CNP compounds

Abstract

The present application relates to novel natriuretic peptide type C (CNP) compounds, pharmaceutical compositions comprising these compounds and these compounds for use as medicaments.

Description

Technical Field

[0001] This application relates to novel C-type natriuretic peptide (CNP) compounds, pharmaceutical compositions comprising these compounds, and these compounds for use as medicaments. Background Art

[0002] Natriuretic peptides are a family of three structurally related hormones that play unique roles in the cardiovascular system. Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) are expressed in the heart and are released in response to volume-induced atrial and ventricular stretch, respectively. Their physiological effects include regulation of heart structure, blood pressure, and blood volume.

[0003] C-type natriuretic peptide (CNP) is highly expressed in endothelial cells and is constitutively released in endothelial cells. Other cells within the cardiovascular system, including cardiomyocytes and fibroblasts, also produce CNP, although to a lesser extent. CNP has direct effects on inflammation, fibrosis, cardiac contractility, endothelial function, angiogenesis, and blood pressure.

[0004] There are three known receptors for natriuretic peptides. Natriuretic peptide receptor-1 (NPR1) is a particulate guanylate cyclase that catalyzes the synthesis of cGMP upon binding of ANP or BNP. NPR1 is expressed in the kidney, lung, adipose tissue, adrenal gland, brain, heart, testis, and vascular smooth muscle tissue. NPR2 is expressed in bone, brain, fibroblasts, heart, kidney, liver, lung, uterus, and vascular smooth muscle tissue, is homologous to NPR1, but is selectively activated by CNP. In contrast, NPR3 contains only an intracellular domain consisting of 37 residues and lacks guanylate cyclase activity. It controls local natriuretic peptide concentration through receptor-mediated internalization and degradation, but there is increasing evidence that NPR3 has a signaling function. All three natriuretic peptides bind to NPR3 with high affinity, and this receptor is the most widely expressed and abundant of the three receptors. All three of these receptors also have very high conservation, and in agreement with CNP, NPR2 has the highest conservation.

[0005] Clinical and preclinical data have demonstrated the important role of CNP and its two receptors in cardio-renal-metabolic functions, and there is also a growing body of data supporting the therapeutic potential of targeting this system in a wide range of cardiovascular, renal, and metabolic diseases (Int. J. Mol. Sci. 2019, 20, 2281; Cardiovasc. Res. 2022, 118, 2085–2102; Cardiovasc. Res., August 25, 2022; cvac125). Similarly, human genetics as well as preclinical and clinical data support NPR2 and CNP as regulators of bone growth and have therapeutic potential in a wide range of short stature indications related to NPR2 and CNP (Lancet, 396:684; J Pharmacol Exp Ther, 370:459; Hisado-Oliva (2018) Genet Med, 20:91) and in skeletal dysplasias related to FGFR3 (Sabir and Cole Orphanet Journal of Rare Diseases (2019) 14:300; Krakow and Rimoin, Genetics in medicine (2010) 12:6, Decker et al., Human Molecular Genetics (2011), 20:8), as well as in RASopathies (Adv Cancer Res. 153:305). Currently, two CNP compounds are in clinical development for achondroplasia, a type of dwarfism (BMN111 (Vosoritide) and TransCon CNP). Mayo clinic has previously explored a CNP compound designed to activate both NPR2 and NPR1 in heart failure (J Am Coll Cardiol. July 1, 2008; 52(1):60–68).

[0006] The clearance of CNP in human plasma is very rapid, with a calculated half-life of a few minutes. Given this short half-life, it would be beneficial to develop new long-acting CNP compounds that can be administered less frequently while maintaining acceptable clinical properties.

[0007] To provide a longer action profile, a variety of different methods have been employed to modify the structure of CNP. WO 2013 / 058833 discloses a fusion peptide of CNP and an Fc domain. WO 2018 / 175534 discloses a fatty acid-modified NPR1 agonist. SUMMARY OF THE INVENTION

[0008] The present invention provides CNP compounds with improved pharmaceutical properties.

[0009] In one aspect, the present invention provides a CNP compound comprising a CNP peptide and a modifying group, wherein the compound has a net charge of 0 or negative at physiological pH, and wherein the CNP peptide comprises an amino acid sequence according to Formula I:

[0010] AA 01 -AA 02 -AA 03 -AA 04 -AA 05 -AA 06 -AA 07 -AA 08 -AA 09 -AA 10 -AA 11 -AA 12 -AA 13 -AA 14 -AA 15 -AA 16 -AA 17 -AA 18 -AA 19 -AA 20 -AA 21 -AA 22 -AA 23 -AA 24 -AA 25 -AA 26 -AA 27 -AA 28 -AA 29 -AA 30 -AA 31 -AA 32 -AA 33 -AA 34 -AA 35 -AA 36 -AA 37

[0011] wherein

[0012] AA 01 is Gln or absent,

[0013] AA 02 is Glu or absent,

[0014] AA 03 is His or absent,

[0015] AA 04 is Pro or absent,

[0016] AA05 is Asn or Gln or Glu or absent,

[0017] AA 06 is Ala or absent,

[0018] AA 07 is Arg or His or Ala or absent,

[0019] AA 08 is Lys or Ser or His or absent,

[0020] AA 09 is Tyr or Glu or absent

[0021] AA 10 is Lys or Glu or Gln or His or absent,

[0022] AA 11 is Gly,

[0023] AA 12 is Ala,

[0024] AA 13 is Gln or Asn or Glu,

[0025] AA 14 is Lys or His or Glu,

[0026] AA 15 is Lys or Ser or Glu or Thr or His,

[0027] AA 16 is Gly,

[0028] AA 17 is Leu or Gly or Ser or Val,

[0029] AA 18 is Ser or His

[0030] AA 19 is Gln or Ser or Lys or His,

[0031] AA 20 is Gly,

[0032] AA 21 is Cys,

[0033] AA 22 is Phe,

[0034] AA 23 is Gly,

[0035] AA24 is Leu,

[0036] AA 25 is Pro or Lys,

[0037] AA 26 is Leu,

[0038] AA 27 is Asp or Glu,

[0039] AA 28 is Arg,

[0040] AA 29 is Ile,

[0041] AA 30 is Gly,

[0042] AA 31 is Ser,

[0043] AA 32 is Leu or Nle or Met,

[0044] AA 33 is Ser,

[0045] AA 34 is Gly,

[0046] AA 35 is Leu,

[0047] AA 36 is Gly, and

[0048] AA 37 is Cys;

[0049] wherein the modifying group includes Chemical Formula A, Chemical Formula B and Chemical Formula C;

[0050] wherein Chemical Formula A is selected from:

[0051]

[0052] wherein p is an integer in the range of 14 - 20, and

[0053] wherein * represents an amide bond connecting Chemical Formula A and Chemical Formula B; and

[0054] wherein Chemical Formula B is selected from:

[0055]

[0056]

[0057] wherein q is an integer in the range of 1 - 8,

[0058] wherein * represents an amide bond connecting chemical formula A- and chemical formula B-,

[0059] wherein ** represents an amide bond connecting chemical formula B- and chemical formula C-; and

[0060] wherein chemical formula C is selected from:

[0061]

[0062] wherein r is an integer in the range of 0 - 4,

[0063] wherein s is an integer in the range of 0 - 3,

[0064] wherein t is an integer in the range of 0 - 1,

[0065] wherein ** represents an amide bond connecting chemical formula B- and chemical formula C-,

[0066] wherein *** represents an amide bond connecting chemical formula C- and the N-terminal α-amine of the CNP peptide.

[0067] In one embodiment, the present invention provides a CNP compound, wherein the CNP peptide has any of the following amino acid sequences:

[0068] GAQKKGSSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:136),

[0069] ARKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:77),

[0070] YKGAQKKGGSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:88),

[0071] YKGAQKKGLSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:103),

[0072] QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC (SEQ ID NO:67), and

[0073] QEHPQARKYKGAQKKGLSSGCFGLPLERIGSLSGLGC (SEQ ID NO:104).

[0074] On the other hand, the present invention provides a pharmaceutical composition comprising a compound according to the present invention and one or more pharmaceutically acceptable excipients.

[0075] In one aspect, the present invention also provides the use of the compounds of the present invention as a medicament for the treatment or prevention of cardio-renal metabolic diseases including heart failure and achondroplasia.

[0076] The short half-life of wt CNP of about two minutes renders CNP unsuitable for pharmaceutical use. In the present invention, the half-life of CNP is extended by promoting albumin-binding fatty acylation. However, the combination of the positively charged CNP peptide (at physiological pH) and the modification with a fatty acid albumin conjugate (wherein the CNP compound is overall positively charged) results in significant injection site reactions and low bioavailability upon subcutaneous injection. This is surprising because the positively charged CNP peptide without attached fatty acid albumin conjugate does not show observable injection site reactions. To address these issues, amino acid substitutions were introduced into the CNP peptide and negative charges were introduced into the modifying group (including the fatty acid). The combination of these modifications resulted in a compound that is overall negatively charged at physiological pH, which shows good subcutaneous bioavailability and no or only minor subcutaneous injection site reactions.

[0077] Surprisingly, the addition of negative charges also somehow reduces the in vitro and in vivo potency of the CNP compound, such that higher net negative charges are generally associated with lower in vivo potency. To maintain sufficient potency of the CNP compound, only a small amount of net negative charge was introduced. However, when formulated for subcutaneous administration at physiologically relevant pH, the small amount of net negative charge reduces the solubility and biophysical stability of the CNP compound. The present disclosure has achieved a balance in these parameters and surprisingly provides CNP compounds that have sufficient potency and also sufficient solubility and biophysical stability for liquid formulations.

[0078] Furthermore, wt CNP does not exhibit the required chemical stability in liquid formulations for convenient drug administration. In the present invention, selected amino acid substitutions of the wt CNP sequence were introduced to promote chemical stability in the formulation.

[0079] The present disclosure provides such CNP compounds that have an extended half-life and good subcutaneous administration tolerance, while retaining in vivo potency sufficient for therapeutic applications and high chemical and biophysical stability in liquid formulations.

[0080] The present invention can also solve other problems that will be apparent from the disclosure of the exemplary embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] Figure 1 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 0776 to rats as described in Example 11.

[0082] Figure 2 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 0312 to rats as described in Example 11.

[0083] Figure 3 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1225 to rats as described in Example 11.

[0084] Figure 4 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1233 to rats as described in Example 11.

[0085] Figure 5 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1227 to rats as described in Example 11.

[0086] Figure 6 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1241 to rats as described in Example 11.

[0087] Figure 7 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1351 to rats as described in Example 11.

[0088] Figure 8 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1354 to rats as described in Example 11.

[0089] Figure 9 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1356 to rats as described in Example 11.

[0090] Figure 10 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1375 to rats as described in Example 11.

[0091] Figure 11 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1376 to rats as described in Example 11.

[0092] Figure 12 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1377 to rats as described in Example 11.

[0093] Figure 13 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1352 to rats as described in Example 11.

[0094] Figure 14 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1379 to rats as described in Example 11.

[0095] Figure 15 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1434 to rats as described in Example 11.

[0096] Figure 16 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9384 to rats as described in Example 11.

[0097] Figure 17 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9407 to rats as described in Example 11.

[0098] Figure 18 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1420 to rats as described in Example 11.

[0099] Figure 19 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1378 to rats as described in Example 11.

[0100] Figure 20 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1381 to rats as described in Example 11.

[0101] Figure 21 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1386 to rats as described in Example 11.

[0102] Figure 22 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1389 to rats as described in Example 11.

[0103] Figure 23 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1392 to rats as described in Example 11.

[0104] Figure 24 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1426 to rats as described in Example 11.

[0105] Figure 25 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9435 to rats as described in Example 11.

[0106] Figure 26 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1235 to rats as described in Example 11.

[0107] Figure 27 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9482 to rats as described in Example 11.

[0108] Figure 28 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 0312 to minipigs as described in Example 12.

[0109] Figure 29 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 0776 to minipigs as described in Example 12.

[0110] Figure 30 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9384 to minipigs as described in Example 12.

[0111] Figure 31 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9407 to minipigs as described in Example 12.

[0112] Figure 32 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9435 to minipigs as described in Example 12.

[0113] Figure 33 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9480 to minipigs as described in Example 12.

[0114] Figure 34 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9482 to minipigs as described in Example 12.

[0115] Figure 35 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 9483 to minipigs as described in Example 12.

[0116] Figure 36 Shows the pharmacodynamic response (cGMP) after subcutaneous administration of Compound ID 0312 to minipigs as described in Example 12.

[0117] Figure 37 Shows the pharmacodynamic response (cGMP) after subcutaneous administration of Compound ID 0776 to minipigs as described in Example 12.

[0118] Figure 38 Shows the pharmacodynamic response (cGMP) after subcutaneous administration of Compound ID 9384 to minipigs as described in Example 12.

[0119] Figure 39 Shows the pharmacodynamic response (cGMP) after subcutaneous administration of Compound ID 9407 to minipigs as described in Example 12.

[0120] Figure 40 Shows the pharmacodynamic response (cGMP) after subcutaneous administration of Compound ID 9435 to minipigs as described in Example 12.

[0121] Figure 41 Shows the pharmacodynamic response (cGMP) after subcutaneous administration of Compound ID 9480 to minipigs as described in Example 12.

[0122] Figure 42 Shows the pharmacodynamic response (cGMP) after subcutaneous administration of Compound ID 9482 to minipigs as described in Example 12.

[0123] Figure 43 Shows the pharmacodynamic response (cGMP) after subcutaneous administration of Compound ID 9483 to minipigs as described in Example 12.

[0124] Figure 44 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 0312 to domestic LYD pigs as described in Example 12.

[0125] Figure 45 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 0776 to domestic LYD pigs as described in Example 12.

[0126] Figure 46 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1225 to domestic LYD pigs as described in Example 12.

[0127] Figure 47Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1227 to domestic LYD pigs as described in Example 12.

[0128] Figure 48 Shows the pharmacodynamic response (cGMP) after intravenous administration of Compound ID 1235 to domestic LYD pigs as described in Example 12. Detailed Description

[0129] The present invention provides CNP compounds having improved pharmaceutical properties. The present invention also provides pharmaceutical compositions and the use of the compounds and compositions of the present invention as medicaments for treating diseases.

[0130] C-Type Natriuretic Peptide

[0131] C-type natriuretic peptide (CNP) (GenBank accession number NP_077720) is a small single-chain peptide in the peptide family (ANP, BNP, CNP), having a disulfide ring structure of 17 amino acid residues and playing an important role in various biological processes. CNP interacts with natriuretic peptide receptor 2 (NPR2, NPR-B, GC-B), stimulating the production of cyclic guanosine monophosphate (cGMP). CNP is more widely expressed in tissues including the central nervous system, reproductive tract, bone, and vascular endothelium. CNP is cleared rapidly in human plasma with a half-life of several minutes.

[0132] Natural CNP genes and polypeptides have been previously described. U.S. Patent No. 5,352,770 discloses CNP-22 isolated and purified from pig brain, which is identical in sequence to human CNP (human wild-type CNP-22: GLSKGCFGLKLDRIGSMSGLGC (SEQ ID NO: 01)). U.S. Patent No. 6,034,231 discloses the human gene and polypeptide of preproCNP (126 amino acids) and the human CNP-53 gene and peptide. Human wild-type CNP-37 has the sequence QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC (SEQ ID NO: 02).

[0133] CNP Compound

[0134] In one aspect, the present invention provides CNP compounds comprising a CNP peptide and a modifying group, wherein the compound has a net charge of 0 or negative at physiological pH.

[0135] The CNP compounds as described herein have been engineered to achieve compounds that are overall neutral or have a net negative charge. This is accomplished by introducing amino acid substitutions in the CNP peptide and introducing negative charges into the modifying groups. Reducing the charge of the CNP compound while still maintaining the biological activity of the CNP compound towards the NPR2 receptor is not an easy task.

[0136] As used herein, the term "net charge" refers to the sum of all negative and positive charges on the CNP compound at a given pH, where the charge is defined solely by the acid dissociation constants of each ionizable group in the CNP compound.

[0137] In one embodiment, as determined as described in Example 4, the CNP compound has a negative net charge at physiological pH.

[0138] In one embodiment, as determined as described in Example 4, the CNP compound has a net charge of 0 to -4 at physiological pH.

[0139] As used herein, the term "physiological pH" refers to a pH in the range of 7.35 to 7.45 (inclusive), more typically an average of 7.4.

[0140] In one aspect, the CNP compounds described herein have an intramolecular disulfide bond (a disulfide bond between two cysteine residues), thereby forming a cyclic structure.

[0141] CNP Peptide

[0142] As used herein, the term "CNP peptide" refers to a peptide that is 37 to 22 amino acids in length and that comprises the CNP-22 amino acid sequence (SEQ ID NO:01) and its amino acid sequences that include one or more amino acid modifications (such as one or more amino acid substitutions, additions, and / or deletions). As used herein, the term "CNP peptide" also encompasses CNP variants.

[0143] As used herein, the term "CNP variant" refers to a CNP peptide that is an amino acid variant of the CNP-37 sequence (SEQ ID NO:02). In other words, a CNP variant is a CNP peptide that contains one or more amino acid modifications, i.e., at least one amino acid has been changed compared to SEQ ID NO:02. These modifications can independently represent one or more amino acid substitutions, additions, and / or deletions.

[0144] CNP variants can be described by reference to the number of altered amino acid residues (i.e., the corresponding positions in the CNP-37 sequence (SEQ ID NO:02)) and the changes (e.g., the identity of the amino acid residue at a given position in the variant). That is, unless otherwise stated, numerical references to specific amino acid residues of the CNP peptide refer to the CNP-37 sequence (SEQ ID NO:2) (i.e., residue 1 is glutamine (Q1) and residue 37 is cysteine (C37)). The following are non-limiting examples of suitable variant nomenclature: des1-4,5Q,13Q,32Nle CNP variant refers to a CNP peptide sequence having the following amino acid changes compared to CNP-37: deletion of amino acid residues at positions 1-4, and replacement of asparagine (N) at position 5 with glutamine (Q), replacement of asparagine (N) at position 13 with glutamine (Q), and replacement of methionine (M) at position 32 with norleucine.

[0145] The term "amino acid" refers to any amino acid that is either naturally occurring (including the 20 standard amino acids encoded by the standard genetic code in humans) or non-naturally occurring. Amino acid residues can be represented by their full name, their single-letter code, and / or their three-letter code, as is common in the art. These three ways are completely equivalent. The term "non-coding amino acid" refers to all amino acids that do not belong to the 20 standard amino acids encoded by the standard genetic code in humans. Non-coding amino acids can be present in nature or be purely synthetic. Non-limiting examples of non-coding amino acids are D-isomers of coding amino acids, and glycine residues in which the side chain is attached to a nitrogen atom rather than an α-carbon atom. The D-isomers of coding amino acids can be represented as (i) "D-" followed by the full name, single-letter code, or three-letter code of the amino acid, or (ii) the lower-case single-letter code of the amino acid. Other abbreviations for non-coding amino acids used in this application are shown below.

[0146]

[0147] In one aspect of the invention, the CNP peptide comprises an amino acid sequence according to Formula I:

[0148] AA 01 -AA 02 -AA 03 -AA 04 -AA 05 -AA 06 -AA 07 -AA 08 -AA 09 -AA 10 -AA 11 -AA 12 -AA 13 -AA 14 -AA15 -AA 16 -AA 17 -AA 18 -AA 19 -AA 20 -AA 21 -AA 22 -AA 23 -AA 24 -AA 25 -AA 26 -AA 27 -AA 28 -AA 29 -AA 30 -AA 31 -AA 32 -AA 33 -AA 34 -AA 35 -AA 36 -AA 37

[0149] wherein

[0150] AA 01 is Gln or absent,

[0151] AA 02 is Glu or absent,

[0152] AA 03 is His or absent,

[0153] AA 04 is Pro or absent,

[0154] AA 05 is Asn or Gln or Glu or absent,

[0155] AA 06 is Ala or absent,

[0156] AA 07 is Arg or His or Ala or absent,

[0157] AA 08 is Lys or Ser or His or absent,

[0158] AA 09 is Tyr or Glu or absent

[0159] AA 10 is Lys or Glu or Gln or His or absent,

[0160] AA 11 is Gly,

[0161] AA 12 is Ala,

[0162] AA 13 is Gln or Asn or Glu,

[0163] AA 14 is Lys or His or Glu,

[0164] AA 15 is Lys or Ser or Glu or Thr or His,

[0165] AA 16 is Gly,

[0166] AA 17 is Leu or Gly or Ser or Val,

[0167] AA 18 is Ser or His

[0168] AA 19 is Gln or Ser or Lys or His,

[0169] AA 20 is Gly,

[0170] AA 21 is Cys,

[0171] AA 22 is Phe,

[0172] AA 23 is Gly,

[0173] AA 24 is Leu,

[0174] AA 25 is Pro or Lys,

[0175] AA 26 is Leu,

[0176] AA 27 is Asp or Glu,

[0177] AA 28 is Arg,

[0178] AA 29 is Ile,

[0179] AA 30 is Gly,

[0180] AA 31 is Ser,

[0181] AA 32is Leu or Nle or Met,

[0182] AA 33 is Ser,

[0183] AA 34 is Gly,

[0184] AA 35 is Leu,

[0185] AA 36 is Gly, and

[0186] AA 37 is Cys.

[0187] In certain embodiments of the present invention, amino acid substitutions are introduced into the CNP peptide to reduce the positive charge of the CNP peptide. For example, Lys25 (AA 25 ) is replaced with Pro and Lys19 (AA 19 ) is replaced with Gln or Ser to remove the positive charge.

[0188] In certain embodiments of the present invention, the AA 13 of the CNP peptide is Gln; AA 19 is Gln or Ser; AA 25 is Pro; and AA 32 is Leu.

[0189] In certain embodiments of the present invention, the AA 5 of the CNP peptide is Gln; AA 13 is Gln; AA 19 is Gln or Ser; AA 25 is Pro; and AA 32 is Leu.

[0190] In certain embodiments of the present invention, the CNP peptide has any of the following amino acid sequences:

[0191] GAQKKGSSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:136),

[0192] ARKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:77),

[0193] YKGAQKKGGSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:88),

[0194] YKGAQKKGLSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:103),

[0195] QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC (SEQ ID NO:67), and

[0196] QEHPQARKYKGAQKKGLSSGCFGLPLERIGSLSGLGC (SEQ ID NO:104).

[0197] Modifying Group

[0198] In one aspect of the present invention, the CNP compound comprises a modifying group covalently linked to an amino acid residue of the CNP peptide.

[0199] In one aspect of the present invention, the modifying group is capable of forming a non-covalent association with albumin, thereby promoting the circulation of the CNP compound in the bloodstream, which has the effect of prolonging the plasma exposure and plasma half-life of the CNP compound compared to the plasma half-lives of CNP-22 (SEQ ID NO:01) and CNP-37 (SEQ ID NO:02). Therefore, the modifying group having the effect of prolonging the action time of the CNP compound may also be referred to as an extending group.

[0200] In some embodiments of the present invention, the modifying group is covalently linked to the N-terminal amino acid of the CNP peptide.

[0201] In some embodiments of the present invention, the modifying group is covalently linked to the N-terminal α-amine of the CNP peptide.

[0202] In one aspect of the present invention, the modifying group comprises Chemical Formula A, Chemical Formula B, and Chemical Formula C; wherein Chemical Formula A is selected from:

[0203]

[0204] wherein p is an integer in the range of 14 - 20, and

[0205] wherein * represents an amide bond connecting Chemical Formula A and Chemical Formula B; and

[0206] wherein Chemical Formula B is selected from:

[0207]

[0208] wherein q is an integer in the range of 1 - 8,

[0209] wherein * represents an amide bond connecting Chemical Formula A- and Chemical Formula B-,

[0210] wherein ** represents an amide bond connecting Chemical Formula B- and Chemical Formula C-; and

[0211] Among them, chemical formula C is selected from:

[0212]

[0213]

[0214] wherein r is an integer in the range of 0 - 4,

[0215] wherein s is an integer in the range of 0 - 3,

[0216] wherein t is an integer in the range of 0 - 1,

[0217] wherein u is an integer in the range of 0 - 3,

[0218] wherein ** represents an amide bond connecting chemical formula B- and chemical formula C-,

[0219] wherein *** represents an amide bond connecting chemical formula C- and the N-terminal α-amine of the CNP peptide.

[0220] The abbreviations of the modifying group elements used in this application are shown below:

[0221]

[0222]

[0223] In some embodiments of the present invention, the modifying group is selected from the following non-limiting examples: chemical formula E, chemical formula F, chemical formula G, chemical formula H, chemical formula I, and chemical formula J (where the dotted line defines the connection to the CNP peptide through an amide bond).

[0224]

[0225]

[0226] The modifying groups of the present invention can exist in different stereoisomeric forms, which have the same molecular formula and sequence of bonded atoms, differing only in the three-dimensional orientation of their atoms in space. In the experimental section, standard nomenclature is used to illustrate the stereoisomerism of the modifying groups exemplified in the present invention in terms of both name and structure. Unless otherwise specified, the present invention relates to all stereoisomeric forms of the claimed modifying groups.

[0227] Functional Property

[0228] In some aspects of the present invention, the CNP compounds have desirable biophysical properties. The CNP compounds exhibit an extended in vivo half-life. The CNP compounds of the present invention also have biological activity. Further, the compounds of the present invention have desirable stability. In addition, the compounds of the present invention have desirable solubility. Moreover, the compounds of the present invention can be administered subcutaneously without causing an unacceptable degree of local tissue reaction such as necrosis.

[0229] Biological Activity

[0230] In one aspect of the present invention, the compounds have in vitro biological activity, which can be determined based on their ability to activate the NPR2 receptor in vitro in a cell line expressing the NPR2 receptor, and this ability can be determined as their ability to increase the production and concentration of cGMP in the cells, which represents receptor activation downstream of NPR2. The in vitro biological activity can be measured, for example, as described in Example 2.

[0231] In one aspect of the present invention, the compounds have in vivo biological activity, which can be determined based on their ability to increase the concentration of cGMP in plasma after intravenous or subcutaneous administration to rats or minipigs or domestic LYD pigs.

[0232] Rats are an example of a suitable animal model, and the activity can be measured in such rats as described in Example 11.

[0233] Minipigs or domestic LYD pigs are other examples of suitable animal models, and the activity can be measured in such pigs as described in Example 12.

[0234] Plasma Half-Life

[0235] In one aspect of the present invention, the CNP compounds have extended plasma exposure and an extended in vivo plasma half-life relative to native CNP, which can be determined in a suitable in vivo pharmacokinetic study. The extended plasma exposure can be determined as the plasma half-life (T / 1 / 2 ) after intravenous or subcutaneous administration to animals such as rats,

[0236] In some embodiments of the present invention, as measured in Example 11, the plasma half-life of the CNP compound after intravenous administration to rats is at least 4 hours, more preferably at least 8 hours, or most preferably at least 10 hours.

[0237] In some embodiments of the present invention, as measured in Example 12, the CNP compound is administered intravenously or subcutaneously to The in vivo plasma half-life in minipigs is at least 10 hours, more preferably at least 40 hours, or most preferably at least 60 hours.

[0238] Bioavailability

[0239] In one aspect of the invention, the compound has suitable bioavailability after subcutaneous injection. Bioavailability (F%) can be determined in any suitable animal model as is known in the art.

[0240] Minipigs or domestic LYD pigs are examples of suitable animal models and bioavailability can be determined in such minipigs or domestic LYD pigs as described in Example 12.

[0241] In some embodiments of the invention, the bioavailability of the CNP compound after subcutaneous administration to minipigs is at least 40%, 50%, 60% or 70%.

[0242] Stability

[0243] In one aspect of the invention, the compound has suitable physical and chemical stability. Lack of physical or chemical stability can lead to changes in the compound structure, resulting in the formation of chemical degradation products which may have reduced biological activity, reduced solubility and / or enhanced immunogenic effects compared to the intact compound.

[0244] Physical stability can be evaluated by measuring the tendency to form fibrils, for example using the ThT assay described in Example 6.

[0245] Chemical stability can be evaluated by measuring the amount of chemical degradation products (such as isomers, isoAsp and hydrolysis products) at different time points after exposure to different conditions such as high temperature, for example, as described in Example 8.

[0246] Stability can also be evaluated by studying the ability of the molecule to form covalent dimers and multimers - called HMWP - for example, as described in Example 7.

[0247] Solubility

[0248] According to the functional aspect of the invention, the CNP compound has desirable solubility. Low solubility of CNP can severely impede its pharmaceutical formulation properties and therapeutic use, and thus development of CNP compounds with high solubility in relevant formulations will improve therapeutic utility.

[0249] Solubility is measured as described in Example 5 herein.

[0250] In certain embodiments, the solubility of the CNP compounds of the present invention is at least 2000 μM, 3000 μM or 4000 μM at pH 4 or pH 6.5.

[0251] Pharmaceutical Composition

[0252] In one aspect, the present invention relates to a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable excipient. The composition is suitable for parenteral administration.

[0253] In one embodiment, the concentration of the compound in the formulation is from about 0.1 mg / ml to about 50 mg / ml. In another aspect, the concentration of the compound in the formulation is from about 10 mg / ml to about 30 mg / ml.

[0254] In another embodiment, the pH of the formulation is from 3.0 to 8.0. In another embodiment, the pH of the formulation is from 3.5 to 5.5. In a further embodiment, the pH of the formulation is from 6.0 to 7.0. The pharmaceutical composition containing the compound of the present invention can be prepared by conventional techniques, for example, as described in Remington’s Pharmaceutical Sciences, 1985 or Remington: The Science and Practice of Pharmacy, 19th Edition, 1995.

[0255] The formulation may further comprise a buffer system, preservatives, isotonic agents, chelating agents, stabilizers and / or surfactants. The use of such excipients in pharmaceutical compositions is well known to those skilled in the art. For convenience, reference is made to Remington: The Science and Practice of Pharmacy, 19th Edition, 1995.

[0256] In one embodiment, the pharmaceutical formulation is an aqueous formulation, i.e., a formulation containing water. Such formulations are typically solutions or suspensions. In a further embodiment of the present invention, the pharmaceutical formulation is an aqueous solution. The term “aqueous formulation” is defined as a formulation containing at least 50% w / w water. Similarly, the term “aqueous solution” is defined as a solution containing at least 50% w / w water, and the term “aqueous suspension” is defined as a suspension containing at least 50% w / w water.

[0257] In another embodiment, the pharmaceutical formulation is a lyophilized formulation to which a solvent and / or diluent is added by a physician or patient prior to use. In another embodiment, the pharmaceutical formulation is a dry formulation (e.g., lyophilized or spray-dried) and can be used without any prior dissolution. The so-called "dry form" refers to the drying of a liquid pharmaceutical composition or formulation by lyophilization (i.e., freeze-drying; see, e.g., Williams and Polli (1984) J. Parenteral Sci. Technol. 38:48-59), spray-drying (see Masters (1991), Spray-Drying Handbook (5th ed.; Longman Scientific and Technical, Essez, U.K.), pp. 491-676; Broadhead et al. (1992) Drug Devel. Ind. Pharm. 18:1169-1206; and Mumenthaler et al. (1994) Pharm. Res. 11:12-20) or air-drying (Carpenter and Crowe (1988) Cryobiology 25:459-470; and Roser (1991) Biopharm. 4:47-53).

[0258] In a further embodiment of the invention, the buffer is selected from acetate, carbonate, citrate, glycylglycine, histidine, glycine, lysine, arginine, dihydrogen phosphate, hydrogen phosphate, phosphate, and tris(hydroxymethyl)aminomethane, bicine (N-bis[2-hydroxyethyl]glycine), tricine (N-[tris(hydroxymethyl)methyl]glycine), malic acid, lactic acid, succinate, maleic acid, fumaric acid, tartaric acid, aspartic acid, or a mixture thereof. Each of these specific buffers constitutes an alternative embodiment of the invention.

[0259] In another embodiment of the invention, the formulation further comprises a pharmaceutically acceptable preservative. In a further embodiment of the invention, the formulation further comprises an isotonic agent such as propylene glycol, mannitol, or glycerol. In a further embodiment of the invention, the formulation further comprises a chelating agent.

[0260] In another embodiment of the invention, the formulation further comprises a stabilizer. The use of stabilizers in pharmaceutical compositions is well known to those skilled in the art. For convenience, reference is made to Remington: The Science and Practice of Pharmacy, 19th ed., 1995.

[0261] The pharmaceutical composition of the present invention may further comprise an amount of amino acid base sufficient to reduce aggregate formation of the peptide during storage of the composition.

[0262] As used herein, "amino acid base" refers to an amino acid or combination of amino acids, where any given amino acid is present in its free base form or salt form. When using a combination of amino acids, all amino acids can be present in their free base form, all amino acids can be present in their salt form, or some amino acids can be present in their free base form while other amino acids are present in their salt form. In one embodiment, the amino acids used to prepare the compositions of the present invention are amino acids with charged side chains, such as arginine, lysine, aspartic acid, and glutamic acid. Any stereoisomer (i.e., L-form, D-form, or a mixture thereof) of a particular amino acid (e.g., methionine, histidine, imidazole, arginine, lysine, isoleucine, aspartic acid, tryptophan, threonine, and mixtures thereof) or a combination of these stereoisomers may be present in the pharmaceutical composition of the present invention, provided that the particular amino acid is present in its free base form or its salt form. In one embodiment, the L-stereoisomer is used. The compositions of the present invention may also be formulated with derivatives of these amino acids. Suitable arginine derivatives include, for example, aminoguanidine, ornithine, and N-monoethyl-L-arginine, suitable methionine derivatives include ethionine and buthionine, and suitable cysteine derivatives include S-methyl-L-cysteine. Like other amino acids, the amino acid derivatives are incorporated into the composition in their free base form or their salt form. In another embodiment of the present invention, the amino acid or its amino acid derivative is used at a concentration sufficient to prevent or delay protein aggregation.

[0263] In another embodiment of the present invention, the formulation further comprises a surfactant. In another embodiment of the present invention, the formulation further comprises a protease inhibitor. The use of protease inhibitors is particularly useful in pharmaceutical compositions containing zymogens of proteases to inhibit autocatalysis.

[0264] Other components may also be present in the pharmaceutical formulations of the present invention. Such additional components may include wetting agents, emulsifying agents, antioxidants, fillers, tonicity regulators, chelating agents, metal ions, oily vehicles, proteins (e.g., human serum albumin, gelatin, or proteins), and zwitterions (e.g., amino acids such as betaine, taurine, arginine, glycine, lysine, and histidine). Of course, such additional components should not adversely affect the overall stability of the pharmaceutical formulations of the present invention.

[0265] The pharmaceutical composition of the present invention can be administered to a patient in need of such treatment at several sites, for example, at a local site, such as the skin and mucosal sites, at sites of bypass absorption, such as in arteries, veins, the heart, and at sites involving absorption, such as in the skin, subcutaneously, intramuscularly, or abdominally.

[0266] The pharmaceutical composition of the present invention can be administered by various routes of administration, for example, to a patient in need of such treatment by tongue, sublingual, buccal, in the mouth, oral, in the stomach and intestine, nasal, lung (e.g., via bronchioles and alveoli or a combination thereof), epidermal, dermal, transdermal, vaginal, rectal, eye (e.g., via the conjunctiva), ureter, and parenterally.

[0267] The composition of the present invention can be administered in various dosage forms, for example, as a solution, suspension, emulsion, microemulsion, multiple emulsion, foam, ointment, paste, plaster, unguent, tablet, coated tablet, rinse, capsule (e.g., hard gelatin capsule and soft gelatin capsule), suppository, rectal capsule, drops, gel, spray, powder, aerosol, inhalant, eye drops, ophthalmic ointment, ophthalmic rinse, vaginal suppository, vaginal ring, vaginal ointment, injection solution, in-situ conversion solution (e.g., in-situ gelling, in-situ solidification, in-situ precipitation, in-situ crystallization), infusion solution, and implant.

[0268] The composition of the present invention can be further incorporated or attached to drug carriers, drug delivery systems, and advanced drug delivery systems, for example, by covalent, hydrophobic, and electrostatic interactions, in order to further enhance the stability of its compounds, improve bioavailability, increase solubility, reduce adverse reactions, achieve chronotherapy well-known to those skilled in the art, and improve patient compliance or any combination thereof.

[0269] The composition of the present invention can be used to formulate solids, semi-solids, powders, and solutions for pulmonary administration of the CNP compound of the present invention using, for example, metered-dose inhalers, dry powder inhalers, and nebulizers, all of which are devices well-known to those skilled in the art.

[0270] The composition of the present invention can be used to formulate controlled, sustained, extended, delayed, and slow-release drug delivery systems.

[0271] Parenteral administration can be carried out by injection subcutaneously, intramuscularly, intraperitoneally, or intravenously with a syringe, optionally with a pen syringe. Alternatively, parenteral administration can also be carried out with an infusion pump. A further option is a composition which can be a solution or suspension for administering the CNP compound of the present invention in the form of a nasal or lung spray. As a still further option, the pharmaceutical composition containing the peptide of the present invention is also suitable for transdermal administration, for example, by needleless injection or patch, optionally an iontophoretic patch, or transmucosal administration, for example, buccal administration.

[0272] The CNP compounds of the present invention can be administered via the pulmonary route in a vehicle in the form of a solution, suspension or dry powder using any known type of device suitable for pulmonary drug delivery. Examples of these include, but are not limited to, the three general types of aerosol generating devices for pulmonary drug delivery, and may include jet or ultrasonic nebulizers, metered dose inhalers or dry powder inhalers (see Yu J, Chien YW. Pulmonary drug delivery: Physiologic and mechanistic aspects. Crit Rev Ther Drug Carr Sys 14(4)(1997)395 - 453).

[0273] In one embodiment of the present invention, the pharmaceutical formulation comprising the compound of the present invention is stable when used for more than 6 weeks and stored for more than 3 years. In another embodiment of the present invention, the pharmaceutical formulation comprising the compound of the present invention is stable when used for more than 2 weeks and stored for more than 2 years.

[0274] In one aspect, a method of preparing a pharmaceutical composition comprising a compound of the present invention comprises mixing the compound of the present invention with at least one pharmaceutically acceptable excipient.

[0275] Medical Indication

[0276] The present invention also relates to compounds for use as medicaments.

[0277] As used herein, the term "treatment" refers to medical treatment of any human subject in need thereof. Depending on the health status of the subject, the timing and purpose of said treatment may vary from individual to individual. The treatment can be prophylactic, palliative, symptomatic and / or curative.

[0278] In one aspect, the compounds of the present invention can be used to treat cardio - renal metabolic diseases. Cardio - renal metabolic diseases include, but are not limited to, hypertension, arteriosclerosis, atherosclerosis, restenosis, acute myocardial infarction, pulmonary hypertension, acute decompensated heart failure, congestive heart failure, cardiac edema, renal edema, hepatic edema*, acute renal insufficiency, chronic renal insufficiency, insulin resistance and type 2 diabetes.

[0279] In one embodiment, the compounds of the present invention are used to treat congestive heart failure.

[0280] In one embodiment, the compounds of the present invention are used to treat acute decompensated heart failure.

[0281] In one aspect, the compounds of the invention can be used to treat growth disorders, including short stature that may be associated with FGFR3-related skeletal dysplasias, and related comorbidities.

[0282] In some embodiments, the compounds of the invention can be used to treat diseases selected from the group consisting of achondroplasia, hypochondroplasia, lethal dysplasia types 1 and 2, SHOX deficiency, Noonan syndrome, Costello syndrome, LEOPARD syndrome, idiopathic short stature, autosomal dominant short stature, growth hormone deficiency, hypophosphatemic rickets, CNP deficiency, aggrecan deficiency, heterozygous NPR2 mutations, osteoarthritis, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome or Crouzonodermoskeletal syndrome), lacrimal duct-auricular-dental-digital syndrome (LADD), osteoglophonic dysplasia, and SADDAN (severe achondroplasia dysplasia delay acanthosis nigricans).

[0283] In some embodiments, the compounds of the invention can be used to treat diseases selected from osteogenesis imperfecta, achondroplasia, chondrodysplasia punctata, homozygous achondroplasia, rhizomelic chondrodysplasia punctata, congenital spondyloepiphyseal dysplasia, congenital short femur, Langer type mesomelic dysplasia, neurofibromatosis, Legius syndrome, and neurofibromatosis type 1.

[0284] In some embodiments, the compounds of the invention can be used to treat phenotypes associated with growth disorders, including short stature that may be associated with FGFR3-related skeletal dysplasias, selected from growth retardation, cranial deformities, dental orthopedic defects, cervical spinal cord compression, spinal stenosis, pain associated with skeletal dysplasia, hydrocephalus, hearing loss caused by chronic otitis media, cardiovascular diseases, neurological diseases, and obesity.

[0285] Unless otherwise specified in the specification, terms presented in the singular may also include the plural.

[0286] List of embodiments

[0287] The invention is further described by the following non-limiting embodiments:

[0288] Embodiment 1: A CNP compound comprising a CNP peptide and a modifying group, wherein the compound has a net charge of 0 or negative at physiological pH, and wherein the CNP peptide comprises an amino acid sequence according to Formula I:

[0289] AA01 -AA 02 -AA 03 -AA 04 -AA 05 -AA 06 -AA 07 -AA 08 -AA 09 -AA 10 -AA 11 -AA 12 -AA 13 -AA 14 -AA 15 -AA 16 -AA 17 -AA 18 -AA 19 -AA 20 -AA 21 -AA 22 -AA 23 -AA 24 -AA 25 -AA 26 -AA 27 -AA 28 -AA 29 -AA 30 -AA 31 -AA 32 -AA 33 -AA 34 -AA 35 -AA 36 -AA 37

[0290] Among them

[0291] AA 01 is Gln or does not exist,

[0292] AA 02 is Glu or does not exist,

[0293] AA 03 is His or does not exist,

[0294] AA 04 is Pro or does not exist,

[0295] AA 05 is Asn or Gln or Glu or does not exist,

[0296] AA 06 is Ala or does not exist,

[0297] AA 07 is Arg or His or Ala or does not exist,

[0298] AA 08 is Lys or Ser or His or absent,

[0299] AA 09 is Tyr or Glu or absent

[0300] AA 10 is Lys or Glu or Gln or His or absent,

[0301] AA 11 is Gly,

[0302] AA 12 is Ala,

[0303] AA 13 is Gln or Asn or Glu,

[0304] AA 14 is Lys or His or Glu,

[0305] AA 15 is Lys or Ser or Glu or Thr or His,

[0306] AA 16 is Gly,

[0307] AA 17 is Leu or Gly or Ser or Val,

[0308] AA 18 is Ser or His

[0309] AA 19 is Gln or Ser or Lys or His,

[0310] AA 20 is Gly,

[0311] AA 21 is Cys,

[0312] AA 22 is Phe,

[0313] AA 23 is Gly,

[0314] AA 24 is Leu,

[0315] AA 25 is Pro or Lys,

[0316] AA 26 is Leu,

[0317] AA 27is Asp or Glu,

[0318] AA 28 is Arg,

[0319] AA 29 is Ile,

[0320] AA 30 is Gly,

[0321] AA 31 is Ser,

[0322] AA 32 is Leu or Nle or Met,

[0323] AA 33 is Ser,

[0324] AA 34 is Gly,

[0325] AA 35 is Leu,

[0326] AA 36 is Gly, and

[0327] AA 37 is Cys;

[0328] wherein the modifying group includes chemical formula A, chemical formula B and chemical formula C;

[0329] wherein chemical formula A is selected from:

[0330]

[0331] where p is an integer in the range of 14 - 20, and

[0332] where * represents an amide bond connecting chemical formula A and chemical formula B; and

[0333] wherein chemical formula B is selected from: chemical formula B1 and chemical formula B2

[0334]

[0335] where q is an integer in the range of 1 - 8,

[0336] where * represents an amide bond connecting chemical formula A- and chemical formula B-,

[0337] where ** represents an amide bond connecting chemical formula B- and chemical formula C-; and

[0338] wherein chemical formula C is selected from:

[0339]

[0340] (Chemical formula C3)

[0341] where r is an integer in the range of 0 - 4,

[0342] where s is an integer in the range of 0 - 3,

[0343] where t is an integer in the range of 0 - 1, where

[0344] ** represents the amide bond connecting chemical formula B- and chemical formula C-

[0345] where *** represents the amide bond connecting chemical formula C- and the N-terminal α-amine of the CNP peptide.

[0346] Embodiment 2: The CNP compound according to Embodiment 1, wherein the AA of the CNP peptide 13 is Gln; AA 19 is Gln or Ser; AA 25 is Pro; and AA 32 is Leu.

[0347] Embodiment 3: The CNP compound according to Embodiment 1, wherein the AA of the CNP peptide 5 is Gln; AA 13 is Gln; AA 19 is Gln or Ser; AA 25 is Pro; and AA 32 is Leu.

[0348] Embodiment 4: The CNP compound according to Embodiment 1, wherein the CNP peptide comprises one of the following amino acid sequences:

[0349]

[0350]

[0351]

[0352]

[0353]

[0354] Embodiment 5: The CNP compound according to Embodiment 1, wherein the CNP peptide has any of the following amino acid sequences:

[0355]

[0356]

[0357]

[0358]

[0359]

[0360]

[0361] Embodiment 6: The CNP compound according to Embodiment 1, wherein the CNP peptide has any of the following amino acid sequences:

[0362] GAQKKGSSQGCFGLPLDRIGSLSGLGC (SEQ ID NO: 136),

[0363] ARKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC (SEQ ID NO: 77),

[0364] YKGAQKKGGSQGCFGLPLDRIGSLSGLGC (SEQ ID NO: 88), YKGAQKKGLSQGCFGLPLDRIGSLSGLGC (SEQ ID NO: 103),

[0365] QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC (SEQ ID NO: 67), and

[0366] QEHPQARKYKGAQKKGLSSGCFGLPLERIGSLSGLGC (SEQ ID NO: 104).

[0367] Embodiment 7: The CNP compound according to Embodiment 1, wherein the CNP peptide has the following amino acid sequence:

[0368] QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC (SEQ ID NO: 67).

[0369] Embodiment 8: The CNP compound according to any of the preceding embodiments, wherein the modifying group is covalently linked to the N-terminal α-amine of the CNP peptide.

[0370] Embodiment 9: The CNP compound according to any of the preceding embodiments, wherein p in the chemical formula A of the modifying group is 16.

[0371] Embodiment 10: The CNP compound according to Embodiments 1-8, wherein the chemical formula A of the modifying group is Chemical Formula A2

[0372]

[0373] where p is 16.

[0374] Embodiment 11: A CNP compound according to any one of the foregoing embodiments, wherein q in the chemical formula B of the modifying group is 1 - 5.

[0375] Embodiment 12: A CNP compound according to any one of the foregoing embodiments, wherein the modifying group is selected from the following non - limiting examples: chemical formula E, chemical formula F, chemical formula G, chemical formula H, chemical formula I, and chemical formula J (where the dashed line defines the connection to the CNP peptide via an amide bond)

[0376]

[0377]

[0378] Embodiment 13: A CNP compound according to any one of the foregoing embodiments, wherein the modifying group is chemical formula I.

[0379] Embodiment 14: A CNP compound comprising a CNP peptide and a modifying group, the modifying group comprising chemical formula A, chemical formula B, and chemical formula C, wherein the compound is any one of the compounds disclosed in Table 1 (see below).

[0380] Embodiment 15: A CNP compound comprising a CNP peptide and a modifying group, wherein the compound is any one of the compounds disclosed in Table 1a.

[0381] Table 1a:

[0382]

[0383]

[0384] Embodiment 16: A CNP compound comprising a CNP peptide and a modifying group, wherein the CNP peptide has the following amino acid sequence,

[0385] QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC (SEQ ID NO:67), and wherein the modifying group is chemical formula I.

[0386] Embodiment 17: A CNP compound according to Formula II:

[0387]

[0388] Embodiment 18: A CNP compound according to any one of Formulas II, III, IV, V, VI, and VII:

[0389]

[0390]

[0391] Embodiment 19: The compound according to any one of Embodiments 1-18, wherein as determined in Example 4, the net charge of the CNP compound at physiological pH is from 0 to -4.

[0392] Embodiment 20: The compound according to any one of Embodiments 1-18, wherein as determined in Example 12, the compound has an in vivo half-life of at least 10 hours after subcutaneous administration to minipigs.

[0393] Embodiment 21: The compound according to any one of Embodiments 1-18, wherein as determined in Example 12, the compound has an in vivo half-life of at least 20 hours after subcutaneous administration to minipigs.

[0394] Embodiment 22: The compound according to any one of Embodiments 1-18, wherein as determined in Example 12, the compound has an in vivo half-life of at least 30 hours after subcutaneous administration to minipigs.

[0395] Embodiment 23: The compound according to any one of Embodiments 1-18, wherein as determined in Example 12, the compound has an in vivo half-life of at least 40 hours after subcutaneous administration to minipigs.

[0396] Embodiment 24: The compound according to any one of Embodiments 1-18, wherein as determined in Example 12, the compound has an in vivo half-life of at least 50 hours after subcutaneous administration to minipigs.

[0397] Embodiment 25: The compound according to any one of Embodiments 1-18, wherein as determined in Example 12, the compound has a bioavailability of at least 40% after subcutaneous administration to minipigs.

[0398] Embodiment 26: The compound according to any one of Embodiments 1-18, wherein as determined in Example 12, the compound has a bioavailability of at least 50% after subcutaneous administration to minipigs.

[0399] Embodiment 27: The compound according to any one of Embodiments 1-18, wherein as determined in Example 12, the bioavailability of the compound after subcutaneous administration to minipigs is at least 60%.

[0400] Embodiment 28: The compound according to any one of Embodiments 1-18, wherein as determined in Example 12, the bioavailability of the compound after subcutaneous administration to minipigs is at least 70%.

[0401] Embodiment 29: The compound according to any one of Embodiments 1-18, wherein as measured in Example 5, the solubility of the compound at pH 4 or pH 6.5 is at least 2000 μM.

[0402] Embodiment 30: The compound according to any one of Embodiments 1-18, wherein as measured in Example 5, the solubility of the compound at pH 4 or pH 6.5 is at least 3000 μM.

[0403] Embodiment 31: The compound according to any one of Embodiments 1-18, wherein as measured in Example 5, the solubility of the compound at pH 4 or pH 6.5 is at least 4000 μM.

[0404] Embodiment 32: The compound according to any one of Embodiments 1-31, wherein the compound contains an intramolecular disulfide bond, resulting in a cyclic structure.

[0405] Embodiment 33: A pharmaceutical composition comprising the compound according to any one of Embodiments 1-32 and one or more pharmaceutically acceptable excipients.

[0406] Embodiment 34: The compound according to any one of Embodiments 1-32, or the composition according to Embodiment 33, for the treatment or prevention of cardio-renal metabolic diseases.

[0407] Embodiment 35: The compound according to any one of Embodiments 1-32, or the composition according to Embodiment 33, for the treatment or prevention of cardio-renal metabolic diseases, including hypertension, arteriosclerosis, atherosclerosis, restenosis, acute myocardial infarction, pulmonary hypertension, acute decompensated heart failure, congestive heart failure, cardiac edema, renal edema, hepatic edema*, acute renal insufficiency, chronic renal insufficiency, insulin resistance, and type 2 diabetes.

[0408] Embodiment 36: The compound according to any one of Embodiments 1-32, or the composition according to Embodiment 33, for the treatment or prevention of congestive heart failure.

[0409] Embodiment 37: The compound according to any one of Embodiments 1-32, or the composition according to Embodiment 33, for the treatment or prevention of decompensated heart failure.

[0410] Embodiment 38: The compound according to any one of Embodiments 1-32, or the composition according to Embodiment 33, for the treatment or prevention of growth disorders, including short stature possibly associated with FGFR3-related skeletal dysplasia, and related comorbidities.

[0411] Embodiment 39: The compound according to any one of Embodiments 1-32, or the composition according to Embodiment 33, for the treatment or prevention of achondroplasia, chondrodysplasia, lethal dysplasia types 1 and 2, SHOX deficiency, Noonan syndrome, Costello syndrome, LEOPARD syndrome, idiopathic short stature, autosomal dominant short stature, growth hormone deficiency, hypophosphatemic rickets, CNP deficiency, aggrecan deficiency, heterozygous NPR2 mutation, osteoarthritis, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome or Crouzonodermoskeletal syndrome), lacrimal duct - ear - tooth - finger syndrome (LADD), otochondrodysplasia and SADDAN (severe achondroplasia dysplasia delay acanthosis nigricans).

[0412] Embodiment 40: The compound according to any one of Embodiments 1-32, or the composition according to Embodiment 33, for the treatment or prevention of osteogenesis imperfecta, achondroplasia, stippled chondrodysplasia, homozygous achondroplasia, rhizomelic chondrodysplasia punctata, congenital spondyloepiphyseal dysplasia, congenital short femur, Langer type mesomelic dysplasia, neurofibromatosis, Legius syndrome and neurofibromatosis type 1.

[0413] Embodiment 41: The compound according to any one of Embodiments 1-32, or the composition according to Embodiment 33, for the treatment or prevention of phenotypes associated with growth disorders, including short stature possibly associated with FGFR3-related skeletal dysplasia, selected from growth retardation, cranial deformity, dental orthopedic defects, cervical spinal cord compression, spinal stenosis, pain associated with skeletal dysplasia, hydrocephalus, hearing loss caused by chronic otitis media, cardiovascular diseases, neurological diseases and obesity.

[0414] Embodiment 42: A method for treating or preventing cardio-renal-metabolic diseases, which comprises administering to a patient in need thereof an effective amount of a compound according to any one of Embodiments 1-32, or a composition according to Embodiment 33.

[0415] Embodiment 43: A method for treating or preventing cardio-renal-metabolic diseases, said cardio-renal-metabolic diseases including hypertension, arteriosclerosis, atherosclerosis, restenosis, acute myocardial infarction, pulmonary hypertension, acute decompensated heart failure, congestive heart failure, cardiac edema, renal edema, hepatic edema*, acute renal insufficiency, chronic renal insufficiency, insulin resistance and type 2 diabetes, said method comprising administering to a patient in need thereof an effective amount of a compound according to any one of Embodiments 1-32, or a composition according to Embodiment 33.

[0416] Embodiment 44: A method for treating or preventing growth disorders, including short stature and related comorbidities that may be associated with FGFR3-related skeletal dysplasia, said method comprising administering to a patient in need thereof an effective amount of a compound according to any one of Embodiments 1-32, or a composition according to Embodiment 33.

[0417] Embodiment 45: A method for treating or preventing achondroplasia, chondrodysplasia, lethal dysplasia types 1 and 2, SHOX deficiency, Noonan syndrome, Costello syndrome, LEOPARD syndrome, idiopathic short stature, autosomal dominant short stature, growth hormone deficiency, hypophosphatemic rickets, CNP deficiency, aggrecan deficiency, heterozygous NPR2 mutations, osteoarthritis, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome or Crouzonodermoskeletal syndrome), lacrimal duct-ear-dental-finger syndrome (LADD), otochondrodysplasia and SADDAN (severe achondroplasia dysplasia delay acanthosis nigricans), said method comprising administering to a patient in need thereof an effective amount of a compound according to any one of Embodiments 1-32, or a composition according to Embodiment 33.

[0418] Embodiment 46: A method for treating or preventing osteogenesis imperfecta, achondroplasia, chondrodysplasia punctata, homozygous achondroplasia, rhizomelic chondrodysplasia punctata, congenital spondyloepiphyseal dysplasia, congenital short femur, Langer type mesomelic dysplasia, neurofibromatosis, Legius syndrome and neurofibromatosis type 1, said method comprising administering to a patient in need thereof an effective amount of a compound according to any one of Embodiments 1-32, or a composition according to Embodiment 33.

[0419] Embodiment 47: A method for treating or preventing a phenotype associated with a growth disorder, said growth disorder including short stature that may be related to FGFR3-related skeletal dysplasia, selected from growth retardation, cranial deformity, dental orthopedic defects, cervical spinal cord compression, spinal stenosis, pain associated with skeletal dysplasia, hydrocephalus, hearing loss caused by chronic otitis, cardiovascular diseases, neurological diseases, and obesity, said method comprising administering to a patient in need thereof an effective amount of a compound according to any one of Embodiments 1-32, or a composition according to Embodiment 33.

[0420] Example

[0421] Materials and Methods

[0422] List of Abbreviations

[0423] 2-ClTrt 2-chlorotrityl resin

[0424] Aldrithiol-4 4,4′-dipyridyldisulfide

[0425] AUC area under the curve

[0426] BEH ethylene-bridged hybrid

[0427] Boc tert-butoxycarbonyl

[0428] C18d octadecanedioic acid

[0429] C20d eicosanedioic acid

[0430] CAD charged aerosol detector

[0431] cGMP cyclic guanosine monophosphate

[0432] CL clearance rate

[0433] CSH charged surface hybrid

[0434] DCM dichloromethane

[0435] DgGlu D-configured γ-glutamyl

[0436] DIC N,N′-diisopropylcarbodiimide

[0437] DMEM Dulbecco's Modified Eagle Medium

[0438] DMF dimethylformamide

[0439] DMSO dimethyl sulfoxide

[0440] DTT Dithiothreitol

[0441] EDTA Ethylenediaminetetraacetic acid

[0442] ESI Electrospray ionization

[0443] F% Bioavailability

[0444] Fd Faraday

[0445] Fmoc 9-Fluorenylmethyloxycarbonyl

[0446] FPLC Fast protein liquid chromatography

[0447] gGlu γ-Glutamyl

[0448] HEPES 2-[4-(2-Hydroxyethyl)piperazin-1-yl]ethanesulfonic acid

[0449] HFIP 1,1,1,3,3,3-Hexafluoro-2-propanol

[0450] HMWP High molecular weight protein

[0451] HSA Human serum albumin

[0452] i.v. Intravenous

[0453] IPA Isopropyl alcohol

[0454] LCMS Liquid chromatography - mass spectrometry

[0455] LLOQ Lower limit of quantitation

[0456] LYD Landrace, Yorkshire & Duroc

[0457] MeCN Acetonitrile

[0458] MMPX 8-Methoxymethyl-3-isobutyl-1-methylxanthine

[0459] MS Mass spectrometry

[0460] NAD No abnormality detected

[0461] NCA Non-compartmental analysis

[0462] n.d. Not detected

[0463] Nle(X) Norleucine

[0464] NMP N-Methyl-2-pyrrolidone

[0465] OEG Oligo(ethylene glycol)

[0466] Oxyma Pure Ethyl Cyano(hydroxyimino)acetate

[0467] Pbf 2,2,4,6,7-Pentamethyldihydrobenzofuran-5-sulfonyl

[0468] PBS Phosphate Buffered Saline

[0469] PD Pharmacodynamics

[0470] PDA Photodiode Array Detection

[0471] PK Pharmacokinetics

[0472] QC Quality Control

[0473] RP Reverse Phase

[0474] RPM Revolutions Per Minute

[0475] s.c. Subcutaneous

[0476] SEC Size Exclusion Chromatography

[0477] SIL Stable Isotope Labeled

[0478] SPPS Solid Phase Peptide Synthesis

[0479] T 1 / 2 Half-life

[0480] tBu tert-Butyl

[0481] Tetrazole-C18 17-(2H-Tetrazol-5-yl)heptadecanoic acid

[0482] TF Time of Flight

[0483] TFA Trifluoroacetic Acid

[0484] ThT Thioflavin T

[0485] TIPS Triisopropylsilane

[0486] Tris 2-Amino-2-(hydroxymethyl)propane-1,3-diol

[0487] Trt Triphenylmethyl

[0488] TUV Tunable Ultraviolet

[0489] UPLC Ultra Performance Liquid Chromatography

[0490] UV Ultraviolet

[0491] General Preparation Method

[0492] Method A - Solid-Phase Peptide Synthesis

[0493] Peptides can be synthesized by solid-phase peptide synthesis well-known in the art, and the method for synthesizing the exemplified compounds of the present invention is described below.

[0494] The synthesis of the compounds was carried out on a Symphony X solid-phase peptide synthesizer (GYROS Protein Technologies AB, Tucson, AZ).

[0495] Typically, 450 μmol of resin was used for the synthesis, but synthesis using 150 or 300 μmol of resin could also be employed. In a typical synthesis, the resin was washed with DMF. After washing with DMF, the resin was incubated with an Fmoc-protected amino acid (5 equivalents, 0.3 M in a DMF solution of 0.3 M Oxyma Pure, 7.5 mL), DIG (5 equivalents, 0.75 M in DMF, 3.0 mL), and collidine (5 equivalents, 0.75 M in DMF, 3.0 mL) for 30 minutes. Then, additional DIG (5 equivalents, 0.75 M in DMF, 3.0 mL) was added, and the resin was incubated for 90 minutes. After each coupling, the resin was capped by treatment with acetic anhydride (1 M, 7.5 mL) and collidine (0.75 M, 3.0 mL) in DMF for 20 minutes. The fatty acid was coupled for 6 hours using fatty acid monoterbutyl ester (5 equivalents, 0.3 M in 0.3 M Oxyma Pure, 7.5 mL), DIG (5 equivalents, 0.75 M in DMF, 3.0 mL), and collidine (5 equivalents, 0.75 M in DMF, 3.0 mL).

[0496] The Fmoc-protected amino acids used were the recommended standards:

[0497] Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Lys(Boc)-OH, Fmoc-Met-OH, Fmoc-Phe-OH, Fmoc-Pro-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Val-OH and Fmoc-Nle-OH, provided, for example, by Anaspec, Bachem, Iris Biotech or NovabioChem. In the absence of other indication, the naturally occurring L-amino acids are used. In the case where serine or threonine is present in the peptide, pseudoproline dipeptides can be used (see also W.R. Sampson et al., J. Pept. Sci. 1999, 5, 403-409).

[0498] For peptides acylated at any position during the SPPS process, the following appropriately protected building blocks, such as but not limited to Fmoc-8-amino-3,6-dioxaoctanoic acid and Boc-Glu(Fmoc)-OH, are provided by, for example, Anaspec, Bachem, Iris Biotech or Novabiochem.

[0499] Monotert-butyl octadecanedioate and monotert-butyl eicosanedioate are prepared according to methods known in the art, for example, as described in WO 2010102886 A1.

[0500] 17-(2H-Tetrazol-5-yl)heptadecanoic acid (tetrazole-C18) is incorporated by coupling the protected structural unit (S)-4-(17-(1H-tetrazol-5-yl)heptadecanamido)-5-(tert-butoxy)-5-oxopentanoic acid, which is synthesized according to the following procedure: 5-Chloro-1-((dimethylamino)(dimethylimino)methyl)-1H-benzo[d][1,2,3]triazol-3-ium tetrafluoroborate (TCTU, 5.87 g, 16.5 mmol) and N-methylmorpholine (NMM, 8.00 mL, 72.8 mmol) are added successively to a stirred suspension of 17-(1H-tetrazol-5-yl)heptadecanoic acid (5.10 g, 15.0 mmol) in anhydrous DMF (70 mL). The mixture is stirred at room temperature for 30 minutes. H-L-Glu-OtBu (L-glutamic acid 1-tert-butyl ester, 4.58 g, 22.5 mmol) is added in one portion. The suspension turns into a yellow solution within 5 minutes. It is left standing at room temperature for 4 hours and then poured into a vigorously stirred ice-cold aqueous solution of hydrochloric acid (35%, 20 mL) in water (1 L). The crude product is filtered off, washed with water (3 x 100 mL), and immediately dried by suction on a glass frit and then redissolved in DMF (70 mL). The solution is poured into water (1 L), the resulting suspension is stirred for 5 minutes, the solid is filtered off, washed with water (3 x 100 mL), and dried in vacuo. The crude product is dissolved in ethyl acetate (400 mL), filtered through a silica gel (Silicagel 60, 0.040 - 0.063 mm) column (100 g), and the column is washed with ethyl acetate (600 mL). The ethyl acetate is removed in vacuo. The solid residue is boiled in 1,2-dichloroethane (200 mL) with vigorous stirring for 5 minutes, then the emulsion is allowed to cool to room temperature within 5 minutes and is left to crystallize overnight at the same temperature. The waxy crystals are filtered off, washed with 1,2-dichloromethane (50 mL) and n-heptane (100 mL), and air-dried. This purification step is repeated once. Finally, the compound is dissolved in 1,4-dioxane (50 mL) and lyophilized to give (S)-4-(17-(1H-tetrazol-5-yl)heptadecanamido)-5-(tert-butoxy)-5-oxopentanoic acid as an off-white powder.

[0501] For peptides with a C-terminal cysteine, the resin is pre-loaded with H-Cys(Trt)-2-ClTrt (e.g., from Bachem or Novabiochem).

[0502] Method B - Cleavage of Peptide from Resin.

[0503] After synthesis, the resin was washed with DCM and the peptide was cleaved from the resin by treatment with TFA / TIPS / water / DTT (e.g., in a ratio of 90:5:2.5:2.5 or 92.5:2.5:2.5:5) for 2 - 3 hours, and then precipitated with ether. The precipitate was centrifuged and washed several times with ether.

[0504] Method C - Disulfide Bond Formation

[0505] Native CNP contains disulfide bonds, and if it is reduced, the binding to the NPR2 receptor will be eliminated. Methods for forming disulfide bonds in synthetic peptides are well known in the art, and three methods for preparing the compounds exemplified in the present invention are described below.

[0506] Folding Using Aldrithiol-4

[0507] The crude precipitated peptide synthesized from 150 μmol was dissolved in 10 ml of DMSO, then water was added to 700 mL, and 100 mL of MeCN was further added. Finally, 2 mL of 0.5 M NaOAc buffer pH 5.0 was added to the peptide solution. The pH was adjusted to pH 5 - 6.5 with 1 M NaOH until the solution was clear. The amount of peptide in the solution was quantified using CAD, and a fresh stock solution of Aldrithiol - 4 (2 mg / ml in MeOH) was prepared. 1.0 equivalent of Aldrithiol - 4 in MeOH (compared to the peptide content) was added with vigorous stirring within 5 - 10 minutes. The solution was stirred for 10 minutes, and an additional 0.5 or 1.0 equivalent of the Aldrithiol - 4 stock solution was added. Once the reaction was determined to be complete by LCMS, the reaction was quenched with 2 mL of TFA. The peptide was purified by RP - HPLC as described below.

[0508] Folding Using DMSO

[0509] The crude peptide precipitate synthesized from 450 μmol was dissolved in MeCN / water 1:1 (50 mL) and added to 500 mM Tris - HCl pH 8.0 buffer (1 L), diluted with water (1.4 L) and DMSO (600 mL). The solution was gently stirred at room temperature for 16 - 24 hours. The reaction was quenched by adding 5.0 equivalents of iodoacetamide (50 mM aqueous solution), acidified to pH 2 with TFA after 10 minutes, and diluted to 5 L with water. The peptide was purified by RP - HPLC as described below.

[0510] Folding Using Cysteine / Cystine or Cysteamine / Cystamine Redox Buffer

[0511] The crude peptide precipitate synthesized from 150 μmol was dissolved in DMSO (20 mL) and added to a buffer solution of 50 mM Tris-HCl, 3 mM cysteine, 0.3 mM cystine at pH 8.2 (800 mL) or a buffer solution of 50 mM Tris-HCl, 3 mM cysteamine, 0.3 mM cystamine at pH 8.2. The solution was gently stirred at room temperature for 16 - 24 hours. The reaction was quenched by adding 3 mL of TFA, diluted with 100 mL of MeCN, and purified by RP-HPLC as described below.

[0512] Method D - Purification

[0513] Peptide purification methods are well known in the art, and two methods for purifying the compounds exemplified in the present invention are described below.

[0514] Reverse-Phase Preparative HPLC Using Acidic Eluent

[0515] The crude folded peptide was purified by preparative reverse-phase HPLC on a Waters Delta Prep 4000 or Gilson Model 322H2 system equipped with a C18 column, such as Waters Xbridge Prep C18 OBD, 5 μm, 250 x 50 mm or Phenomenex Gemini Axia NX C18, 5 μm, 250 x 30 mm. Using a linear gradient of eluents A (0.1% aqueous TFA solution) and B (0.1% TFA acetonitrile solution), for example, at a constant flow rate of 25 mL / min from 27% to 42% of eluent B within 30 minutes ( column), or at a flow rate of 120 mL / min from 30% to 40% of eluent B within 10 minutes ( column), the peptide was eluted from the column. The peptide fractions with acceptable purity were collected, combined, and lyophilized to obtain the peptide TFA salt as a colorless powder.

[0516] Reverse-Phase Preparative HPLC Using Near-Neutral Eluent

[0517] If further purification is required, the lyophilized peptide TFA salt was purified in a similar preparative HPLC system with a neutral or near-neutral pH eluent, such as eluent A: 100 mM NaOAc pH 6.5, 5% MeCN; eluent B: MeCN. Using a linear gradient of eluents A and B, for example, at a constant flow rate of 60 mL / min from 25% to 55% of eluent B within 30 minutes ( column), the peptide was eluted from the column. The peptide fractions with acceptable purity were collected, combined, and desalted using any of the following methods.

[0518] Method E - Resalting and Desalting

[0519] In some cases, it is necessary to change the counterion to sodium for formulations at pH 6.5. Methods for exchanging the counterion of a peptide are well known in the art, and methods for changing the counterion of some exemplary compounds of the present invention are described below.

[0520] Size Exclusion Desalting

[0521] Counterion exchange is carried out by dissolving the peptide TFA salt at 2 - 3 mg / mL in 400 mM NaOAc pH 6.5 and 20% MeCN. The dissolved peptide is then desalted using a HiPrep 26 / 10 desalting column on a Purifer FPLC system. The column is pre-equilibrated with 20% MeCN, the peptide is injected and eluted with an isocratic 20% MeCN at 6 mL / min for 1.5 CV. The peptide is collected, quantified by CAD analysis, and lyophilized to obtain the sodium salt of the peptide as a colorless powder. General Methods for Detection and Characterization Method:

[0522] To confirm the identity, measure the purity, and quantify the amount of the synthesized peptide prepared, the peptide is characterized using methods well known in the art such as LCMS and reverse-phase LIPLC. Methods for characterizing the exemplary compounds of the present invention are described below.

[0523] Method F - Analytical Chromatography

[0524] The prepared CNP compounds are characterized by RP-UPLC and LCMS, and the amount prepared is quantified by RP-UPLC / CAD.

[0525] LCMS34

[0526] LCMS is carried out on an apparatus consisting of a Waters ACQUITY UPLC system and an Xevo G2-XS QTOF mass spectrometer. The system is equipped with a Waters ACQUITY UPLC BEH C18 column, 1.7 μm, 2.1 mm x 50 mm, and the column temperature is 60 °C. UV detection is carried out at 214 nm. The MS ionization mode is ESI+.

[0527] Eluent A: 0.1% formic acid aqueous solution; Eluent B: 0.1% formic acid acetonitrile solution. The analysis is carried out as follows: An appropriate volume of the sample (preferably 1 - 10 μL) is injected into the column and eluted with a linear gradient of Eluents A and B (from 5% to 95% of Eluent B in 4.0 minutes at a constant flow rate of 0.4 mL / min).

[0528] RP-UPLC107:

[0529] Alternatively, LCMS was performed on an apparatus consisting of a Waters ACQUITY UPLC system and a Waters QDA mass detector. The system was equipped with a Waters ACQUITY UPLC BEH C18 column, 1.7 μm, 2.1 mm x 50 mm, and the column temperature was 40 °C. UV detection was performed at 214 nm.

[0530] Eluent A: 0.05% aqueous TFA solution; Eluent B: 0.05% acetonitrile solution of TFA. The analysis was carried out as follows: An appropriate volume of the sample (preferably 1 - 10 μL) was injected into the column and eluted with a linear gradient of Eluents A and B (from 5% to 60% of Eluent B in 1.6 minutes at a constant flow rate of 0.9 mL / min).

[0531] RP-UPLC02:

[0532] RP - UPLC02 was performed on a Waters ACQUITY UPLC system equipped with a TUV or PDA detector. The system was equipped with a Waters ACQUITY UPLC BEH C18 column, 1.7 μm, 2.1 mm x 150 mm, and the column temperature was 40 °C. UV detection was performed at 214 nm. Eluent A: 0.05% aqueous TFA solution; Eluent B: 0.05% acetonitrile solution of TFA.

[0533] The analysis was carried out as follows: An appropriate volume of the sample (preferably 1 - 10 μL) was injected into the column and eluted with a linear gradient of Eluents A and B (from 5% to 95% of Eluent B in 16.0 minutes at a constant flow rate of 0.4 mL / min).

[0534] RP-UPLC61:

[0535] RP - UPLC61 was performed on a Waters ACQUITY UPLC system equipped with a TUV or PDA detector. The system was equipped with a Waters ACQUITY UPLC BEH Shield C18 column, 1.7 μm, 2.1 mm x 150 mm, and the column temperature was 60 °C. UV detection was performed at 214 nm. Eluent A: 20 mM Na 2 SO 4 ,2 mM Na 2 HPO 4 ,2 mM NaH 2 PO 4Eluent A: aqueous solution / MeCN 9:1, pH 7.2; Eluent B: MeCN / water 7:3. Analysis was carried out as follows: An appropriate volume of the sample (preferably 1 - 10 μL) was injected into the column, and elution was performed with a linear gradient of Eluents A and B (at a constant flow rate of 0.4 mL / min, from 5% to 20% B in 3 minutes, then from 20% to 80% B in 17 minutes, and then from 80% to 90% B in 1 minute).

[0536] CAD02 Peptide Quantification:

[0537] CAD was performed on a Thermo Scientific Vanquish UPLC system equipped with UV - DAD and CAD (charged aerosol detector). The system was equipped with a Waters ACQUITY UPLC CSH C18 column, 1.7 μm, 2.1 mm x 50 mm, and the column temperature was 40 °C. UV detection was carried out at 214 nm. Eluent A: 0.1% v / v aqueous solution of TFA; Eluent B: MeCN solution of 0.1% v / v TFA. Analysis was carried out as follows: An appropriate volume of the sample was injected into the column, and elution was performed with a linear gradient of Eluents A and B (at a constant flow rate of 0.45 mL / min from 0% to 95% B in 4 minutes). Calibration was performed using a standard curve generated by an external standard of 3.55 mg / mL insulin aspart.

[0538] Method G - Preparation of CNP Compound Formulations for In Vivo Experiments

[0539] Preparations for in - vivo studies were prepared according to the principles outlined below.

[0540] Procedure: Preparations for in - vivo studies were prepared by dissolving the lyophilized peptide in a formulation buffer having the same composition as the final preparation (the final preparation composition is described in Examples 9 to 15). The following final preparation compositions have been used:

[0541] · pH 4.0: 5 mM sodium acetate, 250 mM glycerol, pH 4.0

[0542] · pH 4.0: 5 mM sodium acetate, 240 mM propylene glycol, pH 4.0

[0543] · pH 4.0: 5 mM sodium acetate, 250 mM glycerol, 0.007% polysorbate 20, pH 4.0

[0544] · pH 6.0: 20 mM sodium phosphate, 223 mM propylene glycol, pH 6.0

[0545] · pH 6.5: 8 mM sodium phosphate, 250 mM glycerol, pH 6.5

[0546] ·pH 7.4: 8 mM sodium phosphate, 250 mM glycerol, pH 7.4

[0547] ·pH 7.4: 8 mM sodium phosphate, 250 mM glycerol, 0.007% polysorbate 20, pH 7.4

[0548] ·pH 7.5: 8 mM sodium phosphate, 250 mM glycerol, pH 7.5

[0549] ·pH 5.5: 1.33 mM citric acid monohydrate; 3.67 mM trisodium citrate; 52 mg / ml trehalose; 15 mg / ml D - mannose; 0.73 mg / ml L - methionine; 0.05 mg / ml polysorbate 80, pH 5.5

[0550] Due to practical reasons, the formulation procedure for the pH 6.5 preparation is slightly different from that for other pH targets.

[0551] Formulation at pH 6.5: Dissolve the lyophilized peptide in a pH 7.4 buffer (8 mM sodium phosphate, 250 mM glycerol, pH 7.4) to a nominal concentration 20 - 50% higher than the target peptide concentration, and adjust the pH to pH 6.5 using 0.2 N NaOH or 0.2 N HCl. Determine the actual peptide concentration using CAD (method F, CAD02), then add a certain volume of pH 6.5 buffer (8 mM sodium phosphate, 250 mM glycerol, pH 6.5) to reach the nominal concentration corresponding to the target peptide concentration, and adjust the pH to pH 6.5 using 0.2 N NaOH or 0.2 N HCl. Determine the actual peptide concentration using CAD (method F, CAD02). Sterilize - filter the preparation (0.22 μm), determine the final peptide concentration using CAD (method F, CAD02), and then fill the preparation into a sterile container.

[0552] Formulations at pH 4.0, pH 6.0, pH 7.4, and pH 7.5: Dissolve the lyophilized peptide in a formulation buffer whose composition corresponds to that of the final preparation (described in Examples 9 to 15) to a nominal concentration 20 - 50% higher than the target peptide concentration, and adjust the pH to the target value using 0.2 N NaOH or 0.2 N HCl. Determine the actual peptide concentration using CAD (method F, CAD02), and add a certain volume of the same formulation buffer to reach the nominal concentration corresponding to the target peptide concentration, and adjust the pH to the target value using 0.2 N NaOH or 0.2 N HCl. Determine the actual peptide concentration using CAD (method F, CAD02). Sterilize - filter the preparation (0.22 μm), determine the final peptide concentration using CAD (method F, CAD02), and then fill the preparation into a sterile container.

[0553] Example 1 - Synthesis of CNP Compound

[0554] The CNP compounds were synthesized and characterized according to the general preparation method described above. Table 1 summarizes the exemplified CNP compounds and their components. The extender and linker elements in Table 1 together constitute the modifying group of the CNP compounds.

[0555] Table 1: Summary of Exemplary Compound Chemical Formulas 1 - 148

[0556]

[0557]

[0558]

[0559]

[0560]

[0561]

[0562] Chemical formula 1; Compound ID 0065; SEQ ID NO:1

[0563] hCNP22

[0564]

[0565] Molecular weight: 2197.6008. LCMS34: m / 3 calculated: 733.5336; m / 3 measured: 733.7000; m / 4 calculated: 550.4002; m / 4 measured: 550.2800

[0566] Chemical formula 2; Compound ID 1510; SEQ ID NO:2

[0567] hCNP37

[0568]

[0569] Molecular weight: 3948.5589. UPLC107UPLC107: m / 3 calculated: 1317.1863

[0570] Chemical formula 3; Compound ID 0312; SEQ ID NO:3

[0571] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-4,5Q,13Q,32Nle]-hCNP37

[0572]

[0573] Molecular Weight: 4182.9477. LCMS34LCMS34: m / z calculated: 1395.3159; m / z found: 1395.1200; m / z calculated: 1046.7369; m / z found: 1046.600

[0574] Chemical Formula 4; Compound ID 0776; SEQ ID NO:4

[0575] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,13Q,27E,32L]-hCNP37

[0576]

[0577] Molecular Weight: 4068.8451. LCMS34LCMS34: m / z calculated: 1357.2817; m / z found: 1357.2694; m / z calculated: 1018.2113; m / z found: 1018.2089

[0578] Chemical Formula 5; Compound ID 1235; SEQ ID NO:5

[0579] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,10H,13Q,32L]-hCNP37

[0580]

[0581] Molecular Weight: 3674.2904. UPLC107: m / z calculated: 1225.7635; m / z found: 1225.5700

[0582] Chemical Formula 6; Compound ID 0262; SEQ ID NO:6

[0583] [N-Terminal([(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), des1-8, 32Nle]-hCNP37

[0584]

[0585] Molecular Weight: 4040.6131. LCMS34LCMS34: m / 3 calculated: 1347.8710; m / 3 found: 1347.7200; m / 4 calculated: 1011.1533; m / 4 found: 1010.7900

[0586] Chemical Formula 7; Compound ID 0296; SEQ ID NO:7

[0587] [N-Terminal([(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), des1-4, 15S, 19S, 32Nle]-hCNP37

[0588]

[0589] Molecular Weight: 4427.9616. LCMS34LCMS34: m / 3 calculated: 1476.9872; m / 3 found: 1476.9200; m / 4 calculated: 1107.9904; m / 4 found: 1108.1700

[0590] Chemical Formula 8; Compound ID 0313; SEQ ID NO:8

[0591] [N-Terminal([(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), des1-10, 32Nle]-hCNP37

[0592]

[0593] Molecular weight: 3620.1536. LCMS34: m / 3 calculated: 1207.7179; m / 3 measured: 1207.6400; m / 4 calculated: 906.0384; m / 4 measured: 905.9900

[0594] Chemical formula 9; Compound ID 0334; SEQ ID NO:9

[0595] [N-terminus([(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), des1-10,13Q,32Nle]-hCNP37

[0596]

[0597] Molecular weight: 3634.1801. LCMS34: m / 3 calculated: 1212.3934; m / 3 measured: 1212.3100; m / 4 calculated: 909.5450; m / 4 measured: 909.2400

[0598] Chemical formula 10; Compound ID 1221; SEQ ID NO:10

[0599] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,32L]-hCNP37

[0600]

[0601] Molecular weight: 3700.3243. UPLC107: m / 3 calculated: 1234.4414; m / 3 measured: 1234.2700

[0602] Chemical formula 11; Compound ID 1222; SEQ ID NO:11

[0603] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,10E,13Q,32L]-hCNP37

[0604]

[0605] Molecular weight: 3666.2651. LCMS34: m / 3 calculated: 1223.0884; m / 3 measured: 1222.9000;

[0606] Chemical formula 12; Compound ID 1223; SEQ ID NO:12

[0607] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,14E,32L]-hCNP37

[0608]

[0609] Molecular weight: 3701.266. LCMS34: m / 3 calculated: 1234.7553; m / 3 measured: 1234.5800;

[0610] Chemical formula 13; Compound ID 1224; SEQ ID NO:13

[0611] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,19S,32L]-hCNP37

[0612]

[0613] Molecular weight: 3659.2294. LCMS34: m / 3 calculated: 1220.7431; m / 3 measured: 1220.3212; m / 4 calculated: 915.8074; m / 4 measured: 915.7400

[0614] Chemical formula 14; Compound ID 1225; SEQ ID NO:14

[0615] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,25P,32L]-hCNP37

[0616]

[0617] Molecular weight: 3669.2672. UPLC107: m / z calculated: 1224.0891; m / z measured: 1223.9400

[0618] Chemical formula 15; Compound ID 1226; SEQ ID NO: 15

[0619] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8, 10E, 13Q, 14E, 25P, 32L]-hCNP37

[0620]

[0621] Molecular weight: 3670.2089. UPLC107: m / z calculated: 1224.4030; m / z measured: 1224.2000

[0622] Chemical formula 16; Compound ID 1227; SEQ ID NO: 16

[0623] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8, 10E, 13Q, 19S, 25P, 32L]-hCNP37

[0624]

[0625] Molecular weight: 3628.1723. LCMS34: m / z calculated: 1210.3908; m / z measured: 1210.4200; m / z calculated: 908.0431; m / z measured: 908.0900

[0626] Chemical formula 17; Compound ID 1228; SEQ ID NO: 17

[0627] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8, 10E, 13Q, 14E, 19S, 25P, 32L]-hCNP37

[0628]

[0629] Molecular weight: 3629.114. LCMS34: m / z calculated: 1210.7047; m / z measured: 1210.5790; m / z calculated: 908.2785; m / z measured: 908.1400

[0630] Chemical formula 18; Compound ID 1229; SEQ ID NO:18

[0631] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,13Q,14E,32L]-hCNP37

[0632]

[0633] Molecular weight: 3666.2651. UPLC107: m / z calculated: 1223.0884; m / z measured: 1222.9300

[0634] Chemical formula 19; Compound ID 1230; SEQ ID NO:19

[0635] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,13Q,14E,19S,32L]-hCNP37

[0636]

[0637] Molecular weight: 3625.1701. UPLC107: m / z calculated: 1209.3900; m / z measured: 1209.2800

[0638] Chemical formula 20; Compound ID 1231; SEQ ID NO:20

[0639] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,10Q,13Q,14E,32L]-hCNP37

[0640]

[0641] Molecular weight: 3666.222. UPLC107: m / z calculated: 1223.0740; m / z measured: 1222.9700

[0642] Chemical formula 21; Compound ID 1232; SEQ ID NO: 21

[0643] [N-Terminal ([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]), des1-8,13Q,14E,32L]-hCNP37

[0644]

[0645] Molecular weight: 3700.3243. UPLC107: m / z calculated: 1234.4414; m / z measured: 1234.2100

[0646] Chemical formula 22; Compound ID 1233; SEQ ID NO: 22

[0647] [N-Terminal ([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]), des1-8,13Q,19S,25P,32L]-hCNP37

[0648]

[0649] Molecular weight: 3627.2306. UPLC107: m / z calculated: 1210.0769; m / z measured: 1209.9600

[0650] Chemical formula 23; Compound ID 1236; SEQ ID NO: 23

[0651] [N-Terminal ([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]), des1-8,9E,10H,13Q,14E,32L]-hCNP37

[0652]

[0653] Molecular weight: 3675.2321. LCMS34: m / z calculated: 1226.0774; m / z measured: 1225.8900;

[0654] Chemical formula 24; Compound ID 1237; SEQ ID NO:24

[0655] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,14E,32L]-hCNP37

[0656]

[0657] Molecular weight: 3709.2913. UPLC107: m / z calculated: 1237.4304; m / z measured: 1237.3400

[0658] Chemical formula 25; Compound ID 1240; SEQ ID NO:25

[0659] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,14E,25P,32L]-hCNP37

[0660]

[0661] Molecular weight: 3678.2342. UPLC107: m / z calculated: 1227.0781; m / z measured: 1226.9500

[0662] Chemical formula 26; Compound ID 1241; SEQ ID NO:26

[0663] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,19S,25P,32L]-hCNP37

[0664]

[0665] Molecular weight: 3636.1976. LCMS34: m / 3 calculated: 1213.0659; m / 3 measured: 1213.1011; m / 4 calculated: 910.0494; m / 4 measured: 910.0900

[0666] Chemical formula 27; Compound ID 1242; SEQ ID NO:27

[0667] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,14E,19S,25P,32L]-hCNP37

[0668]

[0669] Molecular weight: 3637.1393. UPLC107: m / 3 calculated: 1213.3798; m / 3 measured: 1213.2100

[0670] Chemical formula 28; Compound ID 1274; SEQ ID NO:28

[0671] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13E,19Q,25P,32L]-hCNP37

[0672]

[0673] Molecular weight: 3670.2089. LCMS34: m / 3 calculated: 1224.4030; m / 3 measured: 1224.3150; m / 4 calculated: 918.5522; m / 4 measured: 918.4840

[0674] Chemical formula 29; Compound ID 1287; SEQ ID NO:29

[0675] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-9,10E,13Q,14E,19S,25P,32L]-hCNP37

[0676]

[0677] Molecular weight: 3465.9407. LCMS34: m / 3 calculated: 1156.3136; m / 3 measured: 1156.5100; m / 4 calculated: 867.4852; m / 4 measured: 867.3800

[0678] Chemical formula 30; Compound ID 1288; SEQ ID NO:30

[0679] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-9,10E,13Q,15E,19S,25P,32L]-hCNP37

[0680]

[0681] Molecular weight: 3465.9407. LCMS34: m / 3 calculated: 1156.3136; m / 3 measured: 1155.8400; m / 4 calculated: 867.4852; m / 4 measured: 867.3800

[0682] Chemical formula 31; Compound ID 1289; SEQ ID NO:31

[0683] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,15E,19S,25P,32L]-hCNP37

[0684]

[0685] Molecular weight: 3629.114. LCMS34: m / 3 calculated: 1210.7047; m / 3 measured: 1210.5500; m / 4 calculated: 908.2785; m / 4 measured: 908.1600

[0686] Chemical formula 32; Compound ID 1290; SEQ ID NO:16

[0687] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,19S,25P,32L]-hCNP37

[0688]

[0689] Molecular weight: 3757.2862. LCMS34: m / z calculated: 1253.4287; m / z found: 1253.5700; m / z calculated: 940.3216; m / z found: 940.1700

[0690] Chemical formula 33; Compound ID 1302; SEQ ID NO: 32

[0691] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,15T,19S,25P,32L]-hCNP37

[0692]

[0693] Molecular weight: 3609.1292. LCMS34: m / z calculated: 1204.0431; m / z found: 1203.7523; m / z calculated: 903.2823; m / z found: 903.3355

[0694] Chemical formula 34; Compound ID 1303; SEQ ID NO: 33

[0695] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,15T,17V,19S,25P,32L]-hCNP37

[0696]

[0697] Molecular weight: 3595.1026. LCMS34: m / 3 calculated: 1199.3675; m / 3 measured: 1199.3210; m / 4 calculated: 899.7757; m / 4 measured: 899.7320

[0698] Chemical formula 35; Compound ID 1304; SEQ ID NO:34

[0699] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,14H,15T,19S,25P,32L]-hCNP37

[0700]

[0701] Molecular weight: 3618.0962. LCMS34: m / 3 calculated: 1207.0321; m / 3 measured: 1206.7366; m / 4 calculated: 905.5241; m / 4 measured: 905.5760

[0702] Chemical formula 36; Compound ID 1305; SEQ ID NO:35

[0703] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,14H,15T,19S,25P,32L]-hCNP37

[0704]

[0705] Molecular weight: 3609.1292. LCMS34: m / 3 calculated: 1204.0431; m / 3 measured: 1204.0040; m / 4 calculated: 903.2823; m / 4 measured: 903.2470

[0706] Chemical formula 37; Compound ID 1309; SEQ ID NO:36

[0707] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,10H,13Q,25P,32L]-hCNP37

[0708]

[0709] Molecular weight: 3643.2333. LCMS34: m / 3 calculated: 1215.4111; m / 3 measured: 1215.4546; m / 4 calculated: 911.8083; m / 4 measured: 911.8567

[0710] Chemical formula 38; Compound ID 1310; SEQ ID NO: 37

[0711] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,10H,13Q,19S,25P,32L]-hCNP37

[0712]

[0713] Molecular weight: 3602.1383. LCMS34: m / 3 calculated: 1201.7128; m / 3 measured: 1201.7507; m / 4 calculated: 901.5346; m / 4 measured: 901.5770

[0714] Chemical formula 39; Compound ID 1311; SEQ ID NO: 38

[0715] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,10H,13Q,17V,19S,25P,32L]-hCNP37

[0716]

[0717] Molecular weight: 3588.1117. LCMS34: m / 3 calculated: 1197.0372; m / 3 measured: 1197.0725; m / 4 calculated: 898.0279; m / 4 measured: 898.0776

[0718] Chemical formula 40; Compound ID 1312; SEQ ID NO: 39

[0719] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-8,9E,10H,13Q,15T,17V,19S,25P,32L]-hCNP37

[0720]

[0721] Molecular weight: 3561.0433. LCMS34: m / z calculated: 1188.0144; m / z found: 1187.9800; m / z calculated: 891.2608; m / z found: 891.2320

[0722] Chemical formula 41; Compound ID 1322; SEQ ID NO:40

[0723] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-8,10H,13Q,14H,15H,27E,32L]-hCNP37

[0724]

[0725] Molecular weight: 3740.3102. LCMS34: m / z calculated: 1247.7701; m / z found: 1247.6970; m / z calculated: 936.0776; m / z found: 936.0280

[0726] Chemical formula 42; Compound ID 1323; SEQ ID NO:40

[0727] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-8,10H,13Q,14H,15H,27E,32L]-hCNP37

[0728]

[0729] Molecular weight: 3869.4242. LCMS34: m / 3 calculated: 1290.8081; m / 3 measured: 1290.7130; m / 4 calculated: 968.3561; m / 4 measured: 968.2840

[0730] Chemical formula 43; Compound ID 1324; SEQ ID NO:41

[0731] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,14H,15H,19Q,27E,32L]-hCNP37

[0732]

[0733] Molecular weight: 3860.4141. LCMS34: m / 3 calculated: 1287.8047; m / 3 measured: 1287.7150; m / 4 calculated: 966.1035; m / 4 measured: 966.0310

[0734] Chemical formula 44; Compound ID 1338; SEQ ID NO:42

[0735] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,19S,25P,27E,32L]-hCNP37

[0736]

[0737] Molecular weight: 3642.1988. LCMS34: m / 3 calculated: 1215.0663; m / 3 measured: 1214.9827; m / 4 calculated: 911.5497; m / 4 measured: 911.2273

[0738] Chemical formula 45; Compound ID 1339; SEQ ID NO:43

[0739] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-9,10E,13Q,19S,25P,32L]-hCNP37

[0740]

[0741] Molecular weight: 3464.999. LCMS34: m / 3 calculated: 1155.9997; m / 3 measured: 1155.9600; m / 4 calculated: 867.2498; m / 4 measured: 866.9667

[0742] Chemical formula 46; Compound ID 1340; SEQ ID NO:44

[0743] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,14H,15H,19Q,32L]-hCNP37

[0744]

[0745] Molecular weight: 3717.2736. LCMS34: m / 3 calculated: 1240.0912; m / 3 measured: 1240.0320; m / 4 calculated: 930.3184; m / 4 measured: 930.2630

[0746] Chemical formula 47; Compound ID 1341; SEQ ID NO:45

[0747] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,14H,15H,19Q,25P,32L]-hCNP37

[0748]

[0749] Molecular weight: 3686.2165. LCMS34: m / 3 calculated: 1229.7388; m / 3 measured: 1229.6760; m / 4 calculated: 922.5541; m / 4 measured: 922.5020

[0750] Chemical formula 48; Compound ID 1345; SEQ ID NO:46

[0751] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8, 10H, 13Q, 14H, 15H, 32L]-hCNP37

[0752]

[0753] Molecular weight: 3726.2836. LCMS34: m / 3 calculated: 1243.0945; m / 3 measured: 1243.0310; m / 4 calculated: 932.5709; m / 4 measured: 932.5240

[0754] Chemical formula 49; Compound ID 1346; SEQ ID NO:47

[0755] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8, 10H, 13Q, 14H, 15H, 25P, 32L]-hCNP37

[0756]

[0757] Molecular weight: 3695.2265. LCMS34: m / 3 calculated: 1232.7422; m / 3 measured: 1232.6930; m / 4 calculated: 924.8066; m / 4 measured: 924.7340

[0758] Chemical formula 50; Compound ID 1347; SEQ ID NO:48

[0759] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-10, 13Q, 19H, 25P, 32L]-hCNP37

[0760]

[0761] Molecular weight: 3385.947. LCMS34: m / 3 calculated: 1129.6490; m / 3 measured: 1129.2775; m / 4 calculated: 847.4868; m / 4 measured: 847.2063

[0762] Chemical formula 51; Compound ID 1348; SEQ ID NO:49

[0763] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-10,13Q,18H,19S,25P,32L]-hCNP37

[0764]

[0765] Molecular weight: 3385.947. LCMS34: m / 3 calculated: 1129.6490; m / 3 measured: 1129.2775; m / 4 calculated: 847.4868; m / 4 measured: 847.4684

[0766] Chemical formula 52; Compound ID 1350; SEQ ID NO:50

[0767] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,10H,13Q,14H,15T,19S,25P,32L]-hCNP37

[0768]

[0769] Molecular weight: 3973.532. LCMS34: m / 3 calculated: 1325.5107; m / 3 measured: 1325.3463; m / 4 calculated: 994.3830; m / 4 measured: 994.2843

[0770] Chemical formula 53; Compound ID 1351; SEQ ID NO:51

[0771] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,10H,13Q,14H,15T,17V,19S,25P,32L]-hCNP37

[0772]

[0773] Molecular weight: 3959.5054. LCMS34: m / 3 calculated: 1320.8351; m / 3 measured: 1320.6271; m / 4 calculated: 990.8764; m / 4 measured: 990.7383

[0774] Chemical formula 54; Compound ID 1352; SEQ ID NO:33

[0775] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,15T,17V,19S,25P,32L]-hCNP37

[0776]

[0777] Molecular weight: 3740.259. LCMS34: m / 3 calculated: 1247.7530; m / 3 measured: 1247.3048; m / 4 calculated: 936.0648; m / 4 measured: 935.9911

[0778] Chemical formula 55; Compound ID 1353; SEQ ID NO:36

[0779] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,10H,13Q,25P,32L]-hCNP37

[0780]

[0781] Molecular weight: 3788.3897. LCMS34: m / 3 calculated: 1263.7966; m / 3 measured: 1263.6780; m / 4 calculated: 948.0974; m / 4 measured: 947.7735

[0782] Chemical formula 56; Compound ID 1354; SEQ ID NO:52

[0783] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,10H,13Q,15T,17V,19S,25P,32L]-hCNP37

[0784]

[0785] Molecular weight: 4079.6524. LCMS34: m / 3 calculated: 1360.8841; m / 3 measured: 1360.7530; m / 4 calculated: 1020.9131; m / 4 measured: 1020.8160

[0786] Chemical formula 57; Compound ID 1355; SEQ ID NO:22

[0787] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,19S,25P,32L]-hCNP37

[0788]

[0789] Molecular weight: 3772.387. LCMS34: m / 3 calculated: 1258.4623; m / 3 measured: 1258.3439; m / 4 calculated: 944.0968; m / 4 measured: 944.0223

[0790] Chemical formula 58; Compound ID 1356; SEQ ID NO:22

[0791] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,19S,25P,32L]-hCNP37

[0792]

[0793] Molecular Weight: 3901.501. LCMS34: m / z calculated: 1301.5003; m / z found: 1301.3470; m / z calculated: 976.3753; m / z found: 976.2725

[0794] Chemical Formula 59; Compound ID 1357; SEQ ID NO:5

[0795] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,9E,10H,13Q,32L]-hCNP37

[0796]

[0797] Molecular Weight: 3948.5608. LCMS34: m / z calculated: 1317.1869; m / z found: 1317.0300; m / z calculated: 988.1402; m / z found: 988.0348

[0798] Chemical Formula 60; Compound ID 1359; SEQ ID NO:53

[0799] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,9E,10H,13Q,15T,17V,19S,25P,32L]-hCNP37

[0800]

[0801] Molecular weight: 3916.4792. LCMS34: m / 3 calculated: 1306.4931; m / 3 measured: 1306.0170; m / 4 calculated: 980.1198; m / 4 measured: 979.7797

[0802] Chemical formula 61; Compound ID 1360; SEQ ID NO:54

[0803] [N-terminal ([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]), des1-5,9E,10H,13Q,14H,15T,17V,19S,25P,32L]-hCNP37

[0804]

[0805] Molecular weight: 3925.4462. LCMS34: m / 3 calculated: 1309.4821; m / 3 measured: 1309.3323; m / 4 calculated: 982.3616; m / 4 measured: 982.2667

[0806] Chemical formula 62; Compound ID 1375; SEQ ID NO:14

[0807] [N-terminal ([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]), des1-8,10E,13Q,25P,32L]-hCNP37

[0808]

[0809] Molecular weight: 3959.5801. LCMS34: m / 3 calculated: 1320.8600; m / 3 measured: 1320.7200; m / 4 calculated: 990.8950; m / 4 measured: 990.5400

[0810] Chemical formula 63; Compound ID 1376; SEQ ID NO:14

[0811] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8, 10E, 13Q, 25P, 32L]-hCNP37

[0812]

[0813] Molecular weight: 3697.3204. LCMS34: m / z calculated: 1233.4401; m / z measured: 1233.3500; m / z calculated: 925.3301; m / z measured: 925.2600

[0814] Chemical formula 64; Compound ID 1377; SEQ ID NO: 55

[0815] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8, 10E, 13Q, 17G, 25P, 32L]-hCNP37

[0816]

[0817] Molecular weight: 3613.1609. LCMS34: m / z calculated: 1205.3870; m / z measured: 1205.3100; m / z calculated: 904.2902; m / z measured: 903.9800

[0818] Chemical formula 65; Compound ID 1378; SEQ ID NO: 56

[0819] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5, 6G, 7G, 8G, 10E, 13Q, 19S, 25P, 32L]-hCNP37

[0820]

[0821] Molecular weight: 3799.3262. LCMS34: m / z calculated: 1267.4421; m / z measured: 1267.3470; m / z calculated: 950.8316; m / z measured: 950.7240

[0822] Chemical formula 66; Compound ID 1379; SEQ ID NO:57

[0823] [N - terminus([2 - [2 - [2 - [[2 - [2 - [2 - [[(4S)-4 - carboxy - 4 - (17 - carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,8S,10E,13Q,19S,25P,32L]-hCNP37

[0824]

[0825] Molecular weight: 4562.14. LCMS34: m / z calculated: 1521.7133; m / z measured: 1521.3220; m / z calculated: 1141.5350; m / z measured: 1141.2370

[0826] Chemical formula 67; Compound ID 1380; SEQ ID NO:58

[0827] [N - terminus([2 - [2 - [2 - [[2 - [2 - [2 - [[(4S)-4 - carboxy - 4 - (17 - carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5E,8S,10E,13Q,19S,25P,32L]-hCNP37

[0828]

[0829] Molecular weight: 4563.1248. LCMS34: m / z calculated: 1522.0416; m / z measured: 1521.8490; m / z calculated: 1141.7812; m / z measured: 1141.6240

[0830] Chemical formula 68; Compound ID 1381; SEQ ID NO:59

[0831] [N - terminus([2 - [2 - [2 - [[2 - [2 - [2 - [[(4S)-4 - carboxy - 4 - (17 - carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,7H,10H,13Q,14H,15H,17S,19Q,25P,32L]-hCNP37

[0832]

[0833] Molecular weight: 4625.1195. LCMS34: m / 3 calculated: 1542.7065; m / 3 measured: 1542.3070; m / 4 calculated: 1157.2799; m / 4 measured: 1156.9700

[0834] Chemical formula 69; Compound ID 1382; SEQ ID NO:60

[0835] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,7H,13Q,19Q,25P,32L]-hCNP37

[0836]

[0837] Molecular weight: 4882.5268. LCMS34: m / 3 calculated: 1628.5089; m / 3 measured: 1628.2280; m / 4 calculated: 1221.6317; m / 4 measured: 1221.2810

[0838] Chemical formula 70; Compound ID 1383; SEQ ID NO:61

[0839] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,7H,10H,13Q,15H,17S,19Q,25P,32L]-hCNP37

[0840]

[0841] Molecular weight: 4745.2665. LCMS34: m / 3 calculated: 1582.7555; m / 3 measured: 1582.5080; m / 4 calculated: 1187.3166; m / 4 measured: 1186.9970

[0842] Chemical formula 71; Compound ID 9384; SEQ ID NO:62

[0843] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-6,7G,8Q,9A,10P,13Q,17S,19Q,25P,32L]-hCNP37

[0844]

[0845] Molecular Weight: 3704.2302. LCMS34: m / 3 calculated: 1235.7434; m / 3 measured: 1235.5400; m / 4 calculated: 927.0576; m / 4 measured: 926.9010

[0846] Chemical formula 72; Compound ID 1385; SEQ ID NO: 63

[0847] [N-Terminal([(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), des1-5,10E,13Q,25P,32L]-hCNP37

[0848]

[0849] Molecular Weight: 4121.7322. LCMS34: m / 3 calculated: 1374.9107; m / 3 measured: 1374.6120; m / 4 calculated: 1031.4331; m / 4 measured: 1031.2130

[0850] Chemical formula 73; Compound ID 1386; SEQ ID NO: 64

[0851] [N-Terminal([(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), des1-5,13Q,19S,25P,32L]-hCNP37

[0852]

[0853] Molecular weight: 4079.6955. LCMS34: m / 3 calculated: 1360.8985; m / 3 measured: 1360.7680; m / 4 calculated: 1020.9239; m / 4 measured: 1020.8160

[0854] Chemical formula 74; Compound ID 1387; SEQ ID NO:65

[0855] [N-terminus([(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), des1-5,6G,7G,8G,13Q,19S,25P,32L]-hCNP37

[0856]

[0857] Molecular weight: 3895.4136. LCMS34: m / 3 calculated: 1299.4712; m / 3 measured: 1299.3300; m / 4 calculated: 974.8534; m / 4 measured: 974.7500

[0858] Chemical formula 75; Compound ID 1388; SEQ ID NO:66

[0859] [N-terminus([(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), des1-5,6G,7G,8G,13Q,25P,32L]-hCNP37

[0860]

[0861] Molecular weight: 3936.5086. LCMS34: m / 3 calculated: 1313.1695; m / 3 measured: 1313.0250; m / 4 calculated: 985.1272; m / 4 measured: 985.2870

[0862] Chemical formula 76; Compound ID 1389; SEQ ID NO:67

[0863] [N-Terminal([(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]),5Q,13Q,19S,25P,32L]-hCNP37

[0864]

[0865] Molecular Weight: 4699.3224. LCMS34: m / 3 Calculated: 1567.4408; m / 3 Found: 1567.4835; m / 4 Calculated: 1175.8306; m / 4 Found: 1176.1075

[0866] Chemical Formula 77; Compound ID 1390; SEQ ID NO:68

[0867] [N-Terminal([(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]),5Q,10E,13Q,19S,25P,32L]-hCNP37

[0868]

[0869] Molecular Weight: 4700.2641. LCMS34: m / 3 Calculated: 1567.7547; m / 3 Found: 1567.5480; m / 4 Calculated: 1176.0660; m / 4 Found: 1175.8970

[0870] Chemical Formula 78; Compound ID 1391; SEQ ID NO:55

[0871] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-8,10E,13Q,17G,25P,32L]-hCNP37

[0872]

[0873] Molecular weight: 3903.4738. LCMS34: m / 3 calculated: 1302.1579; m / 3 measured: 1302.0100; m / 4 calculated: 976.8685; m / 4 measured: 976.5200

[0874] Chemical formula 79; Compound ID 1392; SEQ ID NO:16

[0875] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,19S,25P,32L]-hCNP37

[0876]

[0877] Molecular weight: 3918.4851. LCMS34: m / 3 calculated: 1307.1617; m / 3 measured: 1307.3500; m / 4 calculated: 980.6213; m / 4 measured: 980.5300

[0878] Chemical formula 80; Compound ID 1393; SEQ ID NO:69

[0879] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,19Q,25P,32L]-hCNP37

[0880]

[0881] Molecular weight: 3959.5371. LCMS34: m / 3 calculated: 1320.8457; m / 3 measured: 1321.0300; m / 4 calculated: 990.8843; m / 4 measured: 990.7800

[0882] Chemical formula 81; Compound ID 1394; SEQ ID NO:70

[0883] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,10E,13Q,19Q,25P,32L]-hCNP37

[0884]

[0885] Molecular weight: 4644.2869. LCMS34: m / z calculated: 1549.0956; m / z found: 1548.7900; m / z calculated: 1162.0717; m / z found: 1161.8500

[0886] Chemical formula 82; Compound ID 1395; SEQ ID NO:71

[0887] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,7A,10E,13Q,19Q,25P,32L]-hCNP37

[0888]

[0889] Molecular weight: 4559.1791. LCMS34: m / z calculated: 1520.7264; m / z found: 1520.4600; m / z calculated: 1140.7948; m / z found: 1140.600

[0890] Chemical formula 83; Compound ID 1396; SEQ ID NO:70

[0891] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,10E,13Q,19Q,25P,32L]-hCNP37

[0892]

[0893] Molecular weight: 4773.4009. LCMS34: m / z calculated: 1592.1336; m / z found: 1592.1600; m / z calculated: 1194.3502; m / z found: 1194.3700

[0894] Chemical formula 84; Compound ID 1398; SEQ ID NO:72

[0895] [N - terminal([2 - [2 - [2 - [[2 - [2 - [2 - [[(4S)-4 - carboxy - 4 - [[(4S)-4 - carboxy - 4 - [[(4S)-4 - carboxy - 4 - [[(4S)-4 - carboxy - 4 - (17 - carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,8H,13Q,17S,19Q,25P,32L]-hCNP37

[0896]

[0897] Molecular weight: 5013.5738. LCMS34: m / 3 calculated: 1672.1913; m / 3 measured: 1672.1528; m / 4 calculated: 1254.3935; m / 4 measured: 1254.1224

[0898] Chemical formula 85; Compound ID 1399; SEQ ID NO:72

[0899] [N - terminal([2 - [2 - [2 - [[2 - [2 - [2 - [[(4S)-4 - carboxy - 4 - [[(4S)-4 - carboxy - 4 - [[(4S)-4 - carboxy - 4 - [[(4S)-4 - carboxy - 4 - (19 - carboxynonadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,8H,13Q,17S,19Q,25P,32L]-hCNP37

[0900]

[0901] Molecular weight: 5041.627. LCMS34: m / 3 calculated: 1681.5423; m / 3 measured: 1681.5050; m / 4 calculated: 1261.4068; m / 4 measured: 1261.1331

[0902] Chemical formula 86; Compound ID 1400; SEQ ID NO:73

[0903] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,7H,13Q,17G,19Q,25P,32L]-hCNP37

[0904]

[0905] Molecular weight: 4955.5344. LCMS34: m / z calculated: 1652.8448; m / z found: 1652.8279; m / z calculated: 1239.8836; m / z found: 1239.8716

[0906] Chemical formula 87; Compound ID 1401; SEQ ID NO:74

[0907] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,13Q,32L]-hCNP37

[0908]

[0909] Molecular weight: 5578.2432. LCMS34: m / z calculated: 1860.4144; m / z found: 1860.2654; m / z calculated: 1395.5608; m / z found: 1395.4623

[0910] Chemical formula 88; Compound ID 1402; SEQ ID NO:75

[0911] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,13Q,19Q,25P,32L]-hCNP37

[0912]

[0913] Molecular weight: 5288.9151. LCMS34: m / 3 calculated: 1763.9717; m / 3 found: 1763.8729; m / 4 calculated: 1323.2288; m / 4 found: 1323.1718

[0914] Chemical formula 89; Compound ID 1403; SEQ ID NO:76

[0915] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,13Q,17G,19Q,25P,32L]-hCNP37

[0916]

[0917] Molecular weight: 5232.8088. LCMS34: m / 3 calculated: 1745.2696; m / 3 found: 1744.8611; m / 4 calculated: 1309.2022; m / 4 found: 1309.1547

[0918] Chemical formula 90; Compound ID 1404; SEQ ID NO:75

[0919] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,13Q,19Q,25P,32L]-hCNP37

[0920]

[0921] Molecular weight: 5579.228. LCMS34: m / z calculated: 1860.7427; m / z measured: 1860.6008; m / z calculated: 1395.8070; m / z measured: 1395.7069

[0922] Chemical formula 91; Compound ID 1405; SEQ ID NO:75

[0923] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(19-carboxynonadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,13Q,19Q,25P,32L]-hCNP37

[0924]

[0925] Molecular weight: 5316.9683. LCMS34: m / z calculated: 1773.3228; m / z measured: 1773.2192; m / z calculated: 1330.2421; m / z measured: 1330.1785

[0926] Chemical formula 92; Compound ID 1406; SEQ ID NO:75

[0927] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,13Q,19Q,25P,32L]-hCNP37

[0928]

[0929] Molecular weight: 5450.114. LCMS34: m / z calculated: 1817.7047; m / z measured: 1817.5808; m / z4 calculated: 1363.5285; m / z4 measured: 1363.4484

[0930] Chemical formula 93; Compound ID 9407; SEQ ID NO:77

[0931] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,13Q,19Q,25P,32L]-hCNP37

[0932]

[0933] Molecular weight: 4830.4871. LCMS34: m / z calculated: 1611.1624; m / z measured: 1610.8330; m / z4 calculated: 1208.6218; m / z4 measured: 1208.3823

[0934] Chemical formula 94; Compound ID 1419; SEQ ID NO:51

[0935] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5, 10H, 13Q, 14H, 15T, 17V, 19S, 25P, 32L]-hCNP37

[0936]

[0937] Molecular Weight: 4088.6194. LCMS34: m / z calculated: 1363.8731; m / z found: 1363.6902; m / z calculated: 1023.1549; m / z found: 1023.0371

[0938] Chemical Formula 95; Compound ID 1420; SEQ ID NO:78

[0939] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5, 10H, 13Q, 14H, 15T, 17V, 25P, 32L]-hCNP37

[0940]

[0941] Molecular Weight: 4258.8284. LCMS34: m / z calculated: 1420.6095; m / z found: 1420.3875; m / z calculated: 1065.7071; m / z found: 1065.5448

[0942] Chemical Formula 96; Compound ID 1421; SEQ ID NO:79

[0943] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5, 13Q, 15T, 17V, 19S, 25P, 32L]-hCNP37

[0944]

[0945] Molecular weight: 4199.7994. LCMS34: m / 3 calculated: 1400.9331; m / 3 measured: 1400.7268; m / 4 calculated: 1050.9499; m / 4 measured: 1050.8062

[0946] Chemical formula 97; Compound ID 1422; SEQ ID NO:80

[0947] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5, 10H, 13Q, 17V, 19S, 25P, 32L]-hCNP37

[0948]

[0949] Molecular weight: 4235.8348. LCMS34: m / 3 calculated: 1412.9449; m / 3 measured: 1412.7313; m / 4 calculated: 1059.9587; m / 4 measured: 1059.8075

[0950] Chemical formula 98; Compound ID 1423; SEQ ID NO:81

[0951] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5, 10H, 13Q, 15T, 17V, 25P, 32L]-hCNP37

[0952]

[0953] Molecular weight: 4249.8614. LCMS34: m / 3 calculated: 1417.6205; m / 3 measured: 1417.3965; m / 4 calculated: 1063.4654; m / 4 measured: 1063.3148

[0954] Chemical formula 99; Compound ID 1424; SEQ ID NO:82

[0955] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5, 13Q, 15T, 17V, 25P, 32L]-hCNP37

[0956]

[0957] Molecular weight: 4370.0084. LCMS34: m / z calculated: 1457.6695; m / z found: 1457.4309; m / z calculated: 1093.5021; m / z found: 1093.5833

[0958] Chemical formula 100; Compound ID 1425; SEQ ID NO: 83

[0959] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8, 10H, 13Q, 15T, 17V, 25P, 32L]-hCNP37

[0960]

[0961] Molecular weight: 4055.6244. LCMS34: m / z calculated: 1352.8748; m / z found: 1352.6941; m / z calculated: 1014.9061; m / z found: 1014.5325

[0962] Chemical formula 101; Compound ID 1426; SEQ ID NO: 84

[0963] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[17-(1H-tetrazol-5-yl)heptadecanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,10H,13Q,15S,19Q,25P,32L]-hCNP37

[0964]

[0965] Molecular weight: 4144.7323. LCMS34: m / z calculated: 1382.5774; m / z measured: 1382.3732; m / z calculated: 1037.1831; m / z measured: 1037.0341

[0966] Chemical formula 102; Compound ID 1431; SEQ ID NO:85

[0967] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10E,13Q,17G,19H,25P,32L]-hCNP37

[0968]

[0969] Molecular weight: 3912.4408. LCMS34: m / z calculated: 1305.1469; m / z measured: 1304.9080; m / z calculated: 979.1102; m / z measured: 978.9250

[0970] Chemical formula 103; Compound ID 1432; SEQ ID NO:86

[0971] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,10E,13Q,14H,17G,19Q,25P,32L]-hCNP37

[0972]

[0973] Molecular weight: 4396.9476. LCMS34: m / 3 calculated: 1466.6492; m / 3 measured: 1466.3500; m / 4 calculated: 1100.2369; m / 4 measured: 1100.0354

[0974] Chemical formula 104; Compound ID 1434; SEQ ID NO: 87

[0975] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,17G,19Q,25P,32L]-hCNP37

[0976]

[0977] Molecular weight: 4040.57. LCMS34: m / 3 calculated: 1347.8567; m / 3 measured: 1347.9714; m / 4 calculated: 1011.1425; m / 4 measured: 1011.2499

[0978] Chemical formula 105; Compound ID 9435; SEQ ID NO: 88

[0979] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,17G,19Q,25P,32L]-hCNP37

[0980]

[0981] Molecular weight: 4031.603. LCMS34: m / 3 calculated: 1344.8677; m / 3 measured: 1344.7600; m / 4 calculated: 1008.9008; m / 4 measured: 1008.8210

[0982] Chemical formula 106; Compound ID 1436; SEQ ID NO:89

[0983] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,17V,19Q,25P,32L]-hCNP37

[0984]

[0985] Molecular weight: 4073.6828. LCMS34: m / 3 calculated: 1358.8943; m / 3 measured: 1358.6617; m / 4 calculated: 1019.4207; m / 4 measured: 1019.2703

[0986] Chemical formula 107; Compound ID 1437; SEQ ID NO:90

[0987] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,17V,19Q,25P,32L]-hCNP37

[0988]

[0989] Molecular weight: 4082.6498. LCMS34: m / 3 calculated: 1361.8833; m / 3 measured: 1361.7580; m / 4 calculated: 1021.6625; m / 4 measured: 1021.5750

[0990] Chemical formula 108; Compound ID 1448; SEQ ID NO:91

[0991] [N-Terminal([(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]),5Q,10E,13Q,25P,32L]-hCNP37

[0992]

[0993] Molecular Weight: 4741.359. LCMS34: m / z calculated: 1581.4530; m / z measured: 1581.2060; m / z calculated: 1186.3398; m / z measured: 1186.0240

[0994] Chemical Formula 109; Compound ID 1449; SEQ ID NO:51

[0995] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,10H,13Q,14H,15T,17V,19S,25P,32L]-hCNP37

[0996]

[0997] Molecular Weight: 4217.7334. LCMS34: m / z calculated: 1406.9111; m / z measured: 9407.0500; m / z calculated: 1055.4334; m / z measured: 1055.300

[0998] Chemical Formula 110; Compound ID 1450; SEQ ID NO:52

[0999] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,10H,13Q,15T,17V,19S,25P,32L]-hCNP37

[1000]

[1001] Molecular weight: 4208.7664. LCMS34: m / 3 calculated: 1403.9221; m / 3 measured: 1403.7330; m / 4 calculated: 1053.1916; m / 4 measured: 1053.0130

[1002] Chemical formula 111; Compound ID 1451; SEQ ID NO:92

[1003] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,10H,13Q,17V,19S,25P,27E,32L]-hCNP37

[1004]

[1005] Molecular weight: 4249.8614. LCMS34: m / 3 calculated: 1417.6205; m / 3 measured: 1417.4290; m / 4 calculated: 1063.4654; m / 4 measured: 1063.3310

[1006] Chemical formula 112; Compound ID 1452; SEQ ID NO:77

[1007] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,13Q,19Q,25P,32L]-hCNP37

[1008]

[1009] Molecular weight: 4685.3307. LCMS34: m / 3 calculated: 1562.7769; m / 3 measured: 1562.3981; m / 4 calculated: 1172.3327; m / 4 measured: 1172.3179

[1010] Chemical formula 113; Compound ID 1453; SEQ ID NO:77

[1011] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,13Q,19Q,25P,32L]-hCNP37

[1012]

[1013] Molecular weight: 4540.1743. LCMS34: m / 3 calculated: 1514.3914; m / 3 measured: 1514.0494; m / 4 calculated: 1136.0436; m / 4 measured: 1135.8064

[1014] Chemical formula 114; Compound ID 1454; SEQ ID NO:77

[1015] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,13Q,19Q,25P,32L]-hCNP37

[1016]

[1017] Molecular weight: 4701.3732. LCMS34: m / 3 calculated: 1568.1244; m / 3 measured: 1567.7415; m / 4 calculated: 1176.3433; m / 4 measured: 1176.0765

[1018] Chemical formula 115; Compound ID 1455; SEQ ID NO:77

[1019] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,13Q,19Q,25P,32L]-hCNP37

[1020]

[1021] Molecular weight: 4556.2167. LCMS34: m / z calculated: 1519.7389; m / z found: 1519.3893; m / z+1 calculated: 1140.0542; m / z+1 found: 1139.8102

[1022] Chemical formula 116; Compound ID 1456; SEQ ID NO:77

[1023] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,13Q,19Q,25P,32L]-hCNP37

[1024]

[1025] Molecular weight: 4411.0603. LCMS34: m / z calculated: 1471.3534; m / z found: 1471.0409; m / z+1 calculated: 1103.7651; m / z+1 found: 1103.5541

[1026] Chemical formula 117; Compound ID 1457; SEQ ID NO:93

[1027] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),-2G,-1Q,2P,3G,4Q,5A,6P,7G,8Q,9A,10P,13Q,19Q,25P,32L]-hCNP37

[1028]

[1029] Molecular weight: 4410.9775. LCMS34: m / 3 calculated: 1471.3258; m / 3 measured: 1471.0251; m / 4 calculated: 1103.7444; m / 4 measured: 1103.7853

[1030] Chemical formula 118; Compound ID 1458; SEQ ID NO: 94

[1031] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-2,3G,4Q,5A,6P,7G,8Q,9A,10P,13Q,17S,19Q,25P,32L]-hCNP37

[1032]

[1033] Molecular weight: 4057.6038. LCMS34: m / 3 calculated: 1353.5346; m / 3 measured: 1353.3113; m / 4 calculated: 1015.4010; m / 4 measured: 1015.2496

[1034] Chemical formula 119; Compound ID 1459; SEQ ID NO: 95

[1035] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1,2G,3Q,4A,5P,13Q,17S,19Q,25P,32L]-hCNP37

[1036]

[1037] Molecular weight: 4738.3536. LCMS34: m / 3 calculated: 1580.4512; m / 3 measured: 1580.0658; m / 4 calculated: 1185.5884; m / 4 measured: 1185.5701

[1038] Chemical formula 120; Compound ID 1460; SEQ ID NO:96

[1039] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,13Q,25P,32L]-hCNP37

[1040]

[1041] Molecular weight: 4669.3313. LCMS34: m / 3 calculated: 1557.4438; m / 3 found: 1557.4031; m / 4 calculated: 1168.3328; m / 4 found: 1168.0756

[1042] Chemical formula 121; Compound ID 1461; SEQ ID NO:97

[1043] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,10H,13Q,19Q,25P,32L]-hCNP37

[1044]

[1045] Molecular weight: 4096.6763. LCMS34: m / 3 calculated: 1366.5588; m / 3 found: 1366.3214; m / 4 calculated: 1025.1691; m / 4 found: 1025.0148

[1046] Chemical formula 122; Compound ID 1462; SEQ ID NO:96

[1047] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,13Q,25P,32L]-hCNP37

[1048]

[1049] Molecular weight: 4830.5302. LCMS34: m / z calculated: 1611.1767; m / z measured: 1611.1900; m / z4 calculated: 1208.6326; m / z4 measured: 1208.6500

[1050] Chemical formula 123; Compound ID 1463; SEQ ID NO:98

[1051] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,13Q,19Q,32L]-hCNP37

[1052]

[1053] Molecular weight: 4861.5442. LCMS34: m / z calculated: 1621.5147; m / z measured: 1621.1760; m / z4 calculated: 1216.3861; m / z4 measured: 1216.1420

[1054] Chemical formula 124; Compound ID 1464; SEQ ID NO:99

[1055] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,19Q,25P,32L]-hCNP37

[1056]

[1057] Molecular weight: 4816.4606. LCMS34: m / z calculated: 1606.4869; m / z found: 1606.1700; m / z calculated: 1205.1152; m / z found: 1204.8820

[1058] Chemical formula 125; Compound ID 1465; SEQ ID NO:100

[1059] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,13Q,19Q,25P]-hCNP37

[1060]

[1061] Molecular weight: 4848.5256. LCMS34: m / z calculated: 1617.1752; m / z found: 1616.8130; m / z calculated: 1213.1314; m / z found: 1212.8620

[1062] Chemical formula 126; Compound ID 1470; SEQ ID NO:96

[1063] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-5,13Q,25P,32L]-hCNP37

[1064]

[1065] Molecular weight: 4540.2173. LCMS34: m / z calculated: 1514.4058; m / z found: 1514.1500; m / z calculated: 1136.0543; m / z found: 1135.8500

[1066] Chemical formula 127; Compound ID 1471; SEQ ID NO:75

[1067] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), 5Q,13Q,19Q,25P,32L]-hCNP37

[1068]

[1069] Molecular weight: 5159.8011. LCMS34: m / z calculated: 1720.9337; m / z found: 1720.8900; m / z calculated: 1290.9503; m / z found: 1290.9200

[1070] Chemical formula 128; Compound ID 1472; SEQ ID NO:75

[1071] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,13Q,19Q,25P,32L]-hCNP37

[1072]

[1073] Molecular weight: 5030.6872. LCMS34: m / z calculated: 1677.8957; m / z measured: 1677.8700; m / z calculated: 1258.6718; m / z measured: 1258.6600

[1074] Chemical formula 129; Compound ID 1473; SEQ ID NO:67

[1075] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,13Q,19S,25P,32L]-hCNP37

[1076]

[1077] Molecular weight: 4989.6352. LCMS34: m / z calculated: 1664.2117; m / z measured: 1664.2000; m / z calculated: 1248.4088; m / z measured: 1248.400

[1078] Chemical formula 130; Compound ID 1474; SEQ ID NO:101

[1079] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),5Q,13Q,25P,32L]-hCNP37

[1080]

[1081] Molecular weight: 5159.8442. LCMS34: m / z calculated: 1720.9481; m / z measured: 1720.8900; m / z calculated: 1290.9611; m / z measured: 1290.9200

[1082] Chemical formula 131; Compound ID 1475; SEQ ID NO:77

[1083] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(2S)-4-carboxy-2-[[(2S)-4-carboxy-2-[[(2S)-4-carboxy-2-[[(2S)-4-carboxy-2-[[(2S)-4-carboxy-2-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,13Q,19Q,25P,32L]-hCNP37

[1084]

[1085] Molecular weight: 4830.4871. LCMS34: m / z calculated: 1611.1624; m / z measured: 1610.7510; m / z calculated: 1208.6218; m / z measured: 1208.5814

[1086] Chemical formula 132; Compound ID 1476; SEQ ID NO:77

[1087] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(2S)-4-Carboxy-2-[[2-[[(2S)-4-carboxy-2-[[2-[[(2S)-4-carboxy-2-[[2-[[(2S)-4-carboxy-2-[[2-[[(2S)-4-carboxy-2-(17-carboxyheptadecanoyl amino)butanoyl]amino]acetyl]amino]butanoyl]amino]acetyl]amino]butanoyl]amino]acetyl]amino]butanoyl]amino]acetyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,13Q,19Q,25P,32L]-hCNP37

[1088]

[1089] Molecular Weight: 4768.3795. LCMS34: m / 3 calculated: 1590.4598; m / 3 found: 1590.4137; m / 4 calculated: 1193.0949; m / 4 found: 1193.0793

[1090] Chemical Formula 133; Compound ID 1477; SEQ ID NO:102

[1091] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,13Q,19Q,25P,27E,32L]-hCNP37

[1092]

[1093] Molecular Weight: 4844.5137. LCMS34: m / 3 calculated: 1615.8379; m / 3 found: 1615.4194; m / 4 calculated: 1212.1284; m / 4 found: 1212.0844

[1094] Chemical Formula 134; Compound ID 1478; SEQ ID NO:103

[1095] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,19Q,25P,32L]-hCNP37

[1096]

[1097] Molecular weight: 4087.7093. LCMS34: m / z calculated: 1363.5698; m / z found: 1363.3427; m / z calculated: 1022.9273; m / z found: 1022.7753

[1098] Chemical formula 135; Compound ID 9480; SEQ ID NO:103

[1099] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,19Q,25P,32L]-hCNP37

[1100]

[1101] Molecular weight: 4216.8233. LCMS34: m / z calculated: 1406.6078; m / z found: 1406.6766; m / z calculated: 1055.2058; m / z found: 1055.0295

[1102] Chemical formula 136; Compound ID 1481; SEQ ID NO:103

[1103] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(2S)-4-carboxy-2-[[(2S)-4-carboxy-2-[[(2S)-4-carboxy-2-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,19Q,25P,32L]-hCNP37

[1104]

[1105] Molecular weight: 4216.8233. LCMS34: m / z calculated: 1406.6078; m / z found: 1406.3542; m / z calculated: 1055.2058; m / z found: 1055.0295

[1106] Chemical formula 137; Compound ID 9482; SEQ ID NO:67

[1107] [N-terminus([(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), 5Q,13Q,19S,25P,32L]-hCNP37

[1108]

[1109] Molecular weight: 4828.4363. LCMS34: m / z calculated: 1610.4788; m / z found: 1610.4060; m / z calculated: 1208.1091; m / z found: 1207.8275

[1110] Chemical formula 138; Compound ID 9483; SEQ ID NO:104

[1111] [N-terminus([(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]butanoyl]amino]butanoyl]), 5Q,13Q,19S,25P,27E,32L]-hCNP37

[1112]

[1113] Molecular weight: 4713.3489. LCMS34: m / z calculated: 1572.1163; m / z measured: 1572.1500; m / z calculated: 1179.3372; m / z measured: 1179.1200

[1114] Chemical formula 139; Compound ID 1484; SEQ ID NO:105

[1115] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-6,7G,8Q,9A,10P,13Q,19Q,25P,32L]-hCNP37

[1116]

[1117] Molecular weight: 3730.3106. LCMS34: m / z calculated: 1244.4369; m / z measured: 1244.7042; m / z calculated: 933.5777; m / z measured: 933.5259

[1118] Chemical formula 140; Compound ID 1486; SEQ ID NO:106

[1119] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-6,7G,8Q,9A,10P,13Q,17S,19Q,25P,27E,32L]-hCNP37

[1120]

[1121] Molecular weight: 3718.2568. LCMS34: m / z calculated: 1240.4189; m / z measured: 1240.3220; m / z calculated: 930.5642; m / z measured: 930.4920

[1122] Chemical formula 141; Compound ID 1487; SEQ ID NO:107

[1123] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),-2G,-1Q,2P,3G,4Q,5A,6P,7G,8Q,9A,10P,13Q,19Q,25P,32L]-hCNP37

[1124]

[1125] Molecular Weight: 4437.0578. LCMS34: m / 3 calculated: 9480.0193; m / 3 found: 1479.8291; m / 4 calculated: 1110.2645; m / 4 found: 1110.3766

[1126] Chemical formula 142; Compound ID 1488; SEQ ID NO:107

[1127] [N-Terminal([(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]),-2G,-1Q,2P,3G,4Q,5A,6P,7G,8Q,9A,10P,13Q,19Q,25P,32L]-hCNP37

[1128]

[1129] Molecular Weight: 4146.7449. LCMS34: m / 3 calculated: 1383.2483; m / 3 found: 1383.4402; m / 4 calculated: 1037.6862; m / 4 found: 1037.5803

[1130] Chemical formula 143; Compound ID 1489; SEQ ID NO:108

[1131] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-8,13Q,17G,19Q,25P,27E,32L]-hCNP37

[1132]

[1133] Molecular weight: 4045.6296. LCMS34: m / 3 calculated: 1349.5432; m / 3 measured: 1349.3710; m / 4 calculated: 1012.4074; m / 4 measured: 1012.2860

[1134] Chemical formula 144; Compound ID 1493; SEQ ID NO:109

[1135] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,19Q,25P,27E,32L]-hCNP37

[1136]

[1137] Molecular weight: 4101.7359. LCMS34: m / 3 calculated: 1368.2453; m / 3 measured: 1368.0680; m / 4 calculated: 1026.4340; m / 4 measured: 1026.3130

[1138] Chemical formula 145; Compound ID 1511; SEQ ID NO:62

[1139] [N-terminus([2-[2-[2-[[2-[2-[2-[[(2S)-4-carboxy-2-(17-carboxyheptadecanoyl)amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-6,7G,8Q,9A,10P,13Q,17S,19Q,25P,32L]-hCNP37

[1140]

[1141] Molecular weight: 3704.2302. LCMS34: m / 3 calculated: 1235.7434; m / 3 measured: 1235.6700; m / 4 calculated: 927.0576; m / 4 measured: 927.0010

[1142] Chemical formula 146; Compound ID 1512; SEQ ID NO:88

[1143] [N-terminus([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(2S)-4-carboxy-2-[[(2S)-4-carboxy-2-(17-carboxyheptadecanoyl)amino]butanoyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]), des1-8,13Q,17G,19Q,25P,32L]-hCNP37

[1144]

[1145] Molecular weight: 4031.603. LCMS34: m / z calculated: 1344.8677; m / z measured: 1344.7600; m / z calculated: 1008.9008; m / z measured: 1008.8080

[1146] Chemical formula 147; Compound ID 1513; SEQ ID NO: 110

[1147] [N-terminus(17-carboxyheptadecanoyl), -5E, -4E, -3E, -2E, -1E, 5Q, 13Q, 19S, 25P, 32L]-hCNP37

[1148]

[1149] Molecular weight: 4828.4363. LCMS34: m / z calculated: 1610.4788; m / z measured: 1610.2260; m / z calculated: 1208.1091; m / z measured: 1207.9160

[1150] Chemical formula 148; Compound ID 1514; SEQ ID NO: 111

[1151] [N-terminus(17-carboxyheptadecanoyl), -4E, -3E, -2E, -1E, 5Q, 13Q, 19S, 25P, 27E, 32L]-hCNP37

[1152]

[1153] Molecular weight: 4713.3489. LCMS34: m / z calculated: 1572.1163; m / z measured: 1571.8940; m / z calculated: 1179.3372; m / z measured: 1179.1550

[1154] Chemical formula 149; Compound ID 1265; SEQ ID NO: 233

[1155] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4R)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),des1-5,6a,7r,8k,9y,10k,12a,13q,14k,15k,17l,18s,19k,21c,22f,24l,25k,26l,27e,28r,29i,31s,32l,33s,35l,37c]-hCNP37

[1156]

[1157] Molecular weight: 4068.8451. LCMS34: m / 4 calculated: 1018.2113; m / 4 measured: 1018.1958

[1158] Chemical formula 150; Compound ID 0106; SEQ ID NO: 234

[1159] [-2P, -1G]-hCNP37

[1160]

[1161] Molecular weight: 4102.7254. LCMS34: m / 3 calculated: 1368.5751; m / 3 measured: 1368.362; m / 4 calculated: 1026.6814; m / 4 measured: 1026.529

[1162] Chemical formula 151; Compound ID 0089; SEQ ID NO: 1

[1163] [N-terminus([2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl])]-hCNP22

[1164]

[1165] Molecular weight: 2913.4725. LCMS34: m / 3 calculated: 972.1575; m / 3 measured: 971.844; m / 4 calculated: 729.3681; m / 4 measured: 729.131

[1166] Chemical formula 152; Compound ID 0230; SEQ ID NO: 235

[1167] [N-Terminal([2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]),-5G,-4Q,-3A,-2P,-1G,2A,3P,4G,5Q,7P,8G,9Q,10A,11P,12G,13Q,14A,15P]-hCNP37

[1168]

[1169] Molecular weight: 4680.3406. LCMS34: m / z calculated: 1561.11; m / z found: 1560.773; m / z calculated: 1171.0852; m / z found: 1170.834

[1170] Chemical formula 153; Compound ID 0231; SEQ ID NO:1

[1171] [N-Terminal([2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl])]-hCNP22

[1172]

[1173] Molecular weight: 3784.4112. LCMS34: m / z calculated: 1262.4704; m / z

[1174] found: 1262.303; m / z calculated: 947.1028; m / z found: 946.995

[1175] Example 2 - In Vitro hNPR2 Activity Assay

[1176] To evaluate the in vitro activities of CNP22 (0065; chemical formula 1) and the CNP compounds of the present invention, their abilities to induce the production of cyclic guanosine monophosphate (cGMP) in reporter cells expressing human natriuretic peptide receptor 2 (hNPR2) were tested.

[1177] Cell Culture:To generate the reporter cell line, HEK293 ( CRL-1573 TM ) cells were stably transfected with the pGloSensor-42F reporter plasmid (GloSensor cGMP reporter gene - luciferase, Promega) and an expression plasmid encoding the hNPR2 receptor. Single cell clones were isolated as the reporter cell line. When the reporter cells are exposed to an hNPR2 activating compound, hNPR2 causes the production of cGMP. The GloSensor luciferase produced by the pGloSensor-42F plasmid binds to cGMP, and this binding induces a conformational change in the GloSensor luciferase, thereby activating the originally inactive enzyme. The active GloSensor luciferase can convert luciferin to oxyluciferin, and this process produces bioluminescence. After cell lysis and providing the luciferase substrate, the luciferase activity can be quantified by detecting the luminescence. Thus, adding an hNPR2 activating compound to the reporter cells and subsequently adding the detection reagent produces luminescence in a dose-dependent manner. By testing multiple different concentrations of the hNPR2 activating compound, the half maximal effective concentration (EC50) can be determined. This measurement is reported for all tested compounds and is also known as the in vitro potency. In all in vitro activity experiments, CNP22 was used as the positive in-plate control for hNPR2 activation. To reduce the differences between assays, we also report the potency of each compound relative to the potency of the CNP22 control on each plate.

[1178] Procedure: HEK293-hNPR2 / GloSensor clone 35 cells were cultured continuously in selection medium (DMEM containing 10% FCS, 1% P / S, 200 μg / mL G418, and 100 μg / mL hygromycin). For the assay, the cells were detached with TrypLE Express (Gibco#12604013), passed through a 70 μm cell strainer, counted, and seeded at a density of 10,000 cells / well in 30 μL / well of assay medium (DMEM without phenol red but containing 1% P / S) in white 384-well plates (Perkin Elmer#6007680) overnight. The next day, the test compounds were serially diluted in dilution medium (DMEM without phenol red, containing 1% OVA (Sigma#A5505 lot04M7001V), 0.01% Tween 20 (Roche#33766700), and 1% P / S), and then 10 μL / well of the diluted test compounds were added to the cells. For each compound, ten different concentrations in the range of 0.25 pM–10 μM were tested. At 37 °C and 5% CO 2After incubation for 30 minutes, 40 μL / well of Bright-Glo assay reagent (Promega #E2650) was added. Immediately or 15 minutes later, luminescence was measured using an EnVision multimode plate reader (PerkinElmer). To determine the in vitro potency or EC50 of each test compound, four-parameter logistic regression was performed on the raw data of each test compound using the Python package SciPy optimize. The average EC50 values are listed in Table 2 as "hNPR2 0% HSA. [EC50 (nM)]". Relative potency was calculated by the following expression: EC50 (CNP22 intraplate control) / EC50 (test compound) * 100, which is shown in Table 2 as "hNPR2 0% HSA. [EC50% relative to CNP]". At least two replicate measurements were performed for each test compound. The reported values are the averages of the replicate measurements and are listed in Table 2.

[1179] Table 2 - In Vitro Human NPR2 Potency of CNP Compounds:

[1180]

[1181]

[1182]

[1183]

[1184]

[1185] These data demonstrate that the tested CNP compounds cover a wide range of in vitro activation levels of NPR2, and negatively charged CNP compounds generally exhibit reduced in vitro NPR2 potency relative to positively charged CNP compounds.

[1186] Example 3 - In Vitro Neutral Neprilysin Stability of CNP Compounds

[1187] It has been reported that native CNP can be metabolized by neutral lysosomes (neutral endopeptidase, NEP), a protease widely present in the body. Therefore, an in vitro neutral lysosome stability assay was established to test the degradation of selected CNP compounds mediated by neutral lysosomes.

[1188] Procedure:Briefly, the assay buffer consisted of PBS buffer (pH 7.4, ThermoFisher Scientific) containing 0.005% v / v Tween 20 (Sigma - Aldrich). Recombinant human Neprilysin (rhNEP; R&D Systems) and the CNP compound were dissolved in the assay buffer. Each day of incubation included native CNP22 (0065, Chemical Formula 1) as a positive control. First, the CNP compound (final concentration 1000 nM) was pre - incubated in the assay buffer at 37 °C for 10 minutes. The assay was started by adding rhNEP (final concentration 2 μg protein / mL) or the assay buffer (adsorption experiment) and incubated at 37 °C. The reaction was terminated at selected time points (0.5, 5, 15, 30, 60, 90, and 120 minutes) by adding one volume of the incubation mixture to three volumes of ethanol (containing 1% v / v formic acid). The mixture was centrifuged at 13000 rpm for 20 minutes at 4 °C. After centrifugation, one volume of the supernatant was mixed with one volume of Milli - Q water.

[1189] Analysis by LCMS: The mixture was analyzed by LCMS using an Acquity UPLC PeptideCSH C18 analytical column from Waters operating at 60 °C ( 1.7 μm, 1 * 50 mm). A Nexera UHPLC system (Shimadzu) was used with gradient elution using mobile phase A (consisting of Milli - Q water containing 0.1% formic acid) and mobile phase B (consisting of acetonitrile containing 0.1% formic acid). The flow rate was 0.3 ml / min. A zenoTOF 7600 mass spectrometer (Sciex) was used as the detector and operated in positive electrospray ionization mode. Data were recorded in full scan mode (m / z 300 - 1700).

[1190] Calibration curves were made in the assay buffer and treated as study samples. These samples were used to calculate the concentration of the relevant CNP compound in the in vitro samples. Quality control samples were included. For at least 75% of the standards and QCs, the deviation between the nominal concentration and the calculated concentration was less than 20%.

[1191] All incubations were performed in duplicate. Data were reported as the percentage remaining at 90 minutes (based on the calculated concentration) compared to the average concentration in the sample at the 0.5 - minute time point. Examples of seven compounds and CNP22 (0065, Chemical Formula 1) are shown in Table 3.

[1192] Table 3 - Percentage Remaining of CNP Compounds after 90 Minutes of Incubation with rhNEP

[1193] Compound Remaining of Parent after 90 Minutes (%) 0065*(CNP22) 1.7 0106 95.1 9384 87.8 9407 96.6 9435 93.2 9480 98.3 9482 96.3 9483 105.6

[1194] *: Run as a positive control within each experiment

[1195] These data demonstrate that, compared to the rapidly degraded CNP22, the CNP compounds of the present invention are substantially stable in the presence of recombinant human neutrophil lysosomes.

[1196] Example 4 - Peptide Net Charge Calculation

[1197] The charge of the ionizable groups in a peptide at a given pH can be calculated using the Henderson - Hasselbalch equation, where the acid dissociation constants (pKa) of each amino acid residue or modified group, determined empirically, are used, as is well known in the art, for example, by the methods described by B. Skoog and A. Wichman (Trends in Analytical Chemistry, 1986, vol. 5, pp. 82 - 83) and L. Kozlowski (Biology Direct, 2016, 11:55). Negatively charged amino acids or modified groups are shown in Equation 1, while positively charged amino acids and modified groups are shown in Equation 2, where pKn is the pKa of the negatively charged group, pKp is the pKa of the positively charged group, and where the pH in the equation is the target pH. Thus, the net charge of the peptide at a given pH is the sum of the negative and positive charges of all the ionizable groups in the peptide.

[1198] Equation 1:

[1199]

[1200] Equation 2:

[1201]

[1202] For natural amino acid residues, the pKa constants listed in the table are used for the ionizable sites in the free termini and side chains. The pKa values of modified amino acids and modified groups are estimated by the ACD / Labs ver. 12 software package and are also shown in Table 4.

[1203] For example, to calculate the net charge of compound 9482 at pH 7.4, when using the pKa values shown in Table 4, the sum of the negative charges carried by positions Cys37 (C-terminal acid), Asp27, Glu2, gGlu(1), gGlu(2), gGlu(3), gGlu(4), gGlu(5), and C18d fatty acid is -8.995 Fd. The sum of the positive charges carried by positions Arg28, Lys15, Lys14, Lys10, Lys8, Arg7, and His3 is 6.162 Fd. Thus, the net charge is the sum of the above items, which is -2.833.

[1204] The charge values for the CNP compounds of the present invention calculated using the above principle for selected pH values (pH 7.4 and pH 6.5) are shown in Table 5.

[1205] Table 4 - Amino Acids and Modifying Group Terminal and Side Chain pKa Values Used When Calculating the pI and Net Charge of CNP Compounds pKa Value

[1206] Residue N-Terminal C-Terminal Side Chain Ala 7.8 3.7 N / A Arg 7.4 3.6 13.3 Asn 6.1 3.5 N / A Asp 7.4 3.5 4.2 Cys 6.1 3.3 9.1 Gln 6.9 3.5 N / A Glu 8.4 3.5 4.4 Gly 7.3 3.7 N / A His 6.1 3.1 6.7 Ile 7.9 3.7 N / A Leu 7.9 3.7 N / A Lys 7.2 3.6 10.4 Met 6.9 3.5 N / A Phe 7.0 3.6 N / A Pro 8.9 3.4 N / A Ser 6.3 3.3 N / A Thr 6.3 3.3 N / A Trp 7.7 3.7 N / A Tyr 7.3 3.7 9.8 Val 7.9 3.7 N / A Cys in Disulfide 6.1 3.3 N / A C18d N / A N / A 4.8 C20d N / A N / A 4.8 Tetrazole-C18 N / A N / A 5.1 gGlu N / A N / A 3.5

[1207] Table 5 - Calculated Net Charge (Q) Values of CNP Compounds at pH 7.4 and Calculated Net Charge Values at pH 6.5

[1208]

[1209]

[1210]

[1211]

[1212]

[1213] Example 5 - Solubility of CNP Compounds

[1214] To evaluate the suitability of the CNP compounds for soluble liquid formulations, the solubility of the CNP compounds in drug-related buffers was measured. Since pH is expected to affect solubility, the solubility was examined at two different pH levels (pH 4.0 and pH 6.5).

[1215] Preparation of pH 4.0 Samples: To prepare the pH 4.0 sample of the CNP compound, the lyophilized peptide was mixed with 5 mM sodium acetate, 250 mM glycerol buffer at pH 4.0 to a nominal concentration of at least 4000 - 5000 nmol / mL at room temperature. The pH of the sample was measured and adjusted to pH 4.0. If the sample did not dissolve completely, it was allowed to equilibrate at room temperature until the next day.

[1216] Preparation of pH 6.5 Samples:A pH 6.5 sample of the CNP compound was prepared by dissolving the freeze-dried peptide in water. While stirring, a portion of the peptide solution was transferred to a buffer solution (pH 7.5 - 8.0) consisting of 41 mM sodium phosphate and 118 mg / mL glycerol. Subsequently, dilute sodium hydroxide was added, followed by another portion of the peptide solution, and further dilute sodium hydroxide was added. This process was repeated until all of the peptide solution was added, at which point a nominal concentration of at least 4000 - 5000 nmol / mL was reached and the buffer solution consisted of 8 mM sodium phosphate and 23 mg / mL glycerol. The pH of the sample was measured and adjusted to pH 6.5. If the sample did not dissolve completely, it was allowed to equilibrate at room temperature until the next day.

[1217] Procedure: A sample of pH 4.0 was prepared as described above, and the compound concentration in the supernatant was determined using CAD (Method F, CAD02). The compound concentration at pH 4.0 represents the solubility at pH 4.0, and the measured values are given in Table 6.

[1218] Table 6 - Solubility of Formulated CNP Compounds at pH 4.0

[1219] Compound ID (Chemical Formula Reference) Solubility at pH 4.0 (μM) 1351 6168 1381 8995 1389 8896 1422 8286 1426 11242 9435 7416 1470 6476 1473 5832 1474 6376 1478 6507 9480 23 9482 99 9483 4964 1484 0 1489 5932

[1220] A sample of pH 6.5 was prepared as described above, and the peptide concentration in the supernatant was determined by CAD (Method F, CAD02). The peptide concentration at pH 6.5 represents the solubility at pH 6.5, and the measured values are given in Table 7.

[1221] Table 7 – Solubility of CNP Compound Formulations at pH 6.5

[1222]

[1223]

[1224] Empirically, only samples with a content exceeding 4000 μM were further tested as described below.

[1225] These results demonstrate that the CNP compounds of the present invention can be formulated in a pharmaceutically relevant buffer at a concentration suitable for subcutaneous administration.

[1226] Example 6 - Thioflavin T (ThT) Assay

[1227] The ThT assay was used to evaluate the physical stability of the CNP compound and its suitability for liquid formulations. The low physical stability of peptides can lead to the formation of amyloid fibrils, which are observed in samples as well-ordered linear macromolecular structures that ultimately result in gel formation. Traditionally, this has been tested by visual inspection of the samples. However, such measurements are highly subjective and depend on the observer. Therefore, the application of small molecule indicator probes is more advantageous. Thioflavin T (ThT) is such a probe that has unique fluorescence characteristics when bound to fibrils [Naiki et al. (1989) Anal. Biochem 177, 244 - 249; LeVine (1999) Methods. Enzymol. 09, 274 - 284].

[1228] Procedure: Thioflavin T was added to the supernatant samples (pH 4.0 and 6.5) from Example 5 using an aqueous ThT stock solution until the final concentration in the sample was approximately 1 μM. 150 μL sample aliquots were placed in a 96-well microtiter plate (Packard OptiPlateTM-96, white polystyrene). Typically, two to three replicates of each sample were applied to the plate. The plate was sealed and placed in a fluorescence plate reader (Fluoroskan Ascent FL) and incubated at 37 °C with orbital shaking (960 rpm; 1 mm amplitude). Fluorescence measurements were taken every 20 minutes using excitation through a 444 nm filter and emission through a 485 nm filter. Between each measurement, the plate was shaken and heated as described above, and the assay was terminated after 45 hours. The fluorescence measurements for each microtiter plate well were plotted against time, and the lag time (the time until an increase in ThT fluorescence was observed) was estimated.

[1229] To evaluate the loss of the dissolved peptide after 45 hours of incubation, the peptide recovery was measured as the ratio of the total peak area after incubation to that before incubation using the RP-UPLC method described below.

[1230] The lag time and peptide recovery % were measured using RP-UPLC: RP-UPLC was performed using an Acquity UPLC BEH C18 1.7 μm (2.1 x 30 mm) Waters column (eluent A: 0.1% v / v% aqueous TFA solution; eluent B: acetonitrile / water (4:1) solution of 0.1% v / v TFA) with gradient elution (0 min: 95% A; 2 min: 20% A; 2.3 min: 20% A; 2.4 min: 95% A; 9 min: 95% A) at a flow rate of 0.9 mL / min and a column temperature of 30 °C. UV detection at 215 nm was used to evaluate the total area of the peptide.

[1231] Prepare the sample at pH 4.0 as described in Example 5 and perform the ThT assay as described above. The lag time and peptide recovery values are given in Table 8.

[1232] Table 8 – ThT Assay Lag Time and Peptide Recovery of CNP Compounds at pH 4.0

[1233]

[1234] Prepare the sample at pH 6.5 as described in Example 5 and perform the ThT assay as described above. The lag time and peptide recovery values are given in Table 9.

[1235] Table 9 – ThT Assay Lag Time and Peptide Recovery of CNP Compounds at pH 6.5

[1236]

[1237]

[1238] In summary, this experiment demonstrates that most of the CNP compounds of the present invention can be formulated in pharmaceutically relevant buffers and have a low tendency to form fibrils.

[1239] Example 7 - Measurement of High Molecular Weight Protein (HMWP)

[1240] To evaluate the tendency of the CNP compounds to form covalent dimers or oligomers upon formulation, the high molecular weight protein content of the CNP compound formulations was determined at pH 4.0 and 6.5.

[1241] Procedure: The relative amount of covalent HMWP was evaluated using SEC-LIPLC. SEC-UPLC was performed using a Waters UPLC Protein BEH SEC column, 4.6 x 150 mm, 1.7 μm with isocratic elution (0.3 M NaCl, 10 mM NaH 2 PO 4 and 5 mM H 3 PO 4 , 50% (v / v) isopropanol, pH 2.4) at a flow rate of 0.3 mL / min, column temperature of 50 °C, and UV detection at 215 nm. The total area of the peaks eluting before the monomer main peak was defined as HMWP and was given as a percentage relative to the total peptide peak area.

[1242] Results:HMWP formation after static incubation at 37°C for 2 weeks: Samples with pH 4.0 were prepared as described in Example 5, and the amount of HMWP was determined as described above. The samples were incubated at 37°C for two weeks, and the amount of HMWP was measured again. The increase in HMWP after two weeks of incubation at 37°C is reported in Table 10 (if the incubation period deviates from two weeks, the increase in HMWP is normalized to two weeks assuming a constant formation rate).

[1243] Table 10 – 2 - Week HMWP Formation at pH 4.0 (%)

[1244]

[1245]

[1246] Samples with pH 6.5 were prepared as described in Example 5, and the amount of HMWP was determined as described above. The samples were incubated at 37°C for two weeks, and the amount of HMWP was measured again. The increase in HMWP after two weeks of incubation at 37°C is reported in Table 11 (if the incubation period deviates from two weeks, the increase in HMWP is normalized to two weeks assuming a constant formation rate).

[1247] Table 11 - 2 - Week HMWP Formation at pH 6.5 (%)

[1248]

[1249] In summary, this experiment demonstrated that the CNP compounds of the present invention can be formulated in pharmaceutically relevant buffers and have a low tendency to form HMWP.

[1250] Example 8 - Accelerated Chemical Stability of CNP Compounds

[1251] To determine the chemical stability of the CNP compounds in the formulation, samples of the CNP compounds in buffer were exposed to accelerated chemical degradation by heating. The stability of a given peptide was evaluated by quantitatively determining the recovery of the intact peptide by RP-UPLC. To evaluate the chemical degradation pattern, the samples were also analyzed by LC-MS.

[1252] Sample Preparation: Samples for the 2-week study were prepared according to the principles described in Example 5.

[1253] Samples for the 6-week study were prepared as follows: The lyophilized peptide was dissolved in buffer (8 mM phosphate, 250 mM glycerol, pH 7.4) to a nominal peptide concentration of 5000 nmol / ml, and then the pH was adjusted to pH 6.5 with 0.1 N HCl.

[1254] Incubation:For the 2-week study, time zero (TZ) samples were taken and stored at -18 °C, and the remaining volume was incubated at 37 °C. After two weeks of incubation, samples were taken and stored at -18 °C. The frozen samples were thawed, diluted to approximately 200 nmol / mL, and the extent of chemical degradation was determined using LCMS as described.

[1255] For the 6-week study, time zero (TZ) samples were taken and stored at -18 °C, and the remaining volume was incubated at 37 °C. Samples were taken weekly and stored at -18 °C until the last sampling time point at 6 weeks. The frozen samples were thawed, diluted to approximately 200 nmol / mL, and the extent of chemical degradation was determined using RP-UPLC and LCMS as described.

[1256] Analysis of Peptide Recovery by RP - UPLC: The RP-UPLC method was implemented on a Waters ACQUITY H-CLASS UPLC system equipped with a PDA detector. Separation was carried out on an ACQUITY UPLC BEH C8 column ( 1.7 μm, 2.1 mm X 150 mm) using eluent A (19.59 mM NH 4 H 2 PO 4 , 41.71 mM (NH 4 ) 2 HPO 4 , pH 6.5 and MeCN 9:1 v:v) and eluent B (80% aqueous MeCN). UV detection was carried out at 214 nm. Analysis was carried out as follows: 3 μL of the sample was injected onto the column and eluted with a 1-step linear gradient of eluents A and B (from 27% to 30% B in 20 minutes at a constant flow rate of 0.3 mL / min, followed by washing with 95% B for 2 minutes, followed by a 7-minute re-equilibration phase). This resulted in an analysis time of 30 minutes for each sample. The area of the peak corresponding to the intact CNP compound was given as a percentage of the total peptide peak area.

[1257] Analysis of Chemical Degradation Pattern by LCMS: A Waters Acquity C18 reversed-phase column was used. 1.7 μM particles, 1 mm x 150 mm, samples were analyzed on a VANQUISH UPLC from Thermo Scientific. Compounds were separated by a linear gradient from 5% to 55% of solvent B in 48 minutes using solvent A: water with 0.1% formic acid containing 0.02% TFA, and solvent B: acetonitrile with 0.1% formic acid containing 0.02% TFA. Mass spectra were acquired on a Q ExactivePlus hybrid quadrupole - orbitrap mass spectrometer (BioPharma option) (Thermo Scientific) interfaced with an H-ESI II ion source. Data acquisition was performed with a scan range of m / z 500 to 2000 and a resolution of 30000.

[1258] The data lists were summarized into a compound summary table. The header definitions are as follows:

[1259] "Recovery": The recovery of the compound peak (i.e., the intensity of the main peak with the single isotope mass of the compound at time zero) was normalized relative to the compound peak at a given time point; "Isomers": The total intensity of the peaks with isobaric mass to the main compound, which are separated in the RT dimension; "18 amu Loss": I.e., the total intensity of the peaks 18 amu lower in mass than the CNP compound peak, which are separated in the RT dimension; Fragments: The total intensity of the peaks 50 amu lower in mass than the compound peak and the peaks 18 amu higher in mass than the compound mass, which are separated in the RT dimension and develop over time, assumed to originate from hydrolysis of the peptide backbone cleavage; Finally, "Others": Includes the total intensity of the peaks with a 32 amu increase or the dimer of the main peak, which develop over time.

[1260] Results: The LCMS analysis results of the 2-week accelerated degradation study for several CNP compounds of the present invention are summarized in Table 12. The RP-UPLC and LCMS analysis results of the 6-week accelerated degradation study for CNP compound 9482 (Chemical Formula 137) are summarized in Table 13 and Table 14 respectively.

[1261] Table 12 – Recovery and Chemical Degradation Pattern of CNP Compounds after 2 - Week Incubation at 37°C

[1262]

[1263]

[1264] Table 13 – RP - UPLC Analysis of Peptide Recovery of CNP Compound

[9482] after 6 Weeks

[1265]

[1266] Table 14 – LCMS Analysis of Recovery and Chemical Degradation Pattern of CNP Compound 9482 (Chemical Formula 137) after 6 Weeks

[1267]

[1268] These data indicate that the CNP compounds of the present invention exhibit considerable chemical stability in the formulation.

[1269] Example 9 - Local Tolerance of CNP Compounds in Rats

[1270] The purpose of this study was to evaluate the local tissue reactions to subcutaneous injection of CNP compounds in rats.

[1271] Procedure: The CNP compounds were formulated according to the principles described in General Preparation Method - Method G. 120 hours before tissue collection, 250 or 350 μL of the same compound (formulation concentration 1000 nmol / mL) was administered subcutaneously to each Sprague Dawley rat (male, 10 - 11 weeks old at arrival, Janvier, France), with each animal receiving a total of two injections (Table 15). Within each group and time point, the injection site was alternated between animals (flank or neck).

[1272] Table 15 – Experimental Timeline of Local Tolerance Study in Rats

[1273] Time Point Study Activity Day 0 Shave (e.g., neck), followed by administration of the first dose to the shaved area Day 3 Shave (e.g., flank) Day 4 Administer the second dose to the shaved area (e.g., flank) Day 5 Observe skin, tissue collection, and termination

[1274] Animals had free access to tap water and food (Altromin 1324). For the vehicle and positive control, the group size was n = 3, and for the test compounds, the group size was n = 4. As shown in Table 16, the vehicle and compounds were formulated in 5 mM sodium acetate, 250 mM glycerol (pH 4.0) or 8 mM sodium phosphate, 250 mM glycerol (pH 7.4).

[1275] The animals were anesthetized with isoflurane. Using an electric clipper, a square patch of hair (2 cm x 2 cm) was removed from the rat's neck or flank area. For administration, a new injection needle (25G cannula, 1 ml syringe) was placed in the middle of the square boundary closest to the animal's head, and the needle was inserted into the skin from the boundary towards the center of the square. Thus, the tip of the needle was located at the center of the square, the position was marked with a non - fading marker, and the formulation was inserted into the center of the square. On the day of tissue collection, the rats were deeply anesthetized with isoflurane and euthanized by exsanguination. The injection sites were examined and scored for pathological lesions. The injection sites were evaluated, and any changes (position, color, shape, and size) were recorded, and samples were taken for histological preparation.

[1276] The injection sites were removed and fixed in 4% phosphate - buffered neutral formaldehyde in a sealed plastic container.

[1277] Subsequently, each injection site was cut into three slices at 0.8 cm, 1.0 cm, and 1.2 cm from the square boundary where the injection needle was placed. These slices represented the injection site and adjacent tissues respectively. The tissues were processed, embedded in paraffin, cut into thin sections of 2 - 4 μm, stained with hematoxylin and eosin (H&E), and microscopically evaluated using an optical microscope. Histopathological changes were graded on a 5 - point scale (Minimal (Min), Mild (Mil), Moderate (Mod), Marked (Mar), and Severe (Sev)). These studies were summarized in Table 16.

[1278] Table 16 - Overview of Compounds, Sampling Time Points, and Formulations

[1279]

[1280] Results: The results of the local tolerance study are summarized in Table 17.

[1281] Table 17 – Severity and Incidence of Local Subcutaneous Reactions* after Injection of Different Compounds in Rats 4 or 5 Days Later

[1282]

[1283] *Necrosis was used as a marker for local subcutaneous reaction; **NAD = No abnormalities detected

[1284] Conclusion: Positive net charge control 0776 showed moderate to marked local subcutaneous necrosis in rats 5 days after subcutaneous injection.

[1285] In contrast, compounds 1351, 9384, 9407, and 1420 did not show or showed mild local subcutaneous necrosis in rats 5 days after subcutaneous injection and were all evaluated as acceptable for subcutaneous administration in humans.

[1286] Example 10 - Local Tolerance Study of CNP in LYD Pigs

[1287] The purpose of this study was to evaluate the local tissue reaction to subcutaneous injection of CNP compounds in pigs.

[1288] Procedure: In domestic pigs, histopathological examination of the injection site was performed after subcutaneous administration. The injection sites were located on the mid - back between the shoulders and the hips on both sides of the mid - dorsal line, and one or two days before injection, the 2 x 2 cm injection sites were shaved and marked with indelible ink under light anesthesia.

[1289] Each injection consisted of 100 or 200 μl of a formulation at a concentration of approximately 4400 μM. The CNP compound formulation was prepared according to the principles in General Preparation Method - Method G and was A) 8 mM sodium phosphate, 250 mM glycerol, pH 6.5, B) 5 mM sodium acetate, 250 mM glycerol, pH 4.0, or C) 5 mM sodium acetate, 240 mM propylene glycol, pH 4.0, and included physiological saline as a negative control.

[1290] On the first day, the pigs were lightly anesthetized. Using an insulin pen (NovoPen 4) and an insulin needle (NovoTwist 32G / 5 mm) (the needle perpendicular to the skin), the CNP formulation was precisely deposited into the subcutaneous adipose tissue.

[1291] On day 5 (4 days after injection) or day 6 (5 days after injection), the pigs were euthanized, the injection sites were removed and fixed in 4% phosphate - buffered neutral formaldehyde in a sealed plastic container. The tissue blocks were pre - fixed in 10% buffered formalin for 2 - 4 hours, cut into 2 - mm - thick slabs with a multi - purpose knife, macroscopic changes were examined, and finally fixed in the cassette overnight. Two tissue slabs were selected for optical microscopy evaluation, where the tissue reaction was the greatest or in the central part of the injection site. Three to four 2 - 4 - μm sections were cut at 100 - μm intervals from three levels, stained with hematoxylin and eosin (H&E), and evaluated under an optical microscope. Histopathological changes were graded on a 5 - point scale (Min (minimal), Mil (mild), Mod (moderate), Mar (marked), and Sev (severe)). These studies are summarized in Table 18.

[1292] Table 18 - Overview of Compounds, Sampling Time Points, and Formulations:

[1293]

[1294]

[1295] Results: The results are summarized in Table 19. *Necrosis was used as a marker for local subcutaneous reaction;

[1296] **NAD: No abnormalities were detected

[1297] Table 19 – Severity and Incidence of Local Subcutaneous Reactions after Injection of Different Compounds in Pigs 4 or 5 Days Later

[1298]

[1299] In summary, the positively charged control compound 0776 showed moderate to marked local subcutaneous necrosis in pigs 5 days after subcutaneous injection, while the positively charged CNP peptide 0106 without attached fatty acid albumin conjugate did not show observable injection site reactions.

[1300] In contrast, compounds 9480 and 9482 did not show local subcutaneous necrosis in pigs after 5 days of subcutaneous injection; compounds 9384 and 9407 did not show or showed minimal local subcutaneous necrosis in pigs after 5 days of subcutaneous injection; compounds 9435 and 1235 showed minimal to mild local subcutaneous necrosis in pigs after 5 days of subcutaneous injection, while compound 9483 showed no to moderate local subcutaneous necrosis in pigs after 5 days of subcutaneous injection. Compounds 9384, 9407, 9435, 9480 and 9482 were all evaluated as acceptable for human subcutaneous administration.

[1301] Example 11 - PK / PD of CNP Compounds after Intravenous and Subcutaneous Administration to Rats

[1302] To evaluate the intravenous and subcutaneous PK / PD profiles of the CNP compounds of the present invention, formulations of the CNP compounds were administered to rats, blood samples were collected, and exposure and cGMP responses were measured.

[1303] cGMP Biomarker Assay: The biological activity of the CNP compounds in vivo was evaluated by measuring cGMP in plasma samples according to plasma protein precipitation and then quantifying it by LC-MS / MS.

[1304] To evaluate the cyclic guanosine monophosphate (cGMP) biomarker response in biological samples, after administration of the CNP compounds to the study animals, a plasma protein precipitation procedure was applied, followed by liquid chromatography with tandem mass spectrometry (LC-MS / MS) analysis. In principle, a standard curve prepared using a stable isotope-labeled (SIL) surrogate analyte ( 13 C 10 15 N 5 cGMP) as a reference was used to determine the cGMP concentration in the study plasma samples. The surrogate analyte was spiked into the pooled true blank matrix to cover an analytical range of 5 nM - 2000 nM. 8-Methoxymethyl-3-isobutyl-1-methylxanthine (MMPX) was used as an internal standard. The plasma protein precipitation procedure was performed using an organic solvent to precipitate plasma proteins, leaving the supernatant containing cGMP molecules, which was then analyzed on an LC-MS / MS instrument.

[1305] To prepare the standard samples, thaw the frozen authentic blank plasma matrix and then centrifuge at 4000 RPM for 5 minutes at 4°C. Prepare the 2000 nM standard in blank plasma using the SIL-cGMP stock solution and then transfer it to a 1 mL Eppendorf 96-well plate (Patent 8,636,965). Place it in a liquid handler (TECAN Fluent 78) and prepare the standards and QCs (5, 10, 20, 50, 100, 200, 500, 1000, 2000 nM) by serial dilution in blank plasma in a 96-well microplate (Chimney Well 651201). Prepare the blank plasma samples in the same plate. Ventilate and thaw the study samples at ambient temperature for 10 minutes, shake on a bench top shaker at ambient temperature for 5 minutes, and then centrifuge at 4000 RPM for 5 minutes at 4°C. Then place the samples in the liquid handler for the plasma protein precipitation procedure. A certain volume of the standards, QCs, zero samples, and study samples are precipitated with 4 volumes of acetonitrile ( LC / MS grade) containing 30 nM MMPX (Sigma#M2547). Then shake the samples on a bench top shaker at ambient temperature for 4 minutes, then centrifuge at 4000 RPM for 30 minutes at 4°C, and then transfer the supernatant to a new 96-well microplate. Subsequently, use an Eppendorf concentrator plus TM to evaporate the plate at 30°C for 60 minutes. Reconstitute the residue in a certain volume of 5% acetonitrile and 0.1% formic acid (RATHBURN LC / MS grade), shake on a bench top shaker at ambient temperature for 2 minutes, and centrifuge at 4°C for 10 minutes. Then analyze the prepared samples on an LC-MS / MS instrument (Sciex ExionLC coupled with a Sciex 6500+ triple quadrupole TM ). Use 0.1% formic acid as mobile phase A and 95% acetonitrile and 0.1% formic acid as mobile phase B for gradient elution on a reversed-phase LC. Use a Waters Acquity CSH TM Fluoro-Phenyl 1.7 μm, 2.1 x 100 mm column as the stationary phase and thermostat at 60°C. Detect SIL-cGMP, endogenous cGMP, and MMPX in positive ion mode in MS / MS by selective reaction monitoring.

[1306] Acceptance Criteria for Analysis Runs:An SIL-cGMP standard curve was established with 1 / x2 weighting and evaluated in terms of linearity, precision, and accuracy. When calculating backwards, at least 75% of the calibration standards must meet the following criteria: The calibration standards must fall within ±15% of the nominal concentration of their respective calibration standards, except for the LLOQ, which must fall within ±20% of the nominal concentration of the LLOQ standard. Values outside these limits must be discarded as long as the established model is not changed. If discarding one calibration standard causes another calibration standard to go out of bounds, then that calibration standard is also discarded.

[1307] At least two-thirds of the QC samples must be within 15% of their respective nominal values. One-third of the QC samples may exceed 15% of their respective nominal values, but never by more than 50% of the QC samples of the same nominal concentration.

[1308] Quantitative Plasma Analysis of CNP Compounds: The plasma concentration of the CNP compound was determined by plasma protein precipitation and analyzed by turboflow liquid chromatography mass spectrometry (TF-LC-MS). Calibrators were prepared by spiking relevant CNP compounds in the range of 0.5 to 2000 nM into blank rat or minipig plasma. Protein precipitation was performed to prepare calibrators, plasma blanks, or study samples for TF-LC-MS by adding three or four volumes of ethanol or methanol containing 20 nM internal standard to one volume of the sample. For some compounds, formic acid (final concentration 1% v / v) was added to the precipitation reagent. After adding the precipitation reagent, the mixture was centrifuged at 6200 rpm for 30 minutes at 4°C. After centrifugation, one volume of the supernatant was mixed with two volumes of water (containing 1% v / v formic acid). A Cyclone turboflow column (0.5 x 50 mm, ThermoFisher Scientific) and XBridge Protein BEH C4 3.5μm or XBridge Protein BEH C18 3.5μm The analytical columns (both 50 x 2.1 mm, obtained from Waters) were used to analyze the mixture by TF-LC-MS. Gradient elution was performed using mobile phase A (consisting of Milli-Q water containing 1% formic acid and 5% methanol / acetonitrile (50 / 50)) and mobile phase B (consisting of methanol / acetonitrile (50 / 50) containing 1% formic acid and 5% Milli-Q water). The mass spectrometer was operated in positive ionization mode. A TSQ Altis mass spectrometer (ThermoFisher Scientific) was used as the detector in selected reaction monitoring mode (specific transitions were optimized for each CNP compound). Concentrations in plasma samples were calculated using a linear calibration curve (weight 1 / x2). Quality control samples were included. The deviation between the nominal and calculated concentrations of the calibrators and quality control samples was less than 15%.

[1309] Animals and Administration: For each compound and route of administration, CNP compounds were administered to groups of 4 - 6 rats (male, Sprague Dawley, 350 g upon arrival, Janvier, France), formulated in either A) 8 mM sodium phosphate, 250 mM glycerol, 0.007% polysorbate 20, pH 7.4, or B) 5 mM sodium acetate, 250 mM glycerol, 0.007% polysorbate 20, pH 4.0, depending on the respective compound. The subcutaneous dose was 300 nmol / kg at a concentration of 1000 nmol / L, resulting in a dosing volume of 0.3 mL / kg, while the intravenous dose was 100 nmol / kg at a concentration of 100 nmol / L, resulting in a dosing volume of 1 mL / kg.

[1310] Sampling: At baseline (1 h before dosing) and at 5 min (intravenous dosing group only), 2 h, 6 h, 24 h, 48 h, and 72 h (for exposure analysis only) after dosing, 250 μL of sublingual blood samples were collected from unanesthetized rats and placed in EDTA-coated vials (Microvette 600 K3E reference number 15.1673.100 Sarstedt). The blood samples were rapidly kept on ice and centrifuged within 10 minutes after collection (4 °C, 8000 RPM, 5 minutes). 50 μL of plasma samples were transferred to each micronic tube for exposure or cGMP analysis, kept on dry ice and stored in the freezer (-20 °C). The rats had free access to food (Altromin) and tap water.

[1311] Analysis:For each animal, the PK parameters were calculated by non-compartmental analysis (NCA) using the software Phoenix WinNonlin (version 8.1 or higher, Certara). The clearance (Cl) was calculated as: Dose / AUCinf (linear up, log down). The half-life (t 1 / 2 ) was calculated as ln(2) / λ, where λ was estimated by linear regression of time-log concentration, typically in the time range between 2 and 48 hours (e.g., 6 to 48 hours). The bioavailability was calculated as dose-normalized AUC: (AUC(sc) / Dose(sc)) / (AUC(iv) / Dose(iv)). The values in the table are the mean of individual animals (n = 4 - 6 individual animals). Subsequently, the plot of cGMP plasma concentration against time was the mean cGMP concentration after intravenous administration (n = 4 - 6 individual animals) on the (Y-axis) and the nominal plasma sampling time on the (X-axis).

[1312] Results: The PK data including clearance (CL), half-life T 1 / 2 and bioavailability (F%) are summarized in Table 20. The plasma cGMP concentration of the CNP compound after intravenous administration is shown in Figures 1 to 27 and the doses are shown in Table 20. Plasma cGMP was reported as the mean value.

[1313] Table 20 – Pharmacokinetic Profiles of CNP Compounds Administered Intravenously or Subcutaneously in Rats

[1314]

[1315] In summary, this experiment showed that the CNP compounds of the present invention exhibited an extended half-life and reduced clearance in an in vivo rat model. Additionally, this experiment showed that the CNP compounds are biologically active as they are able to elicit a cGMP response in the dosed animals ( Figures 1 to 27 ).

[1316] Example 12 - PK / PD of CNP Compounds after Intravenous and Subcutaneous Administration to Mini - Pigs and LYD Pigs

[1317] The purpose of this study was to investigate the pharmacokinetic and pharmacodynamic properties after intravenous (i.v.) and subcutaneous (s.c.) administration to minipigs and domestic LYD pigs and to estimate the bioavailability after subcutaneous administration. Quantitative plasma analysis of CNP and cGMP levels was performed as described in Example 11.

[1318] Animals and Administration: At CRO Minerva Imaging or Novo Nordisk A / S respectively for Pharmacokinetic and pharmacodynamic studies were conducted in miniature pigs or domestic LYD pigs. The CNP compound was administered to normal female 1) miniature pigs at 6 - 12 months of age with an average body weight of 22 kg or 2) domestic LYD pigs at 4 - 5 months of age with an average body weight of 80 kg. At least one week before dosing, a permanent central venous catheter was implanted for blood sampling and intravenous (i.v.) administration. For subcutaneous (s.c.) administration, the pigs were scanned by ultrasound, and the optimal injection area on the side of the neck was marked with a permanent tattoo. The CNP preparation was accurately deposited into the subcutaneous adipose tissue using an insulin pen (NovoPen Echo or NovoPen 4) and an insulin needle (NovoFine 32G / 4mm or NovoTwist 32G / 5mm) (the needle perpendicular to the skin). The CNP compound was formulated according to the principles in General Preparation Method - Method G and was A) 8 mM sodium phosphate, 250 mM glycerol, pH 6.5, B) 5 mM sodium acetate, 250 mM glycerol, pH 4.0, C) 5 mM sodium acetate, 240 mM propylene glycol, pH = 4.0, D) 8 mM sodium phosphate, 250 mM glycerol, pH 7.5, or E) 20 mM sodium phosphate, 223 mM propylene glycol, pH 6.0.

[1319] For intravenous administration, using formulations A - E, the CNP compound was administered to miniature pigs at 30 - 60 nmol / kg and to domestic pigs at 15 - 43 nmol / kg. For subcutaneous administration, using formulations A - D, the CNP compound was administered to miniature pigs at 55 - 62 nmol / kg and to domestic pigs at 20 - 30 nmol / kg.

[1320]

[1321] Sampling: Approximately 1 mL of blood samples were collected before dosing and up to 14 days after dosing and placed in EDTA - coated tubes (8 mM) for cGMP and CNP compound analysis by LC - MS. In a few cases, domestic LYD pigs were analyzed for cGMP only up to 48 hours after intravenous administration. The blood samples were stored on wet ice for up to 30 minutes until centrifuged at 4°C and at least 1500 G for 10 minutes, and the plasma samples were stored at - 20°C until analysis.

[1322] ​Phoenix WinNonlin 8.1, Pharsight Inc., Mountain View, CA, USA was used to analyze the plasma concentration-time curve by non-compartmental PK analysis. The area under the plasma concentration-time curve (AUC) was calculated based on the "linear up-log down" method and uniform weighting was used to estimate the terminal rate constant (λz). Subcutaneous bioavailability (F) was calculated as the dose-normalized AUC (AUC / dose) after subcutaneous administration divided by the AUC / dose after intravenous administration.

[1323] Results: The PK data of minipigs are summarized in Table 21, including clearance (CL), half-life T 1 / 2 and bioavailability (F%). The PK data of LYD domestic pigs are summarized in Table 22, including clearance (CL), half-life T 1 / 2 and bioavailability (F%). Figures 28 to 48 The plasma cGMP concentrations after intravenous or subcutaneous administration of the CNP compound to minipigs and domestic pigs are shown, with the doses as shown in Tables 21 and 22. Plasma cGMP was reported as the mean of n = 2 - 4 over 14 days for minipigs and the mean of n = 2 - 4 over 2 days for domestic LYD pigs.

[1324] Table 21. Pharmacokinetic Parameters of CNP Compounds after Intravenous and Subcutaneous Administration to Mini - Pigs

[1325]

[1326] Table 22 - Pharmacokinetic Parameters of CNP Compounds after Intravenous and Subcutaneous Administration to Domestic LYD Pigs

[1327]

[1328] Half-life (T 1 / 2 ) and subcutaneous bioavailability (F) are given as means in Tables 21 and 22.

[1329] Conclusion: This experiment shows that the CNP compounds of the present invention exhibit an extended half-life and reduced clearance in both minipigs and domestic LYD pigs. In addition, this experiment shows that the CNP compounds are biologically active as they are able to elicit a cGMP response in the dosed animals ( Figures 28 to 48 ).

[1330] In addition, this experiment shows that the subcutaneous bioavailability (F) of the compounds of the present invention (Tables 21 and 22) is higher than 40%. While the positively charged non-working examples (Compound IDs 0776 and 0312) showed low subcutaneous bioavailability (7-13%). The non-working example with a substantially neutral charge (Compound ID 1235) showed a moderate subcutaneous bioavailability of 25%.

[1331] Example 13 - Telemetry of Heart Rate and Blood Pressure

[1332] The purpose of this study was to investigate the effects of acute administration of CNP compound 9482 (chemical formula 137) on mean arterial blood pressure (MAP) and heart rate (HR) in conscious rats.

[1333] Procedure: The transducer was implanted into Sprague Dawley rats (Charles River), and the rats were allowed a 2-3 week recovery period. Before the surgery, the device was registered in the software by placing it on the receiving board and turning on the power. Sterile saline was added to the transducer catheter before the start of the surgery, and then the transmitter was turned on. The transmitter was placed in the abdomen, and the sensor was located in the abdominal aorta. Isoflurane was used as an anesthetic, and during and after the surgery, the rats received Temgesic (0.05 mg / kg subcutaneously), Norodyl (5 mg / kg subcutaneously), and Baytril (10 mg / kg subcutaneously) as pain relievers and to prevent bacterial infection.

[1334] Study Design: Two rats were housed per cage (one with the device and one as a social partner). Throughout the study, all animals had free access to water and food (altromin). The rats were housed according to a light cycle of 6 am to 6 pm light and 6 pm to 6 am dark. According to a crossover study design, 8 rats were divided into two groups.

[1335] Telemetry: Each rat was equipped with a transducer (HD-S10 from DSI) that was capable of measuring blood pressure, heart rate, and activity using Ponemah software at a frequency of 100 Hz. During the measurement, the room where the rats were housed should not be disturbed to minimize the variation in the data. Before the start of the study, a 4-hour baseline recording was performed to capture the mean arterial blood pressure (MAP) and heart rate (HR) patterns of untreated and undisturbed rats. The recording period after administration was 24 hours.

[1336] Dosing Regimen:Rats were dosed using a crossover study design, with 4 rats per group. Each rat was given a one-week washout period before the next dosing event. Dosing was intravenous (IV) via the tail vein. In separate experiments, the administration of 9482 was tested intravenously at 30, 100, and 300 nmol / kg. The vehicle consisted of 8 mM phosphate, 250 mM glycerol (pH 7.4).

[1337] Data analysis and statistics were performed in Ponemah software and Graphpad prism. For statistics, a paired t-test was used, with a confidence level of 95%, and P < 0.05 indicating statistical significance.

[1338] Results: Within the first 30 minutes after dosing, no differences in MAP and HR were observed for the CNP compound 9482 at 30, 100, or 300 nmol / kg compared to the vehicle (Tables 23 and 25). The mean values for the first 12 hours showed no differences in MAP between the vehicle and the three test doses of 9482 (Table 24). During the same period, an 8.8 ± 10.1% and 6.4 ± 6.6% increase in HR was detected for 100 and 300 nmol / kg (P < 0.05) (Table 26). No change in HR was observed for the 30 nmol / kg dose (Table 26).

[1339] Table 23 – Mean Arterial Pressure: 30 min

[1340]

[1341]

[1342] Table 24 – Mean Arterial Pressure: 12 h

[1343]

[1344] Table 25 – Heart Rate: 30 min

[1345]

[1346] Table 26 – Heart Rate: 12 h

[1347]

[1348] Conclusion: An acute intravenous infusion of the CNP compound 9482 did not cause any major hemodynamic changes within 12 hours, as evidenced by the unchanged MAP and only a slight increase in HR at the two highest doses.

[1349] Example 14 - Mouse Metatarsal Dissection and Culture:

[1350] The purpose of this study was to evaluate bone growth after administration of CNP compounds in an ex vivo mouse model.

[1351] Procedure: One-day-old mouse pups (NMRI, Javier Labs France) were euthanized and then dissected. All applicable international and domestic Novo Nordisk guidelines regarding animal care and use were followed. Three middle metatarsal bones were dissected from each hind paw. The dissected bones were placed in a 24-well cell culture plate and cultured in 400 μl of α-minimum essential medium supplemented with 0.2% BSA and 1% penicillin / streptomycin at 37 °C and 5% CO 2 2. The bones from all animals were randomly divided into a control group and six CNP compound treatment groups (100 nM; n = 8). The medium was changed every 2 - 3 days. All compounds were formulated according to the principles described in General Preparation Method - Method G. Compounds 9435 and 9483 were formulated in A) 5 mM sodium acetate, 250 mM glycerol, pH 4.0, while compounds 9384, 9407, 9480, and 9482 were formulated in B) 8 mM sodium phosphate, 250 mM glycerol, pH 7.4.

[1352] The vehicle alone did not affect bone growth compared to medium alone.

[1353] On days 0, 4, 7, 11, and 14, digital photographs were taken using a digital camera attached to a Nikon microscope. The length of each metatarsal bone was measured using ImageJ software, and the increase in bone length was expressed as a percentage change (mean (SD)) compared to the length measured on the day of dissection.

[1354] Results: After 11 days of treatment, all CNP compounds significantly increased bone length compared to the control group (p < 0.05; two-way ANOVA). After 14 days of culture, the total increase in bone length after treatment with CNP compounds was 28 - 48% higher compared to the control group. The study results are summarized in Table 27.

[1355] Table 27 - Increase in bone length after administration of CNP compound (100 nM):

[1356]

[1357]

[1358]

[1359] In Table 27, metatarsal bones were isolated from mice and cultured for up to 14 days. Throughout the study, the length was measured every 3 - 4 days and expressed as the percentage change compared to day 0 (mean (SD)). Statistically significant differences compared to the control group were indicated (two-way ANOVA).

[1360] Conclusion: This experiment demonstrated that the CNP compound of the present invention was able to significantly increase bone length in an ex vivo mouse model.

[1361] Example 15 - Mouse growth study

[1362] The purpose of this study was to evaluate the growth of mice after treatment with two doses of the CNP compound.

[1363] Procedure: Mice (C57BL / 6J, male, 4 weeks old; n = 10) received daily subcutaneous injections of compound 9482 or vehicle for 35 days. The formulations were prepared according to the principles described in General Preparation Method - Method G (30 and 70 nmol / kg, 10 mL / kg; formulation buffer 8 mM sodium phosphate, 250 mM glycerol, 0.007% polysorbate 20, pH 7.4). Throughout the study, body weight, tail length, and body length were measured weekly. The increase in growth was expressed as the percentage change compared to day 0 (mean (SD)).

[1364] Results: Compared to the control, the 30 and 70 nmol / kg doses of CNP compound 9482 significantly increased tail length and body length after 7 days and 18 days, respectively (p < 0.05, two-way ANOVA). After 35 days, compared to control animals, the changes in tail length (compared to day 0) increased by 45% and 83% respectively after treatment with 30 and 70 nmol / kg CNP compound. After 35 days of treatment with 30 and 70 nmol / kg CNP compound, body length increased by 30% and 67% respectively compared to the control group. The study results are summarized in Tables 28 and 29.

[1365] Table 28 - Tail length of mice treated with daily injection of CNP compound for 35 days

[1366]

[1367] In Table 28, the increase in tail length was expressed as the percentage change compared to day 0 (mean (SD)). Statistically significant differences compared to the control group were indicated (two-way ANOVA).

[1368] Table 29 - Body length of mice treated with daily injection of CNP compound for 35 days

[1369]

[1370] In Table 29, the increase in body length is expressed as the percentage change (mean (SD)) compared to day 0. Statistically significant differences compared to the control group are indicated (two-way ANOVA).

[1371] Conclusion: This experiment demonstrated that when subcutaneously administered in a pharmaceutically relevant formulation, the CNP compounds of the present invention significantly increased the tail length and body length of mice.

Claims

1. A CNP compound comprising a CNP peptide and a modifying group, wherein the net charge of the compound at physiological pH is 0 or negative, and wherein the CNP peptide comprises an amino acid sequence according to Formula I: AA 01 -AA 02 -AA 03 -AA 04 -AA 05 -AA 06 -AA 07 -AA 08 -AA 09 -AA 10 -AA 11 -AA 12 - AA 13 -AA 14 -AA 15 -AA 16 -AA 17 -AA 18 -AA 19 -AA 20 -AA 21 -AA 22 -AA 23 -AA 24 - AA 25 -AA 26 -AA 27 -AA 28 -AA 29 -AA 30 -AA 31 -AA 32 -AA 33 -AA 34 -AA 35 -AA 36 - AA 37 wherein AA 01 is Gln or absent, AA 02 is Glu or absent, AA 03 is His or does not exist, AA 04 is Pro or does not exist, AA 05 is Asn or Gln or Glu or absent, AA 06 is Ala or absent, AA 07 is Arg or His or Ala or absent, AA 08 is Lys or Ser or His or absent, AA 09 is Tyr or Glu or absent, AA 10 is Lys or Glu or Gln or His or absent, AA 11 is Gly, AA 12 is Ala, AA 13 is Gln or Asn or Glu, AA 14 is Lys or His or Glu, AA 15 is Lys or Ser or Glu or Thr or His, AA 16 is Gly, AA 17 is Leu or Gly or Ser or Val, AA 18 is Ser or His, AA 19 is Gln or Ser or Lys or His, AA 20 is Gly, AA 21 is Cys, AA 22 is Phe, AA 23 is Gly, AA 24 is Leu, AA 25 is Pro or Lys, AA 26 is Leu, AA 27 is Asp or Glu, AA 28 is Arg, AA 29 is Ile, AA 30 is Gly, AA 31 is Ser AA 32 is Leu or Nle or Met, AA 33 is Ser AA 34 is Gly, AA 35 is Leu, AA 36 is Gly, and AA 37 is Cys; wherein the modifying group comprises Chemical Formula A, Chemical Formula B, and Chemical Formula C; wherein Chemical Formula A is selected from: (Chemical Formula A1), and (Chemical Formula A2) where p is an integer in the range of 14 - 20, and where * represents an amide bond connecting Chemical Formula A and Chemical Formula B; and where Chemical Formula B is selected from: where q is an integer in the range of 1 - 8, where * represents an amide bond connecting Chemical Formula A- and Chemical Formula B-, where ** represents an amide bond connecting Chemical Formula B- and Chemical Formula C-; and where Chemical Formula C is selected from: (Chemical Formula C4), where r is an integer in the range of 0 - 4, where s is an integer in the range of 0 - 3, where t is an integer in the range of 0 - 1, where u is an integer in the range of 0 - 3, where ** represents an amide bond connecting Chemical Formula B- and Chemical Formula C-, where *** represents an amide bond connecting Chemical Formula C- and the N-terminal α-amine on the CNP peptide.

2. The CNP compound according to claim 1, wherein the AA of the CNP peptide 13 is Gln; AA 19 is Gln or Ser; AA 25 is Pro; and AA 32 is Leu.

3. The CNP compound according to claim 1, wherein the AA of the CNP peptide 5 is Gln; AA 13 is Gln; AA 19 is Gln or Ser; AA 25 is Pro; and AA 32 is Leu.

4. The CNP compound according to claim 1, wherein the CNP peptide comprises one of the following amino acid sequences: YKGANKKGLSKGCFGLKLDRIGSXSGLGC SEQ ID NO:6 NARKYKGANKSGLSSGCFGLKLDRIGSXSGLGC SEQ ID NO:7 GANKKGLSKGCFGLKLDRIGSXSGLGC SEQ ID NO:8 GAQKKGLSKGCFGLKLDRIGSXSGLGC SEQ ID NO:9 YEGAQKKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:10 EEGAQKKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:11 YEGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:12 YEGAQKKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:13 YEGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:14 YEGAQEKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:15 YEGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:16 YEGAQEKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:17 EKGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:18 EKGAQEKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:19 EQGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:20 YKGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:21 YKGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:22 EHGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:23 YHGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:24 YHGAQEKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:25 YHGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:26 YHGAQEKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:27 YEGAEKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:28 EGAQEKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:29 EGAQKEGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:30 YEGAQKEGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:31 YHGAQKTGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:32 YHGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:33 YHGAQHTGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:34 YKGAQHTGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:35 EHGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:36 EHGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:37 EHGAQKKGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:38 EHGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:39 YHGAQHHGLSKGCFGLKLERIGSLSGLGC SEQ ID NO:40 YKGAQHHGLSQGCFGLKLERIGSLSGLGC SEQ ID NO:41 YEGAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:42 EGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:43 YKGAQHHGLSQGCFGLKLDRIGSLSGLGC SEQ ID NO:44 YKGAQHHGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:45 YHGAQHHGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:46 YHGAQHHGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:47 GAQKKGLSHGCFGLPLDRIGSLSGLGC SEQ ID NO:48 GAQKKGLHSGCFGLPLDRIGSLSGLGC SEQ ID NO:49 ARKYHGAQHTGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:50 ARKYHGAQHTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:51 ARKYHGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:52 ARKEHGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:53 ARKEHGAQHTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:54 YEGAQKKGGSKGCFGLPLDRIGSLSGLGC SEQ ID NO:55 QEHPQARSYEGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:57 QEHPEARSYEGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:58 QEHPQAHKYHGAQHHGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:59 QEHPQAHKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:60 QEHPQAHKYHGAQKHGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:61 ARKYEGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:63 ARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:64 QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:67 QEHPQARKYEGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:68 YEGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:69 QEHPQARKYEGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:70 QEHPQAAKYEGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:71 QEHPQARHYKGAQKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:72 QEHPQAHKYKGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:73 QEHPQARKYKGAQKKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:74 QEHPQARKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:75 QEHPQARKYKGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:76 ARKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:77 ARKYHGAQHTGVSKGCFGLPLDRIGSLSGLGC SEQ ID NO:78 ARKYKGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:79 ARKYHGAQKKGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:80 ARKYHGAQKTGVSKGCFGLPLDRIGSLSGLGC SEQ ID NO:81 ARKYKGAQKTGVSKGCFGLPLDRIGSLSGLGC SEQ ID NO:82 YHGAQKTGVSKGCFGLPLDRIGSLSGLGC SEQ ID NO:83 ARKYHGAQKSGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:84 YEGAQKKGGSHGCFGLPLDRIGSLSGLGC SEQ ID NO:85 ARKYEGAQHKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:86 YHGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:87 YKGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:88 YKGAQKKGVSQGCFGLPLDRIGSLSGLGC SEQ ID NO:89 YHGAQKKGVSQGCFGLPLDRIGSLSGLGC SEQ ID NO:90 QEHPQARKYEGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:91 ARKYHGAQKKGVSSGCFGLPLERIGSLSGLGC SEQ ID NO:92 ARKYKGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:96 YHGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:97 ARKYKGAQKKGLSQGCFGLKLDRIGSLSGLGC SEQ ID NO:98 ARKYKGANKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:99 ARKYKGAQKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:100 QEHPQARKYKGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:101 ARKYKGAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:102 YKGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:103 QEHPQARKYKGAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:104 YKGAQKKGGSQGCFGLPLERIGSLSGLGC SEQ ID NO:108 YKGAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:109 GAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:112 GAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:113 GAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:114 GAQKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:115 GAQKKGLSQGCFGLKLDRIGSLSGLGC SEQ ID NO:116 GAQKKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:117 GAQKKGGSQGCFGLKLDRIGSLSGLGC SEQ ID NO:118 GAQKKGGSSGCFGLKLDRIGSLSGLGC SEQ ID NO:119 GANKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:120 GANKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:121 GANKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:122 GANKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:123 GAQKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:124 GAQKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:125 GAQKKGGSQGCFGLPLDRIGSMSGLGC SEQ ID NO:126 GAQKKGGSSGCFGLPLDRIGSMSGLGC SEQ ID NO:127 GAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:128 GAQKKGGSKGCFGLPLDRIGSLSGLGC SEQ ID NO:129 GAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:130 GAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:131 GANKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:132 GANKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:133 GANKKGLSQGCFGLPLERIGSMSGLGC SEQ ID NO:134 GANKKGLSSGCFGLPLERIGSMSGLGC SEQ ID NO:135 GAQKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:136 GAQKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:137 GAQKKGSSQGCFGLKLDRIGSLSGLGC SEQ ID NO:138 GAQKKGSSSGCFGLKLDRIGSLSGLGC SEQ ID NO:139 GANKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:140 GANKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:141 GAQKKGSSQGCFGLPLDRIGSMSGLGC SEQ ID NO:142 GAQKKGSSSGCFGLPLDRIGSMSGLGC SEQ ID NO:143 GAQKKGSSKGCFGLPLDRIGSLSGLGC SEQ ID NO:144 YKGAQKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:145 YKGAQKKGLSQGCFGLKLDRIGSLSGLGC SEQ ID NO:146 YKGAQKKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:147 YKGAQKKGGSQGCFGLKLDRIGSLSGLGC SEQ ID NO:148 YKGAQKKGGSSGCFGLKLDRIGSLSGLGC SEQ ID NO:149 YKGANKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:150 YKGANKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:151 YKGANKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:152 YKGANKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:153 YKGAQKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:154 YKGAQKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:155 YKGAQKKGGSQGCFGLPLDRIGSMSGLGC SEQ ID NO:156 YKGAQKKGGSSGCFGLPLDRIGSMSGLGC SEQ ID NO:157 YKGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:158 YKGAQKKGGSKGCFGLPLDRIGSLSGLGC SEQ ID NO:159 YKGAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:160 YKGAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:161 YKGANKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:162 YKGANKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:163 YKGANKKGLSQGCFGLPLERIGSMSGLGC SEQ ID NO:164 YKGANKKGLSSGCFGLPLERIGSMSGLGC SEQ ID NO:165 YKGAQKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:166 YKGAQKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:167 YKGAQKKGSSQGCFGLKLDRIGSLSGLGC SEQ ID NO:168 YKGAQKKGSSSGCFGLKLDRIGSLSGLGC SEQ ID NO:169 YKGANKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:170 YKGANKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:171 YKGAQKKGSSQGCFGLPLDRIGSMSGLGC SEQ ID NO:172 YKGAQKKGSSSGCFGLPLDRIGSMSGLGC SEQ ID NO:173 YKGAQKKGSSKGCFGLPLDRIGSLSGLGC SEQ ID NO:174 ARKYKGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:175 ARKYKGAQKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:176 ARKYKGAQKKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:177 ARKYKGAQKKGGSQGCFGLKLDRIGSLSGLGC SEQ ID NO:178 ARKYKGAQKKGGSSGCFGLKLDRIGSLSGLGC SEQ ID NO:179 ARKYKGANKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:180 ARKYKGANKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:181 ARKYKGANKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:182 ARKYKGANKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:183 ARKYKGAQKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:184 ARKYKGAQKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:185 ARKYKGAQKKGGSQGCFGLPLDRIGSMSGLGC SEQ ID NO:186 ARKYKGAQKKGGSSGCFGLPLDRIGSMSGLGC SEQ ID NO:187 ARKYKGAQKKGGSKGCFGLPLDRIGSLSGLGC SEQ ID NO:188 ARKYKGAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:189 ARKYKGAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:190 ARKYKGANKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:191 ARKYKGANKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:192 ARKYKGANKKGLSQGCFGLPLERIGSMSGLGC SEQ ID NO:193 ARKYKGANKKGLSSGCFGLPLERIGSMSGLGC SEQ ID NO:194 ARKYKGAQKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:195 ARKYKGAQKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:196 ARKYKGAQKKGSSQGCFGLKLDRIGSLSGLGC SEQ ID NO:197 ARKYKGAQKKGSSSGCFGLKLDRIGSLSGLGC SEQ ID NO:198 ARKYKGANKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:199 ARKYKGANKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:200 ARKYKGAQKKGSSQGCFGLPLDRIGSMSGLGC SEQ ID NO:201 ARKYKGAQKKGSSSGCFGLPLDRIGSMSGLGC SEQ ID NO:202 ARKYKGAQKKGSSKGCFGLPLDRIGSLSGLGC SEQ ID NO:203 GAQKKGSSQGCFGLPLERIGSLSGLGC SEQ ID NO:232。 5. The CNP compound according to claim 1, wherein the CNP peptide has any of the following amino acid sequences: YKGANKKGLSKGCFGLKLDRIGSXSGLGC SEQ ID NO:6 NARKYKGANKSGLSSGCFGLKLDRIGSXSGLGC SEQ ID NO:7 GANKKGLSKGCFGLKLDRIGSXSGLGC SEQ ID NO:8 GAQKKGLSKGCFGLKLDRIGSXSGLGC SEQ ID NO:9 YEGAQKKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:10 EEGAQKKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:11 YEGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:12 YEGAQKKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:13 YEGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:14 YEGAQEKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:15 YEGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:16 YEGAQEKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:17 EKGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:18 EKGAQEKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:19 EQGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:20 YKGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:21 YKGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:22 EHGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:23 YHGAQEKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:24 YHGAQEKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:25 YHGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:26 YHGAQEKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:27 YEGAEKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:28 EGAQEKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:29 EGAQKEGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:30 YEGAQKEGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:31 YHGAQKTGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:32 YHGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:33 YHGAQHTGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:34 YKGAQHTGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:35 EHGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:36 EHGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:37 EHGAQKKGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:38 EHGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:39 YHGAQHHGLSKGCFGLKLERIGSLSGLGC SEQ ID NO:40 YKGAQHHGLSQGCFGLKLERIGSLSGLGC SEQ ID NO:41 YEGAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:42 EGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:43 YKGAQHHGLSQGCFGLKLDRIGSLSGLGC SEQ ID NO:44 YKGAQHHGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:45 YHGAQHHGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:46 YHGAQHHGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:47 GAQKKGLSHGCFGLPLDRIGSLSGLGC SEQ ID NO:48 GAQKKGLHSGCFGLPLDRIGSLSGLGC SEQ ID NO:49 ARKYHGAQHTGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:50 ARKYHGAQHTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:51 ARKYHGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:52 ARKEHGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:53 ARKEHGAQHTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:54 YEGAQKKGGSKGCFGLPLDRIGSLSGLGC SEQ ID NO:55 QEHPQARSYEGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:57 QEHPEARSYEGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:58 QEHPQAHKYHGAQHHGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:59 QEHPQAHKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:60 QEHPQAHKYHGAQKHGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:61 ARKYEGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:63 ARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:64 QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:67 QEHPQARKYEGAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:68 YEGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:69 QEHPQARKYEGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:70 QEHPQAAKYEGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:71 QEHPQARHYKGAQKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:72 QEHPQAHKYKGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:73 QEHPQARKYKGAQKKGLSKGCFGLKLDRIGSLSGLGC SEQ ID NO:74 QEHPQARKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:75 QEHPQARKYKGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:76 ARKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:77 ARKYHGAQHTGVSKGCFGLPLDRIGSLSGLGC SEQ ID NO:78 ARKYKGAQKTGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:79 ARKYHGAQKKGVSSGCFGLPLDRIGSLSGLGC SEQ ID NO:80 ARKYHGAQKTGVSKGCFGLPLDRIGSLSGLGC SEQ ID NO:81 ARKYKGAQKTGVSKGCFGLPLDRIGSLSGLGC SEQ ID NO:82 YHGAQKTGVSKGCFGLPLDRIGSLSGLGC SEQ ID NO:83 ARKYHGAQKSGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:84 YEGAQKKGGSHGCFGLPLDRIGSLSGLGC SEQ ID NO:85 ARKYEGAQHKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:86 YHGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:87 YKGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:88 YKGAQKKGVSQGCFGLPLDRIGSLSGLGC SEQ ID NO:89 YHGAQKKGVSQGCFGLPLDRIGSLSGLGC SEQ ID NO:90 QEHPQARKYEGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:91 ARKYHGAQKKGVSSGCFGLPLERIGSLSGLGC SEQ ID NO:92 ARKYKGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:96 YHGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:97 ARKYKGAQKKGLSQGCFGLKLDRIGSLSGLGC SEQ ID NO:98 ARKYKGANKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:99 ARKYKGAQKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:100 QEHPQARKYKGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:101 ARKYKGAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:102 YKGAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:103 QEHPQARKYKGAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:104 YKGAQKKGGSQGCFGLPLERIGSLSGLGC SEQ ID NO:108 YKGAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:109 GAQKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:112 GAQKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:113 GAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:114 GAQKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:115 GAQKKGLSQGCFGLKLDRIGSLSGLGC SEQ ID NO:116 GAQKKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:117 GAQKKGGSQGCFGLKLDRIGSLSGLGC SEQ ID NO:118 GAQKKGGSSGCFGLKLDRIGSLSGLGC SEQ ID NO:119 GANKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:120 GANKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:121 GANKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:122 GANKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:123 GAQKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:124 GAQKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:125 GAQKKGGSQGCFGLPLDRIGSMSGLGC SEQ ID NO:126 GAQKKGGSSGCFGLPLDRIGSMSGLGC SEQ ID NO:127 GAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:128 GAQKKGGSKGCFGLPLDRIGSLSGLGC SEQ ID NO:129 GAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:130 GAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:131 GANKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:132 GANKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:133 GANKKGLSQGCFGLPLERIGSMSGLGC SEQ ID NO:134 GANKKGLSSGCFGLPLERIGSMSGLGC SEQ ID NO:135 GAQKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:136 GAQKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:137 GAQKKGSSQGCFGLKLDRIGSLSGLGC SEQ ID NO:138 GAQKKGSSSGCFGLKLDRIGSLSGLGC SEQ ID NO:139 GANKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:140 GANKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:141 GAQKKGSSQGCFGLPLDRIGSMSGLGC SEQ ID NO:142 GAQKKGSSSGCFGLPLDRIGSMSGLGC SEQ ID NO:143 GAQKKGSSKGCFGLPLDRIGSLSGLGC SEQ ID NO:144 YKGAQKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:145 YKGAQKKGLSQGCFGLKLDRIGSLSGLGC SEQ ID NO:146 YKGAQKKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:147 YKGAQKKGGSQGCFGLKLDRIGSLSGLGC SEQ ID NO:148 YKGAQKKGGSSGCFGLKLDRIGSLSGLGC SEQ ID NO:149 YKGANKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:150 YKGANKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:151 YKGANKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:152 YKGANKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:153 YKGAQKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:154 YKGAQKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:155 YKGAQKKGGSQGCFGLPLDRIGSMSGLGC SEQ ID NO:156 YKGAQKKGGSSGCFGLPLDRIGSMSGLGC SEQ ID NO:157 YKGAQKKGLSKGCFGLPLDRIGSLSGLGC SEQ ID NO:158 YKGAQKKGGSKGCFGLPLDRIGSLSGLGC SEQ ID NO:159 YKGAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:160 YKGAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:161 YKGANKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:162 YKGANKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:163 YKGANKKGLSQGCFGLPLERIGSMSGLGC SEQ ID NO:164 YKGANKKGLSSGCFGLPLERIGSMSGLGC SEQ ID NO:165 YKGAQKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:166 YKGAQKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:167 YKGAQKKGSSQGCFGLKLDRIGSLSGLGC SEQ ID NO:168 YKGAQKKGSSSGCFGLKLDRIGSLSGLGC SEQ ID NO:169 YKGANKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:170 YKGANKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:171 YKGAQKKGSSQGCFGLPLDRIGSMSGLGC SEQ ID NO:172 YKGAQKKGSSSGCFGLPLDRIGSMSGLGC SEQ ID NO:173 YKGAQKKGSSKGCFGLPLDRIGSLSGLGC SEQ ID NO:174 ARKYKGAQKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:175 ARKYKGAQKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:176 ARKYKGAQKKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:177 ARKYKGAQKKGGSQGCFGLKLDRIGSLSGLGC SEQ ID NO:178 ARKYKGAQKKGGSSGCFGLKLDRIGSLSGLGC SEQ ID NO:179 ARKYKGANKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:180 ARKYKGANKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:181 ARKYKGANKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:182 ARKYKGANKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:183 ARKYKGAQKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:184 ARKYKGAQKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:185 ARKYKGAQKKGGSQGCFGLPLDRIGSMSGLGC SEQ ID NO:186 ARKYKGAQKKGGSSGCFGLPLDRIGSMSGLGC SEQ ID NO:187 ARKYKGAQKKGGSKGCFGLPLDRIGSLSGLGC SEQ ID NO:188 ARKYKGAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:189 ARKYKGAQKKGLSSGCFGLPLERIGSLSGLGC SEQ ID NO:190 ARKYKGANKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:191 ARKYKGANKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:192 ARKYKGANKKGLSQGCFGLPLERIGSMSGLGC SEQ ID NO:193 ARKYKGANKKGLSSGCFGLPLERIGSMSGLGC SEQ ID NO:194 ARKYKGAQKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:195 ARKYKGAQKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:196 ARKYKGAQKKGSSQGCFGLKLDRIGSLSGLGC SEQ ID NO:197 ARKYKGAQKKGSSSGCFGLKLDRIGSLSGLGC SEQ ID NO:198 ARKYKGANKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:199 ARKYKGANKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:200 ARKYKGAQKKGSSQGCFGLPLDRIGSMSGLGC SEQ ID NO:201 ARKYKGAQKKGSSSGCFGLPLDRIGSMSGLGC SEQ ID NO:202 ARKYKGAQKKGSSKGCFGLPLDRIGSLSGLGC SEQ ID NO:203 QEHPQARKYKGAQKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:204 QEHPQARKYKGAQKKGLSQGCFGLKLDRIGSLSGLGC SEQ ID NO:205 QEHPQARKYKGAQKKGLSSGCFGLKLDRIGSLSGLGC SEQ ID NO:206 QEHPQARKYKGAQKKGGSQGCFGLKLDRIGSLSGLGC SEQ ID NO:207 QEHPQARKYKGAQKKGGSSGCFGLKLDRIGSLSGLGC SEQ ID NO:208 QEHPNARKYKGANKKGLSQGCFGLPLDRIGSLSGLGC SEQ ID NO:209 QEHPNARKYKGANKKGLSSGCFGLPLDRIGSLSGLGC SEQ ID NO:210 QEHPNARKYKGANKKGGSQGCFGLPLDRIGSLSGLGC SEQ ID NO:211 QEHPNARKYKGANKKGGSSGCFGLPLDRIGSLSGLGC SEQ ID NO:212 QEHPQARKYKGAQKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:213 QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:214 QEHPQARKYKGAQKKGGSQGCFGLPLDRIGSMSGLGC SEQ ID NO:215 QEHPQARKYKGAQKKGGSSGCFGLPLDRIGSMSGLGC SEQ ID NO:216 QEHPQARKYKGAQKKGGSKGCFGLPLDRIGSLSGLGC SEQ ID NO:217 QEHPQARKYKGAQKKGLSQGCFGLPLERIGSLSGLGC SEQ ID NO:218 QEHPNARKYKGANKKGLSQGCFGLPLDRIGSMSGLGC SEQ ID NO:219 QEHPNARKYKGANKKGLSSGCFGLPLDRIGSMSGLGC SEQ ID NO:220 QEHPNARKYKGANKKGLSQGCFGLPLERIGSMSGLGC SEQ ID NO:221 QEHPNARKYKGANKKGLSSGCFGLPLERIGSMSGLGC SEQ ID NO:222 QEHPQARKYKGAQKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:223 QEHPQARKYKGAQKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:224 QEHPQARKYKGAQKKGSSQGCFGLKLDRIGSLSGLGC SEQ ID NO:225 QEHPQARKYKGAQKKGSSSGCFGLKLDRIGSLSGLGC SEQ ID NO:226 QEHPNARKYKGANKKGSSQGCFGLPLDRIGSLSGLGC SEQ ID NO:227 QEHPNARKYKGANKKGSSSGCFGLPLDRIGSLSGLGC SEQ ID NO:228 QEHPQARKYKGAQKKGSSQGCFGLPLDRIGSMSGLGC SEQ ID NO:229 QEHPQARKYKGAQKKGSSSGCFGLPLDRIGSMSGLGC SEQ ID NO:230 QEHPQARKYKGAQKKGSSKGCFGLPLDRIGSLSGLGC SEQ ID NO:231 GAQKKGSSQGCFGLPLERIGSLSGLGC SEQ ID NO:232。 6. The CNP compound according to claim 1, wherein the CNP peptide has any one of the following amino acid sequences: GAQKKGSSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:136), ARKYKGAQKKGLSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:77), YKGAQKKGGSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:88), YKGAQKKGLSQGCFGLPLDRIGSLSGLGC (SEQ ID NO:103), QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC (SEQ ID NO:67), and QEHPQARKYKGAQKKGLSSGCFGLPLERIGSLSGLGC (SEQ ID NO:104).

7. The CNP compound according to claim 1, wherein the CNP peptide has the following amino acid sequence: QEHPQARKYKGAQKKGLSSGCFGLPLDRIGSLSGLGC (SEQ ID NO:67).

8. The CNP compound according to any one of the preceding claims, wherein q in the chemical formula B of the modifying group is 1 - 5.

9. The CNP compound according to any one of the preceding claims, wherein the modifying group is selected from the following: chemical formula E, chemical formula F, chemical formula G, chemical formula H, chemical formula I and chemical formula J (where the dashed line defines the connection via an amide bond to the N-terminal α-amine of the CNP peptide) 10. The CNP compound according to formula II:

11. The CNP compound according to any one of formulas II, III, IV, V, VI and VII:

12. A CNP compound comprising a CNP peptide and a modifying group, wherein the compound is any one of the compounds disclosed in Table 1: Table 1: Summary of compound chemical formulas 6 - 148 13. A pharmaceutical composition comprising the compound according to any one of claims 1 - 12 and one or more pharmaceutically acceptable excipients.

14. The compound according to any one of claims 1 - 12 or the composition according to claim 13, for the treatment or prevention of cardio-renal metabolic diseases including heart failure and growth disorders including achondroplasia.

15. A method for treating or preventing heart failure, which comprises administering to a patient in need an effective amount of the compound according to any one of claims 1 - 12, optionally in combination with one or more additional therapeutically active compounds.

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