CNP prodrug having a large carrier moiety

Abstract

To provide methods of a more convenient and safer CNP treatment with a reduced risk of hypotension.SOLUTION: The present invention relates to a CNP prodrug or a pharmaceutically acceptable salt thereof comprising: a CNP moiety -D; and a carrier moiety -Z that is conjugated through a moiety -L2- to a reversible prodrug linker moiety -L1-, where the reversible prodrug linker moiety -L1- is covalently and reversibly conjugated to -D; where -L2- is a chemical bond or a spacer, and -Z is a polymer having a molecular weight of at least 10 kDa. The present invention further relates to pharmaceutical compositions comprising the CNP prodrug or pharmaceutically acceptable salt thereof, to their use as a medicament, and to methods of treatment.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a CNP prodrug, or a pharmaceutically acceptable salt thereof, which comprises a CNP moiety -D; and a carrier moiety -Z, wherein the carrier moiety is -L 2 - is reversibly coupled to the prodrug linker moiety -L via the - moiety 1 - is coupled to - 1 - is reversibly covalently coupled to -D; -L 2 - is a chemical bond or a spacer; and -Z is a polymer having a molecular weight of at least 10 kDa, and relates to the above-mentioned CNP prodrug, or a pharmaceutically acceptable salt thereof. The present invention further relates to a pharmaceutical composition containing the CNP prodrug or a pharmaceutically acceptable salt thereof, its use as a medicine, and a method of treatment.

Background Art

[0002] Gain-of-function mutations in FGFR3 result in achondroplasia (ACH), hypochondroplasia (HCH), and thanatophoric dysplasia (TD). All of these diseases are caused by increased signaling of fibroblast growth factor receptor 3 (FGFR3), which is characterized by disproportionate rhizomelic dwarfism, but vary in severity, ranging from mild (HCH) to severe (ACH) and lethal (TD). FGFR3 is a major regulator of endochondral bone formation and signals through several intracellular pathways, including the signal transducer and activator of transcription (STAT) and mitogen-activated protein kinase (MAPK) pathways. Constitutive activation of FGFR3 impairs the proliferation and terminal differentiation of growth plate chondrocytes and the synthesis of the extracellular matrix. Activation of FGFR3 is associated with increased phosphorylation of the STAT and MAPK pathways. The MAPK signaling pathway is regulated by C-type natriuretic peptide (CNP). Binding of CNP to its receptor, natriuretic peptide receptor B (NPR-B), inhibits downstream signaling of FGFR3 and thus causes endochondral growth and skeletal overgrowth, as observed in both mice and humans overexpressing CNP. Overproduction of CNP in cartilage or continuous administration of CNP by intravenous (iv) injection normalizes the dwarfism of achondroplasia mice, suggesting that administration of supra-physiological levels of CNP is a strategy for treating ACH.

[0003] However, considering the short half-life of CNP-22 (2 minutes after intravenous (iv) administration), CNP as a therapeutic agent requires continuous infusion, which is difficult in the pediatric population. Moreover, since CNP is completely inactivated in subcutaneous tissue, intravenous injection is required.

[0004] Potter (FEBS Journal 278 (2011) 1808-1817) describes the clearance of CNP that occurs via two degradation pathways, namely receptor-mediated degradation and degradation by extracellular proteases. CNP is degraded by the action of neutral endopeptidase 24.11 (NEP) and removed from the systemic circulation by the natriuretic peptide clearance receptor NPR-C, which binds CNP and accumulates it within lysosomes where CNP is degraded.

[0005] Reducing the degradation by these two clearance pathways, or either of them, is thought to help extend the half-life of CNP.

[0006] Due to the limited size of the cavity in its active site, NEP selects and recognizes substrates smaller than about 3 kDa. U.S. Patent No. 8,377,884 B2 describes variants of CNP that are optionally and permanently conjugated with PEG polymers to enhance resistance to NEP cleavage. However, the addition of PEG to wild-type CNP, even for PEG as small as about 0.6 kDa, decreases CNP activity, and the addition of PEG of more than about 2 or 3 kDa to CNP or its variants has been found to decrease the functional activity of CNP in a size-dependent manner. Therefore, adding PEG molecules of more than 2 - 3 kDa to reduce NEP degradation is accompanied by a loss of activity, which may reduce the therapeutic potential of such molecules.

[0007] The attachment of PEG or other polymers to CNP may, in addition to adversely affecting the activity of the peptide, also prevent its effective distribution to the growth plate. Farnum et al. (Anat Rec A Discov Mol Cell Evol Biol. 2006 January; 288(1): 91-103) demonstrated that the distribution of molecules from the systemic vasculature to the growth plate is size-dependent, and that small molecules (less than 10 kDa) can be distributed to the growth plate, while molecular sizes of 40 kDa and above impede their translocation to the growth plate.

[0008] International Application WO 2009 / 156481 A1 relates to reversible PEG conjugates of BNP, which term was defined to include all members of the natriuretic peptide family. This application focused only on the cardiovascular effects of peptides of this class, which are mediated by the natriuretic peptide receptor A (NPR-A). WO 2009 / 156481 A1 did not disclose the specific properties of CNP regarding the control of growth, proliferation and differentiation of chondrocytes in the growth plate cartilage, which are mediated by the activation of the natriuretic peptide receptor B (NPR-B).

[0009] Different approaches to creating NEP-resistant CNP molecules to enable subcutaneous administration are described in The American Journal of Human Genetics 91, 1108-1114. BMN-111 is a modified recombinant human C-type natriuretic peptide (CNP) with 17 amino acids added to form a 39-amino acid CNP pharmacological analog. BMN-111 mimics the pharmacological activity of CNP in the growth plate and has a longer half-life as a result of resistance to neutral endopeptidase (NEP), which enables once-daily subcutaneous (SC) administration. Since BMN-111 is a non-natural peptide, the risk of inducing an immune response is higher compared to natural peptides, and an immune response to BMN-111 was observed in animal experiments as described by Martz in "sFGFR for achondroplasia" (SciBx, Biocentury October 2013), although the presence of antibodies did not affect the pharmacological activity of this drug. However, since the half-life of BMN-111 is only 20 minutes, even with daily administration, the duration of exposure to effective levels of the drug is short.

[0010] To increase exposure to effective levels of the drug, the dosage of the drug with CNP activity can be increased. Since natriuretic peptides are a family of hormones that can affect blood volume and blood pressure, an increase in dosage may be associated with cardiovascular side effects. Studies of BMN-111 in animals and humans have demonstrated that as the dosage increases, arterial pressure decreases and heart rate increases. In healthy individuals, administration of BMN-111 at a dosage of 15 μg / kg or less resulted in mild hypotension. Therefore, increasing the dosage of a drug with CNP activity to increase drug exposure may result in unacceptable cardiovascular side effects such as hypotension.

[0011] In WO2009 / 0676639A2 and WO2010 / 135541A2, the use of PEGylated CNP has been considered. However, the authors consider the retention of functionality of such PEGylated CNP as an essential characteristic of their conjugates. The authors teach that the addition of PEG larger than about 2 or 3 kDa may size-dependently reduce CNP functional activity and to avoid the use of larger PEG molecules.

[0012] In summary, there is a need for a more accessible and safer CNP treatment with a low risk of hypotension.

Prior Art Documents

Patent Documents

[0013]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Non-Patent Documents

[0014]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Summary of the Invention

Problems to be Solved by the Invention

[0015] Therefore, an object of the present invention is to at least partially overcome the above-mentioned drawbacks.

Means for Solving the Problems

[0016] This object is achieved by CNP moiety-D; and moiety-L 2 - via a reversible prodrug linker moiety-L 1 - conjugated to a carrier moiety-Z, wherein the reversible prodrug linker moiety-L 1 - is covalently and reversibly bound to -D; [wherein, -L 2 - is a chemical bond or a spacer; -Z is a polymer having a molecular weight of at least 10 kDa.] This is achieved by a CNP prodrug containing the same or a pharmaceutically acceptable salt thereof.

Brief Description of the Drawings

[0017]

Figure 1

Modes for Carrying Out the Invention

[0018] Surprisingly, it has been found that the binding of a polymer-Z having a molecular weight of at least 10 kDa reduces the affinity of the prodrug for NPR-B, which reduces the risk of hypotension. Since the binding between the CNP moiety and the polymer-Z is reversible, the NPR-B binding is restored when released from the CNP prodrug, and the released CNP can be effectively distributed to the growth plate.

[0019] Initial inactivity towards NPR-B and subsequent controlled release from the prodrug conjugate such that the CNP moiety is converted to a fully active drug with respect to NPR-B binding provides several advantages, such as reduced risk of hypotension, the option of high-dose administration, and reduced dosing frequency, which enhance patient compliance and convenience.

[0020] Furthermore, surprisingly, sustained release of CNP from a release control system, such as the prodrugs of the present invention, has been found to be more effective than once-daily bolus administration.

[0021] In the present invention, the following terms are used with the following meanings.

[0022] As used herein, the term "CNP" preferably refers to all CNP polypeptides derived from mammalian species, more preferably human and mammalian species, more preferably human and murine species, as well as variants, analogs, orthologs, homologs and derivatives thereof, and fragments thereof, which are characterized by control of the growth, proliferation and differentiation of chondrocyte growth plate cartilage. Preferably, the term "CNP" refers to the CNP polypeptide of SEQ ID NO: 24, as well as variants, homologs and derivatives thereof that exhibit substantially the same biological activity, namely control of the growth, proliferation and differentiation of chondrocyte growth plate cartilage. More preferably, the term "CNP" refers to the polypeptide of SEQ ID NO: 24.

[0023] In another preferred embodiment, the term "CNP" refers to the polypeptide of SEQ ID NO: 20.

[0024] In another preferred embodiment, the term "CNP" refers to the polypeptide of SEQ ID NO: 21.

[0025] In another preferred embodiment, the term "CNP" refers to the polypeptide of SEQ ID NO: 22.

[0026] In another preferred embodiment, the term "CNP" refers to the polypeptide of SEQ ID NO: 23.

[0027] In another preferred embodiment, the term "CNP" refers to the polypeptide of SEQ ID NO: 30.

[0028] Naturally occurring CNP-22 (SEQ ID NO: 1) has the following sequence: GLSKGCFGLKLDRIGSMSGLGC and the cysteines at positions 6 and 22 are linked by a disulfide bridge as shown in Figure 1.

[0029] SEQ ID NO: 24 has the following sequence: LQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC and the cysteines at positions 22 and 38 are linked by a disulfide bridge.

[0030] The term "CNP" also includes all CNP variants, analogs, orthologs, homologs and derivatives, and fragments thereof, described in WO 2009 / 067639 A2 and WO 2010 / 135541 A2, which are incorporated herein by reference.

[0031] Thus, the term "CNP" preferably also represents the following peptide sequences: SEQ ID NO: 2 (CNP-53): DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 3 (G-CNP-53): GDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 4 (M-CNP-53): MDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 5 (P-CNP-53): PDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 6 (CNP-53 M48N): DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC; Accession number 7 (CNP-53 Δ15-31): DLRVDTKSRAAWARGLSKGCFGLKLDRIGSMSGLGC; Accession number 8 (CNP-52): LRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 9 (CNP-51): RVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 10 (CNP-50): VDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 11 (CNP-49): DTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 12 (CNP-48): TKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 13 (CNP-47): KSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 14 (CNP-46): SRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 15 (CNP-45): RAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 16 (CNP-44): AAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 17 (CNP-44 Δ14-22): AAWARLLQEHPNAGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 18 (CNP-44 Δ15-22): AAWARLLQEHPNARGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 19 (CNP-43): AWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 20 (CNP-42): WARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 21 (CNP-41): ARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 22 (CNP-40): RLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 23 (CNP-39): LLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 24 (CNP-38): LQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 25 (CNP-37): QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 26 (CNP-37 Q1pQ, where pQ = pyroglutamic acid in the sequence): pQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 27 (G-CNP-37): GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 28 (P-CNP-37): PQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 29 (M-CNP-37): MQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 30 (PG-CNP-37): PGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 31 (MG-CNP-37): MGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 32 (CNP-37 M32N): QEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC; SEQ ID NO: 33 (G-CNP-37 M32N): GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC; SEQ ID NO: 34 (G-CNP-37 K14Q): GQEHPNARKYKGANQKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 35 (G-CNP-37 K14P): GQEHPNARKYKGANPKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 36 (G-CNP-37 K14Q, Δ15): GQEHPNARKYKGANQGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 37 (G-CNP-37 K14Q, K15Q): GQEHPNARKYKGANQQGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 38 (CNP-36): EHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 39 (CNP-35): HPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 40 (CNP-34): PNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 41 (CNP-33): NARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 42 (CNP-32): ARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 43 (CNP-31): RKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 44 (CNP-30): KYKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 45 (CNP-29): YKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 46 (CNP-28): KGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 47 (GHKSEVAHRF-CNP-28): GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 48 (CNP-27): GANKKGLSKGCFGLKLDRIGSMSGLGC; Accession number 49 (CNP-27 K4Q, K5Q): GANQQGLSKGCFGLKLDRIGSMSGLGC; Accession number 50 (CNP-27 K4R, K5R): GANRRGLSKGCFGLKLDRIGSMSGLGC; Accession number 51 (CNP-27 K4P, K5R): GANPRGLSKGCFGLKLDRIGSMSGLGC; Accession No. 52 (CNP-27 K4S, K5S): GANSSGLSKGCFGLKLDRIGSMSGLGC; Accession No. 53 (CNP-27 K4P, K5R): GANGANPRGLSRGCFGLKLDRIGSMSGLGC; Accession No. 54 (CNP-27 K4R, K5R, K9R): GANRRGLSRGCFGLKLDRIGSMSGLGC; Accession No. 55 (CNP-27 K4R, K5R, K9R, M22N): GANRRGLSRGCFGLKLDRIGSNSGLGC; Accession No. 56 (P-CNP-27 K4R, K5R, K9R): PGANRRGLSRGCFGLKLDRIGSMSGLGC; Accession No. 57 (M-CNP-27 K4R, K5R, K9R): MGANRRGLSRGCFGLKLDRIGSMSGLGC; Accession No. 58 (HSA fragment-CNP-27): GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLG; Accession No. 59 (HSA fragment-CNP-27 M22N): GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSNSGLGC; Accession No. 60 (M-HSA fragment-CNP-27): MGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession No. 61 (P-HSA fragment-CNP-27): PGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC; Accession No. 62 (CNP-26): ANKKGLSKGCFGLKLDRIGSMSGLGC; Accession No. 63 (CNP-25): NKKGLSKGCFGLKLDRIGSMSGLGC; Accession No. 64 (CNP-24): KKGLSKGCFGLKLDRIGSMSGLGC; Accession No. 65 (CNP-23): KGLSKGCFGLKLDRIGSMSGLGC; Accession No. 66 (R-CNP-22): RGLSKGCFGLKLDRIGSMSGLGC; Accession No. 67 (ER-CNP-22): ERGLSKGCFGLKLDRIGSMSGLGC; Accession No. 68 (R-CNP-22 K4R): RGLSRGCFGLKLDRIGSMSGLGC; Accession No. 69 (ER-CNP-22 4KR): ERGLSRGCFGLKLDRIGSMSGLGC; Accession No. 70 (RR-CNP-22): RRGLSRGCFGLKLDRIGSMSGLGC; Accession No. 71 (HRGP fragment-CNP-22): GHHSHEQHPHGANQQGLSKGCFGLKLDRIGSMSGLGC; Accession No. 72 (HRGP fragment-CNP-22): GAHHPHEHDTHGANQQGLSKGCFGLKLDRIGSMSGLGC; Accession No. 73 (HRGP fragment-CNP-22): GHHSHEQHPHGANPRGLSKGCFGLKLDRIGSMSGLGC; Accession No. 74 (IgG 1 (F c ) fragment-CNP-22): GQPREPQVYTLPPSGLSKGCFGLKLDRIGSMSGLGC; Accession No. 75 (HSA fragment-CNP-22): GQHKDDNPNLPRGANPRGLSKGCFGLKLDRIGSMSGLGC; Accession No. 76 (HSA fragment-CNP-22): GERAFKAWAVARLSQGLSKGCFGLKLDRIGSMSGLGC; Accession No. 77 (osteocrin NPR C inhibitor fragment - CNP22): FGIPMDRIGRNPRGLSKGCFGLKLDRIGSMSGLGC; Accession No. 78 (FGF2 heparin-binding domain fragment - CNP22): GKRTGQYKLGSKTGPGPKGLSKGCFGLKLDRIGSMSGLGC; Accession No. 79 (IgG 1 (F c ) fragment - CNP - 22 K4R): GQPREPQVYTGANQQGLSRGCFGLKLDRIGSMSGLGC; Accession No. 80 (HSA fragment - CNP - 22 K4R): GVPQVSTSTGANQQGLSRGCFGLKLDRIGSMSGLGC; Accession No. 81 (fibronectin fragment - CNP - 22 K4R): GQPSSSSQSTGANQQGLSRGCFGLKLDRIGSMSGLGC; Accession No. 82 (fibronectin fragment - CNP - 22 K4R): GQTHSSGTQSGANQQGLSRGCFGLKLDRIGSMSGLGC; Accession No. 83 (fibronectin fragment - CNP - 22 K4R): GSTGQWHSESGANQQGLSRGCFGLKLDRIGSMSGLGC; Accession No. 84 (Zn finger fragment - CNP - 22 K4R): GSSSSSSSSSGANQQGLSRGCFGLKLDRIGSMSGLGC; Accession No. 85 (CNP - 21): LSKGCFGLKLDRIGSMSGLGC; Accession No. 86 (CNP - 20): SKGCFGLKLDRIGSMSGLGC; Accession No. 87 (CNP - 19): KGCFGLKLDRIGSMSGLGC; SEQ ID NO: 88 (CNP-18): GCFGLKLDRIGSMSGLGC; SEQ ID NO: 89 (CNP-17): CFGLKLDRIGSMSGLGC; SEQ ID NO: 90 (BNP fragment - CNP-17 - BNP fragment): SPKMVQGSGCFGLKLDRIGSMSGLGCKVLRRH; SEQ ID NO: 91 (CNP-38 L1G): GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 92 (Ac-CNP-37; Ac = acetyl in the sequence): Ac-QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC。

[0032] More preferably, the term "CNP" refers to the sequences of SEQ ID NOs: 2, 19, 20, 21, 22, 23, 24, 25, 26, 30, 32, 38, 39, 40, 41, 42, 43, 91, 92. Even more preferably, the term "CNP" refers to the sequences of SEQ ID NOs: 23, 24, 25, 26, 38, 39, 91, and 92. In a particularly preferred embodiment, the term "CNP" refers to the sequence of SEQ ID NO: 24. In an equally preferred embodiment, the term "CNP" refers to the sequence of SEQ ID NO: 30. In an equally preferred embodiment, the term "CNP" refers to the sequence of SEQ ID NO: 20. In an equally preferred embodiment, the term "CNP" refers to the sequence of SEQ ID NO: 21. In an equally preferred embodiment, the term "CNP" refers to the sequence of SEQ ID NO: 22. In an equally preferred embodiment, the term "CNP" refers to the sequence of SEQ ID NO: 23.

[0033] In one embodiment, the term "CNP" refers to the sequence of SEQ ID NO: 93 QEHPNARX 1 YX 2 GANX 3 X 4 GLSX5 GCFGLX 6 LDRIGSMSGLGC refers to X in this array 1 X 2 X 3 X 4 X 5 and X 6 are each independently selected from the group consisting of K, R, P, S, and Q, provided that X 1 X 2 X 3 X 4 X 5 and X 6 at least one of which is selected from the group consisting of R, P, S, and Q; preferably, X 1 X 2 X 3 X 4 X 5 and X 6 are selected from the group consisting of K and R, provided that X 1 X 2 X 3 X 4 X 5 and X 6 at least one of which is R; More preferably, the term "CNP" refers to the sequence of SEQ ID NO: 94 QEHPNARKYKGANX 1 X 2 GLSX 3 GCFGLX 4 LDRIGSMSGLGC refers to X in this array 1 X 2 X 3 and X 4 are each independently selected from the group consisting of K, R, P, S, and Q, provided that X 1 X 2 X 3 and X 4 at least one of which is selected from the group consisting of R, P, S, and Q; preferably, X 1 X 2 X 3 and X 4is selected from K and R, provided that X 1 X 2 X 3 and X 4 at least one of is R; More preferably, the term "CNP" refers to the sequence of SEQ ID NO: 95 QEHPNARKYKGANX 1 X 2 GLSKGCFGLKLDRIGSMSGLGC and in this sequence X 1 X 2 is selected from the group consisting of KR, RK, KP, PK, SS, RS, SR, QK, QR, KQ, RQ, RR and QQ.

[0034] It is understood that in SEQ ID NOs: 2-95, those corresponding to the cysteines at positions 22 and 38 of SEQ ID NO: 24 are linked by disulfide bridges.

[0035] The present invention is also understood to include CNP variants, in which any one or more, up to all, of the residues that are susceptible to amide hydrolysis or amide hydrolysis-like reactions (e.g., isomerization) can be converted to other residues by amide hydrolysis or amide hydrolysis-like reactions, to an extent of up to 100% conversion for each converted residue. In certain embodiments, the present specification includes CNP variants, in which: (1) Any one or some, up to all, of the asparagine (Asn / N) residues can be converted to aspartic acid or aspartate, and / or isoaspartic acid or isoaspartate, by amide hydrolysis, to an extent of about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion for each converted residue; or (2) Any one or several, and at most all, of the glutamine (Gln / Q) residues may be converted to glutamic acid or glutamate, and / or isoglutamic acid or isoglutamate, by amidolysis, up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion for each residue being converted; or (3) Any one or several, and at most all, of the aspartic acid or aspartate (Asp / D) residues may be converted to isoaspartic acid or isoaspartate by an amidolysis-like reaction (also referred to as isomerization), up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion for each residue being converted; or (4) Any one or several, and at most all, of the glutamic acid or glutamate (Glu / E) residues may be converted to isoglutamic acid or isoglutamate by an amidolysis-like reaction (also referred to as isomerization), up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion for each residue being converted; or (5) The N-terminal glutamine, if present, may be converted to pyroglutamate up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion; or (6) The above combinations.

[0036] As used herein, the term "CNP polypeptide variant" refers to a polypeptide derived from the same species that is different from the reference CNP polypeptide. Preferably, such a reference CNP polypeptide sequence is the sequence of SEQ ID NO: 24. Generally, since the differences are limited, the amino acid sequences of the reference and the variant are very similar overall and identical in many regions. Preferably, the CNP polypeptide variant is at least 70%, 80%, 90% or 95% identical to the reference CNP polypeptide, preferably the CNP polypeptide of SEQ ID NO: 24. A polypeptide having a query amino acid sequence and, for example, at least 95% "identical" amino acid sequence means that the amino acid sequence of the subject polypeptide is identical to the query sequence, except that it may contain five or fewer amino acid changes per 100 amino acids of the query amino acid sequence. Such changes in the reference sequence may occur at the amino-terminal (N-terminal) or carboxy-terminal (C-terminal) positions of the reference amino acid sequence, but may occur anywhere between such terminal positions, scattered individually among the residues within the reference sequence, or scattered as one or more adjacent groups within the reference sequence. The query sequence may be the complete amino acid sequence of the reference sequence or any of the specified fragments described herein. Preferably, the query sequence is the sequence of SEQ ID NO: 24.

[0037] Such CNP polypeptide variants can be naturally occurring variants, such as naturally occurring allelic variants encoded by one of several alternative forms of CNP that occupy a particular locus on a chromosome or in an organism, or isoforms encoded by naturally occurring splice variants derived from a single primary transcript. Alternatively, the CNP polypeptide variant may be a naturally occurring unknown variant that can be generated by mutagenesis techniques known in the art.

[0038] It is known in the art that one or more amino acids can be deleted from the N-terminus or C-terminus of a bioactive peptide or protein without substantially losing its biological function. Such N-terminal and / or C-terminal deletions are also included in the term CNP polypeptide variant.

[0039] It is also recognized by those skilled in the art that some amino acid sequences of the CNP polypeptide can be altered without significantly affecting the structure or function of the peptide. Such variants contain deletions, insertions, inversions, repeats, and substitutions selected according to principles known in the art so as to have little effect on activity. For example, guidelines for making phenotypically silent amino acid substitutions are provided in Bowie et al. (1990), Science 247:1306-1310, which is hereby incorporated by reference in its entirety, where the authors show that there are two main approaches for examining the tolerance of amino acid sequence changes.

[0040] The term CNP polypeptide also includes all CNP polypeptides encoded by CNP analogs, orthologs, and / or species homologs. As used herein, the term "CNP analog" refers to a CNP from a different unrelated organism that performs the same function in each organism but does not originate from the ancestral structure commonly shared by the ancestors of those organisms. Instead, the analog CNP evolved separately and then evolved to perform the same or similar functions. In other words, an analog CNP polypeptide is a polypeptide having quite different amino acid sequences that achieve the same biological activity, namely, the control of the growth, proliferation, and differentiation of chondrocytes in the cartilage growth plate.

[0041] As used herein, the term "CNP ortholog" refers to CNP present in two different species, the sequences of which are related to each other through a common homologous CNP in the ancestral species but have evolved to be different from each other.

[0042] As used herein, the term "CNP homolog" refers to CNP from different organisms, which performs the same function in each organism and is derived from an ancestral structure commonly possessed by the ancestors of those organisms. In other words, a homologous CNP polypeptide is a polypeptide having very similar amino acid sequences that achieve the same biological activity, i.e., control of the growth, proliferation, and differentiation of chondrocytes in the growth plate of cartilage. Preferably, a CNP polypeptide homolog can be defined as a polypeptide that exhibits at least 40%, 50%, 60%, 70%, 80%, 90%, or 95% identity to a reference CNP polypeptide, preferably the CNP polypeptide of SEQ ID NO: 24.

[0043] Thus, the CNP polypeptide of the present invention can be, for example, (i) a CNP polypeptide in which at least one of the amino acid residues is substituted by a conservative or non-conservative amino acid residue, preferably a conservative amino acid residue, and such substituted amino acid residues may or may not be encoded by the genetic code; and / or (ii) a CNP polypeptide in which at least one of the amino acid residues contains a substituent; and / or (iii) a CNP polypeptide in which the CNP polypeptide is fused to another compound, and the compound is, for example, a compound that prolongs the half-life of the polypeptide (such as polyethylene glycol); and / or (iv) an additional amino acid is fused to the CNP polypeptide, and it is, for example, an IgG Fc fusion region peptide, or a leader sequence, or a secretion sequence, or a sequence used for the purification of the polypeptide in the above form, or a preprotein sequence.

[0044] As used herein, the term "CNP polypeptide fragment" refers to any peptide that includes a continuous range of a part of the amino acid sequence of a CNP polypeptide, preferably the polypeptide of SEQ ID NO: 24.

[0045] More specifically, the CNP polypeptide fragment contains at least 6 consecutive amino acids of the CNP polypeptide, more preferably the polypeptide of SEQ ID NO: 24, for example, contains at least 8, at least 10, or at least 17 consecutive amino acids. The CNP polypeptide fragment can further be described as a subclass of the CNP polypeptide containing at least 6 amino acids, where "at least 6" is defined as any integer between 6 and the integer representing the C-terminal amino acid of the CNP polypeptide, and this CNP polypeptide is preferably the polypeptide of SEQ ID NO: 24. Further, the molecular species as described above, which are CNP polypeptide fragments with a length of at least 6 amino acids as described above and further specified for the N-terminal and C-terminal positions, are included. Also, the term "CNP polypeptide fragment" includes all CNP polypeptide fragments with a length of at least 6 amino acids as described above that can be specifically identified by the N-terminal and C-terminal positions as individual molecular species. That is, fragments with a length of at least 6 consecutive amino acid residues that can occupy any combination of N-terminal and C-terminal positions on any given amino acid sequence of the CNP polypeptide, preferably the CNP polypeptide of SEQ ID NO: 24, are included in the present invention.

[0046] The term "CNP" also includes poly(amino acid) conjugates having the above-described sequence but having a backbone containing both amide bonds and non-amide bonds such as ester bonds, for example, depsipeptides. A depsipeptide is a chain of amino acid residues whose backbone contains both amide (peptide) bonds and ester bonds. Thus, the term "side chain" as used herein refers to the portion attached to the α-carbon of the amino acid moiety when the amino acid moiety is linked by an amine bond as in a polypeptide, or, for example, in the case of a depsipeptide, any carbon atom-containing portion attached to the backbone of the poly(amino acid) conjugate. Preferably, the term "CNP" represents a polypeptide having a backbone formed by amide (peptide) bonds.

[0047] Since the term "CNP" includes the above variants, analogs, orthologs, homologs, derivatives and fragments of CNP, any reference to a specific position within the reference sequence includes the equivalent position in variants, analogs, orthologs, homologs, derivatives and fragments of the CNP moiety, even if not explicitly mentioned.

[0048] As used herein, the term "loop portion" refers to a continuous series of contiguous amino acid residues of a CNP drug or CNP moiety that are located between two cysteine residues that form an intramolecular disulfide bridge or between homologous amino acid residues that are linked via a chemical crosslinking agent. Preferably, the loop portion is between two cysteine residues that form an intramolecular disulfide bridge. These two cysteines correspond to the cysteines at positions 22 and 38 in the sequence of CNP-38 (SEQ ID NO: 24). Thus, when a CNP drug or CNP moiety has the sequence of CNP-38, amino acids 23 to 37 are located in the aforementioned loop portion.

[0049] Regardless of the length of the CNP moiety, the sequence of the loop portion of wild-type CNP is FGLKLDRIGSMSGLG (SEQ ID NO: 96).

[0050] As noted above, the term "CNP" relates to CNP drugs or CNP moieties having varying numbers of amino acids. Those skilled in the art will recognize that the positions of corresponding amino acids vary in CNP drugs or CNP moieties of different lengths, and will have no difficulty identifying the two cysteines that form the disulfide bridge or the two homologous amino acid residues that are linked to each other via a chemical linker in longer, shorter, and / or otherwise modified versions of CNP.

[0051] The term CNP includes the above-mentioned variants, analogs, orthologs, homologs, derivatives, and fragments of CNP. Thus, the term "ring portion" also includes the corresponding variants, analogs, orthologs, homologs, derivatives, and fragments of the sequence of SEQ ID NO:96. Accordingly, any reference to a specific position within the reference sequence also includes the equivalent position in the variants, analogs, orthologs, homologs, derivatives, and fragments of the CNP portion, even if not explicitly mentioned.

[0052] As used herein, the term "random coil" refers to a peptide or protein that adopts / has / forms a three-dimensional structure that is substantially devoid of a defined secondary and tertiary structure, as determined by circular dichroism spectroscopy in an aqueous buffer at pH 7.4 at room temperature. Preferably, room temperature (ambient temperature) is about 20 °C, i.e., 18 °C to 22 °C, and most preferably, room temperature is 20 °C.

[0053] As used herein, the term "drug (agent)" refers to a substance used for the treatment, cure, prevention, or diagnosis of a disease, or a substance used to enhance physical or mental health. When a drug D-H is conjugated to another moiety, the -D moiety of the resulting product originating from that drug is called the "bioactive moiety".

[0054] As used herein, the term "prodrug" refers to a bioactive moiety reversibly and covalently linked to a specific protecting group via a reversible prodrug linker moiety, where the reversible prodrug linker is a linker moiety that includes a reversible bond with the bioactive moiety, whereby the specific protecting group modifies or removes undesirable properties of the parent molecule. This includes enhancing the desirable properties of the drug as well as suppressing undesirable properties. A non-toxic specific protecting group is called a "carrier". The prodrug releases the reversibly and covalently linked bioactive moiety -D in the form of its corresponding drug D-H. In other words, a prodrug is a conjugate containing a bioactive moiety, where the bioactive moiety is covalently and reversibly conjugated to a carrier moiety via a reversible prodrug linker moiety, and the covalent and reversible bond between the carrier and the reversible prodrug linker moiety is either direct or via a spacer such as -L 2 - etc. Such a conjugate releases the bioactive moiety that was originally conjugated in the form of a free drug.

[0055] A "biodegradable bond" or "reversible bond" is a bond that is degradable, i.e., cleavable, by hydrolysis without an enzyme under physiological conditions (aqueous buffer at pH 7.4, 37 °C), and has a half-life of from 1 hour to 6 months, preferably from 1 hour to 4 months, more preferably from 1 hour to 3 months, even more preferably from 1 hour to 2 months, and even more preferably from 1 hour to 1 month. Thus, a "stable bond" is a bond that has a half-life of more than 6 months under physiological conditions (aqueous buffer at pH 7.4, 37 °C).

[0056] Thus, a "reversible prodrug linker moiety" is a moiety that is covalently conjugated to a bioactive moiety such as CNP by a reversible bond and is also covalently conjugated to a carrier moiety such as -Z, where the covalent bond with the carrier moiety is either direct or via a spacer moiety such as -L 2 - etc. -Z and -L 2The bond with - is preferably a stable bond.

[0057] As used herein, the term "non - leaving - trace prodrug linker" means a reversible prodrug linker that releases the drug in its free form upon cleavage. The "free form" of the drug as used herein means the unmodified pharmacologically active form of the drug.

[0058] As used herein, the term "pharmaceutical composition" contains one or more active ingredients, such as a drug or a prodrug, specifically the CNP prodrug of the present invention herein, and optionally one or more additives. In addition, any product directly or indirectly obtained as a result of the blending, complex formation, or aggregation of any two or more of the components of the composition, or as a result of the dissociation of one or more components, or as a result of other types of reactions or interactions of one or more components, is also included. Thus, the pharmaceutical composition of the present invention includes any composition prepared by mixing one or more CNP prodrugs of the present invention and, optionally, pharmaceutically acceptable additives.

[0059] As used herein, the term "additive (excipient)" refers to a diluent, adjuvant, or vehicle that is used together with a therapeutic agent such as a drug or a prodrug. Such pharmaceutical additives can be sterile liquids, such as water and oils, and the oils include those of petroleum, animal, plant, or synthetic origin, including but not limited to peanut oil, soybean oil, mineral oil, sesame oil, etc. When the pharmaceutical composition is administered orally, water is a preferred additive. When the pharmaceutical composition is administered intravenously, physiological saline and glucose aqueous solution are preferred additives. As liquid additives for injections, it is preferred to use physiological saline, glucose aqueous solution, and glycerol solution. Suitable pharmaceutical additives include starch, glucose, lactose, sucrose, mannitol, trehalose, gelatin, malt, rice, cereal flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene glycol, water, ethanol, etc. The pharmaceutical composition may optionally contain a small amount of wetting agent or emulsifier, pH buffer, such as acetate, succinate, tris, carbonate, phosphate, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), MES (2-(N-morpholino)ethanesulfonic acid), etc., and may also contain surfactants, such as Tween, poloxamer, poloxamine, CHAPS, Igepal, etc., or amino acids, such as glycine, lysine, or histidine, etc. Such pharmaceutical compositions can take forms such as solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release preparations, etc. The pharmaceutical composition may also be formulated as a suppository using conventional binders and additives, such as triglycerides. Oral formulations can contain standard additives, such as pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Such compositions will contain a therapeutically effective amount of a drug or bioactive moiety together with a suitable amount of additives to provide a form for proper administration to a patient. The formulation should be suitable for the method of administration.

[0060] As used herein, the term "liquid composition" refers to a mixture containing a water-soluble CNP prodrug and one or more solvents such as water.

[0061] As used herein, the term "dry composition" means that the pharmaceutical composition is provided in a dried state. Suitable drying methods are spray drying and freeze drying, i.e., lyophilization. Such a dry composition of the prodrug of the present invention has a residual moisture content of at most 10%, preferably less than 5%, more preferably less than 2% as measured by Karl Fischer. The pharmaceutical composition of the present invention is preferably dried by freeze drying.

[0062] As used herein, the term "micelle" means an aggregate of amphiphilic molecules dispersed in a liquid colloid. In an aqueous solution, a typical micelle forms an aggregate in which the hydrophilic part of the surfactant molecule faces the surrounding solvent and the hydrophobic part of the surfactant molecule faces inward, and is also called a "normal-phase micelle". An "inverse micelle" has the hydrophilic part facing inward and the hydrophobic part facing the surrounding solvent.

[0063] As used herein, the term "liposome" refers to a vesicle having at least one lipid bilayer, preferably a spherical vesicle. Preferably, the liposome contains phospholipids, and more preferably contains phosphatidylcholine. The term "liposome" refers to various structures and sizes, such as multilamellar liposome vesicles (MLV) with two or more concentric lipid bilayers and an average diameter ranging from 100 to 1000 nm, small unilamellar liposome vesicles (SUV) with one lipid bilayer and an average diameter ranging from 25 to 100 nm, large unilamellar liposome vesicles (LUV) with one lipid bilayer and an average diameter of about 1000 μm, and giant unilamellar vesicles (GUV) with one lipid bilayer and an average diameter ranging from 1 to 100 μm. The term "liposome" also includes deformable vesicles such as transferosomes and esosomes.

[0064] As used herein, the term "aquasome" refers to spherical nanoparticles having a diameter of 60 to 300 nm and having at least three layers of self-assembled structures, which are solid-phase nanocrystalline cores coated with oligomeric films that adsorb drug molecules, either by modifying the drug or without modification.

[0065] As used herein, the term "ethosome" refers to lipid vesicles containing phospholipids, ethanol and / or isopropanol at relatively high concentrations, and water, and refers to said lipid vesicles having a size in the range of several tens of nanometers to micrometers.

[0066] As used herein, the term "LeciPlex" refers to a vesicle system based on positively charged phospholipids, which contains soybean PC, a cationic substance, and a biocompatible solvent, such as PEG 300, PEG 400, diethylene glycol monoethyl ether, tetrahydrofurfuryl alcohol polyethylene glycol ether, or 2-pyrrolidone or N-methyl-2-pyrrolidone.

[0067] As used herein, the term "niosome" refers to a unilamellar or multilamellar vesicle containing a nonionic surfactant.

[0068] As used herein, the term "pharmacosome" refers to ultrafine, micellar or hexagonal aggregates composed of lipids covalently bonded to a bioactive moiety.

[0069] As used herein, the term "proniosome" refers to a dry preparation of a surfactant-coated carrier that, upon rehydration and gentle agitation, gives niosomes.

[0070] As used herein, the term "polymersome" refers to an artificial spherical vesicle that contains a membrane formed from an amphiphilic synthetic block copolymer and can optionally contain an aqueous solution within its core. Polymersomes have diameters in the range of 50 nm to 5 μm or more. This term includes syntosomes, which are polymersomes engineered to contain channels that allow specific chemicals to pass through the membrane and enter and exit the vesicle.

[0071] As used herein, the term "sphingosome" refers to a concentric bilayer vesicle in which an aqueous solution is completely enclosed by a lipid bilayer mainly composed of natural or synthetic sphingolipids.

[0072] As used herein, the term "transferosome" refers to a highly deformable lipid vesicle containing an aqueous core, formed from a mixture of common polar, appropriately end-functionalized lipids, which mixture facilitates the formation of highly curved bilayers that make transferosomes very deformable.

[0073] As used herein, the term "ufasome" refers to a vesicle containing unsaturated fatty acids.

[0074] As used herein, the term "reagent" means a compound containing at least one functional group for reacting with a functional group of another compound or drug. Drugs containing functional groups (such as primary or secondary amines or hydroxyl functional groups) are also understood to be reagents.

[0075] As used herein, the term "moiety (substructure)" means a part of a molecule that has one or more fewer atoms compared to the corresponding reagent. For example, when a reagent of the formula "H-X-H" reacts with another reagent and becomes part of the reaction product, the corresponding moiety of the reaction product has the structure "H-X-" or "-X-", where each "-" represents a bond to another moiety. Thus, the biologically active moiety is released as a drug from the prodrug.

[0076] When an atomic group is provided with an arrangement or chemical structure in which it is bonded to two moieties or inserted into one moiety, it is understood that, unless otherwise clearly stated, the said arrangement or chemical structure may be bonded to the two moieties in any orientation. For example, the moiety "-C(O)N(R 1 )-" may be bonded to two moieties as "-C(O)N(R 1 )-" or as "-N(R 1 )C(O)-", or may be inserted into one moiety. Similarly, the moiety

[0077]

Chem.

[0078]

Chem.

[0079]

Chem.

[0080] As used herein, the term "functional group" means an atomic group that can react with other atomic groups. Functional groups include, but are not limited to, the following groups: carboxylic acid (-(C=O)OH), primary or secondary amine (-NH 2, -NH-), maleimide, thiol (-SH), sulfonic acid (-(O=S=O)OH), carbonate, carbamate (-O(C=O)N<), hydroxyl (-OH), aldehyde (-(C=O)H), ketone (-(C=O)-), hydrazine (>N-N<), isocyanate, isothiocyanate, phosphoric acid (-O(P=O)OHOH), phosphonic acid (-O(P=O)OHH), haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine, disulfide, sulfonamide, sulfonic acid, vinyl sulfone, vinyl ketone, diazoalkane, oxirane and aziridine.

[0081] When the prodrug of the present invention contains one or more acidic or basic groups, the present invention also includes their corresponding pharmaceutically or toxicologically acceptable salts, particularly their pharmaceutically available salts. Thus, a prodrug of the present invention containing an acidic group can be used according to the present invention, for example, as an alkali metal salt, an alkaline earth metal salt or an ammonium salt. More detailed examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts, or salts with ammonia or organic amines (e.g., ethylamine, ethanolamine, triethanolamine, etc.) or amino acids. There may also be present a prodrug of the present invention containing one or more basic groups, i.e., groups that can be protonated, and such prodrugs can be used according to the present invention in the form of their addition salts with inorganic or organic acids. Examples of suitable acids include hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, and other acids known to those skilled in the art. Further methods for converting basic groups to cations, such as alkylation of amine groups to provide appropriate counterions for positively charged ammonium groups and their salts, are known to those skilled in the art. When the prodrug of the present invention contains both acidic and basic groups simultaneously, the present invention also includes inner salts or betaines (zwitterions) in addition to the salt forms mentioned. Each salt can be obtained by conventional methods known to those skilled in the art, for example, by contacting these prodrugs with organic or inorganic acids or bases in a solvent or dispersion, or by anion exchange or cation exchange with other salts. The present invention also includes all salts of the prodrugs of the present invention, which, because of their low physiological compatibility, are not directly suitable for use in pharmaceuticals, but can be used, for example, as intermediates in chemical reactions or in the preparation of pharmaceutically acceptable salts.

[0082] The term "pharmaceutically acceptable" means a substance that does not cause harm when administered to a patient, and preferably means that it has been approved for use in animals, preferably for use in humans, by a regulatory agency, such as the EMA (Europe) and / or the FDA (USA) and / or any other national regulatory agency.

[0083] The term "about" as used herein in combination with a numerical value means, in addition to the numerical value itself, a range of plus / minus 10% or less of said numerical value, more preferably plus / minus 8% or less of said numerical value, even more preferably plus / minus 5% or less of said numerical value, and most preferably plus / minus 2% or less of said numerical value. For example, the phrase "about 200" means a range of 200+ / -10%, i.e., a range of 180 to 220, preferably a range of 200+ / -8%, i.e., a range of 184 to 216, even more preferably a range of 200+ / -5%, i.e., a range of 190 to 210, and most preferably a range of 200+ / -2%, i.e., a range of 196 to 204. It is understood that the percentage indicated as "about 20%" does not mean a range of "20%+ / -10%", i.e., 10 to 30%, but rather that "about 20%" means a range of plus / minus 10% of the numerical value of 20, i.e., 18 to 22.

[0084] As used herein, the term "polymer" means a molecule containing repeating structural units, i.e., monomers, linked by chemical bonds in a linear, cyclic, branched, crosslinked, or dendrimer-like fashion or combinations thereof, which may be of synthetic origin, of biological origin, or a combination of both. It is understood that a polymer may also contain one or more other chemical groups and / or moieties, such as for example one or more functional groups. Preferably, a soluble polymer has a molecular weight of at least 0.5 kDa, for example at least 1 kDa, at least 2 kDa, at least 3 kDa, or at least 5 kDa. When the polymer is soluble, it preferably has a molecular weight of up to 1000 kDa, for example up to 750 kDa, for example up to 500 kDa, for example up to 300 kDa, for example up to 200 kDa, for example up to 100 kDa. It is understood that a protein is also a polymer in which the amino acids are the repeating structural units, even though the side chains of each amino acid may be different.

[0085] As used herein, the term "polymeric" means a reagent or moiety containing one or more polymers or polymer moieties. A polymeric reagent or moiety may optionally also contain one or more other moieties, such other moieties preferably being selected from the group consisting of: · C 1-50 alkyl, C 2-50 alkenyl, C 2-50 alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, phenyl, naphthyl, indenyl, indanyl, and tetralinyl, and · a bond selected from the group consisting of:

Chemical formula

[0086] Those skilled in the art will understand that the polymerization products obtained from the polymerization reaction do not all have the same molecular weight, but rather exhibit a molecular weight distribution. Therefore, the molecular weight ranges, molecular weights, monomer number ranges in the polymer, and monomer numbers in the polymer used herein refer to the number average molecular weight and the average number of monomers, that is, the arithmetic average of the molecular weights of the polymer or polymer portion, and the arithmetic average of the number of monomers in the polymer or polymer portion.

[0087] Thus, in the case of a polymer portion containing "x" monomer units, any integer applied to "x" corresponds to the arithmetic average number of monomers. Any range of integers applied to "x" provides a range of integers in which the arithmetic average number of monomers exists. The integer "x" indicated as "about x" means that the arithmetic average number of monomers exists within the integer range of x+ / -10%, preferably x+ / -8%, more preferably x+ / -5%, and most preferably x+ / -2%.

[0088] The term "number average molecular weight" as used herein means the ordinary arithmetic average of the molecular weights of the individual polymers.

[0089] The term "water-soluble" as used herein with respect to a carrier means that when such a carrier is part of the CNP prodrug of the present invention, at least 1 g of the CNP prodrug containing such a water-soluble carrier can be dissolved in 1 liter of water at 20 °C to form a homogeneous solution. Thus, the term "water-insoluble" with respect to a carrier means that when such a carrier is part of the CNP prodrug of the present invention, less than 1 g of the CNP prodrug containing such a water-insoluble carrier can be dissolved in 1 liter of water at 20 °C to form a homogeneous solution.

[0090] As used herein, the term "PEG-based" with respect to a moiety or reagent means that the moiety or reagent contains PEG. Preferably, the PEG-based moiety or reagent contains at least 10% (w / w) PEG, such as at least 20% (w / w) PEG, such as at least 30% (w / w) PEG, such as at least 40% (w / w) PEG, such as at least 50% (w / w), such as at least 60 (w / w) PEG, such as at least 70% (w / w) PEG, such as at least 80% (w / w) PEG, such as at least 90% (w / w), such as at least 95% (w / w) PEG. The remaining weight percentage of the PEG-based moiety or reagent is preferably other moieties selected from the following moieties and linkages: ·C 1-50 alkyl, C 2-50 alkenyl, C 2-50 alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, phenyl, naphthyl, indenyl, indanyl and tetralinyl, and ·linkages selected from the group consisting of:

Chemical formula

[0091] As used herein, the term "PEG-based containing at least X% PEG" with respect to a moiety or reagent means that the moiety or reagent contains at least X% (w / w) ethylene glycol units (-CH 2 CH 2means containing O-), and the ethylene glycol units may be arranged in an alternating block pattern or randomly distributed within the moiety or reagent. Preferably, all the ethylene glycol units of the moiety or reagent are present within one block, and the remaining weight percentage of the PEG-based moiety or reagent is other moieties preferably selected from the following moieties and linkages: ·C 1-50 alkyl, C 2-50 alkenyl, C 2-50 alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, phenyl, naphthyl, indenyl, indanyl, and tetralinyl, and ·linkages selected from the group consisting of the following:

Chemical formula

[0092] The term "hyaluronic acid-based containing at least X% hyaluronic acid" is used as appropriate.

[0093] As used herein, the term "substituted" means that one or more -H atoms of a molecule or moiety are replaced by another atom or atomic group, and the said another atom or atomic group is referred to as a "substituent".

[0094] Preferably, one or more additional optional substituents are independently of each other halogen, -CN, -COOR x1 , -OR x1 , -C(O)R x1 , -C(O)N(R x1 R x1a ), -S(O) 2 N(R x1 R x1a ), -S(O)N(Rx1 R x1a )、 -S(O) 2 R x1 、 -S(O)R x1 、 -N(R x1 )S(O) 2 N(R x1a R x1b )、 -SR x1 、 -N(R x1 R x1a )、 -NO 2 、 -OC(O)R x1 、 -N(R x1 )C(O)R x1a 、 -N(R x1 )S(O) 2 R x1a 、 -N(R x1 )S(O)R x1a 、 -N(R x1 )C(O)OR x1a 、 -N(R x1 )C(O)N(R x1a R x1b )、 -OC(O)N(R x1 R x1a )、 -T 0 、 C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl selected from the group consisting of, where -T 0 、 C 1-50 alkyl, C 2-50 alkenyl and C 2-50 alkynyl are optionally substituted with one or more -R x2 and C 1-50 alkyl, C 2-50 alkenyl and C 2-50 alkynyl may contain -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O) 2 N(R x3 )-, -S(O)N(R x3 )-, -S(O) 2 -, -S(O)-, -N(R x3 )S(O) 2 N(R x3a )-, -S-, -N(R x3)-, -OC(OR x3 )(R x3a )-, -N(R x3 )C(O)N(R x3a )-, and -OC(O)N(R x3 )- may optionally interrupt, -R x1 , -R x1a , -R x1b are, independently of each other, -H, -T 0 , C 1-50 alkyl, C 2-50 alkenyl and C 2-50 alkynyl, and said -T 0 , C 1-50 alkyl, C 2-50 alkenyl and C 2-50 alkynyl may optionally be substituted with one or more of the same or different -R x2 ), and said C 1-50 alkyl, C 2-50 alkenyl and C 2-50 alkynyl may optionally contain -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 ), -, -S(O) 2 N(R x3 ), -, -S(O)N(R x3 ), -; -S(O) 2 -, -S(O)-, -N(R x3 ), S(O) 2 N(R x3a ), -, -S-, -N(R x3 ), -, -OC(OR x3 )(R x3a ), -, -N(R x3 ), C(O)N(R x3a ), and -OC(O)N(R x3 ), may optionally interrupt, each T 0 is, independently, selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclic, and each said T 0is independently one or more of the same or different -R x2 optionally substituted with each R x2 is independently halogen, -CN, oxo(=O), -COOR x4 , -OR x4 , -C(O)R x4 , -C(O)N(R x4 R x4a ), -S(O) 2 N(R x4 R x4a ), -S(O)N(R x4 R x4a ), -S(O) 2 R x4 , -S(O)R x4 , -N(R x4 ), -N(R 2 R x4a R x4b ), -SR x4 , -N(R x4 R x4a ), -NO 2 , -OC(O)R x4 , -N(R x4 ), -N(R x4a ), -N(R x4 ), -N(R 2 R x4a ), -N(R x4 ), -N(R x4a ), -N(R x4 ), -N(R x4a ), -N(R x4 ), -OC(O)N(R x4a R x4b ), and C x4 alkyl selected from the group consisting of, said C x4a alkyl is optionally substituted with one or more of the same or different halogen, 1-6 each -R 1-6 , -R , -R x3 , -R x3a , -R x4 , -R x4a , -R x4b is independently selected from the group consisting of -H and C 1-6 alkyl, said C 1-6The alkyl is optionally substituted with one or more of the same or different halogens.

[0095] More preferably, one or more additional optional substituents are, independently of each other, halogen, -CN, -COOR x1 , -OR x1 , -C(O)R x1 , -C(O)N(R x1 R x1a ), -S(O) 2 N(R x1 R x1a ), -S(O)N(R x1 R x1a ), -S(O) 2 R x1 , -S(O)R x1 , -N(R x1 ), -N(R 2 )S(O) x1a N(R x1b ), -SR x1 , -N(R x1 R x1a ), -NO 2 , -OC(O)R x1 , -N(R x1 ), -N(R x1a ), -N(R x1 ), -N(R 2 R x1a ), -N(R x1 ), -N(R x1a ), -N(R x1 ), -N(R x1a ), -N(R x1 ), -N(R x1a R x1b ), -OC(O)N(R x1 R x1a ), -T 0 , C 1-10 alkyl, C 2-10 alkenyl, and C 2-10 alkynyl, where -T 0 , C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl are optionally substituted with one or more of the same or different -R x2 , and C 1-10Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl may contain one or more groups selected from the group consisting of -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O) 2 N(R x3 )-, -S(O)N(R x3 )-, -S(O) 2 -, -S(O)-, -N(R x3 )S(O) 2 N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(R x3 )C(O)N(R x3a )-, and -OC(O)N(R x3 )- optionally intervening, Each -R x1 , -R x1a , -R x1b , -R x3 , -R x3a is independently selected from the group consisting of -H, halogen, C 1-6 alkyl, C 2-6 alkenyl and C 2-6 alkynyl, Each T 0 is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclic, and each said T 0 is optionally substituted with one or more -R x2 which are the same or different, Each R x2 is independently halogen, -CN, oxo (=O), -COOR x4 , -OR x4 , -C(O)R x4 , -C(O)N(R x4 R x4a ), -S(O) 2 N(R x4 R x4a ), -S(O)N(R x4 Rx4a )、 -S(O) 2 R x4 、 -S(O)R x4 、 -N(R x4 )S(O) 2 N(R x4a R x4b )、 -SR x4 、 -N(R x4 R x4a )、 -NO 2 、 -OC(O)R x4 、 -N(R x4 )C(O)R x4a 、 -N(R x4 )S(O) 2 R x4a 、 -N(R x4 )S(O)R x4a 、 -N(R x4 )C(O)OR x4a 、 -N(R x4 )C(O)N(R x4a R x4b )、 -OC(O)N(R x4 R x4a )、 and C 1-6 selected from the group consisting of alkyl, wherein said C 1-6 alkyl is optionally substituted with one or more identical or different halogens, each -R x4 、 -R x4a 、 -R x4b is independently selected from the group consisting of -H, halogen, C 1-6 alkyl, C 2-6 alkenyl and C 2-6 alkynyl.

[0096] More preferably, one or more additional optional substituents are, independently of each other, halogen, -CN, -COOR x1 、 -OR x1 、 -C(O)R x1 、 -C(O)N(R x1 R x1a )、 -S(O) 2 N(R x1 R x1a )、 -S(O)N(R x1 R x1a )、 -S(O) 2 Rx1 、 -S(O)R x1 、 -N(R x1 )S(O) 2 N(R x1a R x1b )、 -SR x1 、 -N(R x1 R x1a )、 -NO 2 、 -OC(O)R x1 、 -N(R x1 )C(O)R x1a 、 -N(R x1 )S(O) 2 R x1a 、 -N(R x1 )S(O)R x1a 、 -N(R x1 )C(O)OR x1a 、 -N(R x1 )C(O)N(R x1a R x1b )、 -OC(O)N(R x1 R x1a )、 -T 0 、 C 1-6 alkyl, C 2-6 alkenyl, and C 2-6 alkynyl selected from the group consisting of, where -T 0 、 C 1-6 alkyl, C 2-6 alkenyl and C 2-6 alkynyl may be optionally substituted with one or more -R x2 and the C 1-6 alkyl, C 2-6 alkenyl and C 2-6 alkynyl may contain -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O) 2 N(R x3 )-, -S(O)N(R x3 )-, -S(O) 2 -, -S(O)-, -N(R x3 )S(O) 2 N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(Rx3 )C(O)N(R x3a )-, and -OC(O)N(R x3 )- from the group consisting of one or more groups selected may optionally intervene, each -R x1 , -R x1a , -R x1b , -R x2 , -R x3 , -R x3a is independently, -H, halogen, C 1-6 alkyl, C 2-6 alkenyl and C 2-6 alkynyl selected from the group consisting of, each T 0 is independently, phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclic selected from the group consisting of, said each T 0 is independently, one or more -R x2 optionally substituted with.

[0097] Preferably, up to 6 -H atoms of the optionally substituted molecule are independently replaced by substituents, for example, 5 -H atoms are independently replaced by substituents, 4 -H atoms are independently replaced by substituents, 3 -H atoms are independently replaced by substituents, 2 -H atoms are independently replaced by substituents, or 1 -H atom is replaced by a substituent.

[0098] The term "intervening (interrupted)" means that the moiety is inserted between two carbon atoms, or if the insertion is at the end of the moiety, the moiety is inserted between a carbon or heteroatom and a hydrogen atom.

[0099] The term "C 1-4 alkyl" as used herein alone or in combination means a straight-chain or branched alkyl moiety having 1 to 4 carbon atoms. When present at the end of a molecule, straight-chain or branched C1-4 Examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. When two portions of a molecule are bonded by C 1-4 alkyl, such C 1-4 Examples of alkyl groups include -CH 2 -, -CH 2 -CH 2 -, -CH(CH 3 )-, -CH 2 -CH 2 -CH 2 -, -CH(C 2 H 5 )-, -C(CH 3 ) 2 -. Each hydrogen of the C 1-4 alkyl carbon may optionally be replaced by a substituent as defined above. Optionally, one or more moieties as defined below may interrupt the C 1-4 alkyl.

[0100] The term "C 1-6 alkyl" as used herein alone or in combination means a straight-chain or branched alkyl moiety having 1 to 6 carbon atoms. When present at the end of a molecule, examples of straight-chain and branched C 1-6 alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethylpropyl. When two portions of a molecule are bonded by a C 1-6 alkyl group, examples of such C 1-6 alkyl groups include -CH 2 -, -CH 2 -CH 2 -, -CH(CH 3 )-, -CH 2 -CH 2 -CH 2 -, -CH(C 2 H 5 )- and -C(CH3 ) 2 - is C 1-6 Each hydrogen atom of carbon may optionally be replaced by a substituent as defined above. Optionally, C 1-6 One or more moieties as defined below may be inserted into the alkyl.

[0101] Thus, "C 1-10 alkyl", "C 1-20 alkyl" or "C 1-50 alkyl" each means an alkyl chain having 1 to 10, 1 to 20, or 1 to 50 carbon atoms, respectively, and C 1-10 , C 1-20 or C 1-50 Each hydrogen atom of carbon may optionally be replaced by a substituent as defined above. Optionally, C 1-10 or C 1-50 One or more moieties as defined below may be inserted into the alkyl.

[0102] The term "C 2-6 alkenyl" as used herein alone or in combination means a straight-chain or branched hydrocarbon moiety having 2 to 6 carbon atoms and containing at least one carbon-carbon double bond. When present at the end of the molecule, examples are -CH=CH 2 , -CH=CH-CH 3 , -CH 2 -CH=CH 2 , -CH=CHCH 2 -CH 3 and -CH=CH-CH=CH 2 . When two moieties of the molecule are joined by C 2-6 alkenyl, an example of such C 2-6 alkenyl is -CH=CH-. Each hydrogen atom of the C 2-6 alkenyl moiety may optionally be replaced by a substituent as defined above. Optionally, C 2-6 One or more moieties as defined below may be inserted into the alkenyl.

[0103] Accordingly, the term "C 2-10 alkenyl", "C 2-20 alkenyl", or "C 2-50 alkenyl", alone or in combination, means a straight-chain or branched hydrocarbon moiety having from 2 to 10, from 2 to 20, or from 2 to 50 carbon atoms and containing at least one carbon-carbon double bond. Each hydrogen atom of the C 2-10 alkenyl, C 2-20 alkenyl, or C 2-50 alkenyl group may optionally be replaced by a substituent as defined above. Optionally, one or more moieties as defined below may interrupt the C 2-10 alkenyl, C 2-20 alkenyl, or C 2-50 alkenyl.

[0104] The term "C 2-6 alkynyl", used herein alone or in combination, means a straight-chain or branched hydrocarbon moiety having from 2 to 6 carbon atoms and containing at least one carbon-carbon triple bond. When present at the end of a molecule, examples are -C≡CH, -CH 2 -C≡CH, CH 2 -CH 2 -C≡CH, and CH 2 -C≡C-CH 3 . When two moieties of a molecule are joined by an alkynyl group, an example is -C≡C-. Each hydrogen atom of the C 2-6 alkynyl group may optionally be replaced by a substituent as defined above. Optionally, one or more double bonds may be present. Optionally, one or more moieties as defined below may interrupt the C 2-6 alkynyl.

[0105] Accordingly, the terms "C 2-10 alkynyl", "C 2-20 alkynyl", or "C 2-50 alkynyl", used herein alone or in combination, each mean a straight-chain or branched hydrocarbon moiety having from 2 to 10, from 2 to 20, or from 2 to 50 carbon atoms and containing at least one carbon-carbon triple bond. C2-10 alkynyl, C 2-20 alkynyl or C 2-50 Each hydrogen atom of the alkynyl group may optionally be replaced by a substituent as defined above. Optionally, one or more double bonds may be present. Optionally, C 2-10 alkynyl, C 2-20 alkynyl or C 2-50 one or more moieties as defined below may be inserted into the alkynyl.

[0106] As described above, C 1-4 alkyl, C 1-6 alkyl, C 1-10 alkyl, C 1-20 alkyl, C 1-50 alkyl, C 2-6 alkenyl, C 2-10 alkenyl, C 2-20 alkenyl, C 2-50 alkenyl, C 2-6 alkynyl, C 2-10 alkynyl, C 2-20 alkenyl or C 2-50 one or more moieties may optionally be inserted into the alkynyl, and the one or more moieties are preferably

Chemical formula

[0107] As used herein, the term "C 3-10 cycloalkyl" means a cyclic alkyl chain having 3 to 10 carbon atoms, which may be saturated or unsaturated, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl. C 3-10Each hydrogen atom of the cycloalkyl carbon may be replaced by a substituent as defined above. The term "C 3-10 cycloalkyl" also includes bridged bicyclics such as norbornane or norbornene.

[0108] The term "8- to 30-membered carbopolycyclic" or "8- to 30-membered carbon polycyclic" means a cyclic moiety having 8 to 30 ring atoms, in which two adjacent rings share at least one ring atom, and which can contain up to the maximum number of double bonds (aromatic or non-aromatic rings that are fully saturated, partially saturated, or unsaturated). Preferably, 8- to 30-membered carbopolycyclic means a cyclic moiety of 2, 3, 4, or 5 rings, more preferably 2, 3, or 4 rings.

[0109] As used herein, the term "3- to 10-membered heterocyclyl" or "3- to 10-membered heterocyclic" has 3, 4, 5, 6, 7, 8, 9, or 10 ring atoms, and at least one ring atom, up to a maximum of 4 ring atoms, is sulfur (-S(O)-, -S(O) 2a ring replaced by a heteroatom selected from the group consisting of oxygen, and nitrogen (including =N(O)-), which can contain up to the maximum number of double bonds (a completely saturated, partially saturated, or unsaturated, aromatic or non-aromatic ring), and means a ring bonded to the remainder of the molecule by a carbon or nitrogen atom. Examples of 3- to 10-membered heterocycles include, but are not limited to, aziridine, oxirane, thiirane, azirine, oxirene, thiirene, azetidine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazole, isothiazoline, thiadiazole, thiadiazoline, tetrahydrofuran, tetrahydrothiophene, pyrrolidine, imidazolidine, pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, thiadiazolidine, sulfolane, pyran, dihydropyran, tetrahydropyran, imidazolidine, pyridine, pyridazine, pyrazine, pyrimidine, piperazine, piperidine, morpholine, tetrazole, triazole, triazolidine, tetrazolidine, diazepane, azepine and homopiperazine. Each hydrogen atom of a 3- to 10-membered heterocyclyl or 3- to 10-membered heterocyclic group may optionally be replaced by a substituent as defined below.

[0110] As used herein, the term "8- to 11-membered heterobicyclic" or "8- to 11-membered heterobicyclic ring" refers to a bicyclic heterocyclic moiety having 8 to 11 ring atoms, which can contain up to the maximum number of double bonds (a completely saturated, partially saturated, or unsaturated, aromatic or non-aromatic ring), wherein at least one ring atom is shared by both rings, and at least one ring atom, up to a maximum of 6 ring atoms, is sulfur (-S(O)-, -S(O) 2is replaced by a heteroatom selected from the group consisting of oxygen and nitrogen (including =N(O)-), and the ring is attached to the remainder of the molecule by a carbon or nitrogen atom, meaning a heterocyclic moiety. Examples of 8- to 11-membered heterobicycles are indole, indoline, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzisothiazole, benzimidazole, benzimidazoline, quinoline, quinazoline, dihydroquinazoline, quinoline, dihydroquinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, decahydroisoquinoline, tetrahydroisoquinoline, dihydroisoquinoline, benzazepine, purine and pteridine. The term 8- to 11-membered heterobicycle also includes bicyclic spiro structures such as 1,4-dioxa-8-azaspiro[4.5]decane, or bridged heterocycles such as 8-aza-bicyclo[3.2.1]octane. Each hydrogen atom of the 8- to 11-membered heterobicyclic or 8- to 11-membered heterobicyclic carbon may optionally be replaced by a substituent as defined below.

[0111] Similarly, the term "8- to 30-membered hetero-polycyclic" or "8- to 30-membered hetero-polycycle" means a heterocyclic moiety having 8 to 30 ring atoms and capable of containing up to the maximum number of double bonds (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic rings), of more than two rings, preferably 3, 4 or 5 rings, wherein two adjacent rings share at least one ring atom, and at least one ring atom, up to a maximum of 10 ring atoms, is replaced by a heteroatom selected from the group consisting of sulfur (-S(O)-, -S(O) 2 -containing), oxygen, and nitrogen (including =N(O)-), and the ring is attached to the remainder of the molecule by a carbon or nitrogen atom.

[0112] Structure:

Chemical Structure

[0113] Structure: [Chemical formula] Regarding the moiety of x / R y The phrase "form ring A together with the atoms to which they are attached" is also understood to mean that R x and R y form the following structure: [Chemical formula]

[0114] As used herein, the term "terminal alkyne" means the following moiety: [Chemical formula]

[0115] As used herein, "halogen" means fluoro, chloro, bromo or iodo. Halogen is generally preferably fluoro or chloro.

[0116] Generally, the term "comprising" or "comprises" also encompasses "consisting of" or "consists of".

[0117] Preferably, -Z is water-soluble.

[0118] Preferably, -Z has a molecular weight in the range of 10 to 500 kDa. Even more preferably, -Z has a molecular weight in the range of 10 to 250 kDa, even more preferably 10 to 150 kDa, even more preferably 12 to 100, and most preferably 15 to 80. In certain preferred embodiments, -Z has a molecular weight of about 20 kDa. In another preferred embodiment, -Z has a molecular weight of about 40 kDa.

[0119] -Z preferably contains a polymer selected from the group consisting of 2-methacryloyloxyethyl phosphorylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amide amines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyl oxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organic phosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinyl methyl ethers), poly(vinyl pyrrolidones), silicones, celluloses, carboxymethyl celluloses, hydroxypropyl methyl celluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other hydrocarbon-based polymers, xylans, and copolymers thereof.

[0120] The moiety -Z can be a linear, branched, multi-branched or dendritic polymer moiety.

[0121] In one embodiment, -Z is a linear polymer moiety.

[0122] In another embodiment, -Z is a multi-branched polymer moiety.

[0123] In another embodiment, -Z is a dendritic polymer moiety.

[0124] In a preferred embodiment, -Z is a branched polymer moiety.

[0125] In one embodiment, -Z comprises a protein. Preferred proteins are the carboxyl-terminal peptide of chorionic gonadotropin as described in US2012 / 0035101A1 (incorporated herein by reference); albumin; the XTEN sequence as described in WO2011123813A2 (incorporated herein by reference); the proline / alanine random coil sequence as described in WO2011 / 144756A1 (incorporated herein by reference); the proline / alanine / serine random coil sequence as described in WO2008 / 155134A1 and WO2013 / 024049A1 (incorporated herein by reference); and Fc fusion proteins, and are selected from the group consisting of.

[0126] In one embodiment, -Z is polysarcosine.

[0127] In another preferred embodiment, -Z comprises poly(N-methylglycine).

[0128] In a particularly preferred embodiment, -Z comprises a random coil protein moiety.

[0129] In a preferred embodiment, -Z comprises one random coil protein moiety.

[0130] In another preferred embodiment, -Z comprises two random coil protein moieties.

[0131] In another preferred embodiment, -Z comprises three random coil protein moieties.

[0132] In another preferred embodiment, -Z comprises four random coil protein moieties.

[0133] In another preferred embodiment, -Z comprises five random coil protein moieties.

[0134] In another preferred embodiment, -Z comprises six random coil protein moieties.

[0135] In another preferred embodiment, -Z comprises seven random coil protein moieties.

[0136] In another preferred embodiment, -Z comprises eight random coil protein moieties.

[0137] Preferably, such random coil protein moieties contain at least 25 amino acid residues and at most 2000 amino acids. Even more preferably, such random coil protein moieties contain at least 30 amino acid residues and at most 1500 amino acid residues. Even more preferably, such random coil protein moieties contain at least 50 amino acid residues and at most 500 amino acid residues.

[0138] In a preferred embodiment, -Z comprises a random coil protein portion in which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming the random coil protein portion are selected from alanine and proline. Even more preferably, at least 10% but less than 75%, preferably less than 65% of the total number of amino acid residues of such a random coil protein portion are proline residues. Preferably, such a random coil protein portion is as described in WO2011 / 144756A1 (the entire content of which is incorporated herein by reference). Even more preferably, -Z comprises at least one portion selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 51 and SEQ ID NO: 61 disclosed in WO2011 / 144756 (incorporated herein by reference). A portion containing such a random coil protein containing alanine and proline is referred to as "PA" or "PA portion".

[0139] Accordingly, -Z comprises a PA portion.

[0140] In a similarly preferred embodiment, -Z comprises a random coil protein portion in which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming the random coil protein portion are selected from alanine, serine and proline. Even more preferably, at least 4% but less than 40% of the total number of amino acid residues of such a random coil protein portion are proline residues. Preferably, such a random coil protein portion is as described in WO2008 / 155134A1 (incorporated herein by reference in its entirety). Even more preferably -Z comprises at least one portion selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54 and SEQ ID NO:56 as disclosed in WO2008 / 155134A1 (incorporated herein by reference). A portion comprising such a random coil protein portion containing alanine, serine and proline is referred to as a "PAS" or "PAS portion".

[0141] Accordingly, -Z comprises a PAS portion.

[0142] In a similarly preferred embodiment, -Z comprises a random coil protein portion in which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming the random coil protein portion are selected from alanine, glycine and proline. A portion comprising such a random coil protein portion containing alanine, glycine and proline is referred to as a "PAG" or "PAG portion".

[0143] Accordingly, -Z contains a PAG moiety.

[0144] Similarly, in a preferred embodiment, -Z contains a random coil protein moiety in which at least 80%, preferably at least 85%, more preferably at least 90%, more preferably at least 95%, more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming the random coil protein moiety are selected from proline and glycine. Such a moiety containing a random coil protein containing proline and glycine is referred to as a "PG" or "PG moiety".

[0145] Preferably, such a PG moiety contains a moiety of the following formula (a-0). [(Gly) p -Pro-(Gly) q r (a-0); In the formula, p is selected from the group consisting of 0, 1, 2, 3, 4 and 5; q is selected from the group consisting of 0, 1, 2, 3, 4 and 5; r is an integer in the range of 10 to 1000 (including both ends); provided that at least one of p and q is at least 1.

[0146] Preferably, p of the formula (a-0) is selected from the group consisting of 1, 2 and 3.

[0147] Preferably, q of the formula (a-0) is selected from 0, 1 and 2.

[0148] Even more preferably, the PG moiety contains the sequence of SEQ ID NO: 97: GGPGGPGPGGPGGPGPGGPG.

[0149] Even more preferably, the PG moiety contains the sequence of the following formula (a-0-a). (GGPGGPGPGGPGGPGPGGPG)​v (a-0-a) [wherein, v is an integer in the range of 1 to 50 (including both ends)].

[0150] The array of formula (a-0-a) is understood to include v repetitions of the array of array number 97.

[0151] Therefore, -Z includes the PG moiety.

[0152] In a similarly preferred embodiment, -Z comprises a random coil protein moiety in which at least 80%, preferably at least 85%, more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming the random coil protein moiety are selected from alanine, glycine, serine, threonine, glutamic acid and proline. Preferably, such a random coil protein moiety is as described in WO2010 / 091122A1 (incorporated herein by reference). Even more preferably -Z comprises at least one moiety selected from the group consisting of SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184; SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 211, SEQ ID NO: 212, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO: 221, SEQ ID NO: 759, SEQ ID NO: 760, SEQ ID NO: 761, SEQ ID NO: 762, SEQ ID NO: 763, SEQ ID NO: 764, SEQ ID NO: 765, SEQ ID NO: 766, SEQ ID NO: 767, SEQ ID NO: 768, SEQ ID NO: 769, SEQ ID NO: 770, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 773, SEQ ID NO: 774, SEQ ID NO: 775, SEQ ID NO: 776, SEQ ID NO: 777, SEQ ID NO: 778, SEQ ID NO: 779, SEQ ID NO: 1715, SEQ ID NO: 1716, SEQ ID NO: 1718, SEQ ID NO: 1719, SEQ ID NO: 1720, SEQ ID NO: 1721 and SEQ ID NO: 1722 as disclosed in WO2010 / 091122A1 (incorporated herein by reference).Such a portion containing a random coil protein portion containing alanine, glycine, serine, threonine, glutamic acid and proline is referred to as "XTEN" or "XTEN portion" in accordance with the designation in WO2010 / 091122A1.

[0153] Therefore, -Z contains an XTEN portion.

[0154] In another preferred embodiment, -Z is a hyaluronic acid-based polymer.

[0155] In one embodiment, -Z is a carrier disclosed in WO2013 / 024047A1 (incorporated herein by reference).

[0156] In another embodiment, -Z is a carrier disclosed in WO2013 / 024048A1 (incorporated herein by reference).

[0157] In another preferred embodiment, -Z is a PEG-based polymer.

[0158] In a preferred embodiment, -Z is a branched polymer. In one embodiment, -Z is a branched polymer having 1, 2, 3, 4, 5 or 6 branch points. Preferably, -Z is a branched polymer having 1, 2 or 3 branch points. In one embodiment, -Z is a branched polymer having 1 branch point. In another embodiment, -Z is a branched polymer having 2 branch points. In another embodiment, -Z is a branched polymer having 3 branch points.

[0159] Each branch point is preferably independently selected from the group consisting of -N<, -CH< and C<.

[0160] Preferably, such a branched portion -Z is PEG-based.

[0161] In one embodiment, such a branched portion -Z has a molecular weight in the range of 10 kDa to 500 kDa (including both ends), more preferably in the range of 10 kDa to 250 kDa (including both ends), even more preferably in the range of 10 kDa to 150 kDa (including both ends), even more preferably in the range of 12 kDa to 100 kDa (including both ends), and most preferably in the range of 15 kDa to 80 kDa (including both ends).

[0162] In one embodiment, the molecular weight of such a branched portion -Z is about 10 kDa. In another embodiment, the molecular weight of such a branched portion -Z is about 20 kDa. In another embodiment, the molecular weight of such a branched portion -Z is about 30 kDa. In another embodiment, the molecular weight of such a branched portion -Z is about 40 kDa. In another embodiment, the molecular weight of such a branched portion -Z is about 50 kDa. In another embodiment, the molecular weight of such a branched portion -Z is about 60 kDa. In another embodiment, the molecular weight of such a branched portion -Z is about 70 kDa. In another embodiment, the molecular weight of such a branched portion -Z is about 80 kDa. Most preferably, such a branched portion -Z has a molecular weight of about 40 kDa.

[0163] The applicant 1 -L 2It was found that the N-terminal binding of -Z was significantly more effective with respect to NEP stability compared to the binding at internal sites, and that the binding site with the lowest effectiveness with respect to NEP stability was the loop portion of the CNP moiety. However, the present inventors surprisingly found that this drawback of the binding to the loop with respect to NEP stability can be compensated for by using a branched moiety-Z having a molecular weight of at least 10 kDa, such as at least 12 kDa, such as at least 15 kDa, such as at least 18 kDa, such as at least 20 kDa, such as at least 24 kDa, such as at least 25 kDa, such as at least 27 kDa, such as at least 30 kDa. Preferably, such a branched moiety-Z has a molecular weight of 500 kDa or less, preferably 250 kDa or less, preferably 200 Da or less, preferably 150 kDa or less, and most preferably 100 kDa or less. Most preferably, such a branched moiety-Z has a molecular weight of about 40 kDa. Furthermore, surprisingly, it was found that the binding of -Z to the loop portion of -D decreases the affinity for NPR-B. As a result, the use of such a branched moiety-Z at the loop portion of the CNP moiety not only results in an increase in NEP stability, but also combines an increase in NEP stability and a decrease in NPR-B binding related to binding to the loop.

[0164] Surprisingly, it was also found that the binding of the 5 kDa carrier to the loop portion has no significant effect on NPR-C affinity, even if the loop portion is involved in NPR-C binding. Furthermore, surprisingly, even when the total molecular weight is the same, the branched carrier having a 4×10 kDa carrier bound to the loop portion, i.e., a branched carrier having four 10 kDa arms, was found to be more effective in decreasing NPR-C affinity than the 2×20 kDa carrier, i.e., a branched carrier having two 20 kDa arms. Therefore, it is not only the total molecular weight of the carrier bound to the loop portion but also the specific branching pattern of the carrier that affects the NPR-C binding affinity.

[0165] This finding is also supported by the NPR-C affinity measured with a 4-arm 40 kDa carrier having a different branching pattern that also showed high NPR-C affinity.

[0166] In summary, surprisingly, it was found that the NPR-C affinity can be effectively reduced with a multi-branched carrier that binds to a ring moiety having a first branch point close to the CNP moiety of less than 300 atoms, preferably less than 200 atoms, more preferably less than 100 atoms, even more preferably less than 50 atoms, even more preferably less than 25 atoms, and most preferably less than 10 atoms from the CNP moiety.

[0167] Even more preferably, one or more additional branch points are located within less than 500 atoms, even more preferably less than 300 atoms, even more preferably less than 200 atoms, even more preferably less than 100 atoms, even more preferably less than 75 atoms, even more preferably less than 50 atoms, even more preferably less than 40 atoms, and most preferably less than 35 atoms from the CNP moiety.

[0168] Furthermore, it was also found that such a branching pattern is useful for the in vivo stability of the CNP moiety, i.e., protection against proteolysis. Surprisingly, it was found that N-terminal degradation is stronger when using a 2×20 kDa carrier compared to a 4×10 kDa carrier. Similarly, even stronger N-terminal degradation was shown by using a 4-arm 40 kDa carrier having a different branching pattern.

[0169] Preferably, -Z contains the following moiety.

Chemical formula

[0170] In a similarly preferred embodiment, -Z contains an amide bond.

[0171] In one embodiment, -Z includes the portion of the following formula (a). [Chemical formula] In the formula, The dashed line indicates the bond to -L 2 - or the remainder of -Z; BP a is a branch point selected from the group consisting of -N<, -CR<, and C<; -R is selected from the group consisting of -H and C 1-6 alkyl; a is 0 when BP a is -N< or -CR<, and a is 1 when BP a is >C<; -S a -, -S a′ -, -S a″ - and -S a″′ - are each independently a chemical bond or are selected from the group consisting of C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl; C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl may be substituted with one or more identical or different -R 1 and C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl may be -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 ), -S(O) 2 N(R 2 ), -S(O)N(R 2 ), -S(O) 2 -, -S(O)-, -N(R 2 )S(O) 2 N(R 2a ), -S-, -N(R 2 ), -OC(OR 2 )(R 2a ), -N(R 2 ), C(O)N(R2a )-, and -OC(O)N(R 2 )- may optionally be interrupted by one or more groups selected from the group consisting of; Each -T- is independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 Selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbocyclic polycyclic, and 8- to 30-membered heterocyclic polycyclic; each -T- may independently be substituted with one or more identical or different -R 1 ; Each -R 1 is independently halogen, -CN, oxo (=O), -COOR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 R 3a ), -S(O) 2 N(R 3 R 3a ), -S(O)N(R 3 R 3a ), -S(O) 2 R 3 , -S(O)R 3 , -N(R 3 ),S(O) 2 N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO 2 , -OC(O)R 3 , -N(R 3 ),C(O)R 3a , -N(R 3 ),S(O) 2 R 3a , -N(R 3 ),S(O)R 3a , -N(R 3 ),C(O)OR 3a , -N(R 3 ),C(O)N(R 3a R 3b ), -OC(O)N(R 3 R 3a ) and C 1-6 Selected from the group consisting of alkyl; C 1-6The alkyl may be substituted with one or more identical or different halogens; Each -R 2 ,-R 2a ,-R 3 ,-R 3a and -R 3b is independently selected from the group consisting of -H and C 1-6 alkyl, and C 1-6 alkyl may be substituted with one or more identical or different halogens; -P a′ ,-P a″ and -P a″′ is independently a polymer moiety.

[0172] Optionally, the moiety of formula (a) is substituted with one or more substituents.

[0173] In one embodiment, BP of formula (a) a is -N<.

[0174] In another embodiment, BP of formula (a) a is -CR<. Preferably, -R is -H. Thus, a in formula (a) is preferably 0.

[0175] In another embodiment, BP of formula (a) a is >C<.

[0176] In one embodiment, -S of formula (a) a - is a chemical bond.

[0177] In another embodiment, -S of formula (a) a - is selected from the group consisting of C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl, and C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl is -C(O)O-, -O-, -C(O)-, -C(O)N(R 4 )-, -S(O) 2 N(R 4)-, -S(O)N(R 4 )-, -S(O) 2 -, -S(O)-, -N(R 4 )S(O) 2 N(R 4a )-, -S-, -N(R 4 )-, -OC(OR 4 )(R 4a )-, -N(R 4 )C(O)N(R 4a )-, and -OC(O)N(R 4 )- may optionally be interrupted by one or more chemical groups selected from the group consisting of; -R 4 and -R 4a are independently selected from the group consisting of -H, methyl, ethyl, propyl and butyl. Preferably, the -S a - of formula (a) is selected from the group consisting of methyl, ethyl, propyl, butyl, and they may optionally be interrupted by one or more chemical groups selected from the group consisting of -O-, -C(O)- and -C(O)N(R 4 ).

[0178] In one embodiment, the -S a′ - of formula (a) is a chemical bond.

[0179] In another embodiment, the -S a′ - of formula (a) is selected from the group consisting of C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl, and C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl are -C(O)O-, -O-, -C(O)-, -C(O)N(R 4 ), -S(O) 2 N(R 4 ), -S(O)N(R 4 ), -S(O) 2 -, -S(O)-, -N(R 4 )S(O) 2 N(R 4a ), -S-, -N(R 4 ), -OC(OR 4 )(R 4a)-, -N(R 4 )C(O)N(R 4a )-, and -OC(O)N(R 4 )- may optionally be interrupted by one or more chemical groups selected from the group consisting of; -R 4 and -R 4a are independently selected from the group consisting of -H, methyl, ethyl, propyl and butyl. Preferably, the -S a′ - of formula (a) is selected from the group consisting of methyl, ethyl, propyl, butyl, and they may optionally be interrupted by one or more chemical groups selected from the group consisting of -O-, -C(O)- and -C(O)N(R 4 ).

[0180] In one embodiment, the -S a″ - of formula (a) is a chemical bond.

[0181] In another embodiment, the -S a″ - of formula (a) is selected from the group consisting of C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl, and C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl are -C(O)O-, -O-, -C(O)-, -C(O)N(R 4 ), -S(O) 2 N(R 4 ), -S(O)N(R 4 ), -S(O) 2 -, -S(O)-, -N(R 4 )S(O) 2 N(R 4a ), -S-, -N(R 4 ), -OC(OR 4 )(R 4a ), -N(R 4 )C(O)N(R 4a ), and -OC(O)N(R 4 )- may optionally be interrupted by one or more chemical groups selected from the group consisting of; -R 4 and -R 4ais independently selected from the group consisting of -H, methyl, ethyl, propyl and butyl. Preferably, -S of formula (a) a″ - is selected from the group consisting of methyl, ethyl, propyl, butyl, and one or more chemical groups optionally intervening selected from the group consisting of -O-, -C(O)- and -C(O)N(R 4 )-.

[0182] In one embodiment, -S of formula (a) a″′ - is a chemical bond.

[0183] In another embodiment, -S of formula (a) a″′ - is selected from the group consisting of C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl, and C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl are -C(O)O-, -O-, -C(O)-, -C(O)N(R 4 )-, -S(O) 2 N(R 4 )-, -S(O)N(R 4 )-,-S(O) 2 -,-S(O)-,-N(R 4 )S(O) 2 N(R 4a )-,-S-,-N(R 4 )-,-OC(OR 4 )(R 4a )-,-N(R 4 )C(O)N(R 4a )- and -OC(O)N(R 4 )-; -R 4 and -R 4a are independently selected from the group consisting of -H, methyl, ethyl, propyl and butyl. Preferably, -S of formula (a) a″′ - is selected from the group consisting of methyl, ethyl, propyl, butyl, and one or more chemical groups optionally intervening selected from the group consisting of -O-, -C(O)- and -C(O)N(R 4)- may optionally be interrupted by one or more chemical groups selected from the group consisting of.

[0184] Preferably, -P of formula (a) a′ , -P a″ and -P a″′independently includes a polymer selected from the group consisting of 2-methacryloyloxyethyl phosphorylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy)polymers, poly(amides), poly(amideamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyl oxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organic phosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinyl methyl ethers), poly(vinyl pyrrolidones), silicones, celluloses, carboxymethyl celluloses, hydroxypropyl methyl celluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other hydrocarbon-based polymers, xylans, and copolymers thereof.

[0185] Preferably, -P of formula (a) a′ , -P a″ and -P a″′independently has a molecular weight in the range of 5 kDa to 50 kDa (including both ends), more preferably in the range of 5 kDa to 40 kDa (including both ends), even more preferably in the range of 7.5 kDa to 35 kDa (including both ends), even more preferably in the range of 7.5 to 30 kDa, and even more preferably in the range of 10 to 30 kDa (including both ends).

[0186] In one embodiment, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of about 5 kDa.

[0187] In another embodiment, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of about 7.5 kDa.

[0188] In another embodiment, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of about 10 kDa.

[0189] In another embodiment, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of about 12.5 kDa.

[0190] In another embodiment, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of about 15 kDa.

[0191] In another embodiment, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of about 20 kDa.

[0192] More preferably, -P of formula (a) a′ , -P a″ and -P a″′independently contains a PEG moiety. Even more preferably, -P of formula (a) a′ , -P a″ and -P a″′ each independently contains a PEG moiety that contains at least 20% PEG, even more preferably at least 30%, even more preferably at least 40% PEG, even more preferably at least 50% PEG, even more preferably at least 60% PEG, even more preferably at least 70% PEG, even more preferably at least 80% PEG, and most preferably at least 90% PEG.

[0193] In a similarly preferred embodiment, -P of formula (a) a′ , -P a″ and -P a″′ each independently contains a random coil protein moiety selected from the group consisting of protein moieties, more preferably random coil protein moieties, and most preferably PA, PAS, PAG, PG, and XTEN moieties.

[0194] In one embodiment, -P of formula (a) a′ , -P a″ and -P a″′ is a PA moiety.

[0195] In another embodiment, -P of formula (a) a′ , -P a″ and -P a″′ is a PAS moiety.

[0196] In another embodiment, -P of formula (a) a′ , -P a″ and -P a″′ is a PAG moiety.

[0197] In another embodiment, -P of formula (a) a′ , -P a″ and -P a″′ is a PG moiety.

[0198] In another embodiment, -P of formula (a) a′ , -P a″and -P a″′ is the XTEN part.

[0199] In one embodiment, -Z includes one part of formula (a).

[0200] In another embodiment, -Z includes two parts of formula (a).

[0201] In another embodiment, -Z includes three parts of formula (a).

[0202] In another embodiment, -Z includes four parts of formula (a).

[0203] In another embodiment, -Z includes five parts of formula (a).

[0204] In another embodiment, -Z includes six parts of formula (a).

[0205] In a preferred embodiment, -Z includes two parts of formula (a).

[0206] In a preferred embodiment, -Z includes a part of the following formula (b).

Chemical formula

[0207] In some cases, the moiety of formula (b) is substituted with one or more substituents.

[0208] Preferably, b3 and b4 in formula (b) are the same integer.

[0209] In a preferred embodiment, both b3 and b4 are integers in the range of 200 to 250, and most preferably b3 and b4 in formula (b) are about 225.

[0210] In another preferred embodiment, both b3 and b4 are integers in the range of 400 to 500, and most preferably b3 and b4 in formula (b) are about 450.

[0211] Preferably, b1 in formula (b) is selected from the group consisting of 0, 1, 2, 3, and 4. More preferably, b1 in formula (b) is selected from the group consisting of 1, 2, and 3. Most preferably, b1 in formula (b) is 2.

[0212] Preferably, b2 in formula (b) is selected from the group consisting of 1, 2, 3, 4, and 5. More preferably, b2 in formula (b) is selected from the group consisting of 2, 3, and 4. Most preferably, b2 in formula (b) is 3.

[0213] In a particularly preferred embodiment, b1 in formula (b) is 2, b2 in formula (b) is 3, and both b3 and b4 are about 450.

[0214] In another particularly preferred embodiment, b1 in formula (b) is 2, b2 in formula (b) is 3, and both b3 and b4 are about 225.

[0215] In one embodiment, -Z contains one moiety of formula (b).

[0216] In another embodiment, -Z contains two moieties of formula (b).

[0217] In another embodiment, -Z contains three moieties of formula (b).

[0218] In another embodiment, -Z contains four moieties of formula (b).

[0219] In another embodiment, -Z contains five moieties of formula (b).

[0220] In another embodiment, -Z contains six moieties of formula (b).

[0221] In a preferred embodiment, -Z contains two moieties of formula (b).

[0222] In an even more preferred embodiment, -Z contains a moiety of formula (c).

Chemical formula

[0223] Optionally, the moiety of formula (c) is substituted with one or more substituents.

[0224] Preferably, both c1 and c2 of formula (c) are the same integer.

[0225] In a certain preferred embodiment, c1 and c2 of formula (c) are in the range of 200 to 250 (including both ends), and most preferably about 225. In another preferred embodiment, c1 and c2 of formula (c) are in the range of 400 to 500 (including both ends), and most preferably about 450.

[0226] In a preferred embodiment, sub - Z is a branched PEG - based polymer containing at least 10% PEG, has one branching point and two PEG - based polymer arms, and has a molecular weight of about 40 kDa. Accordingly, each of the two PEG - based polymer arms has a molecular weight of about 20 kDa. Preferably, the branching point is - CH<.

[0227] In one embodiment, - Z contains one moiety of formula (c).

[0228] In another embodiment, - Z contains two moieties of formula (c).

[0229] In another embodiment, - Z contains three moieties of formula (c).

[0230] In another embodiment, - Z contains four moieties of formula (c).

[0231] In another embodiment, - Z contains five moieties of formula (c).

[0232] In another embodiment, - Z contains six moieties of formula (c).

[0233] In a preferred embodiment, - Z contains two moieties of formula (c).

[0234] In a certain preferred embodiment, sub - Z is of the following formula (d).

Chemical formula

[0235] - Z b - is selected from the group consisting of C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl; C 1-50 alkyl, C 2-50 alkenyl, and C 2-50The alkynyl may be substituted with one or more identical or different -R 1 and may be C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 The alkynyl may optionally be interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 ), -S(O) 2 N(R 2 ), -S(O)N(R 2 ), -S(O) 2 -, -S(O)-, -N(R 2 )S(O) 2 N(R 2a ), -S-, -N(R 2 ), -OC(OR 2 )(R 2a ), -N(R 2 )C(O)N(R 2a ), and -OC(O)N(R 2 ); Each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbocyclic polycycle, and 8- to 30-membered heterocyclic polycycle; each -T- may optionally be substituted with one or more identical or different -R 1 ; Each -R 1 is independently halogen, -CN, oxo (=O), -COOR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 R 3a ), -S(O) 2 N(R 3 R 3a ), -S(O)N(R 3 R 3a ), -S(O) 2 R 3 , -S(O)R 3 , -N(R 3 )S(O) 2 N(R 3a R 3b), -SR 3 , -N(R 3 R 3a ), -NO 2 , -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O) 2 R 3a , -N(R 3 )S(O)R 3a , -N(R 3 )C(O)OR 3a , -N(R 3 )C(O)N(R 3a R 3b ), -OC(O)N(R 3 R 3a ) and C 1-6 alkyl; 1-6 The alkyl may be optionally substituted with one or more identical or different halogens; Each-R 2 , -R 2a , -R 3 , -R 3a and -R 3b are independently -H and C 1-6 alkyl, C 1-6 The alkyl may be optionally substituted with one or more identical or different halogens; -Z a is: [ka] and; During the ceremony, BP a , -S a -, -S a′ -, -S a″ -, -S a″′ -, -P a′ , -P a″ , -P a″′ and a are used as defined for formula (a).

[0236] Optionally, the moiety of formula (d) is substituted with one or more substituents.

[0237] BP of formula (d) a -S a -S a′ -S a″ -S a″′ -P a′ -P a″ -P a″′ The preferred embodiment of is as defined above for formula (a).

[0238] Preferably, -Z of formula (d) a is the same as that of formula (b). The preferred embodiments of b1, b2, b3 and b4 are as described for formula (b).

[0239] Even more preferably, -Z of formula (d) a is the same as that of formula (c). The preferred embodiments for c1 and c2 are as described for formula (c).

[0240] In an even more preferred embodiment, the moiety -Z is of the following formula (e).

Chemical formula

Chemical formula

[0241] Optionally, the moiety of formula (e) is substituted with one or more substituents.

[0242] The preferred embodiments for b1, b2, b3 and b4 of formula (e) are as defined above for formula (b).

[0243] In one embodiment, e in formula (e) is 1. In another embodiment, e in formula (e) is 2. In another embodiment, e in formula (e) is 3. In another embodiment, e in formula (e) is 4. In another embodiment, e in formula (e) is 5. In another embodiment, e in formula (e) is 6. In another embodiment, e in formula (e) is 7. In another embodiment, e in formula (e) is 8. In another embodiment, e in formula (e) is 9. In another embodiment, e in formula (e) is 10. In another embodiment, e in formula (e) is 11. In another embodiment, e in formula (e) is 12. In another embodiment, e in formula (e) is 13. In another embodiment, e in formula (e) is 14. In another embodiment, e in formula (e) is 15.

[0244] Preferably, e in formula (e) is selected from the group consisting of 2, 3, 4, 5, 6, 7, 8 and 9. Even more preferably, e in formula (e) is selected from 3, 4, 5 and 6. Most preferably, e in formula (e) is 5.

[0245] Preferably, e in formula (e) is 5, b1 in formula (e) is 2, b2 in formula (e) is 3, and both b3 and b4 in formula (e) are about 450.

[0246] In a similarly preferred embodiment, Sub-Z is of the following formula (e-i) or (e-i′).

Chemical formula

Chemical formula

[0247] Preferred embodiments for b1, b2, b3 and b4 in formulas (e-i) and (e-i') are as defined above for formula (b).

[0248] Preferred embodiments for e in formulas (e-i) and (e-i') are as described for formula (e).

[0249] Preferably, b1 in formulas (e-i) and (e-i') is 2, b2 in formulas (e-i) and (e-i') is 3, and both b3 and b4 in formulas (e-i) and (e-i') are about 450.

[0250] In a preferred embodiment, -Z is that of formula (e-i).

[0251] In another preferred embodiment, the moiety -Z is a branched PEG-based polymer containing at least 10% PEG, having 3 branch points and 4 PEG-based polymer arms, and having a molecular weight of about 40 kDa. Accordingly, each of the 4 PEG-based polymer arms has a molecular weight of about 10 kDa. Preferably, each of the 3 branch points is -CH<.

[0252] In a preferred embodiment, the moiety -Z is that of the following formula (f).

Chemical formula

[0253] Optionally, the moiety of formula (f) is substituted with one or more substituents.

[0254] BP of formula (f) a , -S a -, -S a′ -, -S a″ -, -S a″′ -, -P a′ , -P a″ and -P a″′ The preferred embodiments of are as defined above for formula (a).

[0255] Preferably, the BP of formula (f) f is -CR< and r is 0. Preferably, -R is -H. Preferably, the -S of formula (f) f - is a chemical bond.

[0256] Preferably, the -Z of formula (f) a′ , -Z a″ and -Z a″′ have the same structure. Preferably, the -Z of formula (f) a′ , -Z a″ and -Z a″′ are those of formula (b).

[0257] The preferred embodiments of b1, b2, b3 and b4 are as described for formula (b).

[0258] Preferably, the -S of formula (f) f - is a chemical bond and the BP of formula (f) a is -CR< and -R is -H. Even more preferably, the -S of formula (f) f - is a chemical bond and the BP of formula (f) a is -CR< and -R is -H and the -Z of formula (f)a′ , -Z a″ and -Z a″′ is of formula (b).

[0259] Even more preferably, -Z is of the following formula (g).

Chemical formula

[0260] , -S g , -S g′ , - and -S g″ , - are independently selected from the group consisting of C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl; C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl may be substituted with one or more identical or different -R 1 , and C 1-50 alkyl, C 2-50 alkenyl and C 2-50 alkynyl may be interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 ), -, -S(O) 2 N(R 2 ), -, -S(O)N(R 2 ), -, -S(O) 2 -, -S(O)-, -N(R 2 )S(O) 2 N(R 2a ), -, -S-, -N(R 2 ), -, -OC(OR 2 )(R 2a ), -, -N(R 2 )C(O)N(R 2a ), and -OC(O)N(R 2 ); each -T- is independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10Selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbocyclic, and 8- to 30-membered heterocyclic; each -T- is independently one or more identical or different -R 1 and may be substituted with; each R 1 is independently halogen, -CN, oxo(=O), -COOR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 R 3a ), -S(O) 2 N(R 3 R 3a ), -S(O)N(R 3 R 3a ), -S(O) 2 R 3 , -S(O)R 3 , -N(R 3 ), S(O) 2 N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO 2 , -OC(O)R 3 , -N(R 3 ), C(O)R 3a , -N(R 3 ), S(O) 2 R 3a , -N(R 3 ), S(O)R 3a , -N(R 3 ), C(O)OR 3a , -N(R 3 ), C(O)N(R 3a R 3b ), -OC(O)N(R 3 R 3a ) and C 1-6 alkyl selected from the group consisting of; C 1-6 alkyl may be substituted with one or more identical or different halogens; each -R 2 , -R 2a , -R 3 , -R 3a and -R 3b is independently -H, and C1-6 selected from the group consisting of alkyl, C 1-6 the alkyl may be substituted with one or more identical or different halogens; -Z a and -Z a′ are each independently,

Chemical formula

[0261] Optionally, the moiety of formula (g) is substituted with one or more substituents.

[0262] BP of formula (g) a , -S a -, -S a′ -, -S a″ -, -S a″′ -, -P a′ -, -P a″ and -P a″′ The preferred embodiments are as defined above for formula (a).

[0263] Preferably, -S g - of formula (g) is selected from the group consisting of C 1-6 alkyl, C 2-6 alkenyl and C 2-6 alkynyl, which may be substituted with one or more identical or different -R 1 and, -R 1 is halogen, oxo (=O), -COOR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 R 3a ), -S(O) 2N(R 3 R 3a )、 -S(O)N(R 3 R 3a )、 -S(O) 2 R 3 、 -S(O)R 3 、 -N(R 3 )S(O) 2 N(R 3a R 3b )、 -SR 3 、 -N(R 3 R 3a )、 -NO 2 、 -OC(O)R 3 、 -N(R 3 )C(O)R 3a 、 -N(R 3 )S(O) 2 R 3a 、 -N(R 3 )S(O)R 3a 、 -N(R 3 )C(O)OR 3a 、 -N(R 3 )C(O)N(R 3a R 3b )、 -OC(O)N(R 3 R 3a )、 and C 1-6 is selected from the group consisting of alkyl; C 1-6 alkyl may be substituted with one or more identical or different halogens; -R 3 、 -R 3a and -R 3b are independently selected from -H, methyl, ethyl, propyl, and butyl.

[0264] Even more preferably, the -S g - of formula (g) is selected from C 1-6 alkyl.

[0265] Preferably, the -S g′ - of formula (g) is selected from the group consisting of C 1-6 alkyl, C 2-6 alkenyl, and C 2-6 alkynyl, which may be substituted with one or more identical or different -R 1 ;

[0266] -R 1 is halogen, oxo(=O), -COOR 3 -OR 3 -C(O)R 3 -C(O)N(R 3 R 3a )、-S(O) 2 N(R 3 R 3a )、-S(O)N(R 3 R 3a )、-S(O) 2 R 3 、-S(O)R 3 、-N(R 3 )S(O) 2 N(R 3a R 3b )、-SR 3 、-N(R 3 R 3a )、-NO 2 、-OC(O)R 3 、-N(R 3 )C(O)R 3a 、-N(R 3 )S(O) 2 R 3a 、-N(R 3 )S(O)R 3a 、-N(R 3 )C(O)OR 3a 、-N(R 3 )C(O)N(R 3a R 3b )、-OC(O)N(R 3 R 3a )、and C 1-6 is selected from the group consisting of alkyl; C 1-6 alkyl may be substituted with one or more identical or different halogens; -R 3 、-R 3a and -R 3b are independently selected from -H, methyl, ethyl, propyl, and butyl.

[0267] Even more preferably, the -S g′ - of formula (g) is selected from C 1-6 alkyl.

[0268] Preferably, the -S of formula (g) g″ - is C 1-6 alkyl, C 2-6 alkenyl, and C 2-6 alkynyl, and is selected from the group consisting thereof, and it may be substituted with one or more identical or different -R 1 ; -R 1 is halogen, oxo (=O), -COOR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 R 3a ), -S(O) 2 N(R 3 R 3a ), -S(O)N(R 3 R 3a ), -S(O) 2 R 3 , -S(O)R 3 , -N(R 3 )S(O) 2 N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO 2 , -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O) 2 R 3a , -N(R 3 )S(O)R 3a , -N(R 3 )C(O)OR 3a , -N(R 3 )C(O)N(R 3a R 3b ), -OC(O)N(R 3 R 3a ), and C 1-6 alkyl, and is selected from the group consisting thereof; C 1-6 alkyl may be substituted with one or more identical or different halogens; -R 3 , -R 3a , and -R 3bindependently selected from -H, methyl, ethyl, propyl and butyl.

[0269] Even more preferably, -S of formula (g) g″ - is C 1-6 selected from alkyl.

[0270] Preferably, -Z of formula (g) a and -Z a′ have the same structure. Preferably, -Z of formula (g) a and -Z a′ are those of formula (b).

[0271] In another even more preferred embodiment, -Z is of the following formula (g-i).

Chemical formula

Chemical formula

Chemical formula

[0272] Optionally, the moiety of formula (g-i) is substituted with one or more substituents.

[0273] Preferably, both -Y a1 - and -Y a1′ - are both

Chemical formula

[0274] BP of formula (g-i) a , -S a -, -S a′ -, -S a″ -, -S a″′ -, -P a′ , -P a″ and -P a″′ The preferred embodiment of is as defined above for formula (a).

[0275] -S of formula (g-i) g -, -S g′ - and -S g″ The preferred embodiment of - and -S is as defined for formula (g).

[0276] Preferably, -Z of formula (g-i) a and -Z a′ have the same structure. Preferably, -Z of formula (g-i) a and -Z a′ are those of formula (b). The preferred embodiments for b1, b2, b3 and b4 are as described for formula (b).

[0277] Even more preferably, -Z is of the following formula (h).

Chemical formula

Chemical formula

[0278] Optionally, the moiety of formula (h) is substituted with one or more substituents.

[0279] Preferably, both c1s in formula (h) are the same.

[0280] Preferably, both c1s in formula (h) are about 225.

[0281] Even more preferably, -Z is the following formula (h-a):

Chemical formula

Chemical formula

Chemical formula

[0282] Optionally, the moiety of formula (h-a) is substituted with one or more substituents.

[0283] Preferably, each k in formula (h-a) is independently selected from the group consisting of 2, 3, 4, 5, 6, and 7. Preferably, both ks in formula (h-a) are the same.

[0284] Preferably, both c1s in formula (h-a) are the same.

[0285] Preferably, both c1s in formula (h-a) are about 225.

[0286] Preferably, -Y of formula (h-a) a1 - and -Y a1′ - are both

Chemical formula

[0287] In an even more preferred embodiment, moiety -Z is of the following formula (h-i).

Chemical formula

Chemical formula

[0288] Optionally, the moiety of formula (h-i) is substituted with one or more substituents.

[0289] Preferably, both c1s in formula (h-i) are the same.

[0290] Preferably, both c1s in formula (h-i) are about 225.

[0291] In another even more preferred embodiment, moiety -Z is of the following formula (h-ia).

Chemical formula

Chem.

Chem.

[0292] Preferably, each k of formula (h-ia) is independently selected from the group consisting of 2, 3, 4, 5, 6 and 7. Preferably, both k of formula (h-ia) are the same.

[0293] Preferably, both c1 of formula (h-ia) are the same.

[0294] Preferably, both c1 of formula (h-ia) are about 225.

[0295] Preferably, -Y of formula (h-ia) a1 - and -Y a1′ - are both,

Chem.

[0296] In equally preferred embodiments, -Z comprises a moiety selected from the group consisting of the following.

Chem.

Chem.

Chem.

Chemical Structure

[0297] In a preferred embodiment, -Y d1 - and -Y d2 - and -Y d1 -,-Y d2 -,-Y d3 - and -Y d4 - are

Chemical formula

[0298] In another preferred embodiment, -Y d1 - and -Y d2 - and -Y d1 -,-Y d2 -,-Y d3 - and -Y d4 - are

Chemical formula

[0299] Preferably, -X f1 、-X f2 、-X f3, -X f4 , -X f5 , -X f6 , -X f7 and -X f8 is -H; -X of formula (j-i) d1 and -X d2 is -OH; -X of formula (j-i) e1 and -X e2 is selected from the group consisting of -H and methyl; s1, s2, s3 and s4 of formula (j-i) are selected from the group consisting of 2, 3, 4, 5 and 6. Even more preferably, -X of formula (j-i) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 and -X f8 is -H; -X of formula (j-i) d1 and -X d2 is -OH; -X of formula (j-i) e1 and -X e2 is -H; s1, s2, s3 and s4 of formula (j-i) are 4.

[0300] Preferably, -X of formula (j-ii) f1 , -X f2 , -X f3 and -X f4 is -H; -X of formula (j-ii) d1 , -X d2 , -X d3 and -X d2 is -OH; -X of formula (j-ii) e1 , -X e2 , -X e3 and -X e4 is selected from the group consisting of -H and methyl; s1, s2, s3, s4 and s5 of formula (j-ii) are selected from the group consisting of 1, 2, 3, 4, 5 and 6. Even more preferably, -X of formula (j-ii) f1 , -X f2 , -X f3 and -X f4 is -H; -X of formula (j-ii) d1 , -X d2 , -X d3and -X d2 is -OH; -X in formula (j-ii) e1 , -X e2 , -X e3 and -X e4 is -H; s1 in formula (j-ii) is 4, and s2, s3, s4 and s5 in formula (j-ii) are 1.

[0301] Preferably, -X in formula (j-iii) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 , -X f8 , -X f9 and -X f10 is -H; -X in formula (j-iii) d1 , -X d2 , -X d3 and -X d4 is -OH; -X in formula (j-iii) e1 , -X e2 , -X e3 and -X e4 is selected from the group consisting of -H and methyl; s1, s2 and s3 in formula (j-iii) are selected from the group consisting of 2, 3, 4, 5 and 6. Even more preferably, -X in formula (j-iii) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 , -X f8 , -X f9 and -X f10 is -H; -X in formula (j-iii) d1 , -X d2 , -X d3 and -X d4 is -OH; -X in formula (j-iii) e1 , -X e2 , -X e3 and -X e4 is -H; s1, s2 and s3 in formula (j-iii) are 4.

[0302] Preferably, -X of formula (j-iv) f1 , -X f2 , -X f3 , -X f4 , -X f5 and -X f6 is -H; s1, s2, s3, s4, s5, s6 and s7 of formula (j-iv) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; -Y d1 - and -Y d2 - is

Chemical formula

[0303] In an even more preferred embodiment, -X of formula (j-iv) f1 , -X f2 , -X f3 , -X f4 , -X f5 and -X f6 is -H; s1 of formula (j-iv) is 3, s2 of formula (j-iv) is 5, s3 of formula (j-iv) is 2, s4 of formula (j-iv) is 4, s5 of formula (j-iv) is 5, s6 of formula (j-iv) is 2, s7 of formula (j-iv) is 4; -Y of formula (j-iv) d1 - and -Y d2 - is

Chemical formula

[0304] In a similarly preferred embodiment, -X of formula (j-iv) f1 , -X f2 , -X f3 , -X f4 , -X f5 and -X f6 is -H; s1 of formula (j-iv) is 3, s2 of formula (j-iv) is 5, s3 of formula (j-iv) is 2, s4 of formula (j-iv) is 4, s5 of formula (j-iv) is 5, s6 of formula (j-iv) is 2, s7 of formula (j-iv) is 4; -Y of formula (j-iv) d1 - and -Yd2 - is

Chem.

[0305] The dashed lines marked with asterisks are respectively -Z d1 , -Z d2 , -Z d3 and -Z d4 in the direction of, and the unmarked dashed line is -L 2 - in the direction of.

[0306] Preferably, -X f1 , -X f2 , -X f3 and -X f4 in formula (j-v) are -H; s1, s2, s3, s4 and s5 in formula (j-v) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; -Y d1 - and -Y d2 - are

Chem.

[0307] In an even more preferred embodiment, -X f1 , -X f2 , -X f3 and -X f4 in formula (j-v) are -H; s1 in formula (j-v) is 3, s2 in formula (j-v) is 2, s3 in formula (j-v) is 1, s4 in formula (j-v) is 2, s5 in formula (j-v) is 1; -Y d1 - and -Y d2 - are

Chem.

[0308] In a similarly preferred embodiment, -X f1 , -X f2 , -X f3and -X f4 is -H; s1 in formula (j-v) is 3, s2 in formula (j-v) is 2, s3 in formula (j-v) is 1, s4 in formula (j-v) is 2, s5 in formula (j-v) is 1; -Y in formula (j-v) d1 - and -Y d2 - is [Chemical formula] ; wherein

[0309] The dashed lines marked with asterisks are each -Z d1 , -Z d2 , -Z d3 and -Z d4 and are directed towards -L 2 - is directed towards.

[0310] Preferably, -X in formula (j-vi) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 , -X f8 , -X f9 and -X f10 is -H; s1, s2, s3, s4, s5, s6, s7, s8 and s9 in formula (j-vi) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; -Y in formula (j-vi) d1 - and -Y d2 - are [Chemical formula] selected from the group consisting of.

[0311] In an even more preferred embodiment, -X in formula (j-vi) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 , -X f8 , -Xf9 and -X f10 is -H; s1 in formula (j-vi) is 4, s2 in formula (j-vi) is 5, s3 in formula (j-vi) is 2, s4 in formula (j-vi) is 4, s5 in formula (j-vi) is 4, s6 in formula (j-vi) is 5, s7 in formula (j-vi) is 2, s8 in formula (j-vi) is 4, s9 in formula (j-vi) is 4; -Y in formula (j-v) d1 - and -Y d2 - is

Chemical formula

[0312] Similarly, in a preferred embodiment, -X in formula (j-vi) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 , -X f8 , -X f9 and -X f10 is -H; s1 in formula (j-vi) is 4, s2 in formula (j-vi) is 5, s3 in formula (j-vi) is 2, s4 in formula (j-vi) is 4, s5 in formula (j-vi) is 4, s6 in formula (j-vi) is 5, s7 in formula (j-vi) is 2, s8 in formula (j-vi) is 4, s9 in formula (j-vi) is 4; -Y in formula (j-v) d1 - and -Y d2 - is

Chemical formula

[0313] Preferably, -X in formula (j-vii)f1 、 -X f2 、 -X f3 、 -X f4 、 -X f5 、 -X f6 、 -X f7 、 -X f8 、 -X f9 、 -X f10 、 -X f11 、 -X f12 、 -X f13 and -X f14 is -H; s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14 and s15 of formula (j-vii) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; -Y of formula (j-vii) d1 -, -Y d2 -, -Y d3 - and -Y d4 - is [Chemical formula] selected from the group consisting of.

[0314] In a further more preferred embodiment, -X of formula (j-vii) f1 、 -X f2 、 -X f3 、 -X f4 、 -X f5 、 -X f6 、 -X f7 、 -X f8 、 -X f9 、 -X f10 、 -X f11 、 -X f12 、 -X f13 and -X f14is -H; is -H; s1 in formula (j-vii) is 4, s2 in formula (j-vii) is 4, s3 in formula (j-vii) is 5, s4 in formula (j-vii) is 2, s5 in formula (j-vii) is 4, s6 in formula (j-vii) is 5, s7 in formula (j-vii) is 2, s8 in formula (j-vii) is 4, s9 in formula (j-vii) is 4, s10 in formula (j-vii) is 5, s11 in formula (j-vii) is 2, s12 in formula (j-vii) is 4, s13 in formula (j-vii) is 5, s14 in formula (j-vii) is 2, s15 in formula (j-vii) is 4; -Y in formula (j-vii) d1 -, -Y d2 -, -Y d3 - and -Y d4 - is [Chemical formula] as follows.

[0315] Similarly, in a preferred embodiment, -X in formula (j-vii) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 , -X f8 , -X f9 , -X f10 , -X f11 , -X f12 , -X f13 and -X f14 is -H; is -H; s1 in formula (j-vii) is 4, s2 in formula (j-vii) is 4, s3 in formula (j-vii) is 5, s4 in formula (j-vii) is 2, s5 in formula (j-vii) is 4, s6 in formula (j-vii) is 5, s7 in formula (j-vii) is 2, s8 in formula (j-vii) is 4, s9 in formula (j-vii) is 4, s10 in formula (j-vii) is 5, s11 in formula (j-vii) is 2, s12 in formula (j-vii) is 4, s13 in formula (j-vii) is 5, s14 in formula (j-vii) is 2, s15 in formula (j-vii) is 4; -Y in formula (j-vii)d1 -, -Y d2 -, -Y d3 - and -Y d4 - is [Chem.] as follows;

[0316] The dashed lines marked with asterisks are respectively -Z d1 , -Z d2 , -Z d3 and -Z d4 and are directed towards -L 2 - is directed towards.

[0317] Preferably, the -Z in formulas (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 have the same structure.

[0318] In one embodiment, the -Z in formulas (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 is a PA moiety.

[0319] In another embodiment, the -Z in formulas (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 is a PAS moiety.

[0320] In another embodiment, the -Z in formulas (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 is a PAG moiety.

[0321] In another embodiment, -Z in formulas (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 is the PG moiety.

[0322] In another embodiment, -Z in formulas (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 is the XTEN moiety.

[0323] Preferably, the CNP prodrug of the present invention is of the following formula (I).

Chemical formula

[0324] The moiety -L 1 - is a reversible prodrug linker from which the drug, i.e., CNP, is released in its free form, i.e., a prodrug linker that leaves no trace. Suitable prodrug linkers, such as those disclosed in WO 2005 / 099768 A2, WO 2006 / 136586 A2, WO 2011 / 089216 A1 and WO 2013 / 024053 A1, which are incorporated herein by reference, are known in the art.

[0325] In another embodiment, -L 1 ​​​​- is a reversible prodrug linker as described in WO 2011 / 012722 A1, WO 2011 / 089214 A1, WO 2011 / 089215 A1, WO 2013 / 024052 A1 and WO 2013 / 160340 A1, which are incorporated herein by reference.

[0326] Particularly preferred moiety -L 1 - is disclosed in WO 2009 / 095479 A2. Thus, in one preferred embodiment, moiety -L 1 - is of formula (II):

[0327] [Chemical formula] (wherein The dashed line indicates the bond to the nitrogen of -D, which is the CNP moiety, by forming an amide bond, and -X- is -C(R 4 R 4a )-, -N(R 4 )-, -O-, -C(R 4 R 4a )-C(R 5 R 5a )-, -C(R 5 R 5a )-C(R 4 R 4a )-, -C(R 4 R 4a )-N(R 6 )-, -N(R 6 )-C(R 4 R 4a )-, -C(R 4 R 4a )-O-, -O-C(R 4 R 4a )-, or -C(R 7 R 7a )-, and X 1 is C, or S(O), and -X 2 - is -C(R 8 R 8a )-, or -C(R 8 R 8a )-C(R 9 R9a ) - and = X 3 is = O, = S, or = N - CN, - R 1 - R 1a - R 2 - R 2a - R 4 - R 4a - R 5 - R 5a - R 6 - R 8 - R 8a - R 9 - R 9a are independently selected from the group consisting of - H and C 1-6 alkyl, - R 3 - R 3a are independently selected from the group consisting of - H and C 1-6 alkyl, provided that when one or both of - R 3 - R 3a are other than - H, they are linked to the N to which they are attached by an SP 3 hybridized carbon atom, - R 7 is - N(R 10 R 10a ) or - NR 10 - (C = O) - R 11 and - R 7a - R 10 - R 10a - R 11 are independently of each other - H or C 1-6 alkyl, optionally, the pair - R 1a / - R 4a - R 1a / - R 5a - R 1a / - R 7a - R 4a / - R 5a - R 8a / - R 9a one or more of which form a chemical bond, optionally, the pair - R 1 / - R 1a - R 2 / - R2a , -R 4 / -R 4a , -R 5 / -R 5a , -R 8 / -R 8a , -R 9 / -R 9a One or more of which, together with the atoms to which they are attached, form a C 3-10 Cycloalkyl or 3- to 10-membered heterocyclyl, Optionally, the pair -R 1 / -R 4 , -R 1 / -R 5 , -R 1 / -R 6 , -R 1 / -R 7a , -R 4 / -R 5 , -R 4 / -R 6 , -R 8 / -R 9 , -R 2 / -R 3 One or more of which, together with the atoms to which they are attached, form Ring A, Optionally, R 3 / R 3a Forms a 3- to 10-membered heterocyclic ring together with the nitrogen atom to which it is attached, A is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclyl and 8- to 11-membered heterobicyclicyl), and has -L 1 - is -L 2 Substituted with -Z, and optionally, -L 1 - is further substituted, provided that the hydrogen atom marked with an asterisk in formula (II) is not replaced by -L 2 -Z or a substituent, -L 2 - is a single chemical bond or a spacer, -Z is a polymer having a molecular weight of at least 10 kDa.

[0328] Preferably, -L of formula (II) 1 - is a single moiety -L 2 -Z is replaced.

[0329] Preferred embodiments of -Z of formula (II) are as described above.

[0330] In one embodiment, -L of formula (II) 1 - is further unsubstituted.

[0331] -R of formula (II) 3 / -R 3a When they combine with the nitrogen atom to which they are attached to form a 3- to 10-membered heterocyclic ring, it is understood that only such 3- to 10-membered heterocyclic rings in which the atom directly bonded to nitrogen is an sp 3 hybridized carbon atom can be formed. In other words, -R 3 / -R 3a Such 3- to 10-membered heterocyclic rings formed by -R

Chemical formula

[0332] It is also understood that the 3- to 10-membered heterocyclic ring may be further substituted.

[0333] -R of formula (II) 3 / -R 3a Exemplary embodiments of suitable 3- to 10-membered heterocyclic rings formed by -R of formula (II) and the nitrogen atom to which they are attached are as follows:

Chemical formula

[0334] Optionally, -L in formula (II) 1 - may be further substituted. Generally, any substituent may be used as long as it does not affect the principle of cleavage, that is, the hydrogen with an asterisk in formula (II) is not replaced, and the part of formula (II)

Chemical formula

[0335] In one embodiment, -R in formula (II) 1 or -R 1a is substituted with -L 2 -Z. In another embodiment, -R in formula (II) 2 or -R 2a is substituted with -L 2 -Z. In another embodiment, -R in formula (II) 3 or -R 3a is substituted with -L 2 -Z. In another embodiment, -R in formula (II) 4 is substituted with -L 2 -Z. In another embodiment, -R in formula (II) 5 or -R 5a is substituted with -L 2 -Z. In another embodiment, -R in formula (II) 6 is substituted with -L 2 -Z. In another embodiment, -R in formula (II) 7 or -R 7a is substituted with -L 2 -Z. In another embodiment, -R in formula (II) 8or -R 8a is -L 2 is replaced with -Z. In another embodiment, -R of formula (II) 9 or -R 9a is -L 2 is replaced with -Z.

[0336] Most preferably, -R of formula (II) 4 is -L 2 is replaced with -Z.

[0337] Preferably, -X- of formula (II) is -C(R 4 R 4a )- or -N(R 4 )-. Most preferably, -X- of formula (II) is -C(R 4 R 4a )-.

[0338] Preferably, X of formula (II) 1 is C.

[0339] Preferably, =X of formula (II) 3 is =O.

[0340] Preferably, -X of formula (II) 2 - is -C(R 8 R 8a )-.

[0341] Preferably, -R of formula (II) 8 and -R 8a are independently selected from the group consisting of -H, methyl and ethyl. More preferably, at least one of -R of formula (II) 8 and -R 8a is -H. Even more preferably, both -R of formula (II) 8 and -R 8a are -H.

[0342] Preferably, -R of formula (II) 1 and -R 1ais independently selected from the group consisting of -H, methyl, and ethyl. More preferably, -R of formula (II) 1 and -R 1a at least one of which is -H. Even more preferably, -R of formula (II) 1 and -R 1a both are -H.

[0343] Preferably, -R of formula (II) 2 and -R 2a are independently selected from the group consisting of -H, methyl, and ethyl. More preferably, -R of formula (II) 2 and -R 2a at least one of which is -H. Even more preferably, -R of formula (II) 2 and -R 2a both are H.

[0344] Preferably, -R of formula (II) 3 and -R 3a are independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. Even more preferably, -R of formula (II) 3 and -R 3a at least one of which is methyl. In a similarly preferred embodiment, -R of formula (II) 3 and -R 3a both are -H. In another similarly preferred embodiment, -R of formula (II) 3 and -R 3a both are methyl.

[0345] Preferably, R of formula (II) 3 is -H, and -R of formula (II) 3a is methyl.

[0346] Preferably, -R of formula (II) 4 and -R 4a are independently selected from the group consisting of -H, methyl, and ethyl. More preferably, -R of formula (II) 4 and -R 4aAt least one of them is -H. Even more preferably, -R of formula (II) 4 and -R 4a are both -H.

[0347] Preferably, the moiety -L 1 - is of formula (IIa):

Chemical formula

[0348] Preferably, -L of formula (IIa) 1 - is substituted with one moiety -L 2 -Z.

[0349] Preferably, the moiety -L of formula (IIa) 1 - is not further substituted.

[0350] Preferred embodiments of -Z of formula (IIa) are as described above.

[0351] In certain embodiments, -R of formula (IIa) 1 or -R 1a is -L 2is replaced by -Z. In another embodiment, -R of formula (IIa) 2 or -R 2a is replaced by -L 2 -Z. In another embodiment, -R of formula (IIa) 3 or -R 3a is replaced by -L 2 -Z. In another embodiment, -R of formula (IIa) 4 is replaced by -L 2 -Z. In another embodiment, -R of formula (IIa) 8 or -R 8a is replaced by -L 2 -Z. In another embodiment, -R of formula (IIa) 9 or -R 9a is replaced by -L 2 -Z.

[0352] Most preferably, -R of formula (IIa) 4 is replaced by -L 2 -Z.

[0353] Preferably, -R of formula (IIa) 1 and -R 1a are independently selected from the group consisting of -H, methyl and ethyl. More preferably, at least one of -R of formula (IIa) 1 and -R 1a is -H. Even more preferably, both -R of formula (IIa) 1 and -R 1a are -H.

[0354] Preferably, -R of formula (IIa) 4 and -R 4a are independently selected from the group consisting of -H, methyl and ethyl. More preferably, at least one of -R of formula (IIa) 4 and -R 4a is -H. Even more preferably, both -R of formula (IIa) 4 and -R 4a are -H.

[0355] Preferably, -X in formula (IIa) 2 - is -C(R 8 R 8a ).

[0356] Preferably, -R in formula (IIa) 8 and -R 8a are independently selected from the group consisting of -H, methyl, and ethyl. More preferably, at least one of -R 8 and -R 8a in formula (IIa) is -H. Even more preferably, both -R 8 and -R 8a in formula (IIa) are -H.

[0357] Preferably, -R in formula (IIa) 2 and -R 2a are independently selected from the group consisting of -H, methyl, and ethyl. More preferably, at least one of -R 2 and -R 2a in formula (IIa) is -H. Even more preferably, both -R 2 and -R 2a in formula (IIa) are H.

[0358] Preferably, -R in formula (IIa) 3 and -R 3a are independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. Even more preferably, at least one of -R 3 and -R 3a in formula (IIa) is methyl. In a similarly preferred embodiment, both -R 3 and -R 3a in formula (IIa) are -H. In another similarly preferred embodiment, both -R 3 and -R 3a in formula (IIa) are methyl.

[0359] Preferably, R in formula (IIa) 3 is -H and -R in formula (IIa) 3a is methyl.

[0360] Preferably, sub-L 1 - is of formula (IIb):

Chemical formula

[0361] Preferably, -L in formula (IIb) 1 - is replaced by one sub-L 2 -Z.

[0362] Preferably, sub-L in formula (IIb) 1 - is not further substituted.

[0363] Preferred embodiments of -Z in formula (IIa) are as described above.

[0364] Preferably, -X in formula (IIb) 2 - is -C(R 8 R 8a )-.

[0365] Preferably, -R in formula (IIb) 8 and -R 8a are independently selected from the group consisting of -H, methyl and ethyl. More preferably, -R in formula (IIb) 8 and -R 8aAt least one of them is -H. More preferably, -R of formula (IIb) 8 and -R 8a are both -H.

[0366] Preferably, -R of formula (IIb) 2 and -R 2a are independently selected from the group consisting of -H, methyl and ethyl. More preferably, -R of formula (IIb) 2 and -R 2a at least one of them is -H. Even more preferably, -R of formula (IIb) 2 and -R 2a are both H.

[0367] Preferably, -R of formula (IIb) 3 and -R 3a are independently selected from the group consisting of -H, methyl, ethyl, propyl and butyl. Even more preferably, -R of formula (IIb) 3 and -R 3a at least one of them is methyl. In a similarly preferred embodiment, -R of formula (IIb) 3 and -R 3a are both -H. In another similarly preferred embodiment, -R of formula (IIb) 3 and -R 3a are both methyl.

[0368] Most preferably, R of formula (IIb) 3 is -H and -R of formula (IIb) 3a is methyl.

[0369] Even more preferably, moiety -L 1 - is of formula (IIb'):

Chemical formula

[0370] Preferably, the moiety -L of formula (IIb') 1 - is not further substituted.

[0371] Preferred embodiments of -Z of formula (IIb') are as described above.

[0372] Preferably, -X of formula (IIb') 2 - is -C(R 8 R 8a )-.

[0373] Preferably, -R of formula (IIb') 8 and -R 8a are independently selected from the group consisting of -H, methyl and ethyl. More preferably, at least one of -R of formula (IIb') 8 and -R 8a is -H. Even more preferably, both -R of formula (IIb') 8 and -R 8a are -H.

[0374] Preferably, -R of formula (IIb') 2 and -R 2a are independently selected from the group consisting of -H, methyl and ethyl. More preferably, at least one of -R of formula (IIb') 2 and -R 2a is -H. Even more preferably, both -R of formula (IIb') 2 and -R 2a are H.

[0375] Preferably, -R in formula (IIb') 3 and -R 3a are independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. Even more preferably, -R in formula (IIb') 3 and -R 3a is at least one of methyl. In a similarly preferred embodiment, -R in formula (IIb') 3 and -R 3a are both -H. In another similarly preferred embodiment, -R in formula (IIb') 3 and -R 3a are both methyl.

[0376] Most preferably, R in formula (IIb') 3 is -H, and -R in formula (IIb') 3a is methyl.

[0377] Preferably, the moiety -L 1 - is of formula (IIc):

Chemical formula

[0378] Preferably, -L in formula (IIc) 1 - is substituted with one moiety -L 2 -Z.

[0379] Preferably, the moiety -L in formula (IIc) 1 - is not further substituted.

[0380] Preferred embodiments of -Z of formula (IIc) are as described above.

[0381] In another preferred embodiment, moiety -L 1 - is of formula (IIc-a):

Chemical formula

[0382] Preferably, -L of formula (IIc-a) 1 - is replaced by one moiety -L 2 -Z.

[0383] Preferably, moiety -L of formula (IIc-a) 1 - is not further substituted.

[0384] Preferred embodiments of -Z of formula (IIc-a) are as described above.

[0385] In another preferred embodiment, moiety -L 1 - is of formula (IIc-b):

Chemical formula

[0386] Preferably, -L in formula (IIc-b) 1 - is replaced by one moiety -L 2 -Z.

[0387] Preferably, the moiety -L in formula (IIc-b) 1 - is not further substituted.

[0388] Preferred embodiments of -Z in formula (IIc-b) are as described above.

[0389] Even more preferably, the moiety -L 1 - is of formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v):

Chemical formula

[0390] Preferably, the moiety -L in formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v) 1 - is not further substituted.

[0391] Preferred embodiments of -Z in formulae (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v) are as described above.

[0392] In a particularly preferred embodiment, moiety -L 1 - is of formula (IIc-ii)

[0393]

Chemical formula

[0394] Preferably, -L in formula (IIc-ii) 1 - is replaced by one moiety -L 2 -Z.

[0395] Preferred embodiments of -Z in formula (IIc-ii) are as described above.

[0396] In a similarly preferred embodiment, moiety -L 1 - is of formulae (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv') and (IIc-v'):

Chemical formula

[0397] Preferably, the moiety -L of formulas (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv') and (IIc-v') 1 - is not further substituted.

[0398] Preferred embodiments of -Z in formulas (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv') and (IIc-v') are as described above.

[0399] In another particularly preferred embodiment, the moiety -L 1 - is

Chemical formula

[0400] Preferably, the -L of formula (IIc-ii') 1 - is substituted by one moiety -L 2 -Z.

[0401] Preferred embodiments of -Z in formula (IIc-ii') are as described above.

[0402] In a similarly preferred embodiment, the moiety -L 1 - is of formulas (IIc-i''), (IIc-ii''), (IIc-iii'') and (IIc-iv''):

Chemical formula

[0403] Preferably, the moiety -L of formulas (IIc-i''), (IIc-ii''), (IIc-iii'') and (IIc-iv'') 1 - is not further substituted.

[0404] Preferred embodiments of -Z in formulas (IIc-i''), (IIc-ii''), (IIc-iii'') and (IIc-iv'') are as described above.

[0405] In another particularly preferred embodiment, the moiety -L 1 - is

Chemical formula

[0406] Preferably, the -L in formula (IIc-ii'') 1 - is replaced by one moiety -L 2 -Z.

[0407] Preferred embodiments of -Z in formula (IIc-ii'') are as described above.

[0408] -L of formula (II), (IIa), (IIb), (IIb'), (IIc), (IIc-a), (IIc-b), (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv), (IIc-v), (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv'), (IIc-v'), (IIc-i''), (IIc-ii''), (IIc-iii''), (IIc-iv'') 1 Any further optional substituents of - are preferably as described above.

[0409] Another particularly preferred moiety -L 1 - is disclosed in WO 2016 / 020373 pamphlet A1. Thus, in another preferred embodiment, moiety -L 1 - is of formula (III):

Chemical formula

[0410] -L of formula (III) 1 Any optional further substituents of - are preferably as described above.

[0411] Preferably, -L of formula (III) 1 - is substituted with one moiety -L 2 -Z.

[0412] Preferred embodiments of -Z of formula (III) are as described above.

[0413] In one embodiment, -L of formula (III) 1 - is not further substituted.

[0414] -L 1- Further preferred embodiments are disclosed in European Patent No. 1536334 B1, WO2009 / 009712 A1, WO2008 / 034122 A1, WO2009 / 143412 A2, WO2011 / 082368 A2, and U.S. Patent No. 8,618,124 B2, the patent documents of which are hereby incorporated by reference in their entirety.

[0415] -L 1 - Further preferred embodiments are disclosed in U.S. Patent No. 8,946,405 B2 and U.S. Patent No. 8,754,190 B2, the patent documents of which are hereby incorporated by reference in their entirety. Thus, the preferred moiety -L 1 - is of formula (IV):

Chemical Formula

[0416] -L of formula (IV) 1 Any optional further substituents of - are preferably as described above.

[0417] Preferably, -L of formula (IV) 1 - is one moiety -L 2 - is substituted with -Z.

[0418] Preferred embodiments of -Z of formula (IV) are as described above.

[0419] In one embodiment, -L of formula (IV) 1 - is not further substituted.

[0420] The terms used solely in connection with formula (IV) have the following meanings:

[0421] As used herein, the term "alkyl" includes straight-chain, branched or cyclic saturated hydrocarbon groups having 1 to 8 carbon atoms, or in some embodiments 1 to 6 or 1 to 4 carbon atoms.

[0422] The term "alkoxy" includes alkyl groups bonded to oxygen, including methoxy, ethoxy, isopropoxy, cyclopropoxy, cyclobutoxy and the like.

[0423] The term "alkenyl" includes non-aromatic unsaturated hydrocarbons having a carbon-carbon double bond.

[0424] The term "alkynyl" includes non-aromatic unsaturated hydrocarbons having a carbon-carbon triple bond.

[0425] The term "aryl" includes aromatic hydrocarbon groups having 6 to 18 carbon atoms, preferably 6 to 10 carbon atoms, including groups such as phenyl, naphthyl and anthracenyl. The term "heteroaryl" includes groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl and the like, containing 3 to 15 carbon atoms and at least one N, O or S atom, preferably containing 3 to 7 carbon atoms and at least one N, O or S atom, and including an aromatic ring.

[0426] In some cases, the alkenyl, alkynyl, aryl or heteroaryl moiety may be coupled to the remainder of the molecule by an alkylene bond. Substituents in these situations are referred to as alkenylalkyl, alkynylalkyl, arylalkyl or heteroarylalkyl, which indicate that the alkylene moiety is between the alkenyl, alkynyl, aryl or heteroaryl moiety and the molecule to which the alkenyl, alkynyl, aryl or heteroaryl is coupled.

[0427] The term "halogen" includes bromo, fluoro, chloro and iodo.

[0428] The term "heterocyclic ring" refers to a 4- to 8-membered aromatic or non-aromatic ring containing 3 to 7 carbon atoms and at least one N, O or S atom. Examples are piperidinyl, piperazinyl, tetrahydropyranyl, pyrrolidine and tetrahydrofuranyl, as well as the preferred example groups provided above for the term "heteroaryl".

[0429] When the ring system is optionally substituted, suitable substituents are selected from the group consisting of alkyl, alkenyl, alkynyl or a further ring, each of which is optionally further substituted. Optional substituents for any of the groups including the above are halo, nitro, cyano, -OR, -SR, -NR 2 ,-OCOR, -NRCOR, -COOR, -CONR2 、 -SOR, -SO 2 R, -SONR 2 、 -SO 2 NR 2 are exemplified, where each R is independently alkyl, alkenyl, alkynyl, aryl or heteroaryl, or two R groups cooperate with the atoms to which they are attached to form a ring.

[0430] -L 1 Further preferred embodiments for - are disclosed in WO2013 / 036857A1, which patent document is hereby incorporated by reference in its entirety. Accordingly, the preferred moiety -L 1 - is of formula (V):

Chemical formula

[0431] Any optional further substituents of -L of formula (V) are preferably as described above. 1 Preferably, -L of formula (V)

[0432] is substituted by one moiety -L 1 -Z. 2

[0433] Preferred embodiments of -Z of formula (V) are as described above.

[0434] In one embodiment, -L of formula (V) 1 is not further substituted.

[0435] The terms used exclusively in connection with formula (V) have the following meanings:

[0436] "Alkyl", "alkenyl" and "alkynyl" include straight-chain, branched or cyclic hydrocarbon groups having 1 to 8, or 1 to 6, or 1 to 4 carbon atoms, where alkyl is a saturated hydrocarbon, alkenyl contains one or more carbon-carbon double bonds, and alkynyl contains one or more carbon-carbon triple bonds. Unless otherwise specified, these contain 1 to 6 carbons.

[0437] "Aryl" includes aromatic hydrocarbon groups having 6 to 18 carbons, preferably 6 to 10 carbons, such as phenyl, naphthyl and anthracenyl. "Heteroaryl" includes groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl and the like, containing 3 to 15 carbons and containing at least one N, O or S atom, preferably containing 3 to 7 carbons and containing at least one N, O or S atom, and including an aromatic ring.

[0438] The term "substituted" means an alkyl, alkenyl, alkynyl, aryl or heteroaryl group containing one or more substituents in place of one or more hydrogen atoms. Substituents generally include halogen (including F, Cl, Br and I), lower alkyl (including straight-chain, branched and cyclic), lower haloalkyl (including fluoroalkyl, chloroalkyl, bromoalkyl and iodoalkyl), OH, lower alkoxy (including straight-chain, branched and cyclic), SH, lower alkylthio (including straight-chain, branched and cyclic), amino, alkylamino, dialkylamino, silyl (including alkylsilyl, alkoxysilyl and arylsilyl), nitro, cyano, carbonyl, carboxylic acid, carboxylic acid ester, carboxylic acid amide, aminocarbonyl, aminoacyl, carbamate, urea, thiocarbamate, thiourea, ketene, sulfone, sulfonamide, aryl (including phenyl, naphthyl and anthracenyl), heteroaryl (including 5-membered heteroaryl such as pyrrole, imidazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, thiadiazole, triazole, oxadiazole and tetrazole, 6-membered heteroaryl such as pyridine, pyrimidine and pyrazine, and fused heteroaryl such as benzofuran, benzothiophene, benzoxazole, benzimidazole, indole, benzothiazole, benzisoxazole and benzisothiazole) and can be selected from.

[0439] -L 1 Further preferred embodiments for - are disclosed in U.S. Patent No. 7,585,837 B2, which patent document is hereby incorporated by reference in its entirety. Thus, the preferred moiety -L 1 - is of formula (VI):

Chemical formula

[0440] Preferred substituents of formula (VI) are alkyl (e.g., C 1-6 alkyl), alkenyl (e.g., C 2-6 alkenyl), alkynyl (e.g., C 2-6 alkynyl), aryl (e.g., phenyl), heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl (e.g., aromatic 4-7 membered heterocycle) or halogen.

[0441] Any optional further substituents of -L 1 - of formula (VI) are preferably as described above.

[0442] Preferably, -L 1 - of formula (VI) is substituted with one moiety -L 2 -Z.

[0443] Preferred embodiments of -Z of formula (VI) are as described above.

[0444] In one embodiment, -L of formula (VI) 1 - is further unsubstituted.

[0445] The terms used exclusively in connection with formula (VI) have the following meanings:

[0446] The terms "alkyl", "alkoxy", "alkoxyalkyl", "aryl", "alkaryl" and "aralkyl" mean an alkyl radical having 1 to 8 carbon atoms, preferably 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl and butyl, and an aryl radical having 6 to 10 carbon atoms, such as phenyl and naphthyl. The term "halogen" includes bromo, fluoro, chloro and iodo.

[0447] -L 1 Further preferred embodiments for -L are disclosed in WO2002 / 089789A1, which patent document is hereby incorporated by reference in its entirety. Accordingly, the preferred moiety -L 1 - is of formula (VII): [Chemical formula] (wherein The dashed line represents a bond to -D which is the CNP moiety, said bond being by the amine functionality of -D, L 1 is a bifunctional linking group,[[]] Y 1 and Y 2 are independently O, S or NR 7 and R 2 R 3 R 4 R 5 R 6 and R 7 are independently hydrogen, C 1-6 alkyl, C 3-12 branched alkyl, C 3-8 cycloalkyl, C 1-6 substituted alkyl, C 3-8Replacement cycloalkyl, aryl, substituted aryl, aralkyl, C 1-6 Heteroalkyl, substituted C 1-6 Heteroalkyl, C 1-6 Alkoxy, phenoxy and C 1-6 Selected from the group consisting of heteroalkoxy, When Ar is included in formula (VII), it is a moiety that forms a polysubstituted aromatic hydrocarbon or a polysubstituted heterocyclic group, X is a chemical bond, or a moiety that is actively transported to the target cell, a hydrophobic moiety, or a combination thereof, Y is 0 or 1) Having, Said -L 1 - is -L 2 -Z is substituted, and optionally, said -L 1 - is further substituted, -L 2 - is a single chemical bond or a spacer moiety, and -Z is a polymer having a molecular weight of at least 10 kDa.

[0448] -L of formula (VII) 1 Any optional further substituents of - are preferably as described above.

[0449] Preferably, -L of formula (VII) 1 - is substituted with one moiety -L 2 -Z.

[0450] Preferred embodiments of -Z of formula (VII) are as described above.

[0451] In one embodiment, -L of formula (VII) 1 - is not further substituted.

[0452] The terms used exclusively in connection with formula (VII) have the following meanings:

[0453] The term "alkyl" includes, for example, alkoxy, C 3-8Linear, branched, substituted C including cycloalkyl or substituted cycloalkyl, etc. 1-12 is understood to include alkyl.

[0454] The term "substituted" is understood to include adding one or more atoms contained in a functional group or compound and replacing with one or more different atoms.

[0455] Substituted alkyl includes carboxyalkyl, aminoalkyl, dialkylamino, hydroxyalkyl and mercaptoalkyl, substituted cycloalkyl includes moieties such as 4-chlorocyclohexyl, aryl includes moieties such as naphthyl, substituted aryl includes moieties such as 3-bromo-phenyl, aralkyl includes moieties such as toluyl, heteroalkyl includes moieties such as ethylthiophene, substituted heteroalkyl includes moieties such as 3-methoxythiophene, alkoxy includes moieties such as methoxy, and phenoxy includes moieties such as 3-nitrophenoxy. Halo- is understood to include fluoro, chloro, iodo and bromo.

[0456] In another preferred embodiment, -L 1 - is a substructure of formula (VIII)

Chemical formula

[0457] -L of formula (VIII) 1 Any optional further substituents of - are preferably as described above.

[0458] Preferably, -L of formula (VIII) 1 - is substituted with one moiety -L 2 -Z.

[0459] Preferred embodiments of -Z of formula (VIII) are as described above.

[0460] In one embodiment, -L of formula (VIII) 1 - is not further substituted.

[0461] In another preferred embodiment, -L 1 - has the substructure of formula (IX)

Chemical formula

[0462] -L of formula (IX) 1 Any optional further substituents of - are preferably as described above.

[0463] Preferably, -L of formula (IX) 1 - is a single -L 2 - is replaced by -Z.

[0464] In one embodiment, -L of formula (IX) 1 - is further unsubstituted.

[0465] Preferred embodiments of -Z of formula (IX) are as described above.

[0466] The -L moiety 2 - can be bonded to -L by replacing any existing -H, unless specifically excluded. 1

[0467] In the prodrug of the present invention, -L 2 - is a chemical bond or a spacer moiety.

[0468] In one embodiment, -L 2 - is a chemical bond.

[0469] In another embodiment, -L 2 - is a spacer moiety.

[0470] -L 2 When -L is a single chemical bond, -L 2 - is preferably -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O) 2 N(R y1 )-, -S(O)N(R y1 )-, -S(O) 2 -, -S(O)-, -N(R y1 )S(O) 2 N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-, -N(R y1 )C(O)N(R y1a )-, -OC(O)N(R y1 )-, C 1-50 ​Alkyl, C 2-50 Alkenyl, and C 2-50 Alkynyl selected from the group consisting of; -T-, C 1-50 Alkyl, C 2-50 Alkenyl, and C 2-50 Alkynyl may be substituted with one or more identical or different -R y2 and C 1-50 Alkyl, C 2-50 Alkenyl, and C 2-50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-, -S(O) 2 N(R y3 )-, -S(O)N(R y3 )-, -S(O) 2 -, -S(O)-, -N(R y3 )S(O) 2 N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-, -N(R y3 )C(O)N(R y3a )-, and -OC(O)N(R y3 )- may optionally interrupt; -R y1 and -R y1a are each independently -H, -T, C 1-50 Alkyl, C 2-50 Alkenyl, and C 2-50 Alkynyl selected from the group consisting of; -T, C 1-50 Alkyl, C 2-50 Alkenyl, and C 2-50 Alkynyl may be substituted with one or more identical or different -R y2 and C 1-50 Alkyl, C 2-50 Alkenyl, and C 2-50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 )-, -S(O) 2 N(R y4 )-, -S(O)N(R y4 )-, -S(O)2 -, -S(O)-, -N(R y4 )S(O) 2 N(R y4a )-, -S-, -N(R y4 )-, -OC(OR y4 )(R y4a ), -N(R y4 ), C(O)N(R y4a ), and -OC(O)N(R y4 )- may optionally be interrupted by one or more groups selected from the group consisting of; Each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbocyclic polycycle, and 8- to 30-membered heterocyclic polycycle; each T may independently be substituted with one or more identical or different -R y2 ; Each -R y2 is independently halogen, -CN, oxo (=O), -COOR y5 , -OR y5 , -C(O)R y5 , -C(O)N(R y5 R y5a ), -S(O) 2 N(R y5 R y5a ), -S(O)N(R y5 R y5a ), -S(O) 2 R y5 ), -S(O)R y5 , -N(R y5 ), S(O) 2 N(R y5a R y5b ), -SR y5 , -N(R y5 R y5a ), -NO 2 , -OC(O)R y5 , -N(R y5 ), C(O)R y5a , -N(R y5 ), S(O) 2 R y5a , -N(R y5 ), S(O)R y5a , -N(R y5)C(O)OR y5a 、 -N(R y5 )C(O)N(R y5a R y5b )、 -OC(O)N(R y5 R y5a )、 and C 1-6 selected from the group consisting of alkyl; C 1-6 alkyl may be substituted with one or more identical or different halogens; Each -R y3 、 -R y3a 、 -R y4 、 -R y4a 、 -R y5 、 -R y5a and -R y5b is independently selected from the group consisting of -H and C 1-6 alkyl, and C 1-6 alkyl may be substituted with one or more identical or different halogens.

[0471] -L 2 - is other than a single chemical bond, -L 2 - is more preferably -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O) 2 N(R y1 )-, -S(O)N(R y1 )-, -S(O) 2 -, -S(O)-, -N(R y1 )S(O) 2 N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-, -N(R y1 )C(O)N(R y1a )-, -OC(O)N(R y1 )-, C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl; -T-, C 1-20 alkyl, C 2-20 alkenyl, and C 2-20 alkynyl may be substituted with one or more identical or different -R y2may be replaced by C 1-20 alkyl, C 2-20 alkenyl and C 2-20 alkynyl may optionally be interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 ), -S(O) 2 N(R y3 ), -S(O)N(R y3 ), -S(O) 2 -, -S(O)-, -N(R y3 )S(O) 2 N(R y3a ), -S-, -N(R y3 ), -OC(OR y3 )(R y3a ), -N(R y3 )C(O)N(R y3a ), and -OC(O)N(R y3 ); -R y1 and -R y1a are each independently selected from the group consisting of -H, -T, C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl; -T, C 1-10 alkyl, C 2-10 alkenyl, and C 2-10 alkynyl may optionally be substituted with one or more identical or different -R y2 and C 1-10 alkyl, C 2-10 alkenyl, and C 2-10 alkynyl are -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 ), -S(O) 2 N(R y4 ), -S(O)N(R y4 ), -S(O) 2 -, -S(O)-, -N(R y4 )S(O) 2 N(R y4a ), -S-, -N(R y4 ), -OC(OR y4 )(R y4a ), -N(R y4 ), -N(Ry4a )-, and -OC(O)N(R y4 )- from the group consisting of may optionally be interrupted; Each T is independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbocyclic, and 8- to 30-membered heterocyclic selected from the group consisting of; each T is independently one or more identical or different -R y2 May be substituted with; -R y2 Is halogen, -CN, oxo (=O), -COOR y5 , -OR y5 , -C(O)R y5 , -C(O)N(R y5 R y5a ), -S(O) 2 N(R y5 R y5a ), -S(O)N(R y5 R y5a ), -S(O) 2 R y5 ), -S(O)R y5 , -N(R y5 ), S(O) 2 N(R y5a R y5b ), -SR y5 , -N(R y5 R y5a ), -NO 2 , -OC(O)R y5 , -N(R y5 ), C(O)R y5a , -N(R y5 ), S(O) 2 R y5a , -N(R y5 ), S(O)R y5a , -N(R y5 ), C(O)OR y5a , -N(R y5 ), C(O)N(R y5a R y5b ), -OC(O)N(R y5 R y5a ), and C 1-6 Selected from the group consisting of alkyl; C 1-6The alkyl may be substituted with one or more identical or different halogens; each -R y3 、-R y3a 、-R y4 、-R y4a 、-R y5 、-R y5a and -R y5b is independently selected from the group consisting of -H and C 1-6 alkyl; C 1-6 alkyl may be substituted with one or more identical or different halogens.

[0472] -L 2 When - is other than a single chemical bond, -L 2 - is more preferably -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O) 2 N(R y1 )-, -S(O)N(R y1 )-, -S(O) 2 -, -S(O)-, -N(R y1 )S(O) 2 N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-, -N(R y1 )C(O)N(R y1a )-, -OC(O)N(R y1 )-, C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl; -T-, C 1-50 alkyl, C 2-50 alkenyl, and C 2-50 alkynyl may be substituted with one or more identical or different -R y2 , and C 1-50 alkyl, C 2-50 alkenyl and C 2-50 alkynyl are -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-, -S(O) 2 N(R y3)-, -S(O)N(R y3 )-, -S(O) 2 -, -S(O)-, -N(R y3 )S(O) 2 N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-, -N(R y3 )C(O)N(R y3a )-, and -OC(O)N(R y3 )- may optionally be interrupted by one or more groups selected from the group consisting of; -R y1 and -R y1a are independently selected from the group consisting of -H, -T, C 1-10 alkyl, C 2-10 alkenyl, and C 2-10 alkynyl; Each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbocyclic polycycle, and 8- to 30-membered heterocyclic polycycle; Each -R y2 is independently selected from the group consisting of halogen and C 1-6 alkyl; Each -R y3 , -R y3a , -R y4 , -R y4a , -R y5 , -R y5a and -R y5b are independently selected from the group consisting of -H and C 1-6 alkyl; C 1-6 alkyl may optionally be substituted with one or more identical or different halogens.

[0473] Even more preferably, -L 2 - is a C 1-20 alkyl chain, which may optionally be interrupted by one or more groups selected from -O-, -T- and -C(O)N(R y1 ); C 1-20The alkyl chain is independently substituted with one or more groups selected from -OH, -T, and -C(O)N(R y6 R y6a )); -R y1 , -R y6 , -R y6a are independently selected from the group consisting of H and C 1-4 alkyl, and T is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbocyclic polycycle, and 8- to 30-membered heterocyclic polycycle.

[0474] Preferably, -L 2 - has a molecular weight in the range of 14 g / mol to 750 g / mol.

[0475] Preferably, -L 2 - is

Chemical formula

[0476] In a certain preferred embodiment, -L 2 - has a chain length of 1 to 20 atoms.

[0477] As used herein, the term "chain length" with respect to the moiety -L 2 - refers to the number of atoms of -L 1 - present in the shortest linkage between -L 2 - and -Z.

[0478] Preferably, -L 2 - is of the following formula (i). [Chem.] In the formula, The dashed line marked with an asterisk is the bond to -L 1 -; The unmarked dashed line is the bond to -Z; -R 1 is selected from the group consisting of -H, C 1-6 alkyl, C 2-6 alkenyl and C 2-6 alkynyl; n is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 and 18; The moiety of formula (i) may be further substituted.

[0479] Preferably, -R in formula (i) 1 is selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. Even more preferably, -R in formula (i) 1 is selected from the group consisting of -H, methyl, ethyl and propyl. Even more preferably, -R in formula (i) 1 is selected from the group consisting of -H and methyl. Most preferably, -R in formula (i) 1 is methyl.

[0480] Preferably, n in formula (i) is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. Even more preferably, n in formula (i) is selected from the group consisting of 0, 1, 2, 3, 4 and 5. Even more preferably, n in formula (i) is selected from the group consisting of 0, 1, 2 and 3. Even more preferably, n in formula (i) is selected from the group consisting of 0 and 1. Most preferably, n in formula (i) is 0.

[0481] In a preferred embodiment, -L 2 - is [Chem.] a moiety selected from the group consisting of; wherein, the dashed line marked with an asterisk is a bond to -L 1 -; the unmarked dashed line is a bond to -Z; moieties (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi), (xii), (xiii), (xiv), (xv), (xvi) and (xvii) may be further substituted.

[0482] In a preferred embodiment, -L 2 - is

Chemical formula

[0483] Even more preferred -L 2 - is

Chemical formula

[0484] Even more preferably, -L 2 - is of the following formula (xvi).

Chemical formula

[0485] In a preferred embodiment, sub-L 1 -L 2 - is

Chem.

[0486] In an even more preferred embodiment, sub-L 1 -L 2 - is of the following formula (IId-ii).

Chem.

[0487] In the most preferred embodiment, sub-L 1 -L 2 - is of the following formula (IId-ii′).

Chem.

[0488] In another preferred embodiment, sub-L 1 -L 2 - is

Chem.

[0489] In an even more preferred embodiment, moiety -L 1 -L 2 - is of the following formula (IId-iia). [Chemical formula] Wherein, The dashed line without a mark indicates the bond to the nitrogen of -D which is the CNP moiety by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0490] In the most preferred embodiment, moiety -L 1 -L 2 - is of the following formula (IId-iia'). [Chemical formula] Wherein, The dashed line without a mark indicates the bond to the nitrogen of -D which is the CNP moiety by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0491] In another preferred embodiment, moiety -L 1 -L 2 - is [Chemical formula] selected from the group consisting of; Wherein, The dashed line without a mark indicates the bond to the nitrogen of -D which is the CNP moiety by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0492] In an even more preferred embodiment, moiety -L1 -L 2 - is of the following formula (IId-iib). [Chemical formula] In the formula, The unmarked dashed line indicates the bond to the nitrogen of -D which is the CNP moiety by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0493] In the most preferred embodiment, the moiety -L 1 -L 2 - is of the following formula (IId-iib'). [Chemical formula] In the formula, The unmarked dashed line indicates the bond to the nitrogen of -D which is the CNP moiety by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0494] Also surprisingly, it has been found that the extension of the CNP sequence length is advantageous with respect to NEP stability: CNP-22 was more sensitive to NEP degradation than CNP-34, and it was more sensitive than CNP-38.

[0495] Preferably, -D has the sequence of SEQ ID NO: 24, SEQ ID NO: 25 or SEQ ID NO: 30.

[0496] In one embodiment, -D has the sequence of SEQ ID NO: 25.

[0497] In another embodiment, -D has the sequence of SEQ ID NO: 20.

[0498] In another embodiment, -D has the sequence of SEQ ID NO: 21.

[0499] In another embodiment, -D has the sequence of SEQ ID NO: 22.

[0500] In another embodiment, -D has the sequence of SEQ ID NO: 23.

[0501] In another embodiment, -D has the sequence of SEQ ID NO: 30.

[0502] In a preferred embodiment, -D has the sequence of SEQ ID NO: 24.

[0503] Preferably, moiety -L 1 - is attached to -D via a functional group of any corresponding CNP drug D-H selected from the group consisting of carboxylic acid, primary and secondary amines, maleimide, thiol, sulfonic acid, carbonate, carbamate, hydroxyl, aldehyde, ketone, hydrazine, isocyanate, isothiocyanate, phosphoric acid, phosphonic acid, haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine, sulfate, disulfide, vinyl sulfone, vinyl ketone, diazoalkane, oxirane, guanidine, and aziridine. Most preferably, moiety -L 1 - is attached to -D via a functional group of any corresponding CNP drug D-H selected from the group consisting of hydroxyl, primary and secondary amines, and guanidine. In certain preferred embodiments, moiety -L 1 - is attached to -D via a primary or secondary amine, preferably via a primary amine group.

[0504] In one embodiment, -L 1 - is linked to -D via a hydroxyl functional group.

[0505] In another embodiment, -L 1 - is linked to -D via a guanidine functional group.

[0506] In a preferred embodiment, -L 1 - is linked to -D via an amine functional group of D-H.

[0507] 1. In one embodiment, -L 1 - is linked to -D via the N-terminal amine functional group of CNP.

[0508] In a more preferred embodiment, -L 1 - is linked to -D via the amine functional groups provided by the lysine side chains of the CNP drug, i.e., the lysines at positions 9, 11, 15, 16, 20, and 26, when -D has the sequence of SEQ ID NO: 24.

[0509] 1. In one embodiment, -D has the sequence of SEQ ID NO: 24, and -L 1 - is linked to -D via the amine functional group provided by the side chain of the lysine at position 9.

[0510] 1. In one embodiment, -D has the sequence of SEQ ID NO: 24, and -L 1 - is linked to -D via the amine functional group provided by the side chain of the lysine at position 11.

[0511] 1. In one embodiment, -D has the sequence of SEQ ID NO: 24, and -L 1 - is linked to -D via the amine functional group provided by the side chain of the lysine at position 15.

[0512] 1. In one embodiment, -D has the sequence of SEQ ID NO: 24, and -L 1 - is linked to -D via the amine functional group provided by the side chain of the lysine at position 16.

[0513] 1. In one embodiment, -D has the sequence of SEQ ID NO: 24, and -L 1 - is linked to -D via the amine functional group provided by the side chain of the lysine at position 20.

[0514] 1. In one embodiment, -D has the sequence of SEQ ID NO: 24, and -L 1 - is linked to -D via the amine functional group provided by the side chain of the lysine at position 26.

[0515] In one embodiment, -D has the sequence of SEQ ID NO: 20, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 13.

[0516] In one embodiment, -D has the sequence of SEQ ID NO: 20, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 15.

[0517] In one embodiment, -D has the sequence of SEQ ID NO: 20, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 19.

[0518] In one embodiment, -D has the sequence of SEQ ID NO: 20, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 20.

[0519] In one embodiment, -D has the sequence of SEQ ID NO: 20, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 24.

[0520] In one embodiment, -D has the sequence of SEQ ID NO: 20, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 30.

[0521] In one embodiment, -D has the sequence of SEQ ID NO: 21, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 12.

[0522] In one embodiment, -D has the sequence of SEQ ID NO: 21, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 14.

[0523] In one embodiment, -D has the sequence of SEQ ID NO: 21, and -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 18.

[0524] In one embodiment, -D has the sequence of SEQ ID NO: 21, and -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 19.

[0525] In one embodiment, -D has the sequence of SEQ ID NO: 21, and -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 23.

[0526] In one embodiment, -D has the sequence of SEQ ID NO: 21, and -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 29.

[0527] In one embodiment, -D has the sequence of SEQ ID NO: 22, and -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 11.

[0528] In one embodiment, -D has the sequence of SEQ ID NO: 22, and -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 13.

[0529] In one embodiment, -D has the sequence of SEQ ID NO: 22, and -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 17.

[0530] In one embodiment, -D has the sequence of SEQ ID NO: 22, and -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 18.

[0531] In one embodiment, -D has the sequence of SEQ ID NO: 22, -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 22.

[0532] In one embodiment, -D has the sequence of SEQ ID NO: 22, -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 28.

[0533] In one embodiment, -D has the sequence of SEQ ID NO: 23, -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 10.

[0534] In one embodiment, -D has the sequence of SEQ ID NO: 23, -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 12.

[0535] In one embodiment, -D has the sequence of SEQ ID NO: 23, -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 16.

[0536] In one embodiment, -D has the sequence of SEQ ID NO: 23, -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 17.

[0537] In one embodiment, -D has the sequence of SEQ ID NO: 23, -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 21.

[0538] In one embodiment, -D has the sequence of SEQ ID NO: 23, -L 1 - is linked to -D via an amine functional group provided by the side chain of the lysine at position 27.

[0539] In one embodiment, -D has the sequence of SEQ ID NO: 30, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 10.

[0540] In one embodiment, -D has the sequence of SEQ ID NO: 30, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 12.

[0541] In one embodiment, -D has the sequence of SEQ ID NO: 30, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 16.

[0542] In one embodiment, -D has the sequence of SEQ ID NO: 30, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 17.

[0543] In one embodiment, -D has the sequence of SEQ ID NO: 30, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 21.

[0544] In one embodiment, -D has the sequence of SEQ ID NO: 30, and -L 1 - is linked to -D via an amine functional group provided by the side chain of lysine at position 27.

[0545] Moiety -L 1 - can be linked to -D via any kind of linker, provided that it is reversible. Preferably, -L 1 - is linked to -D via a linker selected from the group consisting of amide, ester, carbamate, acetal, aminals, imine, oxime, hydrazone, disulfide and acylguanidine. Even more preferably, -L 1- is linked to -D via a linking moiety selected from the group consisting of amide, ester, carbamate and acylguanidine. These linking moieties may not be reversible by themselves, such as an amide linking moiety, for example, -L 1 It is obvious that preferably selected adjacent groups contained in - may make such a linking moiety reversible.

[0546] In one embodiment, -L 1 - is linked to -D via an ester linking moiety.

[0547] In another embodiment, -L 1 - is linked to -D via a carbamate linking moiety.

[0548] In another embodiment, -L 1 - is linked to -D via an acylguanidine linking moiety.

[0549] In a preferred embodiment, -L 1 - is linked to -D via an amide linking moiety.

[0550] -L 1 The amino acid residue of -D to which -L is attached is selected from the group consisting of protein - constituting amino acid residues and non - protein - constituting amino acid residues.

[0551] In one embodiment, the amino acid residue of -D to which -L 1 is attached is a non - protein - constituting amino acid.

[0552] In a preferred embodiment, -L 1The amino acid residue of -D to which - is attached is a protein constituent amino acid. More preferably, the protein constituent amino acid is selected from the group consisting of histidine, lysine, tryptophan, serine, threonine, tyrosine, aspartic acid, glutamic acid and arginine. Even more preferably, the protein constituent amino acid is selected from the group consisting of lysine, aspartic acid, arginine and serine. Even more preferably, the protein constituent amino acid is selected from the group consisting of lysine, arginine and serine.

[0553] In one embodiment, -L 1 - is linked to -D via the guanidine functional group of arginine of the corresponding agent D-H.

[0554] In another embodiment, -L 1 - is linked to -D via the hydroxyl functional group of serine of the corresponding agent D-H.

[0555] In a preferred embodiment, -L 1 - is linked to -D via the amine functional group of lysine of the corresponding agent D-H.

[0556] Surprisingly, it has been found that the attachment of -L 1 - to the ring portion of CNP significantly reduces the affinity of the CNP prodrug for NPR-B as compared to the attachment at the N-terminus or to the acyclic portion of CNP. This reduced affinity for NPR-B then reduces the risk of cardiovascular side effects such as hypotension.

[0557] Therefore, -L 1 - is preferably conjugated to the side chain of the amino acid residue of the ring portion of -D or to the backbone of the ring portion of -D. Preferably, -L 1 - is covalently and reversibly conjugated to the side chain of the amino acid residue of the ring portion of -D.

[0558] In one embodiment, -L 1The amino acid residue of the ring portion of -D conjugated with - is histidine. It is understood that such histidine does not exist in the sequence of SEQ ID NO: 96 and exists only in its variants, analogs, orthologs, homologs and derivatives.

[0559] In one embodiment, -L 1 The amino acid residue of the ring portion of -D conjugated with - is tryptophan. It is understood that such tryptophan does not exist in the sequence of SEQ ID NO: 96 and exists only in its variants, analogs, orthologs, homologs and derivatives.

[0560] In one embodiment, -L 1 The amino acid residue of the ring portion of -D conjugated with - is threonine. It is understood that such threonine does not exist in the sequence of SEQ ID NO: 96 and exists only in its variants, analogs, orthologs, homologs and derivatives.

[0561] In one embodiment, -L 1 The amino acid residue of the ring portion of -D conjugated with - is tyrosine. It is understood that such tyrosine does not exist in the sequence of SEQ ID NO: 96 and exists only in its variants, analogs, orthologs, homologs and derivatives.

[0562] In one embodiment, -L 1 The amino acid residue of the ring portion of -D conjugated with - is glutamic acid. It is understood that such glutamic acid does not exist in the sequence of SEQ ID NO: 96 and exists only in its variants, analogs, orthologs, homologs and derivatives.

[0563] In one embodiment, -L 1 The amino acid residue of the ring portion of -D conjugated with - is lysine. Preferably, the amino acid is lysine at position 4 of SEQ ID NO: 96 corresponding to lysine at position 26 of SEQ ID NO: 24.

[0564] In another embodiment, -L 1 The amino acid residue of the ring portion of -D to which - is conjugated is aspartic acid. Preferably, the amino acid is aspartic acid at position 6 of SEQ ID NO: 96, corresponding to aspartic acid at position 28 of SEQ ID NO: 24.

[0565] In another embodiment, -L 1 The amino acid residue of the ring portion of -D to which - is conjugated is arginine. Preferably, the amino acid is arginine at position 7 of SEQ ID NO: 96, corresponding to arginine at position 29 of SEQ ID NO: 24.

[0566] In another embodiment, -L 1 The amino acid residue of the ring portion of -D to which - is conjugated is serine. Preferably, the amino acid is serine at position 10 or 12 of SEQ ID NO: 96. In one embodiment, the amino acid is serine at position 10 of SEQ ID NO: 96, corresponding to serine at position 32 of SEQ ID NO: 24. In another embodiment, the amino acid is serine at position 12 of SEQ ID NO: 96, corresponding to serine at position 34 of SEQ ID NO: 24.

[0567] In a preferred embodiment, -L 1 The amino acid residue of the ring portion of -D to which - is conjugated is lysine. Most preferably, -D has the sequence of SEQ ID NO: 24, and -L 1 - is conjugated to the lysine at position 26.

[0568] In a preferred embodiment, the CNP prodrug of the present invention has the formula (IIe):

Chemical formula

Chemical formula

[0569] Preferably, c1 of formula (IIe) is about 450.

[0570] In a similarly preferred embodiment, the CNP prodrug of the present invention has formula (IIe-i):

Chemical formula

Chemical formula

[0571] Preferably, c1 of formula (IIe-i) is about 450.

[0572] In another similarly preferred embodiment, the CNP prodrug of the present invention has formula (IIe-ii):

Chemical formula

Chemical formula

[0573] Preferably, c1 of formula (IIe-ii) is about 450.

[0574] In one embodiment, the CNP moiety of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO: 25.

[0575] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO: 30.

[0576] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO: 20.

[0577] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO: 21.

[0578] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO: 22.

[0579] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO: 23.

[0580] In a preferred embodiment, the CNP moiety of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO: 24.

[0581] In one embodiment, the CNP moiety is attached to -L in the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) by the nitrogen of the N-terminal amine functionality of CNP. 1 - is attached.

[0582] In a preferred embodiment, the CNP prodrug of the present invention has c1 of about 450, the CNP moiety has the sequence of SEQ ID NO: 24, and is attached to -L by an amine functional group provided by the side chain of lysine at position 26 1 and has formula (IIe).

[0583] In another preferred embodiment, the CNP prodrug of the present invention has c1 of about 450, the CNP moiety has the sequence of SEQ ID NO: 24, and is attached to -L by an amine functional group provided by the side chain of lysine at position 26 1 and has formula (IIe-i).

[0584] In another preferred embodiment, the CNP prodrug of the present invention has c1 of about 450, the CNP moiety has the sequence of SEQ ID NO: 24, and is attached to -L by an amine functional group provided by the side chain of lysine at position 26 1 and has formula (IIe-ii).

[0585] In a preferred embodiment, the CNP prodrug of the present invention has formula (IIe), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 20, and is attached to -L via an amine functional group provided by the side chain of lysine at position 30 1 and is attached to -L via an amine functional group provided by the side chain of lysine at position 30

[0586] In another preferred embodiment, the CNP prodrug of the present invention has formula (IIe-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 20, and is attached to -L via an amine functional group provided by the side chain of lysine at position 30 1 and is attached to -L via an amine functional group provided by the side chain of lysine at position 30

[0587] In another preferred embodiment, the CNP prodrug of the present invention has formula (IIe-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 20, and is attached to -L via an amine functional group provided by the side chain of lysine at position 30 1 and is attached to -L via an amine functional group provided by the side chain of lysine at position 30

[0588] In a preferred embodiment, the CNP prodrug of the present invention has the formula (IIe), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 21, and is linked to -L via an amine functional group provided by the side chain of lysine at position 29 1 -.

[0589] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-i), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 21, and is linked to -L via an amine functional group provided by the side chain of lysine at position 29 1 -.

[0590] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-ii), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 21, and is linked to -L via an amine functional group provided by the side chain of lysine at position 29 1 -.

[0591] In a preferred embodiment, the CNP prodrug of the present invention has the formula (IIe), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 22, and is linked to -L via an amine functional group provided by the side chain of lysine at position 28 1 -.

[0592] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-i), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 22, and is linked to -L via an amine functional group provided by the side chain of lysine at position 28 1 -.

[0593] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-ii), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 22, and is linked to -L via an amine functional group provided by the side chain of lysine at position 28 1 -.

[0594] In a preferred embodiment, the CNP prodrug of the present invention has the formula (IIe), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 23, and is linked to -L via the amine functional group provided by the side chain of lysine at position 27 1 -.

[0595] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-i), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 23, and is linked to -L via the amine functional group provided by the side chain of lysine at position 27 1 -.

[0596] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-ii), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 23, and is linked to -L via the amine functional group provided by the side chain of lysine at position 27 1 -.

[0597] In a preferred embodiment, the CNP prodrug of the present invention has the formula (IIe), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 30, and is linked to -L via the amine functional group provided by the side chain of lysine at position 27 1 -.

[0598] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-i), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 30, and is linked to -L via the amine functional group provided by the side chain of lysine at position 27 1 -.

[0599] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-ii), where c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 30, and is linked to -L via the amine functional group provided by the side chain of lysine at position 27 1 -.

[0600] Thus, in a preferred embodiment, the CNP prodrug of the present invention has the formula (IIe'): [Chemical formula] (wherein, The unmarked dashed line indicates the bond to the nitrogen provided by the side chain of lysine at position 26 of the CNP moiety of SEQ ID NO: 24 by forming an amide bond, The dashed line with an asterisk indicates the moiety: [Chemical formula] the bond to, wherein, each c1 is independently an integer in the range of 400 to 500).

[0601] Preferably, each c1 of formula (IIe') is about 450.

[0602] In a preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-i'): [Chemical formula] (wherein, The unmarked dashed line indicates the bond to the nitrogen provided by the side chain of lysine at position 26 of the CNP moiety of SEQ ID NO: 24 by forming an amide bond, The dashed line with an asterisk indicates the moiety: [Chemical formula] the bond to, wherein, each c1 is independently an integer in the range of 400 to 500).

[0603] Preferably, each c1 of formula (IIe-i') is about 450.

[0604] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIe-ii'): [Chemical formula] (wherein, The dashed line without a mark indicates a bond to the nitrogen provided by the side chain of lysine at position 26 of the CNP moiety of SEQ ID NO: 24 by forming an amide bond, The dashed line with an asterisk indicates the moiety:

Chemical formula

[0605] Preferably, each c1 of formula (IIe-ii') is about 450.

[0606] In another preferred embodiment, the CNP prodrug of the present invention has formula (IIf):

Chemical formula

Chemical formula

Chemical formula

[0607] Preferably, each c1 of formula (IIf) is about 225.

[0608] In another preferred embodiment, the CNP prodrug of the present invention has formula (IIf-i):

Chemical formula

[0609] Preferably, each c1 of formula (IIf-i) is about 225.

[0610] In another preferred embodiment, the CNP prodrug of the present invention has formula (IIf-ii): [Chemical formula] (wherein the dashed line without a mark indicates the bond to the nitrogen of -D, which is the CNP moiety, by forming an amide bond, the dashed line with an asterisk indicates the structure: [Chemical formula] the bond to -Z having, wherein each -Z a is [Chemical formula] and each c1 in this formula is independently an integer in the range of 200 to 250).

[0611] Preferably, each c1 of formula (IIf-ii) is about 225.

[0612] In a preferred embodiment, the CNP moiety of the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO: 25.

[0613] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO: 30.

[0614] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO: 20.

[0615] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO: 21.

[0616] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO: 22.

[0617] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO: 23.

[0618] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO: 24.

[0619] In one embodiment, the CNP moiety is attached to -L in the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) via the nitrogen of the N-terminal amine functionality of CNP. 1 - is attached.

[0620] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIf'):

Chemical formula

Chemical formula

Chemical formula

[0621] Preferably, each c1 of formula (IIf') is about 225.

[0622] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIf-i'):

Chemical formula

Chemical formula

Chemical formula

[0623] Preferably, each c1 of formula (IIf-i') is about 225.

[0624] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIf-ii'): [Chemical formula] (wherein, The unmarked dashed line indicates the bond to the nitrogen provided by the side chain of lysine at position 26 of the CNP moiety of SEQ ID NO: 24 by forming an amide bond, The dashed line marked with an asterisk represents the structure: [Chemical formula] represents the bond to -Z having, wherein, each Z a is [Chemical formula] and each c1 in this formula is independently an integer in the range of 200 to 250).

[0625] Preferably, each c1 of the formula (IIf-ii') is about 225.

[0626] Similarly, in a preferred embodiment, the CNP prodrug of the present invention has the following formula (IIea). [Chemical formula] wherein, The unmarked dashed line indicates the bond to the nitrogen of -D which is the CNP moiety by forming an amide bond; k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; The dashed line marked with an asterisk represents the following moiety: [Chemical formula] represents the bond to, each c1 is independently an integer in the range of 400 to 500).

[0627] Preferably, c1 of formula (IIea) is about 450.

[0628] Preferably, k of formula (IIea) is selected from the group consisting of 2, 3, 4, 5, 6, and 7. In a similarly preferred embodiment, the CNP prodrug of the present invention is of the following formula (IIea-i). [Chemical formula] In the formula, The unmarked dashed line indicates the bond to the nitrogen of -D, which is the CNP moiety formed by forming an amide bond; k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; The dashed line marked with an asterisk represents the following moiety: [Chemical formula] indicates the bond to, Each c1 is an integer independently in the range of 400 to 500.

[0629] Preferably, k of formula (IIea-i) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[0630] Preferably, c1 of formula (IIea-i) is about 450.

[0631] In another similarly preferred embodiment, the CNP prodrug of the present invention is of the following formula (IIea-ii). [Chemical formula] In the formula, The unmarked dashed line indicates the bond to the nitrogen of -D, which is the CNP moiety formed by forming an amide bond; k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; The dashed line marked with an asterisk represents the following moiety: [Chemical formula] shows the binding to, Each c1 is independently an integer in the range of 400 to 500.

[0632] Preferably, k in formula (IIea-ii) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

[0633] Preferably, c1 in formula (IIea-ii) is about 450.

[0634] In one embodiment, the CNP moiety of the CNP prodrugs of formulas (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO: 25.

[0635] In another embodiment, the CNP moiety of the CNP prodrugs of formulas (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO: 30.

[0636] In another embodiment, the CNP moiety of the CNP prodrugs of formulas (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO: 20.

[0637] In another embodiment, the CNP moiety of the CNP prodrugs of formulas (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO: 21.

[0638] In another embodiment, the CNP moiety of the CNP prodrugs of formulas (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO: 22.

[0639] In another embodiment, the CNP moiety of the CNP prodrugs of formulas (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO: 23.

[0640] In a preferred embodiment, the CNP moiety of the CNP prodrugs of formulas (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO: 24.

[0641] In one embodiment, the CNP moiety is attached to -L in the CNP prodrugs of formula (IIea), (IIea-i) and (IIea-ii) via the nitrogen of the N-terminal amine functionality of the CNP. 1 - is attached.

[0642] In a preferred embodiment, the CNP prodrug of the present invention is of formula (IIea), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 24, and is attached to -L via the amine functionality provided by the side chain of lysine at position 26. 1 - is attached.

[0643] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea-i), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 24, and is attached to -L via the amine functionality provided by the side chain of lysine at position 26. 1 - is attached.

[0644] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea-ii), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 24, and is attached to -L via the amine functionality provided by the side chain of lysine at position 26. 1 - is attached.

[0645] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 20, and is attached to -L via the amine functionality provided by the side chain of lysine at position 30. 1 - is attached.

[0646] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea-i), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 20, and is attached to -L via the amine functionality provided by the side chain of lysine at position 30. 1 - is attached.

[0647] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea-ii), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 20, and is linked to -L via an amine functional group provided by the side chain of the lysine at position 30 1 - is attached.

[0648] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 21, and is linked to -L via an amine functional group provided by the side chain of the lysine at position 29 1 - is attached.

[0649] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea-i), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 21, and is linked to -L via an amine functional group provided by the side chain of the lysine at position 29 1 - is attached.

[0650] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea-ii), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 21, and is linked to -L via an amine functional group provided by the side chain of the lysine at position 29 1 - is attached.

[0651] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 22, and is linked to -L via an amine functional group provided by the side chain of the lysine at position 28 1 - is attached.

[0652] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea-i), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 22, and is linked to -L via an amine functional group provided by the side chain of the lysine at position 28 1 - is attached.

[0653] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIea-ii), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 22, and is -L via the amine functional group provided by the side chain of lysine at position 28 1 - is attached.

[0654] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIea), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 23, and is -L via the amine functional group provided by the side chain of lysine at position 27 1 - is attached.

[0655] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIea-i), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 23, and is -L via the amine functional group provided by the side chain of lysine at position 27 1 - is attached.

[0656] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIea-ii), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 23, and is -L via the amine functional group provided by the side chain of lysine at position 27 1 - is attached.

[0657] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIea), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 30, and is -L via the amine functional group provided by the side chain of lysine at position 27 1 - is attached.

[0658] In another preferred embodiment, the CNP prodrug of the present invention has the formula (IIea-i), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 30, and is -L via the amine functional group provided by the side chain of lysine at position 27 1 - is attached.

[0659] In another preferred embodiment, the CNP prodrug of the present invention is of formula (IIea-ii), c1 is about 450, the CNP moiety has the sequence of SEQ ID NO: 30, and is through the amine functional group provided by the side chain of lysine at position 27 -L 1 - is bonded.

[0660] Therefore, in a preferred embodiment, the CNP prodrug of the present invention is of the following formula (IIea').

Chemical formula

Chemical formula

[0661] Preferably, k in formula (IIea') is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

[0662] Preferably, each c1 in formula (IIea') is about 450.

[0663] In another preferred embodiment, the CNP prodrug of the present invention is of the following formula (IIea-i').

Chemical formula

Chemical formula

[0664] Preferably, k in formula (IIea-i′) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[0665] Preferably, each c1 in formula (IIea-i′) is about 450.

[0666] In another preferred embodiment, the CNP prodrug of the present invention is of the following formula (IIea-ii′).

Chemical formula

Chemical formula

[0667] Preferably, k in formula (IIea-ii′) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[0668] Preferably, each c1 in formula (IIea-ii′) is about 450.

[0669] In another preferred embodiment, the CNP prodrug of the present invention has the following formula (IIfa). [Chemical Formula] In the formula, The unmarked dashed line indicates the bond to the nitrogen of -D, which is the CNP moiety formed by forming an amide bond; The dashed line marked with an asterisk indicates the following structure: [Chemical Formula] represents the bond to -Z having, k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; Each -Z a is [Chemical Formula] and each c1 is an integer independently in the range of 200 to 250.

[0670] Preferably, k in formula (IIfa) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

[0671] Preferably, each c1 in formula (IIfa) is about 225.

[0672] In another preferred embodiment, the CNP prodrug of the present invention has the following formula (IIfa-i). [Chemical Formula] In the formula, The unmarked dashed line indicates the bond to the nitrogen of -D, which is the CNP moiety formed by forming an amide bond; The dashed line marked with an asterisk indicates the following structure: [Chemical Formula] represents the bond to -Z having, k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; each -Z a is

Chemical formula

[0673] Preferably, k of formula (IIfa-i) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

[0674] Preferably, each c1 of formula (IIfa-i) is about 225.

[0675] In another preferred embodiment, the CNP prodrug of the present invention is of the following formula (IIfa-ii).

Chemical formula

Chemical formula

Chemical formula

[0676] Preferably, each c1 of formula (IIfa-ii) is about 225.

[0677] In one embodiment, the CNP moiety of the CNP prodrugs of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO: 25.

[0678] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO: 30.

[0679] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO: 20.

[0680] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO: 21.

[0681] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO: 22.

[0682] In another embodiment, the CNP moiety of the CNP prodrugs of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO: 23.

[0683] In a preferred embodiment, the CNP moiety of the CNP prodrugs of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO: 24.

[0684] In one embodiment, the CNP moiety is attached to -L in the CNP prodrugs of formula (IIfa), (IIfa-i) and (IIfa-ii) via the nitrogen of the N-terminal amine functionality of CNP. 1 - is attached.

[0685] In another preferred embodiment, the CNP prodrug of the present invention is of the following formula (IIfa').

Chemical formula

[0686] Preferably, k of formula (IIfa') is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

[0687] Preferably, each c1 of formula (IIfa') is about 225.

[0688] In another preferred embodiment, the CNP prodrug of the present invention is of the following formula (IIfa-i'). [Chemical formula] In the formula, The dashed line without a mark indicates a bond to the nitrogen provided by the side chain of lysine at position 26 of the CNP moiety of SEQ ID NO: 24 by forming an amide bond; The dashed line marked with an asterisk indicates the following structure: [Chemical formula] represents a bond to -Z having k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; Each Za is [Chem.] and each c1 is an integer independently in the range of 200 to 250.

[0689] Preferably, k of formula (IIfa-i′) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

[0690] Preferably, each c1 of formula (IIfa-i′) is about 225.

[0691] In another preferred embodiment, the CNP prodrug of the present invention is of the following formula (IIfa-ii′). [Chem.] Wherein, The unmarked dashed line indicates a bond to the nitrogen provided by the side chain of lysine at position 26 of the CNP moiety of SEQ ID NO: 24 by forming an amide bond; The dashed line marked with an asterisk indicates the following structure: [Chem.] represents a bond to -Z having k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; each Z a is [Chem.] and each c1 is an integer independently in the range of 200 to 250.

[0692] Preferably, k of formula (IIfa-ii′) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

[0693] Preferably, each c1 of formula (IIfa-ii′) is about 225.

[0694] In a preferred embodiment, the residual activity of the CNP prodrug of the present invention is less than 10%, more preferably less than 1%, even more preferably less than 0.1%, even more preferably less than 0.01%, even more preferably less than 0.001%, and most preferably less than 0.0001%.

[0695] As used herein, the term "residual activity" refers to the activity exhibited by the CNP prodrug of the present invention having a CNP moiety bound to a carrier, in relation to the activity exhibited by the corresponding free CNP. In this context, the term "activity" refers to NPR-B binding. The measurement of the residual activity of the CNP prodrug of the present invention takes time, during which a certain amount of CNP will be released from the CNP prodrug of the present invention, and it is clear that such released CNP will distort the results measured for the CNP prodrug. Thus, as an acceptable practical operation, the residual activity of a prodrug having a drug moiety, in this case CNP, which is very similar in structure to the CNP prodrug for which the residual activity is to be measured and which is irreversibly, i.e., stably, bound to the carrier, is examined.

[0696] A suitable assay for measuring the CNP activity and residual activity of the CNP prodrug of the present invention, preferably in the form of a stable analogue, is described in WO2010 / 135541A1, Example 4, pages 143 / 144.

[0697] During such experiments, since the CNP prodrug may release a certain amount of CNP and the results may be distorted by the released CNP, the measurement of the NPR-B activity of the CNP prodrug is preferably carried out in the form of a stable analogue that does not release CNP.

[0698] Another aspect of the present invention is a pharmaceutical composition comprising at least one CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof and at least one excipient.

[0699] 1. In one embodiment, a pharmaceutical composition comprising a CNP prodrug molecule of the present invention is a mixture of CNP prodrugs in which the CNP moiety is attached to -L via a functional group of a different amino acid residue of the CNP moiety, preferably via an amine functional group. 1 - is included.

[0700] In a preferred embodiment, the CNP moieties of all CNP prodrug molecules contained in the pharmaceutical composition are attached to -L via a functional group of the same amino acid residue of the CNP moiety, preferably via an amine functional group of the same amino acid residue of the CNP moiety. 1 - is attached. In a preferred embodiment, the CNP moieties of all CNP prodrug molecules contained in the pharmaceutical composition are attached to -L via the amine functional group of the side chain of lysine 26 when the CNP moiety has the sequence of SEQ ID NO: 24. 1 - is attached.

[0701] Preferably, a pharmaceutical composition comprising at least one CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof has a pH in the range of pH 3 to pH 8 (including both ends). More preferably, the pharmaceutical composition has a pH in the range of pH 4 to pH 6 (including both ends). Most preferably, the pharmaceutical composition has a pH in the range of pH 4 to pH 5 (including both ends).

[0702] 1. In one embodiment, a pharmaceutical composition comprising at least one CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof and at least one excipient is a liquid formulation.

[0703] In another embodiment, a pharmaceutical composition comprising at least one CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof and at least one excipient is a dry formulation.

[0704] Such liquid or dry pharmaceutical compositions contain at least one excipient. Excipients used in parenteral formulations can be classified, for example, as buffers, isotonicity regulators, preservatives, stabilizers, adsorption inhibitors, antioxidants, thickeners / viscosity increasing agents, or other auxiliaries. However, in some cases, one excipient may have dual or triple functions. Preferably, at least one excipient contained in the pharmaceutical composition of the present invention is selected from the group consisting of:

[0705] (i) Buffer: Physiologically acceptable buffers for maintaining the pH within a desired range, such as sodium phosphate, bicarbonate, succinate, histidine, citrate and acetate, sulfate, nitrate, chloride, pyruvate. Antacids such as Mg(OH) 2 or ZnCO 3 may also be used;

[0706] (ii) Isotonicity regulator: To minimize pain that may result from cell damage due to osmotic pressure differences in injection depots. Glycerol and sodium chloride are examples. The effective concentration can be determined by osmotic pressure measurement using an assumed molar osmotic concentration of 285 - 315 mOsmol / kg for serum;

[0707] (iii) Preservative and / or antibacterial agent: For parenteral formulations for multiple administrations, the addition of a sufficient concentration of a preservative is required to minimize the risk of infection in patients by injection, and corresponding regulatory requirements have been established. Typical preservatives include m-cresol, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, chlorobutanol, benzyl alcohol, phenylmercuric nitrate, thimerosal, sorbic acid, potassium sorbate, benzoic acid, chlorocresol and benzalkonium chloride;

[0708] (iv) Stabilizers: Stabilization is achieved by enhancing the protein-stabilizing power, by destabilizing the denatured state, or by direct binding of the excipient to the protein. Stabilizers can be amino acids such as alanine, arginine, aspartic acid, glycine, histidine, lysine, proline; sugars such as glucose, sucrose, trehalose; polyols such as glycerol, mannitol, sorbitol; salts such as potassium phosphate, sodium sulfate; chelating agents such as EDTA, hexaphosphate; ligands such as divalent metal ions (zinc, calcium, etc.); other salts or organic molecules such as phenolic derivatives. In addition, oligomers or polymers such as cyclodextrin, dextran, dendrimer, PEG or PVP or protamine or HSA may be used;

[0709] (v) Adsorption inhibitors: Mainly ionic or non-ionic surfactants or other proteins or soluble polymers such as poloxamer (Pluronic F-68), PEG dodecyl ether (Brij 35), polysorbates 20 and 80, dextran, polyethylene glycol, PEG-polyhistidine, BSA and HSA and gelatin are used for coating the inner surface of the formulation container or for competitive adsorption to the inner surface. The excipient concentration and type selected depend on the effects to be avoided, but typically a monolayer of surfactant is formed at the interface just above the CMC value;

[0710] (vi) Antioxidants: Antioxidants such as ascorbic acid, ectoine, methionine, glutathione, monothioglycerol, morin, polyethyleneimine (PEI), propyl gallate and vitamin E. Chelating agents such as citric acid, EDTA, hexaphosphate, and thioglycolic acid may also be used;

[0711] (vii) Thickening agent or viscosity increasing agent: It delays the sedimentation of particles in vials and syringes, promotes the mixing and resuspension of particles, and is used to make it easier to inject the suspension agent (i.e., with a small force on the syringe plunger). Suitable thickening agents or viscosity increasing agents are, for example, carbomer thickening agents such as Carbopol 940, Carbopol Ultrez 10; cellulose derivatives such as hydroxypropyl methylcellulose (hypromellose, HPMC) or diethylaminoethyl cellulose (DEAE or DEAE-C); colloidal magnesium silicate (Veegum) or sodium silicate; hydroxyapatite gel; tricalcium phosphate gel; xanthan; carrageenan such as Satia gum UTC 30; aliphatic poly(hydroxy acids) such as poly(D,L- or L-lactic acid) (PLA) and poly(glycolic acid) (PGA) and their copolymers (PLGA), terpolymers of D,L-lactide, glycolide, and caprolactone; poloxamer; hydrophilic poly(oxyethylene) block and hydrophobic poly(oxypropylene) block for forming a triblock of poly(oxyethylene)-poly(oxypropylene)-poly(oxyethylene) (e.g., Pluronic®); polyether ester copolymer such as polyethylene glycol terephthalate / polybutylene terephthalate copolymer; sucrose acetate isobutyrate (SAIB); dextran or its derivatives; combination of dextran and PEG; polydimethylsiloxane; collagen; chitosan; vinyl alcohol (PVA) and derivatives; polyalkylimide; poly(acrylamide-co-diallyldimethylammonium (DADMA)); polyvinylpyrrolidone (PVP); glycosaminoglycan (GAG) such as dermatan sulfate, chondroitin sulfate, keratan sulfate, heparin, heparan sulfate, hyaluronan; ABA triblock or AB block copolymer composed of a hydrophobic A block such as polylactide (PLA) or poly(lactide-co-glycolide) (PLGA) and a hydrophilic B block such as polyethylene glycol (PEG) or polyvinylpyrrolidone.Such block copolymers and the above-mentioned poloxamers can exhibit inverse thermogelation behavior (fluid state at room temperature to facilitate administration and gel state at a temperature higher than the sol-gel transition temperature at body temperature after injection);

[0712] (viii) Spreading or diffusing agents: Regulate the permeability of connective tissue by hydrolysis of components of the extracellular matrix in the interstitial cavity (e.g., hyaluronic acid, a polysaccharide found in the intercellular space of connective tissue, which is not limited thereto). Spreading agents, such as, but not limited to, hyaluronidase, reduce the viscosity of the extracellular matrix and promote the diffusion of the injected drug; and

[0713] (ix) Other adjuvants: Wetting agents, viscosity regulators, antibiotics, hyaluronidase, etc. Acids and bases such as hyaluronic acid and sodium hydroxide are adjuvants necessary for pH adjustment during production.

[0714] Another aspect of the present invention is the use of the CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising at least one CNP prodrug of the present invention as a medicine.

[0715] Preferably, the medicament is used for the treatment of diseases selected from the group consisting of achondroplasia (achondrogenesis), hypochondroplasia (chondrodysplasia), short stature, dwarfism, osteochondrodysplasia (chondro-osteodysplasia), lethal dysplasia (thanatophoric dysplasia), osteogenesis imperfecta, achondrogenesis, stippled chondrodysplasia, homozygous achondroplasia, campomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly syndrome, rhizomelic stippled chondrodysplasia (proximal limb shortening type stippled chondrodysplasia), Jansen type metaphyseal dysplasia, congenital spondylometaphyseal dysplasia, osteogenesis imperfecta, torsional osteodysplasia, congenital femoral shortening, Langer type intermediate limb dysplasia, Nievergelt type intermediate limb dysplasia, Robinow syndrome, Reinhardt syndrome, acroosteolysis, distal osteolysis, Kneist osteodysplasia, fibrochondrogenesis, Roberts syndrome, distal intermediate limb dysplasia, micromelia, Morquio syndrome, Kneist syndrome, dysplastic osteodysplasia, spondylometaphyseal dysplasia, neurofibromatosis, Legius syndrome, Leopard syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, heart-face-skin syndrome, Costello syndrome, SHOX deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysplasia, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzonodermoskeletal syndrome), dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, segmental dysplasia, enchondroma, fibrous osteodysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis, and osteopathia striata.

[0716] In another embodiment, the medicament is used for the treatment of ophthalmic diseases, such as glaucoma and / or ocular hypertension.

[0717] In another embodiment, the medicament is used for the treatment of cancers associated with overactivation of FGFR3, such as multiple myeloma, myeloproliferative syndrome, leukemia, plasmacytic leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer or breast cancer.

[0718] In another embodiment, the medicament is used for the treatment of vascular smooth muscle disorders, preferably selected from the group consisting of hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, renal edema, hepatic edema, acute renal failure and chronic renal failure.

[0719] In another embodiment, the medicament is used for the treatment of hemorrhagic shock.

[0720] Preferably, the medicament is used for the treatment of achondroplasia phenotypes, selected from the group consisting of growth retardation, craniofacial deformities, orthodontic defects, cervical spinal cord compression, spinal stenosis, hydrocephalus, hearing loss due to chronic otitis media, cardiovascular diseases, neurological diseases and obesity.

[0721] Most preferably, the medicament is used for the treatment of achondroplasia.

[0722] Another aspect of the present invention is a pharmaceutical composition for use in a method of treating a disease treatable with CNP, the CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising at least one CNP prodrug of the present invention.

[0723] Preferably, the disease is selected from the group consisting of achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasia, lethal dysplasia, osteogenesis imperfecta, achondrogenesis, stippled chondrodysplasia, homozygous achondroplasia, campomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly syndrome, rhizomelic stippled chondrodysplasia, Jansen metaphyseal dysplasia, congenital spondyloepiphyseal dysplasia, osteogenesis imperfecta, torsional osteodysplasia, congenital femoral shortening, Langer mesomelic dysplasia, Nievergelt mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acroosteolysis, distal osteolysis, Kneist osteochondrodysplasia, fibrochondrogenesis, Roberts syndrome, distal mesomelic dysplasia, micromelia, Morquio syndrome, Kneist syndrome, dysplastic osteodysplasia, spondyloepiphyseal metaphyseal dysplasia, neurofibromatosis, Legius syndrome, Leopard syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, cardiofaciocutaneous syndrome, Costello syndrome, SHOX deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysplasia, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzonodermoskeletal syndrome, polydactyly (finger abnormalities), brachydactyly, camptodactyly, polydactyly, syndactyly, segmental dysplasia dysplasia, enchondroma, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Yaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis and osteopathia striata.

[0724] In another embodiment, the disease is an ophthalmic disease, such as glaucoma and / or ocular hypertension.

[0725] In another embodiment, the disease is cancer, such as FGFR3 overactivation in multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer or breast cancer.

[0726] In another embodiment, the disease is a vascular smooth muscle disorder preferably selected from the group consisting of hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, renal edema, hepatic edema, acute renal failure and chronic renal failure.

[0727] In another embodiment, the disease is hemorrhagic shock.

[0728] Preferably, the disease is a achondroplasia phenotype selected from the group consisting of growth retardation, cranial deformation, orthodontic defect, cervical spinal cord compression, spinal stenosis, hydrocephalus, hearing loss due to chronic otitis media, cardiovascular disease, neurological disease and obesity.

[0729] Most preferably, the disease is achondroplasia.

[0730] In one embodiment, the patient receiving the treatment method of the present invention is a mammalian patient, preferably a human patient. In one embodiment, this human patient is an adult. In a preferred embodiment, the human patient is a pediatric patient.

[0731] Another aspect of the present invention is the use of the CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising at least one CNP prodrug of the present invention for the manufacture of a medicament for treating a disease that can be treated with CNP.

[0732] Preferably, the disease is selected from the group consisting of achondrogenesis, hypochondrogenesis, short stature, dwarfism, osteochondrodysplasia, lethal dysplasia, osteogenesis imperfecta, achondrogenesis, stippled chondrodysplasia, homozygous achondroplasia, campomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly syndrome, rhizomelic stippled chondrodysplasia, Jansen metaphyseal dysplasia, congenital spondyloepiphyseal dysplasia, osteogenesis imperfecta, torsional osteodysplasia, congenital femoral shortening, Langer type intermediate limb dysplasia, Nievergelt type intermediate limb dysplasia, Robinow syndrome, Reinhardt syndrome, acroosteolysis, distal osteolysis, Kneist osteodysplasia, fibrochondrogenesis, Roberts syndrome, distal intermediate limb dysplasia, micromelia, Morquio syndrome, Kneist syndrome, dysplastic osteodysplasia, spondyloepimetaphyseal dysplasia, neurofibromatosis, Legius syndrome, Leopard syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, heart-face-skin syndrome, Costello syndrome, SHOX deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysplasia, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzonodermoskeletal syndrome), syndactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, segmental dysplasia dysplasia, enchondroma, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Yaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis, and osteopathia striata.

[0733] In another embodiment, the disease is an ophthalmic disease, such as glaucoma and / or ocular hypertension.

[0734] In another embodiment, the disease is cancer, such as FGFR3 overactivation associated with multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, or breast cancer.

[0735] In another embodiment, the disease is a vascular smooth muscle disorder preferably selected from the group consisting of hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, renal edema, hepatic edema, acute renal failure and chronic renal failure.

[0736] In another embodiment, the disease is hemorrhagic shock.

[0737] Preferably, the disease is a achondroplasia phenotype selected from the group consisting of growth retardation, cranial deformation, orthodontic defect, cervical spinal cord compression, spinal stenosis, hydrocephalus, hearing loss due to chronic otitis media, cardiovascular disease, neurological disease and obesity.

[0738] Most preferably, the disease is achondroplasia.

[0739] In one embodiment, the disease to be treated with the CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising at least one CNP prodrug of the present invention occurs in mammalian patients, preferably in human patients. In one embodiment, the human patient is an adult. In a preferred embodiment, the human patient is a pediatric patient.

[0740] A further aspect of the present invention is a method of treating, managing, delaying or preventing in a mammalian patient, preferably a human patient, in need of treatment of one or more diseases that can be treated with CNP, the method comprising administering to the patient in need thereof a therapeutically effective amount of the CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the CNP prodrug of the present invention. In one embodiment, the human patient is an adult. In a preferred embodiment, the human patient is a pediatric patient.

[0741] Preferably, one or more diseases that can be treated with the CNP are selected from the group consisting of achondrogenesis, hypochondrogenesis, short stature, dwarfism, osteochondroplasia, lethal dysplasia, osteogenesis imperfecta, achondrogenesis, stippled chondrodysplasia, homozygous achondroplasia, campomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly syndrome, rhizomelic stippled chondrodysplasia, Jansen metaphyseal dysplasia, congenital spondyloepiphyseal dysplasia, osteogenesis imperfecta, torsional dysplasia, congenital femoral shortening, Langer mesomelic dysplasia, Nievergelt mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acroosteolysis, acroosteolysis, Kneist dysplasia, fibrochondrogenesis, Roberts syndrome, distal mesomelic dysplasia, micromelia, Morquio syndrome, Kneist syndrome, dysplastic dysplasia, spondyloepiphyseal metaphyseal dysplasia, neurofibromatosis, Legius syndrome, Leopard syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, heart-face-skin syndrome, Costello syndrome, SHOX deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysplasia, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzonodermoskeletal syndrome), polydactyly, brachydactyly, clinodactyly, syndactyly, segmental dysplasia, enchondroma, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Yaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis, and osteopathia striata.

[0742] In another embodiment, one or more diseases that can be treated with the CNP are ophthalmic diseases, such as glaucoma and / or ocular hypertension.

[0743] In another embodiment, one or more diseases that can be treated with the CNP are cancers, such as multiple myeloma, myeloproliferative syndromes, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, or breast cancer, which are associated with overactivation of FGFR3.

[0744] In another embodiment, one or more diseases that can be treated with said CNP are vascular smooth muscle disorders preferably selected from the group consisting of hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, renal edema, hepatic edema, acute renal failure and chronic renal failure.

[0745] In another embodiment, one or more diseases that can be treated with said CNP is hemorrhagic shock.

[0746] Preferably, one or more diseases that can be treated with CNP is a achondroplasia phenotype selected from the group consisting of growth retardation, cranial deformation, orthodontic defects, cervical spinal cord compression, spinal stenosis, hydrocephalus, hearing loss due to chronic otitis media, cardiovascular diseases, neurological diseases and obesity.

[0747] Most preferably, one or more diseases that can be treated with CNP is achondroplasia.

[0748] A further aspect of the present invention is a method of administering a CNP prodrug, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention, by topical, enteral or parenteral administration, and by intra-articular, peri-articular, intradermal, subcutaneous, intramuscular, intravenous, intraosseous, intraperitoneal, intrathecal, intracapsular, intraorbital, intravitreal, intra-tympanic, intravesical, intracardiac, transtracheal, subepidermal, subcapsular, subarachnoid, intraspinal, intraventricular, intrasternal injection and infusion, a method including the step of administering a CNP prodrug, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention by an external application, injection or infusion method, preferably by subcutaneous injection, including direct delivery to the brain by an implantable device (e.g., an Ommaya Reservoir) that enables delivery of the present invention to the brain tissue or cerebrospinal fluid, direct intraventricular injection or infusion, injection or infusion into the brain or brain-related regions, injection into the subchoroidal space, posterior orbital injection and eye drops.

[0749] In a preferred embodiment, the present invention relates to a CNP prodrug of the present invention or a pharmaceutically acceptable salt or pharmaceutical composition thereof for use in the treatment of achondroplasia by subcutaneous injection.

[0750] In a further aspect, the present invention relates to a pharmaceutical composition comprising at least one CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition comprises at least one further bioactive moiety or agent.

[0751] Said at least one further bioactive moiety or agent can be in its free form (i.e., in the form of the free drug), in the form of a stable conjugate, or in the form of a sustained release compound.

[0752] In one embodiment, said at least one further bioactive moiety or agent is a drug in its free form, i.e., the pharmaceutical composition of the present invention comprises at least one CNP prodrug and at least one further drug.

[0753] Preferably, said at least one further drug is selected from the group consisting of antihistamines; human anti-FGFR3 antibodies; soluble human fibroblast growth factor receptor 3; tyrosine kinase inhibitors; statins; CNP agonists; growth hormone; IGF-1; ANP; BNP; inhibitors of peptidases and proteases; and inhibitors of NPR-C.

[0754] A preferred antihistamine is meclozine.

[0755] A preferred tyrosine kinase inhibitor is NVP-BGJ398.

[0756] A preferred statin is rosuvastatin.

[0757] A preferred CNP agonist for said at least one further drug is vosoritide.

[0758] Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

[0759] Preferred inhibitors of NEP are thiolphan and candoxatril.

[0760] Preferred inhibitors of NPR-C are the fragment of SEQ ID NO: 98 (FGIPMDRIGRNPR) and antibody B701.

[0761] Preferred inhibitors of tyrosine kinase are as disclosed in U.S. Patent Nos. 6329375 and 6344459 (incorporated herein by reference).

[0762] In one embodiment, the at least one additional agent is an antihistamine.

[0763] In another embodiment, the at least one additional agent is a human anti-FGFR3 antibody.

[0764] In another embodiment, the at least one additional agent is a soluble form of human fibroblast growth factor receptor 3 (sFGFR3).

[0765] In another embodiment, the at least one additional agent is a tyrosine kinase inhibitor.

[0766] In another embodiment, the at least one additional agent is a statin.

[0767] In another embodiment, the at least one additional agent is a growth hormone.

[0768] In another embodiment, the at least one additional agent is a CNP agonist.

[0769] In another embodiment, the at least one additional agent is IGF-1.

[0770] In another embodiment, the at least one additional agent is ANP.

[0771] In another embodiment, said at least one further agent is BNP.

[0772] In another embodiment, said at least one further agent is an inhibitor of peptidases and proteases.

[0773] In another embodiment, said at least one further agent is an inhibitor of NPR-C.

[0774] In another embodiment, said at least one further bioactive moiety or agent is in the form of a stable conjugate.

[0775] In one embodiment, said at least one further bioactive moiety in the form of a stable conjugate comprises at least one bioactive moiety covalently bound, directly or via a spacer moiety, via a stable linker to a polymer moiety, preferably a water-soluble polymer moiety.

[0776] Preferably, such a polymer moiety, more preferably a water-soluble polymer moiety, is selected from the group consisting of 2-methacryloyloxyethyl phosphorylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amide amines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyl oxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organic phosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinyl methyl ethers), poly(vinyl pyrrolidones), silicones, celluloses, carboxymethyl celluloses, hydroxypropyl methyl celluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other hydrocarbon-based polymers, xylans, and copolymers thereof.

[0777] In another embodiment, the at least one additional bioactive moiety in the form of the stable conjugate is covalently attached to the albumin-binding moiety via a stable linker. Preferably, the albumin-binding moiety is a C 8-24 alkyl moiety or a fatty acid derivative. Preferred fatty acid derivatives are those disclosed in WO2005 / 027978A2 and WO2014 / 060512A1 (incorporated herein by reference).

[0778] Preferably, the at least one additional bioactive moiety in the form of the stable conjugate comprises a bioactive moiety selected from the group consisting of an antihistamine; a human anti-FGFR3 antibody; a soluble form of human fibroblast growth factor receptor 3 (sFGFR3); a tyrosine kinase inhibitor; statins; a CNP agonist; a growth hormone; IGF-1; ANP; BNP; an inhibitor of peptidases and proteases; and an inhibitor of NPR-C.

[0779] A preferred antihistamine is meclozine.

[0780] A preferred tyrosine kinase inhibitor is NVP-BGJ398.

[0781] A preferred statin is rosuvastatin.

[0782] A preferred CNP agonist for the at least one additional bioactive moiety is vosoritide.

[0783] Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

[0784] Preferred inhibitors for NEP are thiorphan and candoxatril.

[0785] Preferred inhibitors of NPR-C are the fragment of SEQ ID NO: 98 (FGIPMDRIGRNPR) and antibody B701.

[0786] Preferred inhibitors of tyrosine kinase are disclosed in U.S. Patent Nos. 6329375 and 6344459, which are incorporated herein by reference.

[0787] In one embodiment, the at least one additional bioactive moiety in the form of a stable conjugate comprises an antihistamine moiety.

[0788] In another embodiment, the at least one additional bioactive moiety in the form of a stable conjugate comprises a human anti-FGFR3 antibody moiety.

[0789] In another embodiment, the at least one additional bioactive moiety in the form of a stable conjugate comprises a soluble form of human fibroblast growth factor receptor 3 (sFGFR3) moiety.

[0790] In another embodiment, the at least one additional bioactive moiety in the form of a stable conjugate comprises a tyrosine kinase inhibitor moiety.

[0791] In another embodiment, the at least one additional bioactive moiety in the form of a stable conjugate comprises a statin moiety.

[0792] In another embodiment, the at least one additional bioactive moiety in the form of a stable conjugate comprises a growth hormone moiety.

[0793] In another embodiment, the at least one additional bioactive moiety in the form of a stable conjugate comprises a CNP agonist moiety.

[0794] In another embodiment, the at least one additional bioactive moiety in the form of a stable conjugate comprises an IGF-1 moiety.

[0795] In another embodiment, the at least one additional bioactive moiety in the form of a stable conjugate comprises an ANP moiety.

[0796] In another embodiment, said at least one further bioactive moiety in the form of a stable conjugate comprises a BNP moiety.

[0797] In another embodiment, said at least one further bioactive moiety in the form of a stable conjugate comprises inhibitors of peptidases and proteases moieties.

[0798] In another embodiment, said at least one further bioactive moiety in the form of a stable conjugate comprises an inhibitor of the NPR-C moiety.

[0799] In another embodiment, said at least one further bioactive moiety or agent is in the form of a sustained release compound.

[0800] Preferably, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises at least one bioactive moiety or agent selected from the group consisting of antihistamines; human anti-FGFR3 antibodies; soluble form of human fibroblast growth factor receptor 3; statins; CNP agonists; growth hormone; IGF-1; ANP; BNP; inhibitors of peptidases and proteases; inhibitors of tyrosine kinases; and inhibitors of NPR-C.

[0801] A preferred antihistamine is meclozine.

[0802] A preferred tyrosine kinase inhibitor is NVP-BGJ398.

[0803] A preferred statin is rosuvastatin.

[0804] A preferred CNP agonist for said at least one further agent is vosoritide.

[0805] Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

[0806] Preferred inhibitors of NEP are thiolphan and candoxatril.

[0807] Preferred inhibitors of NPR-C are the fragment of SEQ ID NO: 98 (FGIPMDRIGRNPR) and antibody B701.

[0808] Preferred inhibitors of tyrosine kinases are those disclosed in U.S. Patent Nos. 6329375 and 6344459 (incorporated herein by reference).

[0809] In one embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises an antihistamine moiety or agent.

[0810] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises a human anti-FGFR3 antibody moiety or agent.

[0811] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises a soluble form of human fibroblast growth factor receptor 3 (sFGFR3) moiety or agent.

[0812] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises a tyrosine kinase inhibitor moiety or agent.

[0813] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises a statin moiety or agent.

[0814] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises a growth hormone moiety or agent.

[0815] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises a CNP agonist moiety or agent.

[0816] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises an IGF-1 moiety or agent.

[0817] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises an ANP moiety or agent.

[0818] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises a BNP moiety or agent.

[0819] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises inhibitors of peptidases and proteases moieties or agents.

[0820] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound comprises an NPR-C moiety or inhibitor of an agent.

[0821] In one embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound is water-insoluble.

[0822] Preferably, such water-insoluble sustained release compounds are selected from the group consisting of crystals, nanoparticles, microparticles, nanospheres and microspheres.

[0823] In one embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release compound is a crystal comprising at least one agent or bioactive moiety.

[0824] In another embodiment, the at least one additional bioactive moiety or agent in the form of a water-insoluble sustained release compound is nanoparticles comprising at least one agent or bioactive moiety.

[0825] In another embodiment, the at least one additional bioactive moiety or agent in the form of a water-insoluble sustained release compound is microparticles comprising at least one agent or bioactive moiety.

[0826] In another embodiment, the at least one additional bioactive moiety or agent in the form of a water-insoluble sustained release compound is nanospheres comprising at least one agent or bioactive moiety.

[0827] In another embodiment, the at least one additional bioactive moiety or agent in the form of a water-insoluble sustained release compound is microspheres comprising at least one agent or bioactive moiety.

[0828] In one embodiment, the at least one additional bioactive moiety or agent in the form of a water-insoluble sustained release compound is a medium comprising at least one agent or bioactive moiety. Preferably, such a medium comprising at least one agent or bioactive moiety is micelles, liposomes or polymersomes.

[0829] In one embodiment, the at least one additional bioactive moiety or agent in the form of a water-insoluble sustained release compound is micelles comprising at least one agent or bioactive moiety.

[0830] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release compound is a liposome comprising at least one agent or bioactive moiety. Preferably, such liposomes are selected from the group consisting of aquasomes; nonionic surfactant-based vehicles such as niosomes and proniosomes; cationic liposomes such as LeciPlex; transfersomes; ethosomes; ufasomes; sphingosomes; and pharmacosomes.

[0831] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release compound is a polymersome comprising at least one agent or bioactive moiety.

[0832] In another embodiment, the at least one additional bioactive moiety or agent in the form of a water-insoluble sustained release compound comprises at least one bioactive moiety or agent non-covalently embedded in a water-insoluble polymer. Preferably, such water-insoluble polymers include polymers selected from the group consisting of 2-methacryloyl-oxyethyl phosphorylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyl oxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organic phosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinyl methyl ethers), poly(vinyl pyrrolidones), silicones, celluloses, carboxymethyl celluloses, hydroxypropyl methyl celluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other hydrocarbon-based polymers, xylans, and copolymers thereof.

[0833] In a preferred embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release compound comprises at least one agent or bioactive moiety non-covalently embedded in poly(lactic-co-glycolic acid) (PLGA).

[0834] In another embodiment, the at least one additional bioactive moiety or agent in the form of a water-insoluble sustained release compound comprises at least one bioactive moiety covalently and reversibly bound to an insoluble polymer. Preferably, such water-insoluble polymers are 2-methacryloyl-oxyethyl phosphorylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amide amines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyl oxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organic phosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinyl methyl ethers), poly(vinyl pyrrolidones), silicones, celluloses, carboxymethyl celluloses, hydroxypropyl methyl celluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other hydrocarbon-based polymers, xylans, and copolymers thereof.

[0835] Preferably, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release compound comprises at least one bioactive moiety or agent selected from the group consisting of: an antihistamine; a human anti-FGFR3 antibody; a soluble form of human fibroblast growth factor receptor 3; a tyrosine kinase inhibitor; statins; a CNP agonist; growth hormone; IGF-1; ANP; BNP; an inhibitor of peptidases and proteases; and an inhibitor of NPR-C.

[0836] A preferred antihistamine is meclozine.

[0837] A preferred tyrosine kinase inhibitor is NVP-BGJ398.

[0838] A preferred statin is rosuvastatin.

[0839] A preferred CNP agonist for said at least one further agent is vosoritide.

[0840] Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

[0841] Preferred inhibitors for NEP are thiorphan and candoxatril.

[0842] Preferred inhibitors of NPR-C are the fragment of SEQ ID NO: 98 (FGIPMDRIGRNPR) and antibody B701.

[0843] Preferred inhibitors of tyrosine kinases are those disclosed in U.S. Patent Nos. 6329375 and 6344459 (incorporated herein by reference).

[0844] In one embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises an antihistamine moiety or agent.

[0845] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises a human anti-FGFR3 antibody moiety or agent.

[0846] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises a soluble form of human fibroblast growth factor receptor 3 (sFGFR3) moiety or agent.

[0847] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises a tyrosine kinase inhibitor moiety or agent.

[0848] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises a statin moiety or agent.

[0849] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises a growth hormone moiety or agent.

[0850] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises a CNP agonist moiety or agent.

[0851] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises an IGF-1 moiety or agent.

[0852] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises an ANP moiety or agent.

[0853] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises a BNP moiety or agent.

[0854] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises an inhibitor of peptidases and proteases moieties or agents. In another embodiment, said at least one further bioactive moiety or agent in the form of a water-insoluble sustained release agent comprises an inhibitor of NPR-C moieties or agents.

[0855] In another embodiment, said at least one further bioactive moiety or agent in the form of a sustained release compound is water-soluble.

[0856] In one embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release compound comprises at least one bioactive moiety covalently attached to a water-soluble polymer moiety via a reversible linker, either directly or via a spacer moiety.

[0857] Preferably, such a water-soluble polymer moiety comprises a polymer selected from the group consisting of 2-methacryloyloxyethyl phosphorylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy)polymers, poly(amides), poly(amide amines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyl oxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organic phosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinyl methyl ethers), poly(vinyl pyrrolidones), silicones, celluloses, carboxymethyl celluloses, hydroxypropyl methyl celluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other hydrocarbon-based polymers, xylans, and copolymers thereof.

[0858] In another embodiment, said at least one further bioactive moiety in the form of a water-soluble sustained release compound is covalently bound to an albumin-binding moiety via a stable linker. Preferably, said albumin-binding moiety is a C 8-24 alkyl moiety or a fatty acid derivative. Preferred fatty acid derivatives are those disclosed in WO2005 / 027978A2 and WO2014 / 060512A1 (incorporated herein by reference).

[0859] Preferably, said at least one further bioactive moiety in the form of a water-soluble sustained release agent comprises a bioactive moiety selected from the group consisting of an antihistamine; a human anti-FGFR3 antibody; a soluble form of the human fibroblast growth factor receptor 3; a tyrosine kinase inhibitor; statins; a CNP agonist; growth hormone; IGF-1; ANP; BNP; inhibitors of peptidases and proteases; and an inhibitor of NPR-C.

[0860] A preferred antihistamine is meclozine.

[0861] A preferred tyrosine kinase inhibitor is NVP-BGJ398.

[0862] A preferred statin is rosuvastatin.

[0863] A preferred CNP agonist for said at least one further agent is vosoritide.

[0864] Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

[0865] Preferred inhibitors for NEP are thiorphan and candoxatril.

[0866] Preferred inhibitors of NPR-C are the fragment of SEQ ID NO: 98 (FGIPMDRIGRNPR) and antibody B701.

[0867] Preferred inhibitors of tyrosine kinases are those disclosed in U.S. Patent Nos. 6329375 and 6344459 (incorporated herein by reference).

[0868] In one embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises an antihistamine moiety or agent.

[0869] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises a human anti-FGFR3 antibody moiety or agent.

[0870] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises a soluble form of human fibroblast growth factor receptor 3 (sFGFR3) moiety or agent.

[0871] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises a tyrosine kinase inhibitor moiety or agent.

[0872] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises a statin moiety or agent.

[0873] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises a growth hormone moiety or agent. A preferred water-soluble sustained release growth hormone compound is Compound 2 of Example 2 of WO2016 / 079114A1. Accordingly, a preferred water-soluble sustained release growth hormone compound has the following structure.

Chemical formula

[0874] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises a CNP agonist moiety or agent.

[0875] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises an IGF-1 moiety or agent.

[0876] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises an ANP moiety or agent.

[0877] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises a BNP moiety or agent.

[0878] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises inhibitors of peptidases and proteases moieties or agents.

[0879] In another embodiment, said at least one further bioactive moiety or agent in the form of a water-soluble sustained release agent comprises an NPR-C moiety or an inhibitor of an agent.

[0880] Another aspect of the invention is a pharmaceutical composition for use as a medicament, comprising at least one CNP prodrug of the invention and at least one further bioactive moiety or agent of the invention.

[0881] Another aspect of the invention is a pharmaceutical composition for use in the treatment of a patient suffering from a disorder which is responsive to growth stimulation, comprising at least one CNP prodrug of the invention and at least one further bioactive moiety or agent of the invention.

[0882] Preferably, said patient is a mammalian patient, more preferably a human patient.

[0883] Preferably, such disorders for which growth stimulation is effective are selected from the group consisting of achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasia, lethal dysplasia, osteogenesis imperfecta, achondrogenesis, stippled chondrodysplasia, homozygous achondroplasia, campomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly syndrome, rhizomelic stippled chondrodysplasia, Jansen metaphyseal dysplasia, congenital spondyloepiphyseal dysplasia, osteogenesis imperfecta imperfecta, torsional dysplasia, congenital femoral shortening, Langer mesomelic dysplasia, Nievergelt mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acroosteolysis, terminal osteolysis, Kneist dysplasia, fibrochondrogenesis, Roberts syndrome, distal mesomelic dysplasia, micromelia, Morquio syndrome, Kneist syndrome, dysplastic dysplasia, and spondyloepiphyseal metaphyseal dysplasia. Most preferably, the disorder for which growth stimulation is effective is achondroplasia.

[0884] Another aspect of the invention is a method of treating a patient suffering from a disorder for which growth stimulation is effective by administering the pharmaceutical composition of the invention.

[0885] Preferably, the patient is a mammalian patient, more preferably a human patient.

[0886] Preferably, such disorders in which growth stimulation is effective are selected from the group consisting of achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasia, lethal dysplasia, osteogenesis imperfecta, achondrogenesis, stippled chondrodysplasia, homozygous achondroplasia, campomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly syndrome, rhizomelic stippled chondrodysplasia, Jansen metaphyseal dysplasia, congenital spondyloepiphyseal dysplasia, osteogenesis imperfecta imperfecta, torsional dysplasia, congenital femoral shortening, Langer mesomelic dysplasia, Nievergelt mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acroosteolysis, distal osteolysis, Kneist dysplasia, fibrochondrogenesis, Roberts syndrome, distal mesomelic dysplasia, micromelia, Morquio syndrome, Kneist syndrome, dysplastic dysplasia, and spondyloepiphyseal metaphyseal dysplasia. Most preferably, the disorder in which growth stimulation is effective is achondroplasia.

[0887] The polypeptide portion of the CNP prodrug can be produced by standard solid-phase peptide synthesis methods, for example, by Boc Chemistry (R. B. Merrifield, J. Am. Chem. Soc., 85(14):2149-2154(1963)). Alternatively, Fmoc (fluorenylmethoxycarbonyl) chemistry may be used.

[0888] Using methods known in the art, the purity and / or yield can be increased, including the use of pseudoproline and other dipeptide building blocks, fragment coupling, etc. (J. Wade et al., Lett. Pept. Sci., 7(2):107-112 (2000); Y. Fujiwara et al., Chem. Pharm. Bull., 44(7):1326-1331 (1996); P. Cherkupally et al., Eur. J. Org. Chem., 6372-6378(2013)).

[0889] Alternatively, the polypeptide portion of the CNP prodrug can be produced by recombinant synthesis methods.

Example

[0890] Materials and Methods CNP SEQ ID NO:1 was obtained from Bachem AG, Bubendorf, Switzerland (CNP-22, human, catalog number H-1296). CNP-34 SEQ ID NO:40 and CNP-38 SEQ ID NO:24 were obtained from CASLO ApS, Kongens Lyngby, Denmark.

[0891] Side-chain protected CNP-38 (synthesized by the Fmoc strategy) on CTC resin with a Boc-protected N-terminus and an ivDde-protected side-chain of Lys26 was obtained from CASLO ApS, Kongens Lyngby, Denmark.

[0892] Side-chain protected CNP-34 (synthesized by the Fmoc strategy) on TCP Tentagel resin with a Boc-protected N-terminus and an ivDde-protected side-chain of either Lys12, Lys16 or Lys22 was obtained from Peptide Specialty Laboratories GmbH, Heidelberg, Germany. Side-chain protected CNP-38 (synthesized by the Fmoc strategy) on TCP Tentagel resin with a free N-terminus was obtained from Peptide Specialty Laboratories GmbH, Heidelberg, Germany.

[0893] Methoxy PEG amine 5 kDa was obtained from Rapp Polymere GmbH, Tuebingen, Germany. All other PEGs used in this study were obtained from NOF Europe N.V., Grobbendonk, Belgium.

[0894] FmocN-Me-Asp(OtBu)-OH was obtained from Bachem AG, Bubendorf, Switzerland. S-Trityl-6-mercaptohexanoic acid was purchased from Polypeptide, Strasbourg, France. HATU was obtained from Merck Biosciences GmbH, Schwalbach / Ts, Germany.

[0895] 2,4-Dimethylbenzyl alcohol was obtained from abcr GmbH, Karlsruhe, Germany.

[0896] Fmoc-N-Me-Asp(OBn)-OH was obtained from Peptide International Inc., Louisville, KY, USA.

[0897] Neutral endopeptidase (NEP) was obtained from Enzo Life Sciences GmbH, Lorrach, Germany.

[0898] All other chemicals and reagents were purchased from Sigma Aldrich GmbH, Taufkirchen, Germany.

[0899] A syringe equipped with a polyethylene frit (MultiSynTech GmbH, Witten, Germany) was used as a reaction vessel or in the washing step of the peptide resin.

[0900] General procedure for removal of the ivDde protecting group from side-chain protected CNP on resin The resin was pre-swollen in DMF for 30 min and the solvent was discarded. The ivDde group was removed by incubating the resin with DMF / hydrazine hydrate 4 / 1 (v / v, 2.5 mL / g resin) for 8 × 15 min. A fresh DMF / hydrazine hydrate solution was used for each step. Finally, the resin was washed with DMF (10×), DCM (10×) and dried under vacuum.

[0901] RP-HPLC purification: For preparative RP-HPLC, a Waters 600 controller and 2487 Dual Absorbance Detector were used, equipped with the following columns: Waters XBridge™ BEH300 Prep C18 5μm, 150×10mm, flow rate 6 mL / min, or Waters XBridge™ BEH300 Prep C18 10μm, 150×30mm, flow rate 40 mL / min. A linear gradient of solvent system A (water containing 0.1% TFA v / v or 0.01% concentrated HCl v / v) and solvent system B (acetonitrile containing 0.1% TFA v / v or 0.01% concentrated HCl v / v) was used.

[0902] Unless otherwise stated, HPLC fractions containing the product were pooled and lyophilized.

[0903] Flash chromatography Flash chromatography purification was performed on a Biotage KP-Sil silica cartridge using n-heptane and ethyl acetate as eluents on an Isolera One system from Biotage AB, Sweden. The product was detected at 254 nm.

[0904] Analytical methods Ultra high performance LC (UPLC)-MS analysis was performed on a Waters Acquity system equipped with a Waters BEH300 C18 column (2.1×50mm, particle size 1.7μm, flow rate: 0.25 mL / min, solvent A: water containing 0.04% TFA (v / v), solvent B: acetonitrile containing 0.05% TFA (v / v)) connected to a LTQ Orbitrap Discovery mass spectrometer from Thermo Scientific or a Waters Micromass ZQ.

[0905] Size-exclusion chromatography (SEC) was performed using an Amersham Bioscience AEKTAbasic system equipped with a Superdex 200 5 / 150 GL column (Amersham Bioscience / GE Healthcare) fitted with a 0.45 μm inlet filter, unless otherwise stated. 20 mM sodium phosphate, 140 mM NaCl, pH 7.4 was used as the mobile phase.

[0906] -L 1 Due to the reversible nature of the -L to -D linkage, measurements of NEP stability and receptor affinity were performed using a stable analog...

Claims

1. Use of a CNP prodrug or a pharmaceutically acceptable salt of the prodrug or a pharmaceutical composition comprising the CNP prodrug or a pharmaceutically acceptable salt of the prodrug for the manufacture of a medicament for treating a disease treatable with CNP, wherein the CNP prodrug or a pharmaceutically acceptable salt of the prodrug is -CNP moiety-D, where D has a sequence selected from the group consisting of SEQ ID NOs: 20, 21, 22, 23, 24, 25 and 30; and - moiety - L 2 - via a reversible prodrug linker moiety - L 1 - a carrier moiety - Z attached thereto, wherein said reversible prodrug linker moiety - L 1 - is covalently and reversibly - attached to - D; [moiety-L1-has the structure of formula (IIb'): 【Chemical 1】 (wherein the dashed line represents a bond to the side chain of the amino acid residue of the ring moiety of -D, the dashed line with an asterisk represents a bond to -L2-; -R2 and -R2a are independently selected from -H and C1-6 alkyl; -R3 and -R3a are independently selected from -H and C1-6 alkyl, provided that when one or both of -R3, -R3a are other than -H, they are linked to the N to which they are attached by an sp3 hybridized carbon atom; -X2- is -C(R8R8a)- or -C(R8R8a)-C(R9R9a)-; optionally, said -L1- is further substituted, provided that the hydrogen with an asterisk in formula (IIb') is not replaced by a substituent), -L2- is optionally a C1-20 alkyl chain interrupted by one or more groups independently selected from -O-, -T- and -C(O)N(Ry1)-, and the C1-20 alkyl chain is optionally substituted by a group independently selected from -OH, -T and C(O)N(Ry6Ry6a), where Ry1, Ry6, Ry6a are independently groups selected from the group consisting of H and C1-4 alkyl, and T is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbopolycyclic, and 8- to 30-membered heteropolycyclic; -Z is a branched PEG-based polymer having a molecular weight of at least 10 kDa] Use comprising.

2. The use according to claim 1, wherein -Z is a branched PEG-based polymer having 1, 2, 3, 4, 5 or 6 branch points.

3. The use according to claim 2, wherein each branch point is independently selected from the group consisting of -N<, -CH< and >C<. **Claim 4** The use according to any one of claims 1 to 3, wherein -Z has a molecular weight in the range of 10 kDa to 80 kDa. **Claim 5** -Z is the following moiety 【Chemical 2】 The use according to any one of claims 1 to 4, which comprises. **Claim 6** -Z is the following formula (a) [Chemical Formula 3] [wherein, The dashed line indicates a connection to -L 2 to - or to the remainder of -Z; BP a is a branch point selected from the group consisting of -N<, -CR< and >C<; -R is selected from the group consisting of -H and C 1－6 alkyl; BP a When it is -N< or -CR<, a is 0, and BP a When it is >C<, a is 1; -S a - and -S a′ - and -S a’’ - and -S a’’’ - and -S are each independently a chemical bond, C 1－50 alkyl, C 2－50 alkenyl, and C 2－50 alkynyl, selected from the group consisting of; C 1－50 alkyl, C 2－50 alkenyl and C 2－50 alkynyl may be substituted with one or more identical or different -R 1 ; C 1－50 alkyl, C 2－50 alkenyl and C 2－50 alkynyl may be interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 ), -S(O) 2 N(R 2 ), -S(O)N(R 2 ), -S(O) 2 -, -S(O)-, -N(R 2 ), S(O) 2 N(R 2a ), -S-, -N(R 2 ), -OC(OR 2 )(R 2a ), -N(R 2 ), C(O)N(R 2a ), and -OC(O)N(R 2 ); Each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3－10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbocyclic polycycle, and 8- to 30-membered heterocyclic polycycle; each -T- is independently substituted with one or more identical or different -R 1 and may be substituted; Each -R 1 is independently halogen, -CN, oxo(=O), -COOR 3 -OR 3 -C(O)R 3 -C(O)N(R 3 R 3a ), -S(O) 2 N(R 3 R 3a ), -S(O)N(R 3 R 3a ), -S(O) 2 R 3 ), -S(O)R 3 -N(R 3 ), -N(R 2 S(O) 3a N(R 3b R 3 ), -SR 3 -N(R 3a R 2 ), -NO 3 ), -OC(O)R 3 -N(R 3a ), -N(R 3 ), -N(R 2 S(O) 3a R 3 ), -N(R 3a S(O)R 3 ), -N(R 3a ), -N(R 3 ), -N(R 3a R 3b ), -OC(O)N(R 3 R 3a ), and C 1－6 alkyl; C 1－6 alkyl may be substituted with one or more identical or different halogens; Each -R 2 , -R 2a , -R 3 , -R 3a and -R 3b is independently selected from the group consisting of -H and C 1－6 alkyl, and the C 1－6 alkyl may be substituted with one or more identical or different halogens; -P a’ , -P a’’ and -P a’’’ are independently polymer moieties.] The use according to any one of claims 1 to 5, which comprises the moiety of. **Claim 7** -Z is the following formula (d), [Chemical Formula 4] [wherein, The dashed line indicates the connection to -L 2 -; -Z b - is C 1－50 alkyl, C 2－50 alkenyl, and C 2－50 alkynyl selected from the group consisting of; C 1－50 alkyl, C 2－50 alkenyl and C 2－50 alkynyl may be substituted with one or more identical or different -R 1 and C 1－50 alkyl, C 2－50 alkenyl and C 2－50 alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 ), -S(O) 2 N(R 2 ), -S(O)N(R 2 ), -S(O) 2 -, -S(O)-, -N(R 2 ), S(O) 2 N(R 2a ), -S-, -N(R 2 ), -OC(OR 2 )(R 2a ), -N(R 2 ), C(O)N(R 2a ), and -OC(O)N(R 2 ), and one or more groups selected from the group consisting of may optionally intervene; Each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3－10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclic, 8- to 30-membered carbocyclic polycycle, and 8- to 30-membered heterocyclic polycycle; each -T- is independently optionally substituted with one or more identical or different -R 1 groups; Each -R 1 is independently halogen, -CN, oxo(=O), -COOR 3 -OR 3 -C(O)R 3 -C(O)N(R 3 R 3a ), -S(O) 2 N(R 3 R 3a ), -S(O)N(R 3 R 3a ), -S(O) 2 R 3 ), -S(O)R 3 -N(R 3 ), -N(R 2 S(O) 3a R 3b ), -SR 3 -N(R 3 R 3a ), -NO 2 ), -OC(O)R 3 -N(R 3 ), -N(R 3a C(O)R 3 ), -N(R 2 S(O) 3a R 3 ), -N(R 3a S(O)R 3 ), -N(R 3a C(O)OR 3 ), -N(R 3a C(O)N(R 3b ), -OC(O)N(R 3 R 3a ), and C 1－6 alkyl selected from the group consisting of; C 1－6 alkyl may be substituted with one or more identical or different halogens; Each -R 2 , -R 2a , -R 3 , -R 3a and -R 3b is independently selected from the group consisting of -H and C 1－6 alkyl, and the C 1－6 alkyl may be substituted with one or more identical or different halogens; -Z a is [Chemical Formula 5] is, BP a 、 - S a - 、 - S a’ - 、 - S a’’ 、 - S a’’’ - 、 - P a’ 、 - P a’’ 、 - P a’’’ and a is used as defined in claim 6. The use according to any one of claims 1 to 6, which is of. **Claim 8** The use according to any one of claims 1 to 7, wherein -D has the sequence of SEQ ID NO: 24 or SEQ ID NO: 30.

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