least one amino acid, and 
 iii) W is absent or is an amino acid sequence of at least one amino acid. 
   
     
     
         135 . A kit comprising:
 a) a polypeptide comprising the structure V-NP-W, wherein
 i) NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B), 
 ii) V is absent or is an amino acid sequence of at least one amino acid, and 
 iii) W is absent or is an amino acid sequence of at least one amino acid; and 
   b) instructions for administering said polypeptide to a patient diagnosed with or at risk of developing a neurocutaneous syndrome.   
     
     
         136 . A kit comprising:
 a) a first polypeptide comprising the structure A-sALP-B, wherein
 i) sALP is the extracellular domain of an alkaline phosphatase, 
 ii) A is absent or is an amino acid sequence of at least one amino acid, and 
 iii) B is absent or is an amino acid sequence of at least one amino acid; and 
   b) a second polypeptide comprising the structure V-NP-W, wherein
 i) NP is a natriuretic peptide that is an agonist of natriuretic peptide receptor B (NPR-B), 
 ii) V is absent or is an amino acid sequence of at least one amino acid, and 
 iii) W is absent or is an amino acid sequence of at least one amino acid. 
   
     
     
         137 . The kit of  claim 136 , wherein said first polypeptide and said second polypeptide are formulated together. 
     
     
         138 . The kit of  claim 136 , wherein said first polypeptide and said second polypeptide are formulated separately and in individual dosage amount.