Dry pharmaceutical formulations of CNP conjugates

Abstract

To provide suitable formulations of CNP conjugates comprising CNP covalently linked via a reversible linker to a polymeric moiety, where the peptide will exhibit an acceptable impurity profile and limited premature CNP release even after extended storage.SOLUTION: A dry pharmaceutical formulation is provided. The pharmaceutical formulation comprises a CNP conjugate, a buffering agent, and a bulking agent. The CNP conjugate comprises a CNP moiety that is covalently and reversibly conjugated to a polymeric moiety.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a dry pharmaceutical formulation comprising a CNP conjugate, a buffering agent and a bulking agent. [Background technology]

[0002] Achondroplasia (ACH) is a genetic disorder caused by an autosomal dominant mutation in the fibroblast growth factor receptor 3 (FGFR3) gene, resulting in abnormalities in cartilage formation and dwarfism. C-type natriuretic peptide (CNP) is a hormone that binds to and activates peptide receptor B (NPR-B), resulting in inhibition of FGFR3 downstream signaling. This in turn induces cartilage growth and skeletal overgrowth, as observed in both CNP-overexpressing mice and humans. Overproduction of CNP in cartilage or continuous delivery of CNP via intravenous (iv) infusion normalizes dwarfism in achondroplasia mice, suggesting that administration of CNP at supraphysiological levels is a therapeutic strategy for ACH.

[0003] Human preproCNP, which contains 126 amino acids, is further cleaved by furin to yield CNP-53. CNP-53 is bioactive but is typically processed by an unknown mechanism to a 22-amino acid form, CNP-22, that is bioactive in the circulation. CNP's bioactivity is tightly regulated, and its clearance from plasma is very rapid. Therefore, given CNP's short in vivo half-life (2 minutes after intravenous injection), its use as a therapeutic agent is challenging in the pediatric population, as it would require continuous infusion.

[0004] Various approaches to prolonging the in vivo half-life of CNP have been investigated. For example, Lorget et al. (Am. J. Hum. Genet. 91, 1108-1114, 2012) described recombinant CNP-39 (also called BMN111), which consists of the 37 C-terminal amino acids of human CNP-53 plus glycine and proline added to the N-terminus of the peptide, mimicking the pharmacological activity of human CNP. BMN111 is more resistant to NEP cleavage, but its half-life of only 20 minutes is associated with a short period of exposure to effective drug levels when administered once daily. Summary of the Invention [Problem to be solved by the invention]

[0005] Expanding the scope of in vivo half-life extension of CNPs based on conjugation to water-soluble carrier moieties such as PEG via reversible prodrug linkers has been investigated in WO2016 / 110577A1, WO2017 / 118693A1, WO2017 / 118698A1, WO2017 / 118700A1, WO2017 / 118703A1, WO2017 / 118704A1, and WO2017 / 118707A1. However, no information is provided regarding formulations that allow stable storage of these reversible conjugates.

[0006] Pharmaceutical formulations of such CNP conjugates, in which a polymer is attached to the CNP via a reversible linkage, must provide sufficient CNP conjugate stability to avoid premature CNP release during storage. If the reversible linkage between the polymer and the CNP deteriorates during storage, the concentration of the active drug may increase, potentially creating a risk of overdose upon administration, which may lead to hypotension.

[0007] Furthermore, any drug released during storage will undergo rapid renal clearance when administered to a patient, resulting in a shorter time period during which a long-acting composition provides a therapeutically relevant amount of drug, thereby creating a risk of unmet medical need.

[0008] Furthermore, it is known that CNP or its variants, conjugates or derivatives may undergo degradation reactions upon storage, which may result in impurities / peptide damage in the corresponding formulations, as described below. Degradation products resulting from the oxidation of methionine (Met / M) residues to methionine sulfoxide and methionine sulfone; Degradation products resulting from the isomerization of aspartic acid or aspartate residues (Asp / D) to isoaspartic acid or isoaspartate, for example via a succinimide intermediate; Degradation products resulting from the deamidation of asparagine residues (Asn / N) to aspartic acid or aspartate and / or to isoaspartic acid or isoaspartate, for example via a succinimide intermediate; Degradation products resulting from the deamidation of glutamine residues (Gln / Q) to glutamic acid or isoglutamic acid, for example via glutarimide intermediates; Degradation products resulting from the isomerization of glutamic acid or glutamate residues (Glu / E) to isoglutamic acid or isoglutamate via a glutarimide intermediate; and · Aggregates resulting from peptide aggregation.

[0009] It is desirable to reduce the formation of such degradation products or oligomers during storage, as these products can impair the biological activity of the CNP moiety. Furthermore, the reversible linkage between the CNP moiety and the polymer moiety makes it difficult to store pharmaceutical formulations containing CNP conjugates.

[0010] Therefore, it is important to identify suitable formulations of CNP conjugates comprising CNP covalently linked to a polymer moiety via a reversible linker, where the peptide exhibits an acceptable impurity profile and limited premature CNP release, even after long-term storage.

[0011] SUMMARY OF THE INVENTION It is therefore an object of the present invention to at least partially overcome the above-mentioned drawbacks. [Means for solving the problem]

[0012] This objective is achieved by a dry pharmaceutical formulation comprising a CNP conjugate, a buffer, and a bulking agent, the CNP conjugate comprising a CNP moiety covalently and reversibly bound to a polymer moiety. [Effects of the Invention]

[0013] Surprisingly, it has been found that the dry pharmaceutical formulations of the present invention are capable of stable long-term storage. DETAILED DESCRIPTION OF THE INVENTION

[0014] Within the meaning of the present invention, the terms are used as follows:

[0015] The term "about" used in conjunction with a numerical value herein refers to the numerical value itself, plus or minus a range of up to 10%, in certain embodiments up to 8%, in certain embodiments up to 5%, and in certain embodiments up to 2% of the numerical value. For example, the phrase "about 200" refers to a range of 200 + / - 10%, i.e., 180 to 220, in certain embodiments up to 200 + / - 8%, i.e., 184 to 216, in certain embodiments up to 200 + / - 5%, i.e., 190 to 210, and in certain embodiments up to 200 + / - 2%, i.e., 196 to 204. It is understood that a percentage indicated as "about 20%" does not mean "20% + / - 10%, i.e., a range of 10 to 30%, but rather "about 20%" refers to a range of 18 to 22%, i.e., plus or minus 10% of the numerical value 20.

[0016] As used herein, the term "antimicrobial agent" refers to chemicals, such as chemicals that kill or inhibit the growth of microorganisms, such as bacteria, fungi, yeasts, protozoa, molds, and / or destroy viruses.

[0017] As used herein, the term "anti-adsorption agent" refers primarily to an ionic or non-ionic surfactant, protein, or soluble polymer used to coat or competitively adsorb to the interior surface of a container containing the formulation. The concentration and type of excipient selected will depend on the effect to be avoided, but typically a monolayer of surfactant will form at the interface just above the critical micelle concentration (CMC).

[0018] As used herein, the term "bulking agent" refers to a compound that provides structure to a dried or lyophilized formulation. Bulking agents are added to a formulation to increase the total mass of the dried or lyophilized formulation. In addition to providing a pharmaceutically acceptable formulation, bulking agents can also modify the lyophilization process conditions by altering the collapse temperature, further enhancing peptide stability over long-term storage. Thus, during drying of a mixture containing a bulking agent, the bulking agent can act as a lyoprotectant. During reconstitution of a dried pharmaceutical formulation into a reconstituted pharmaceutical formulation, the bulking agent can act as an isotonicity adjuster.

[0019] As used herein, the term "buffer" or "buffering agent" refers to a compound that maintains pH within a desired range. Physiologically tolerated buffers include, for example, sodium phosphate, succinate, histidine, bicarbonate, citrate, acetate, sulfate, nitrate, chloride, and pyruvate. Antacids such as Mg(OH) or ZnCO can also be used.

[0020] As used herein, the term "CNP" refers, in certain embodiments, to all CNP polypeptides derived from mammalian species, such as humans and mammalian species, particularly humans and murine species, as well as variants, analogs, orthologs, homologs, and derivatives thereof, and fragments thereof, which are characterized by regulating the growth, proliferation, and differentiation of cartilage growth plate chondrocytes. The term "CNP" also includes all CNP variants, analogs, orthologs, homologs, derivatives, and fragments thereof. The CNP variants, analogs, orthologs, homologs, derivatives, and fragments thereof disclosed in WO2009 / 067639A2 and WO2010 / 135541A2 are incorporated herein by reference.

[0021] As used herein, the term "CNP polypeptide variant" refers to a polypeptide derived from the same species that differs from a reference CNP polypeptide. Generally, differences are limited so that the amino acid sequences of the reference and variant are closely similar overall and, in many regions, identical. In certain embodiments, a CNP polypeptide variant is at least 70%, 80%, 90%, or 95% identical to a reference CNP polypeptide. A polypeptide having an amino acid sequence that is, for example, at least 95% "identical" to a query amino acid sequence means that the amino acid sequence of the subject polypeptide is identical to the query sequence, except that the subject polypeptide may contain no more than five amino acid changes per 100 amino acids of the query amino acid sequence. These reference sequence changes may occur at the amino-terminal (N-terminal) or carboxy-terminal (C-terminal) positions of the reference amino acid sequence, or anywhere between these terminal positions, either individually interspersed among residues in the reference sequence or 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. Such CNP polypeptide variants can be naturally occurring variants, e.g., naturally occurring allelic variants encoded by one of several alternative forms of CNP occupying a given genetic locus in a chromosome or organism, or isoforms encoded by naturally occurring splice variants derived from a single primary transcript. Alternatively, CNP polypeptide variants can be variants not known to occur naturally and can be produced by mutation techniques known in the art. It is known in the art that one or more amino acids can be deleted from the N-terminus or C-terminus of a biologically active peptide or protein without significant loss of physiological function. Such N-terminal and / or C-terminal deletions are also encompassed by the term CNP polypeptide variant.

[0022] Those skilled in the art also recognize that some amino acid sequences of CNP polypeptides can be changed 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, guidance on how to make phenotypically silent amino acid substitutions is provided in Bowie et al. (1990), Science 247:1306-1310, which is incorporated herein by reference in its entirety, where the authors show that there are two main approaches to consider the tolerance of amino acid sequence changes.

[0023] As used herein, the term "CNP analog" refers to a CNP from another, unrelated organism that performs the same function in the respective organism but does not originate from an ancestral structure shared by the ancestors of the organisms. Instead, analog CNPs arose separately and subsequently evolved to perform the same or similar functions. In other words, analog CNP polypeptides are polypeptides with significantly different amino acid sequences that achieve the same biological activity, i.e., regulating the growth, proliferation, and differentiation of chondrocytes in the cartilage growth plate.

[0024] As used herein, the term "CNP ortholog" refers to CNPs present in two different species whose sequences are related to each other through a common homologous CNP in an ancestral species, but which have evolved to become distinct from each other.

[0025] As used herein, the term "CNP homolog" refers to CNP from different organisms that perform the same function in each organism and that originate from an ancestral structure shared by the ancestors of those organisms. In other words, a homologous CNP polypeptide is a polypeptide with a highly similar amino acid sequence that achieves the same biological activity, i.e., regulating the growth, proliferation, and differentiation of chondrocytes in the cartilage growth plate. In certain embodiments, 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.

[0026] Thus, a CNP polypeptide can be, for example, (i) a CNP polypeptide in which at least one of the amino acid residues has been substituted with a conserved or non-conserved amino acid residue, and in certain embodiments, with a conserved amino acid residue, where such substituted amino acid residue 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 includes a substituent group; and / or (iii) a CNP polypeptide in which the CNP polypeptide is fused to another compound, such as a compound that increases the half-life of the polypeptide (e.g., polyethylene glycol); and / or (iv) a CNP polypeptide in which additional amino acids have been fused to the CNP polypeptide, such as an IgG Fc fusion region peptide, or a leader sequence, or a secretory sequence, or a sequence used in the purification of such forms of the polypeptide, or a preprotein sequence.

[0027] As used herein, the term "CNP polypeptide fragment" refers to any peptide that includes a contiguous stretch of a portion of the amino acid sequence of a CNP polypeptide.

[0028] More specifically, a CNP polypeptide fragment comprises at least 6 consecutive amino acids of a CNP polypeptide, e.g., at least 8, at least 10, or at least 17 consecutive amino acids. CNP polypeptide fragments can further be described as subgenuses of CNP polypeptides 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. Additionally, CNP polypeptide fragments at least 6 amino acids in length as described above, further specified by their N- and C-terminal positions, are included. The term "CNP polypeptide fragment" also includes all CNP polypeptide fragments at least 6 amino acids in length as described above, which can be specifically specified by their N- and C-terminal positions as individual molecular species. That is, any combination of N- and C-terminal positions that a fragment of at least 6 consecutive amino acid residues in length can occupy on any given amino acid sequence of a CNP polypeptide.

[0029] The term CNP encompasses the above variants, analogs, orthologs, homologs, derivatives and fragments of CNP, and therefore any reference to a specific position in a reference sequence also includes the equivalent position in variants, analogs, orthologs, homologs, derivatives and fragments of the CNP portion, even if not explicitly mentioned.

[0030] Native CNP-22 (SEQ ID NO: 1) has the following sequence: GLSKGCFGLKLDRIGSMSGLGC wherein the cysteines at positions 6 and 22 are linked via a disulfide bridge.

[0031] In certain embodiments, "CNP" refers to the following peptide sequence: SEQ ID NO:2 (CNP-53): DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 3 (G-CNP-53): GDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 4 (M-CNP-53): MDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 5 (P-CNP-53): PDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 6 (CNP-53 M48N): DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC; SEQ ID NO: 7 (CNP-53 Δ15-31): DLRVDTKSRAAWARGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO:8 (CNP-52): LRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 9 (CNP-51): RVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 10 (CNP-50): VDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 11 (CNP-49): DTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 12 (CNP-48): TKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 13 (CNP-47): KSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 14 (CNP-46): SRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 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, where the cysteines at positions 22 and 38 are linked by a disulfide bridge; SEQ ID NO: 25 (CNP-37): QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 26 (CNP-37 Q1pQ, where pQ = pyroglutamic acid): 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; SEQ ID NO: 39 (CNP-35): HPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 40 (CNP-34): PNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 41 (CNP-33): NARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 42 (CNP-32): ARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 43 (CNP-31): RKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 44 (CNP-30): KYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 45 (CNP-29): YKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 46 (CNP-28): KGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 47 (GHKSEVAHRF-CNP-28): GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 48 (CNP-27): GANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 49 (CNP-27 K4Q, K5Q): GANQQGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 50 (CNP-27 K4R, K5R): GANRRGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 51 (CNP-27 K4P, K5R): GANPRGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 52 (CNP-27 K4S, K5S): GANSSGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 53 (CNP-27 K4P, K5R): GANGANPRGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 54 (CNP-27 K4R, K5R, K9R): GANRRGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 55 (CNP-27 K4R, K5R, K9R, M22N): GANRRGLSRGCFGLKLDRIGSNSGLGC; SEQ ID NO: 56 (P-CNP-27 K4R, K5R, K9R): PGANRRGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 57 (M-CNP-27 K4R, K5R, K9R): MGANRRGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 58 (HSA fragment-CNP-27): GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLG; SEQ ID NO: 59 (HSA fragment-CNP-27 M22N): GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSNSGLGC; SEQ ID NO: 60 (M-HSA fragment-CNP-27): MGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 61 (P-HSA fragment-CNP-27): PGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 62 (CNP-26): ANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 63 (CNP-25): NKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 64 (CNP-24): KKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 65 (CNP-23): KGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 66 (R-CNP-22): RGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 67 (ER-CNP-22): ERGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 68 (R-CNP-22 K4R): RGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 69 (ER-CNP-22 4KR): ERGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 70 (RR-CNP-22): RRGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 71 (HRGP fragment-CNP-22): GHHSHEQHPHGANQQGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 72 (HRGP fragment-CNP-22): GAHHPHEHDTHGANQQGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 73 (HRGP fragment-CNP-22): GHHSHEQHPHGANPRGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 74 (IgG1(F c ) fragment-CNP-22): GQPREPQVYTLPSGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 75 (HSA fragment-CNP-22): GQHKDDNPNLPRGANPRGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 76 (HSA fragment-CNP-22): GERAFKAWAVARLSQGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 77 (Osteocrine NPR C inhibitory fragment - CNP22): FGIPMDRIGRNPRGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 78 (FGF2 heparin binding domain fragment-CNP22): GKRTGQYKLGSKTGPGPKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 79 (IgG1(F c ) Fragment-CNP-22 K4R): GQPREPQVYTGANQQGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 80 (HSA fragment-CNP-22 K4R): GVPQVSTSTGANQQGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 81 (Fibronectin fragment-CNP-22 K4R): GQPSSSSQSTGANQQGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 82 (Fibronectin fragment-CNP-22 K4R): GQTHSSGTQSGANQQGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 83 (Fibronectin fragment-CNP-22 K4R): GSTGQWHSESGANQQGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 84 (Zinc finger fragment-CNP-22 K4R): GSSSSSSSSSGANQQGLSRGCFGLKLDRIGSMSGLGC; SEQ ID NO: 85 (CNP-21): LSKGCFGLKLDRIGSMSGLGC; SEQ ID NO: 86 (CNP-20): SKGCFGLKLDRIGSMSGLGC; SEQ ID 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; where Ac = acetyl): Ac-QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC; SEQ ID NO:93: QEHPNARX1YX2GANX3X4GLSX5GCFGLX6LDRIGSMSGLGC, wherein X1, X2, X3, X4, X5, and X6 are independently selected from the group consisting of K, R, P, S, and Q, with the proviso that at least one of X1, X2, X3, X4, X5, and X6 is selected from the group consisting of R, P, S, and Q; in certain embodiments, X1, X2, X3, X4, X5, and X6 are selected from the group consisting of K and R, with the proviso that at least one of X1, X2, X3, X4, X5, and X6 is R; SEQ ID NO:94: QEHPNARKYKGANX1X2GLSX3GCFGLX4LDRIGSMSGLGC, wherein X1, X2, X3, and X4 are independently selected from the group consisting of K, R, P, S, and Q, with the proviso that at least one of X1, X2, X3, and X4 is selected from the group consisting of R, P, S, and Q; in certain embodiments, X1, X2, X3, and X4 are selected from K and R, with the proviso that at least one of X1, X2, X3, and X4 is R; SEQ ID NO:95: QEHPNARKYKGANX1X2GLSKGCFGLKLDRIGSMSGLGC, wherein X1X2 is selected from the group consisting of KR, RK, KP, PK, SS, RS, SR, QK, QR, KQ, RQ, RR and QQ.

[0032] In SEQ ID NOs: 2 to 95, it is understood that the cysteines at positions 22 and 38 of SEQ ID NO: 24 are linked by disulfide bridges.

[0033] The term "CNP" also includes poly(amino acid) conjugates, such as depsipeptides, having the above sequences but with backbones containing both amide bonds as well as non-amide bonds, such as ester bonds. A depsipeptide is a chain of amino acid residues whose backbone contains both amide (peptide) and ester bonds. Thus, the term "side chain," as used herein, refers to the moiety attached to the alpha carbon of an amino acid moiety when the amino acid moieties are linked by amine bonds, as in a polypeptide, or any carbon atom-containing moiety attached to the backbone of a poly(amino acid) conjugate, as in a depsipeptide. In certain embodiments, the term "CNP" refers to a polypeptide having a backbone formed by amide (peptide) bonds.

[0034] The term "ring moiety" as used herein refers to a stretch of consecutive amino acid residues of a CNP drug or CNP moiety located between two cysteine ​​residues that form an intramolecular disulfide bridge or between homologous amino acid residues linked via a chemical cross-linker. Preferably, the ring moiety is located between the 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 ring moiety.

[0035] Regardless of the length of the CNP portion, the sequence of the ring portion of wild-type CNP is FGLKLDRIGSMSGLG (SEQ ID NO: 96).

[0036] As the term CNP includes variants, analogs, orthologs, homologs, derivatives and fragments of CNP as described above, the term "ring moiety" also includes corresponding variants, analogs, orthologs, homologs, derivatives and fragments of the sequence of SEQ ID NO: 96. Thus, any reference to a specific position in a reference sequence also includes the equivalent position in variants, analogs, orthologs, homologs, derivatives and fragments of the CNP moiety, even if not explicitly stated.

[0037] As used herein, the term "cryoprotectant" refers to a compound added to a formulation to protect a drug or drug conjugate during the freezing process.

[0038] As used herein alone or in combination, the term "C 1-4 "Alkyl" means a straight or branched alkyl moiety having 1 to 4 carbon atoms. When present at the terminal end of a molecule, it is 1-4 Examples of alkyl are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. 1-4When bonded by an alkyl, such C 1-4 Examples of alkyl groups are -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -CH(C2H5)-, and -C(CH3)2-. 1-4 Each hydrogen of an alkyl carbon may be optionally replaced by a substituent as defined above. 1-4 The alkyl may be interrupted by one or more moieties as defined below.

[0039] As used herein alone or in combination, the term "C 1-6 "Alkyl" means a straight or branched alkyl moiety having 1 to 6 carbon atoms. When present at the terminal end of a molecule, straight and branched C 1-6 Examples of alkyl groups are 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. 1-6 When bonded by an alkyl group, such C 1-6 Examples of alkyl groups are -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -CH(C2H5)- and -C(CH3)2-. 1-6 Each hydrogen atom of the carbon may optionally be replaced by a substituent as defined above. 1-6 The alkyl may be interrupted by one or more moieties as defined below.

[0040] Therefore, "C 1-10 Alkyl," "C 1-20 Alkyl" or "C 1-50 "Alkyl" means an alkyl chain having 1 to 10, 1 to 20, or 1 to 50 carbon atoms, respectively; C 1-10 , C 1-20 or C 1-50Each hydrogen atom of the carbon may optionally be replaced by a substituent as defined above. 1-10 or C 1-50 The alkyl may be interrupted by one or more moieties as defined below.

[0041] As used herein alone or in combination, the term "C 2-6 "Alkenyl" means a straight or branched hydrocarbon moiety containing at least one carbon-carbon double bond, having 2 to 6 carbon atoms. When present at the end of the molecule, examples are -CH=CH2, -CH=CH-CH3, -CH2-CH=CH2, -CH=CHCH2-CH3, and -CH=CH-CH=CH2. When two parts of the molecule are C 2-6 When bonded by an alkenyl, such C 2-6 An example of an alkenyl is -CH=CH-. 2-6 Each hydrogen atom in the alkenyl moiety may be optionally replaced by a substituent as defined above. 2-6 The alkenyl may be interrupted by one or more moieties as defined below.

[0042] Therefore, the term "C" alone or in combination 2-10 alkenyl," "C 2-20 alkenyl" or "C 2-50 "Alkenyl" means a straight-chain or branched hydrocarbon moiety containing at least one carbon-carbon double bond, having 2 to 10, 2 to 20, or 2 to 50 carbon atoms. 2-10 Alkenyl, C 2-20 Alkenyl or C 2-50 Each hydrogen atom of the alkenyl group may be optionally replaced by a substituent as defined above. 2-10 Alkenyl, C 2-20 Alkenyl or C 2-50 The alkenyl may be interrupted by one or more moieties as defined below.

[0043] As used herein alone or in combination, the term "C 2-6"Alkynyl" means a straight or branched hydrocarbon moiety containing at least one carbon-carbon triple bond, having 2 to 6 carbon atoms. When present at the end of the molecule, examples are -C≡CH, -CH2-C≡CH, CH2-CH2-C≡CH, and CH2-C≡C-CH3. When two parts of the molecule are joined by an alkynyl group, an example is -C≡C-. C 2-6 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 2-6 The alkynyl may be interrupted by one or more moieties as defined below.

[0044] Thus, the term "C" as used herein alone or in combination 2-10 alkynyl", "C 2-20 alkynyl" or "C 2-50 "Alkynyl" means a straight-chain or branched hydrocarbon moiety containing at least one carbon-carbon triple bond, having 2 to 10, 2 to 20, or 2 to 50 carbon atoms, respectively. 2-10 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 The alkynyl may be interrupted by one or more moieties as defined below.

[0045] As mentioned 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, C2-20 Alkenyl or C 2-50 The alkynyl may be optionally interrupted by one or more moieties, which in certain embodiments may be [ka] (In the formula, The dashed line indicates the attachment of the moiety or reagent to the remainder of the -R and -R a are each independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl. is selected from the group consisting of:

[0046] 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, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl. 3-10 Each hydrogen atom of a cycloalkyl carbon may be replaced by a substituent as defined above. 3-10 "Cycloalkyl" also includes bridged bicycles such as norbornane or norbornene.

[0047] The term "8- to 30-membered carbopolycyclyl" or "8- to 30-membered carbopolycycle" refers to a cyclic moiety having 8 to 30 ring atoms, two or more rings, two adjacent rings sharing at least one ring atom, and which may contain up to a maximum number of double bonds (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic rings). Preferably, 8- to 30-membered carbopolycyclyl refers to a cyclic moiety having 2, 3, 4, or 5 rings, more preferably 2, 3, or 4 rings.

[0048] As used herein, the term "3- to 10-membered heterocyclyl" or "3- to 10-membered heterocycle" refers to a ring having 3, 4, 5, 6, 7, 8, 9, or 10 ring atoms, in which at least one ring atom and up to four ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)2-), oxygen, and nitrogen (including =N(O)-), and which may contain up to a maximum number of double bonds (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic rings), and which is attached 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, thiylene, azetidine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazole, isothiazolidine, 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 the 3- to 10-membered heterocyclyl or 3- to 10-membered heterocyclic group may be optionally replaced by a substituent as defined below.

[0049] As used herein, the term "8- to 11-membered heterobicyclyl" or "8- to 11-membered heterobicycle" refers to a two-ring heterocyclic moiety having 8 to 11 ring atoms and which may contain up to a maximum number of double bonds (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic rings), in which at least one ring atom is shared by both rings, and at least one ring atom, up to a maximum of six ring atoms, is replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and the ring is bonded to the remainder of the molecule by a carbon or nitrogen atom. 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 two-ring 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 an 8- to 11-membered heterobicyclyl or an 8- to 11-membered heterobicycle carbon may optionally be replaced by a substituent as defined below.

[0050] Similarly, the terms "8-30 membered heteropolycyclic" or "8-30 membered heteropolycyclic" refer to a heterocyclic moiety of more than two rings, in certain embodiments 3, 4, or 5 rings, having 8 to 30 ring atoms and which may contain up to a maximum number of double bonds (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic rings), wherein two adjacent rings share at least one ring atom, and at least one ring atom, up to 10 ring atoms, is replaced by a heteroatom selected from the group of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and wherein the ring is bonded to the remainder of the molecule by a carbon or nitrogen atom.

[0051] The following structure: [ka] Regarding the part, "Pair R x / R y together with the atoms to which they are attached, C 3-10 The phrase "forms a cycloalkyl or 3- to 10-membered heterocyclyl" refers to R x and R y is understood to mean forming the following structure: [ka] In the formula, R is C 3-10 It is cycloalkyl or 3- to 10-membered heterocyclyl.

[0052] The following structure: [ka] Regarding the part, "Pair R x / R y together with the atoms to which they are attached form a ring A" is x and R y is also understood to mean forming the structure: [ka]

[0053] As used herein, the term "dried pharmaceutical formulation" means that the pharmaceutical formulation is provided in a dry form. Suitable drying methods are spray drying and lyophilization, i.e., freeze-drying. Such dried pharmaceutical formulations containing CNP conjugates have a residual moisture content of up to 10%, in certain embodiments less than 5%, and in certain embodiments less than 2%, as measured by the Karl Fischer method. In certain embodiments, the dried pharmaceutical formulations of the present invention are dried by lyophilization.

[0054] As used herein, the term "drug" refers to a substance used in the treatment, cure, prevention, or diagnosis of disease, or otherwise used to promote physical or mental well-being. When a drug, such as CNP, is attached to another moiety, the portion of the resulting product derived from the drug is referred to as the "drug moiety."

[0055] As used herein, the term "excipient" refers to a compound administered with a drug or pharmaceutical conjugate, such as a buffer, isotonicity adjuster, preservative, stabilizer, anti-adsorption agent, antioxidant, or other auxiliary agent. In some cases, however, a single excipient may have dual or triple functions. The term "excipient" can also refer to a diluent, adjuvant, or carrier used together to administer a drug or pharmaceutical conjugate. Such pharmaceutical additives can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, including, but not limited to, peanut oil, soybean oil, mineral oil, and sesame oil. When the pharmaceutical formulation is administered orally, water is the preferred excipient. When the pharmaceutical formulation is administered intravenously or subcutaneously, saline and aqueous glucose solutions are preferred excipients. In certain embodiments, saline, aqueous glucose solutions, and glycerol solutions are used as liquid excipients for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, mannitol, trehalose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, nonfat dry milk, glycerol, propylene, glycol, water, ethanol, etc. The pharmaceutical formulation may also contain minor amounts of wetting or emulsifying agents, pH buffers such as acetates, succinates, Tris (tris(hydroxymethyl)aminomethane), carbonates, phosphates, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), MES (2-(N-morpholino)ethanesulfonic acid), or surfactants such as Tween®, poloxamer, poloxamine, CHAPS, Igepal®, or amino acids such as glycine, lysine, or histidine. These pharmaceutical formulations can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations, etc. The pharmaceutical formulation can be formulated as a suppository, with traditional binders and excipients such as triglycerides.Oral formulations can contain standard excipients, such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Such formulations will contain a therapeutically effective amount of the drug or drug moiety, together with an appropriate amount of excipients to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

[0056] As used herein, the terms "formulation" or "drug formulation" refer to a preparation containing one or more CNP conjugates and one or more excipients, as well as to a product that results, directly or indirectly, from the combination, complexation, or aggregation of any two or more of the components of the composition, or from the dissociation of one or more of those components, or from any other type of reaction or interaction of one or more of those components. Thus, pharmaceutical formulations of the present invention encompass formulations or compositions made by mixing one or more CNP conjugates and pharmaceutically acceptable excipients, such as buffers and bulking agents.

[0057] As used herein, the term "free form" of a pharmaceutical agent refers to a pharmaceutical agent in its unmodified, pharmacologically fully active form, e.g., after being released from a conjugate.

[0058] As used herein, the term "functional group" means a group of atoms that can react with another group of atoms. Functional groups include, but are not limited to, the following groups: carboxylic acid (-(C=O)OH), primary or secondary amine (-NH, -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 (>NN<), isocyanate, isothiocyanate, phosphate (-O(P=O)OHOH), phosphonate (-O(P=O)OHH), haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine, disulfide, sulfonamide, sulfonic acid, vinyl sulfone, vinyl ketone, diazoalkane, oxirane, and aziridine.

[0059] As used herein, the term "halogen" means fluoro, chloro, bromo, or iodo. It is generally preferred that the halogen is fluoro or chloro.

[0060] As used herein, the term "interrupted" means that a moiety is inserted between two carbon atoms, or, if the insertion is at one of the ends of the moiety, between a carbon or heteroatom and a hydrogen atom, and in certain embodiments, between a carbon and a hydrogen atom.

[0061] As used herein, the term "tonicity adjusting agent" refers to a compound that reduces pain, irritation, and tissue damage that can result from cell damage due to the osmotic pressure difference between the injected solution and the plasma.

[0062] As used herein, the term "lyophilized formulation" means that a formulation containing a CNP conjugate is first frozen, followed by removal of water by vacuum. This term does not exclude an additional drying step that occurs in the manufacturing process before the formulation is filled into the final container.

[0063] As used herein, the terms "lyophilization" or "freeze-drying" are used interchangeably and refer to a dehydration process characterized by freezing a formulation, followed by reduction in ambient pressure and, optionally, application of heat, allowing the frozen water in the formulation to sublime directly from the solid phase to a gas. Typically, the displaced water is recovered by sublimation.

[0064] As used herein, the term "lyoprotectant" refers to a molecule that, when combined with an active ingredient of interest, significantly prevents or reduces the chemical and / or physical instability of the active ingredient during drying in general, lyophilization in particular, and subsequent storage. Exemplary lyoprotectants include sugars, such as sucrose or trehalose; amino acids, such as sodium glutamate, histidine, or arginine; methylamines, such as betaine; lyotropic salts, such as magnesium sulfate; polyhydric alcohols, such as trihydric or higher sugar alcohols, such as glycerin, erythritol, glycerol, arabitol, xylitol, sorbitol, and mannitol; ethylene glycol; propylene glycol; polyethylene glycol; Pluronics; hydroxyalkyl starches, such as hydroxyethyl starch (HES), and combinations thereof. Lyoprotectants can also serve as bulking agents in dry pharmaceutical formulations and as tonicity adjusters in reconstituted pharmaceutical formulations.

[0065] As used herein, the term "moiety" refers to a portion of a molecule that lacks one or more atoms compared to the corresponding drug. For example, when a reagent of the formula "HXH" reacts with another reagent and becomes part of a reaction product, the corresponding moiety in the reaction product has the structure "HX-" or "-X-," where each "-" indicates a bond to another moiety. Thus, a drug moiety, such as a CNP moiety, is released from the conjugate as a drug, such as CNP.

[0066] Where an arrangement or chemical structure of atoms is provided in which the arrangement or chemical structure is attached to two moieties or interrupts one moiety, it is understood that the arrangement or chemical structure may be attached to the two moieties in either orientation, unless expressly stated otherwise. For example, the moiety "-C(O)N(R 1 )-" is "-C(O)N(R 1 )-" or "-N(R 1 )C(O)-" can be attached to two moieties or can interrupt one moiety. Similarly, the moieties: [ka] teeth, [ka] as, or [ka] As such, it can be joined to two parts or interrupted into one part.

[0067] When the CNP moiety contains one or more acidic or basic groups, the pharmaceutical formulation also contains their corresponding pharmaceutically or toxicologically acceptable salts, particularly their pharmaceutically acceptable salts. Thus, CNP moieties containing one or more acidic groups can be present and used, for example, as alkali metal salts, alkaline earth metal salts, or ammonium salts. More specific 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, as well as other salts or amines known to those skilled in the art. CNP moieties containing one or more basic groups, i.e., groups that can be protonated, can also be present and used 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 are known to those skilled in the art, such as alkylation of amine groups to provide a positively charged ammonium group and a suitable counterion for its salt. When the CNP moiety contains both an acidic and a basic group, the pharmaceutical formulation according to the present invention also includes an internal salt or betaine (zwitterion) in addition to the salt forms mentioned above. The respective salts can be obtained by conventional methods known to those skilled in the art, such as by contacting these conjugates with organic or inorganic acids or bases in a solvent or dispersant, or by anion or cation exchange with other salts. The formulations according to the present invention also include all salts of CNP conjugates that are not suitable for direct use in pharmaceuticals due to their low physiological compatibility, but can be used, for example, as intermediates in chemical reactions or for the preparation of pharmaceutically acceptable salts.

[0068] As used herein, the term "antioxidant" or "oxidation protectant" refers to a compound that inhibits the oxidation of other compounds, such as peptides.

[0069] As used herein, the term "pH adjuster" refers to a compound used to adjust the pH of the formulation solution prior to filling and lyophilization.

[0070] As used herein, the term "pharmaceutically acceptable" means a substance that is not harmful when administered to a patient, and preferably means approved for use in animals, preferably for use in humans, by a regulatory authority, such as the EMA (Europe) and / or the FDA (USA) and / or a regulatory authority of any other country.

[0071] As used herein, the term "physiological conditions" refers to aqueous buffered conditions of pH 7.4 and 37°C.

[0072] As used herein, the term "polypeptide" refers to a chain of two to fifty amino acid monomer moieties linked by peptide (amide) linkages. In the case of CNP drugs and CNP moieties only, sequences having more than 50 amino acids are also referred to simply as "polypeptides."

[0073] As used herein, the term "preservative" refers to a chemical that has antimicrobial effects and prevents chemical degradation.

[0074] As used herein, the term "protein" refers to a chain of more than 50 amino acid monomer moieties linked by peptide linkages, preferably 12,000 or fewer amino acid monomers linked by peptide linkages, such as 10,000 or fewer amino acid monomer moieties, 8,000 or fewer amino acid monomer moieties, 5,000 or fewer amino acid monomer moieties, or 2,000 or fewer amino acid monomer moieties.

[0075] As used herein, the term "polymer" refers to a molecule comprising repeating structural units, i.e., monomers, linked by chemical bonds in a linear, cyclic, branched, cross-linked, or dendrimeric fashion, or a combination thereof, and may be of synthetic or biological origin, or a combination of both. It is understood that a polymer may also comprise one or more other chemical groups and / or moieties, such as, for example, one or more functional groups. In certain embodiments, a soluble polymer has a molecular weight of at least 0.5 kDa, e.g., at least 1 kDa, at least 2 kDa, at least 3 kDa, or at least 5 kDa. When a polymer is soluble, in certain embodiments, it has a molecular weight of at most 1000 kDa, e.g., at most 750 kDa, e.g., at most 500 kDa, e.g., at most 300 kDa, e.g., at most 200 kDa, e.g., at most 100 kDa.

[0076] It is understood that a protein or polypeptide is also a polymer in which amino acids are repeating structural units, even though the side chain of each amino acid may be different.

[0077] As used herein, the term "polymeric" or "polymeric moiety" means an agent or moiety that comprises one or more polymers or polymer moieties. A polymeric agent or moiety may optionally also comprise one or more other moieties, which in certain embodiments are 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 heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl, and A linkage selected from the group comprising: [ka] During the ceremony, The dashed line indicates the attachment of the moiety or reagent to the remainder of the -R and -Ra are each independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl, and hexyl.

[0078] Those skilled in the art will appreciate that the polymerization products resulting from a polymerization reaction will not all have the same molecular weight, but rather will exhibit a distribution of molecular weights. Consequently, as used herein, molecular weight ranges, molecular weights, ranges of the number of monomers in a polymer, and numbers of monomers in a polymer refer to number average molecular weights and monomer number averages, i.e., the arithmetic mean of the molecular weights of the polymers or polymer segments and the arithmetic mean of the numbers of monomers in the polymers or polymer segments.

[0079] Thus, for a polymer segment containing "x" monomer units, any integer assigned to "x" corresponds to the arithmetic average number of monomers. Any range of integers assigned to "x" provides a range of integers within which the arithmetic average number of monomers lies. An integer "x" designated as "about x" means that the arithmetic average number of monomers is within the integer range of x + / - 10%, in certain embodiments within the integer range of x + / - 8%, in certain embodiments within the integer range of x + / - 5%, and in certain embodiments within the integer range of x + / - 2%.

[0080] As used herein, the term "PEG-based" with respect to a moiety or reagent means that the moiety or reagent comprises PEG. In certain embodiments, the PEG-based moiety or reagent comprises at least 10% by weight PEG, such as at least 20% by weight PEG, for example at least 30% by weight PEG, such as at least 40% by weight PEG, for example at least 50% by weight, such as at least 60% by weight PEG, for example at least 70% by weight PEG, such as at least 80% by weight PEG, for example at least 90% by weight, such as at least 95% by weight PEG. The remaining weight percentages of the PEG-based moiety or reagent are other moieties selected from the following moieties and linkages: C 1-50 Alkyl, C 2-50 Alkenyl, C2-50 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl, and A linkage selected from the group comprising: [ka] During the ceremony, The dashed line indicates the attachment of the moiety or reagent to the remainder of the -R and -R a are each independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl, and hexyl.

[0081] As used herein, the term "PEG-based comprising at least X% PEG" in reference to a moiety or reagent means that the moiety or reagent comprises at least X% by weight ethylene glycol units (-CHCHO-), which may be arranged in alternating blocks or may be randomly distributed within the moiety or reagent, and in certain embodiments, all of the ethylene glycol units of the moiety or reagent are present in one block, with the remaining weight percentage of the PEG-based moiety or reagent being other moieties selected from the following moieties and linkages in certain embodiments: 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 heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl, and A linkage selected from the group comprising: [ka] During the ceremony, The dashed line indicates the attachment of the moiety or reagent to the remainder of the -R and -R aare each independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl, and hexyl.

[0082] As used herein, the term "a hyaluronic acid system containing at least X% hyaluronic acid" is used arbitrarily.

[0083] Those skilled in the art will also recognize that the conjugates of the present invention are prodrugs. As used herein, the term "prodrug" refers to a pharmaceutical moiety, such as a CNP moiety, reversibly and covalently bonded to a polymer moiety, such as -Z, via a reversible linker moiety. A prodrug releases the reversibly and covalently bonded pharmaceutical moiety in its corresponding pharmaceutical form. That is, a prodrug is a conjugate containing a pharmaceutical moiety, such as a CNP moiety, covalently and reversibly bonded to a polymer moiety via a reversible linker moiety, where the covalent and reversible bond of the polymer moiety to the reversible linker moiety is direct or via a spacer. Such a prodrug or conjugate releases the previously attached pharmaceutical moiety in the form of a free pharmaceutical.

[0084] As used herein, the term "random coil" refers to a peptide or protein that adopts / has / forms, in certain embodiments, a conformation substantially devoid of defined secondary and tertiary structure, as determined by circular dichroism spectroscopy performed in aqueous buffer at pH 7.4 at room temperature. In certain embodiments, room temperature (ambient temperature) is about 20°C, i.e., 18°C ​​to 22°C, while in certain embodiments, ambient temperature is 20°C.

[0085] As used herein, the term "reversible linkage" refers to a linkage that is cleavable in the absence of enzymes under physiological conditions (aqueous buffer at pH 7.4, 37°C) and has a half-life ranging from 1 hour to 6 months, e.g., 1 hour to 4 months, e.g., 1 hour to 3 months, 1 hour to 2 months, or 1 hour to 1 month.

[0086] As used herein, the term "reagent" means a compound that contains at least one functional group for reaction with a functional group of another compound or drug. It is understood that drugs that contain a functional group (e.g., a primary or secondary amine or hydroxyl functionality) are also drugs.

[0087] As used herein, the term "reversible linker moiety" refers to a moiety that is covalently attached to a pharmaceutical moiety, such as a CNP moiety, by a reversible linkage, and that is also covalently attached to a polymer moiety, such as -Z, where the covalent attachment to the polymer moiety is direct or via -L 2 In certain embodiments, -Z and -L 2 - is a stable connection.

[0088] As used herein, the term "reconstitution" means the addition of a liquid to a dry pharmaceutical formulation to return the formulation to its original form, e.g., a liquid.

[0089] As used herein, the term "reconstituted formulation" refers to a formulation obtained upon reconstitution of a dry pharmaceutical formulation by adding a reconstitution solution.

[0090] As used herein, the term "reconstitution solution" refers to a liquid used to reconstitute a dry pharmaceutical formulation prior to administration to a patient in need.

[0091] As used herein, the term "spacer" or "spacer moiety" refers to a moiety suitable for connecting two moieties. Suitable spacers include C 1-50 Alkyl, C 2-50 Alkenyl or C 2-50 alkynyl, and C 1-50 Alkyl, C 2-50 Alkenyl or C 2-50 Alkynyl is -NH-, -N(C 1-4 alkyl)-, -O-, -S-, -C(O)-, -C(O)NH-, -C(O)N(C 1-4It may be interrupted by one or more groups selected from alkyl)-, -OC(O)-, -S(O)-, -S(O)2-, 4- to 7-membered heterocyclyl, phenyl and naphthyl.

[0092] As used herein, the term "substituted" means that one or more -H atoms of a molecule or moiety are replaced by a different atom or group of atoms, referred to as "substituents."

[0093] In certain embodiments, such one or more substituents may be, independently of each other, halogen, —CN, —COOR x1 , -OR x1 , -C(O)R x1 , -C(O)N(R x1 R x1a ), -S(O)2N(R x1 R x1a ), -S(O)N(R x1 R x1a ), -S(O)R x1 , -S(O)R x1 , -N(R x1 )S(O)2N(R x1a R x1b ), -SR x1 , -N(R x1 R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R 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; -T 0 , C 1-50 Alkyl, C2-50 Alkenyl and C 2-50 An alkynyl group may be one or more -R x2 may be substituted with C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(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 )-;

[0094] -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; -T 0 , C 1-50 Alkyl, C 2-50 Alkenyl, and C 2-50 An alkynyl group may be one or more -R x2 may be substituted with C 1-50 Alkyl, C 2-50 Alkenyl, and C 2-50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-;-S(O)2-, -S(O)-, -N(R x3 )S(O)2N(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 )-;

[0095] Each T 0 are independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 is selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; each T 0 are independently one or more -R x2 may be substituted with;

[0096] Each-R x2 are independently halogen, -CN, oxo (=O), -COOR x4 , -OR x4 , -C(O)R x4 , -C(O)N(R x4 R x4a ), -S(O)2N(R x4 R x4a ), -S(O)N(R x4 R x4a ), -S(O)R x4 , -S(O)R x4 , -N(R x4 )S(O)2N(R x4a R x4b ), -SR x4 , -N(R x4 R x4a ), -NO2, -OC(O)R x4 , -N(R x4 )C(O)R x4a , -N(R x4 )S(O)2R 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 alkyl; C 1-6 The alkyl may be optionally substituted with one or more halogens, which may be the same or different;

[0097] Each-R x3 , -R x3a , -R x4 , -R x4a , -R x4b are independently -H and C 1-6 alkyl; C 1-6 The alkyl may be substituted with one or more halogens which may be the same or different.

[0098] In certain embodiments, the one or more 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)2N(R x1 R x1a ), -S(O)N(R x1 R x1a ), -S(O)R x1 , -S(O)R x1 , -N(R x1 )S(O)2N(R x1a R x1b ), -SR x1 , -N(R x1 R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R 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 ), -T0 , C 1-10 Alkyl, C 2-10 Alkenyl, and C 2-10 alkynyl; -T 0 , C 1-10 Alkyl, C 2-10 Alkenyl, and C 2-10 An alkynyl group may be one or more -R x2 may be substituted with C 1-10 Alkyl, C 2-10 Alkenyl, and C 2-10 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(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 )-;

[0099] Each-R x1 , -R x1a , -R x1b , -R x3 , -R x3a are independently -H, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, and C 2-6 alkynyl;

[0100] Each T 0 are independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 is selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; each T 0 are independently one or more -R x2may be substituted with;

[0101] Each-R x2 are independently halogen, -CN, oxo (=O), -COOR x4 , -OR x4 , -C(O)R x4 , -C(O)N(R x4 R x4a ), -S(O)2N(R x4 R x4a ), -S(O)N(R x4 R x4a ), -S(O)R x4 , -S(O)R x4 , -N(R x4 )S(O)2N(R x4a R x4b ), -SR x4 , -N(R x4 R x4a ), -NO2, -OC(O)R x4 , -N(R x4 )C(O)R x4a , -N(R x4 )S(O)2R 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 alkyl; C 1-6 The alkyl may be optionally substituted with one or more halogens, which may be the same or different;

[0102] Each-R x4 , -R x4a , -R x4b are independently -H, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, and C 2-6 alkynyl.

[0103] In certain embodiments, the one or more 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)2N(R x1 R x1a ), -S(O)N(R x1 R x1a ), -S(O)R x1 , -S(O)R x1 , -N(R x1 )S(O)2N(R x1a R x1b ), -SR x1 , -N(R x1 R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R 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; -T 0 , C 1-6 Alkyl, C 2-6 Alkenyl, and C 2-6 An alkynyl group may be one or more -R x2 may be substituted with C 1-6 Alkyl, C 2-6 Alkenyl, and C 2-6 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3)-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(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 )-;

[0104] Each-R x1 , -R x1a , -R x1b , -R x2 , -R x3 , -R x3a are independently -H, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, and C 2-6 alkynyl;

[0105] Each T 0 are independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 is selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; each T 0 are independently one or more -R x2 may be substituted with.

[0106] In certain embodiments, up to six H atoms of an optionally substituted molecule are independently replaced by substituents, e.g., five H atoms are independently replaced by substituents, four H atoms are independently replaced by substituents, three H atoms are independently replaced by substituents, two H atoms are independently replaced by substituents, or one H atom is independently replaced by a substituent.

[0107] As used herein, the terms "stable" and "stability" in relation to a pharmaceutical formulation mean that after a storage period, e.g., 1 month, 2 months, 4 months, 6 months, 8 months, 12 months, 18 months, 24 months, 36 months, 48 ​​months, or 60 months, particularly after the specified storage period, the pharmaceutical formulation contains less than 5% of the drug in free form and less than 20%, e.g., less than 10%, or less than 5% of impurities, such as impurities resulting from aspartic acid or aspartic acid isomerization, methionine oxidation, and peptide aggregation. Impurities can be quantified by RP-HPLC or SEC based on the area of ​​each peak relative to the total area of ​​all CNP conjugate-related peaks in a chromatogram; impurities in the CNP portion of the CNP conjugate can be determined by proteolytic digestion and quantified based on the area of ​​each peak relative to the area of ​​the corresponding unmodified proteolytic peptide.

[0108] As used herein, the term "stabilizer" refers to a compound used to stabilize a pharmaceutical conjugate. Stabilization can be achieved by enhancing the peptide's stabilizing power or by direct binding of an excipient to the pharmaceutical conjugate.

[0109] As used herein, the term "surfactant" refers to a wetting agent that reduces the surface tension of a liquid.

[0110] As used herein, the term "sealing a container" means sealing the container airtight to prevent any gas exchange between the outside and the inside, and to maintain the sterility of the contents.

[0111] As used herein, the term "therapeutically effective amount" refers to an amount sufficient to cure, alleviate, or partially arrest the clinical symptoms of a given disease and its complications. The amount effective for each purpose will depend on the severity of the disease or injury, as well as the weight and general condition of the subject. As will be appreciated, determining the appropriate dosage can be accomplished using routine experimentation by constructing a matrix of values ​​and testing various points within the matrix, all of which is within the ordinary skill of a skilled physician. Within the scope of the present invention, a therapeutically effective amount refers to a dosage that aims to achieve a therapeutic effect over a prolonged period of time, for example, for at least 1 day, e.g., 2 days, e.g., 3 days, e.g., 4 days, e.g., 5 days, e.g., 6 days, e.g., 1 week, or e.g., 2 weeks.

[0112] As used herein, the term "traceless linker" means a reversible linker that, upon cleavage, releases the drug in its free form.

[0113] As used herein, the term "unit dose" means the amount of medication administered to a patient in a single dose.

[0114] As used herein, the term "water-soluble" in reference to a polymer moiety means that when such a polymer moiety is part of a CNP conjugate, at least 1 g of a CNP conjugate comprising such a water-soluble polymer moiety can be dissolved in 1 liter of water at 20°C to form a homogeneous solution.

[0115] In general, the terms "comprise" or "comprising" also encompass "consist of" or "consisting of."

[0116] In certain embodiments, the CNP portion of the CNP conjugate has the sequence of 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:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, or SEQ ID NO:30. In certain embodiments, the CNP portion has the sequence of SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, or SEQ ID NO:25. In certain embodiments, the CNP portion has the sequence of SEQ ID NO:20. In certain embodiments, the CNP portion has the sequence of SEQ ID NO:21. In certain embodiments, the CNP portion has the sequence of SEQ ID NO:22. In certain embodiments, the CNP portion has the sequence of SEQ ID NO:23. In certain embodiments, the CNP portion has the sequence of SEQ ID NO:24. In certain embodiments, the CNP portion has the sequence of SEQ ID NO:25.

[0117] In certain embodiments, the polymeric moiety is defined as variable -Z, which is described in more detail elsewhere herein.

[0118] The dry pharmaceutical formulation according to the present invention comprises a buffer. Examples of buffers can be selected from the group consisting of succinic acid, citric acid, lactic acid, acetic acid, glutamic acid, fumaric acid, aspartic acid, glutaric acid, phosphoric acid, histidine, gluconic acid, tartaric acid, malic acid, and mixtures thereof. It will be apparent to those skilled in the art that the corresponding conjugate bases or salts of the buffers can also be included, such as succinate, citrate, lactate, acetate, glutamate, fumarate, aspartate, glutarate, phosphate, gluconate, tartrate, malate, and mixtures thereof.

[0119] In certain embodiments, the buffering agent is succinic acid. In certain embodiments, the buffering agent is citric acid. In certain embodiments, the buffering agent is lactic acid. In certain embodiments, the buffering agent is acetic acid. In certain embodiments, the buffering agent is glutamic acid. In certain embodiments, the buffering agent is fumaric acid. In certain embodiments, the buffering agent is aspartic acid. In certain embodiments, the buffering agent is glutaric acid. In certain embodiments, the buffering agent is phosphate. In certain embodiments, the buffering agent is histidine. In certain embodiments, the buffering agent is gluconic acid. In certain embodiments, the buffering agent is tartaric acid. In certain embodiments, the buffering agent is malic acid.

[0120] In certain embodiments, the buffering agent has a concentration of about 0.2% to about 3.2% by weight. In certain embodiments, the buffering agent has a concentration of about 0.6% to about 1.6% by weight. In certain embodiments, the buffering agent has a concentration of about 0.9% to about 1.0% by weight. In certain embodiments, the buffering agent has a concentration of about 1.0% by weight.

[0121] The dry pharmaceutical formulation according to the present invention comprises a bulking agent.

[0122] The bulking agent may be selected from the group consisting of trehalose, mannitol, sucrose, raffinose, gelatin, lactose, dipotassium phosphate, sorbitol, xylitol, glycine, histidine, hydroxyethyl starch, dextrose, dextran, Ficoll®, propylene glycol, and mixtures thereof.

[0123] In certain embodiments, the bulking agent may be selected from the group consisting of trehalose, mannitol, sucrose, raffinose, gelatin, lactose, dipotassium phosphate, sorbitol, xylitol, glycine, histidine, hydroxyethyl starch, dextrose, dextran, propylene glycol, and mixtures thereof.

[0124] In certain embodiments, the bulking agent is selected from the group consisting of trehalose, sucrose, and glycine.

[0125] In certain embodiments, the bulking agent is a non-reducing sugar such as trehalose or sucrose.

[0126] In certain embodiments, the bulking agent is trehalose. In certain embodiments, the bulking agent is mannitol. In certain embodiments, the bulking agent is sucrose. In certain embodiments, the bulking agent is raffinose. In certain embodiments, the bulking agent is gelatin. In certain embodiments, the bulking agent is lactose. In certain embodiments, the bulking agent is dipotassium hydrogen phosphate. In certain embodiments, the bulking agent is sorbitol. In certain embodiments, the bulking agent is xylitol. In certain embodiments, the bulking agent is glycine. In certain embodiments, the bulking agent is histidine. In certain embodiments, the bulking agent is hydroxyethyl starch. In certain embodiments, the bulking agent is dextrose. In certain embodiments, the bulking agent is dextran. In certain embodiments, the bulking agent is Ficoll®. In certain embodiments, the bulking agent is propylene glycol.

[0127] As defined herein, the term "trehalose" is intended to encompass all salts and hydrated forms of trehalose, such as trehalose anhydrate or trehalose dihydrate. In certain embodiments, the term "trehalose" refers to trehalose anhydrate. In certain embodiments, the term "trehalose" refers to trehalose dihydrate.

[0128] In certain embodiments, the formulation may include multiple bulking agents.

[0129] In certain embodiments, reducing sugars should be avoided as they may react with the CNP moiety. Thus, in certain embodiments, the pharmaceutical formulation does not include reducing sugars.

[0130] In certain embodiments, the bulking agent has a concentration ranging from about 52.6% to about 98.4% by weight. In certain embodiments, the bulking agent has a concentration ranging from about 62.4% to about 70.4% by weight. In certain embodiments, the bulking agent has a concentration ranging from about 85.7% to about 91.6% by weight. In certain embodiments, the bulking agent has a concentration ranging from about 65.2% by weight.

[0131] The dry pharmaceutical formulation according to the present invention comprises a pH adjusting agent. In certain embodiments, the pH adjusting agent is an acid or an acid salt thereof. The acid may be selected from the group consisting of hydrochloric acid, phosphoric acid, carbonic acid, nitric acid, and mixtures thereof.

[0132] In certain embodiments, the pH adjuster is hydrochloric acid. In certain embodiments, the pH adjuster is phosphoric acid. In certain embodiments, the pH adjuster is carbonic acid. In certain embodiments, the pH adjuster is nitric acid.

[0133] In certain embodiments, the pH adjuster is a base or a basic salt thereof, which may be selected from the group consisting of Tris (tris(hydroxymethyl)aminomethane), sodium hydroxide, potassium hydroxide, lysine, and mixtures thereof.

[0134] In certain embodiments, the pH adjuster is Tris. In certain embodiments, the pH adjuster is sodium hydroxide. In certain embodiments, the pH adjuster is potassium hydroxide. In certain embodiments, the pH adjuster is lysine.

[0135] In certain embodiments, a dry pharmaceutical formulation according to the present invention comprises a mixture of one or more acid and base pH adjusting agents.

[0136] In certain embodiments, the pH adjuster or mixture of pH adjusters has a concentration of about 0.1% to about 5.6% by weight. In certain embodiments, the pH adjuster or mixture of pH adjusters has a concentration of about 0.2% to about 2.8% by weight. In certain embodiments, the pH adjuster or mixture of pH adjusters has a concentration of about 0.5% to about 1.4% by weight. In certain embodiments, the pH adjuster or mixture of pH adjusters has a concentration of about 1.2% by weight. It is understood that in the case of a mixture of pH adjusters, the concentrations provided refer to the combined concentration of all pH adjusters.

[0137] The dry pharmaceutical formulation of the present invention comprises a CNP conjugate.

[0138] In certain embodiments, the CNP conjugate has a concentration of about 23.8% to about 38.7% by weight, about 5.8% to about 12.4% by weight, or about 1.4% to about 3.7% by weight.

[0139] In certain embodiments, the CNP conjugate is of formula (Ia) or (Ib): [ka] During the ceremony, -D is a CNP moiety; -L 1 - is a reversible linker moiety; -L 2 - is a single chemical bond or spacer moiety; -Z is a polymeric moiety; x is an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 and 16; y is an integer selected from the group consisting of 1, 2, 3, 4 and 5.

[0140] -D in formula (Ia) or (Ib) is -L 1 - is covalently and reversibly bound to

[0141] In certain embodiments, x in Formula (Ia) is an integer selected from the group consisting of 1, 2, 3, 4, 6, and 8. In certain embodiments, x in Formula (Ia) is an integer selected from the group consisting of 1, 2, 4, and 6. In certain embodiments, x in Formula (Ia) is an integer selected from the group consisting of 1, 4, and 6, and in certain embodiments, x in Formula (Ia) is 1.

[0142] In certain embodiments, y in Formula (Ib) is an integer selected from the group consisting of 2, 3, 4, and 5. In certain embodiments, y in Formula (Ib) is an integer selected from the group consisting of 2, 3, and 4. In certain embodiments, y in Formula (Ib) is an integer selected from the group consisting of 2 and 3. In certain embodiments, y in Formula (Ib) is an integer selected from the group consisting of 1, 2, and 3. In certain embodiments, y in Formula (Ib) is 1. In certain embodiments, y in Formula (Ib) is 2.

[0143] In certain embodiments, the CNP conjugate is of formula (Ia) where x=1.

[0144] In certain embodiments, -D of Formula (Ia) or (Ib) has the sequence of 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: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, or SEQ ID NO: 30. In certain embodiments, -D of Formula (Ia) or (Ib) has the sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 25.

[0145] In certain embodiments, -D of formula (Ia) or (Ib) has the sequence of SEQ ID NO: 20. In certain embodiments, -D of formula (Ia) or (Ib) has the sequence of SEQ ID NO: 21. In certain embodiments, -D of formula (Ia) or (Ib) has the sequence of SEQ ID NO: 22. In certain embodiments, -D of formula (Ia) or (Ib) has the sequence of SEQ ID NO: 23. In certain embodiments, -D of formula (Ia) or (Ib) has the sequence of SEQ ID NO: 24. In certain embodiments, -D of formula (Ia) or (Ib) has the sequence of SEQ ID NO: 25.

[0146] The moiety -L of formula (Ia) or (Ib) 1 - is attached to a functional group in the side chain of an amino acid residue of -D, to the N-terminal amine functional group or the C-terminal carboxyl functional group of -D, or to a nitrogen atom in the backbone polypeptide chain of -D. Attachment to either the N-terminus or C-terminus can be direct via the corresponding amine or carboxyl functional group, respectively, or a spacer moiety can first be attached to the spacer moiety -L. 1 The bond can be indirect, being attached to an amine or carboxyl functional group to which the - is attached.

[0147] The moiety -L of formula (Ia) or (Ib) 1 - is a reversible prodrug linker whereby the drug, i.e., DH, is released in its free form, i.e., -L 1 - is a traceless linker. Suitable reversible linkers are known in the art, such as the reversible linker moieties 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.

[0148] In certain embodiments, -L 1- is a reversible 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.

[0149] Part-L 1 - can be linked to -D via any type of linkage, provided that it is reversible. In certain embodiments, -L 1 - is linked to -D through a linkage selected from the group consisting of amide, ester, carbamate, acetal, aminal, imine, oxime, hydrazone, disulfide, and acylguanidine. 1 - is linked to -D through a linkage selected from the group consisting of amide, ester, carbamate, and acylguanidine. These linkages may not themselves be reversible, and -L 1 It is understood that the adjacent group contained in - may be one that makes the linkage reversible.

[0150] In certain embodiments, the moiety -L 1 - is linked to -D via an amide linkage.

[0151] Part-L 1 - is disclosed in WO 2009 / 095479 A2. Thus, in certain embodiments, the moiety -L 1 - is a group represented by the formula (II):

[0152] [ka] (In the formula, The dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond, and -X- is -C(R 4 R 4a )-, -N(R 4 )-, -O-, -C(R 4 R 4a )-C(R5 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-, -OC(R 4 R 4a )- or -C(R 7 R 7a )- and X 1 is C or S(O), -X 2 - is -C(R 8 R 8a )- or -C(R 8 R 8a )-C(R 9 R 9a )- 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 -H and C 1-6 is selected from the group consisting of alkyl, -R 3 , -R 3a are independently -H and C 1-6 alkyl, with the proviso that -R 3 , -R 3a If one or both of are other than -H, they may be attached to the N to which they are attached by SP 3 are linked by hybridized carbon atoms, -R7 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 is alkyl, In some cases, 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 the following forms a chemical bond; In some cases, Pair-R 1 / -R 1a , -R 2 / -R 2a , -R 4 / -R 4a , -R 5 / -R 5a , -R 8 / -R 8a , -R 9 / -R 9a One or more of the following, together with the atoms to which they are attached, form a C 3-10 forming a cycloalkyl or a 3- to 10-membered heterocyclyl; In some cases, 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 together with the atoms to which they are attached form ring A, In some cases, R 3 / R 3a together with the nitrogen atom to which they are attached form a 3- to 10-membered heterocyclic ring, A is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; -L 1 -L 2 -Z and optionally -L 1 - is further substituted, provided that the hydrogen with an asterisk in formula (II) is -L 2 is not replaced by -Z or a substituent, -L 2 - is a single chemical bond or spacer, -Z is a water-soluble polymer moiety.

[0153] In certain embodiments, -L of formula (II) 1 - is one part - L 2 Replaced with -Z.

[0154] In certain embodiments, -L of formula (II) 1 - is not further substituted.

[0155] -R in formula (II) 3 / -R 3a However, when they are combined with the nitrogen atom to form a 3- to 10-membered heterocycle, the atom directly bonded to the nitrogen is SP 3 It is understood that only such 3- to 10-membered heterocyclic rings can be formed that are hybridized carbon atoms. 3 / -R 3a and the nitrogen atom to which they are attached have the following structure: [ka] (In the formula, The dashed line is -L 1- indicates the bond to the remainder of the ring contains 3 to 10 atoms, including at least one nitrogen atom, and R # and R ## sp 3 represents a hybridized carbon atom).

[0156] It is also understood that the 3- to 10-membered heterocycle may be further substituted.

[0157] -R in formula (II) 3 / -R 3a and the nitrogen atom to which they are attached are: [ka] (In the formula, The dashed line indicates the bond to the rest of the molecule; -R is -H and C 1-6 alkyl).

[0158] Optionally, -L of formula (II) 1 - may be further substituted. In general, any substituent may be used as long as it does not affect the principle of cleavage, i.e., the hydrogen with an asterisk in formula (II) is not replaced, and the moiety of formula (II) [ka] The nitrogen in remains part of a primary, secondary, or tertiary amine, i.e., -R 3 and -R 3a are each independently -H or SP 3 It is linked to -N< by a hybridized carbon atom.

[0159] In certain embodiments, -R of formula (II) 1 or -R 1a -L 2 In certain embodiments, -R of formula (II) is substituted with -Z. 2 or -R 2a-L 2 In certain embodiments, -R of formula (II) is substituted with -Z. 3 or -R 3a -L 2 In certain embodiments, -R of formula (II) is substituted with -Z. 4 -L 2 In certain embodiments, -R of formula (II) is substituted with -Z. 5 or -R 5a -L 2 In certain embodiments, -R of formula (II) is substituted with -Z. 6 -L 2 In certain embodiments, -R of formula (II) is substituted with -Z. 7 or -R 7a -L 2 In certain embodiments, -R of formula (II) is substituted with -Z. 8 or -R 8a -L 2 In certain embodiments, -R of formula (II) is substituted with -Z. 9 or -R 9a -L 2 Replaced with -Z.

[0160] In certain embodiments, -R of formula (II) 4 -L 2 Replaced with -Z.

[0161] In certain embodiments, -X- in formula (II) is -C(R 4 R 4a )- or -N(R 4 )-.

[0162] In certain embodiments, -X- in formula (II) is -C(R 4 R 4a )-.

[0163] In certain embodiments, X in formula (II) 1 is C.

[0164] In certain embodiments, the =X group of formula (II)3 is =O.

[0165] In certain embodiments, -X in formula (II) 2 - is -C(R 8 R 8a )-.

[0166] In certain embodiments, -R of formula (II) 8 and -R 8a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (II) 8 and -R 8a In certain embodiments, at least one of -R in formula (II) is -H. 8 and -R 8a Both are -H.

[0167] In certain embodiments, -R of formula (II) 1 and -R 1a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (II) 1 and -R 1a In certain embodiments, at least one of -R in formula (II) is -H. 1 and -R 1a Both are -H.

[0168] In certain embodiments, -R of formula (II) 2 and -R 2a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (II) 2 and -R 2a In certain embodiments, at least one of -R in formula (II) is -H. 2 and -R 2a Both are H.

[0169] In certain embodiments, -R of formula (II) 3 and -R 3ais independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. In certain embodiments, -R of formula (II) 3 and -R 3a In certain embodiments, at least one of -R in formula (II) is methyl. 3 and -R 3a are both -H. In certain embodiments, -R of formula (II) 3 and -R 3a In certain embodiments, -R of formula (II) 3 is -H, and -R in formula (II) 3a is methyl.

[0170] In certain embodiments, -R of formula (II) 4 and -R 4a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (II) 4 and -R 4a In certain embodiments, at least one of -R in formula (II) is -H. 4 and -R 4a Both are -H.

[0171] In certain embodiments, the moiety -L 1 - is a group represented by the formula (IIa): [ka] (In the formula, The dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond, and -R 1 , -R 1a , -R 2 , -R 2a , -R 3 , -R 3a , -R 4 , -R 4a and -X 2 - is used as defined in relation to formula (II) and Said-L 1 -L 2-Z, and optionally -L 1 - is further substituted, provided that the hydrogen marked with an asterisk in formula (IIa) is -L 2 -Z or is not replaced by a substituent.

[0172] In certain embodiments, -L of formula (IIa) 1 - is one part - L 2 Replaced with -Z.

[0173] In certain embodiments, the moiety -L of formula (IIa) 1 - is not further substituted.

[0174] In certain embodiments, -R of formula (IIa) 1 and -R 1a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (IIa) 1 and -R 1a In certain embodiments, at least one of -R in formula (IIa) is -H. 1 and -R 1a Both are -H.

[0175] In certain embodiments, -R of formula (IIa) 4 and -R 4a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (IIa) 4 and -R 4a In certain embodiments, at least one of -R in formula (IIa) is -H. 4 and -R 4a Both are -H.

[0176] In certain embodiments, -X in formula (IIa) 2 - is -C(R 8 R 8a )-.

[0177] In certain embodiments, -R of formula (IIa) 8 and -R8a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (IIa) 8 and -R 8a In certain embodiments, at least one of -R in formula (IIa) is -H. 8 and -R 8a Both are -H.

[0178] In certain embodiments, -R of formula (IIa) 2 and -R 2a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (IIa) 2 and -R 2a In certain embodiments, at least one of -R in formula (IIa) is -H. 2 and -R 2a Both are H.

[0179] In certain embodiments, -R of formula (IIa) 3 and -R 3a is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. In certain embodiments, -R of formula (IIa) 3 and -R 3a In certain embodiments, at least one of -R in formula (IIa) is methyl. 3 and -R 3a are both -H. In certain embodiments, -R of formula (IIa) 3 and -R 3a are both methyl. In certain embodiments, -R 3 is -H, and -R in formula (IIa) 3a is methyl.

[0180] In certain embodiments, the moiety -L 1 - is a group represented by the formula (IIb): [ka] (In the formula, The dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond, and -R 2 , -R 2a , -R 3 , -R 3a and -X 2 - is used as defined in relation to formula (II) and Said-L 1 -L 2 -Z, and optionally -L 1 - is further substituted, provided that the hydrogen marked with an asterisk in formula (IIb) is -L 2 -Z or is not replaced by a substituent.

[0181] In certain embodiments, -L of formula (IIb) 1 - is one part - L 2 Replaced with -Z.

[0182] In certain embodiments, the moiety -L of formula (IIb) 1 - is not further substituted.

[0183] In certain embodiments, -X in formula (IIb) 2 - is -C(R 8 R 8a )-.

[0184] In certain embodiments, -R of formula (IIb) 8 and -R 8a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (IIb) 8 and -R 8a In certain embodiments, at least one of -R in formula (IIb) is -H. 8 and -R 8a Both are -H.

[0185] In certain embodiments, -R of formula (IIb) 2 and -R 2ais independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (IIb) 2 and -R 2a In certain embodiments, at least one of -R in formula (IIb) is -H. 2 and -R 2a Both are H.

[0186] In certain embodiments, -R of formula (IIb) 3 and -R 3a is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. In certain embodiments, -R of formula (IIb) 3 and -R 3a In certain embodiments, at least one of -R in formula (IIb) is methyl. 3 and -R 3a are both -H. In certain embodiments, -R of formula (IIb) 3 and -R 3a In certain embodiments, -R in formula (IIb) 3 is -H, and -R in formula (IIb) 3a is methyl.

[0187] In certain embodiments, the moiety -L 1 - is a group represented by the formula (IIb'): [ka] (In the formula, The dashed line indicates the attachment of the CNP moiety, D, to the nitrogen by forming an amide bond; A dashed line with an asterisk indicates -L 2 - indicates a bond to -R 2 , -R 2a , -R 3 , -R 3a , and -X 2 - is used as defined in relation to formula (II) and Optionally, said -L 1- is further substituted, provided that the hydrogen marked with an asterisk in formula (IIb') is not replaced by a substituent.

[0188] In certain embodiments, the moiety -L of formula (IIb') 1 - is not further substituted.

[0189] In certain embodiments, -X in formula (IIb') 2 - is -C(R 8 R 8a )-.

[0190] In certain embodiments, -R 8 and -R 8a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R 8 and -R 8a In certain embodiments, at least one of -R in formula (IIb') is -H. 8 and -R 8a Both are -H.

[0191] In certain embodiments, -R 2 and -R 2a is independently selected from the group consisting of -H, methyl, and ethyl. In certain embodiments, -R of formula (IIb') 2 and -R 2a In certain embodiments, at least one of -R in formula (IIb') is -H. 2 and -R 2a Both are H.

[0192] In certain embodiments, -R 3 and -R 3a is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. In certain embodiments, -R 3 and -R 3a In certain embodiments, at least one of -R in formula (IIb') is methyl. 3 and -R3a are both -H. In certain embodiments, -R 3 and -R 3a are both methyl. In certain embodiments, -R 3 is —H, and —R in formula (IIb′) 3a is methyl.

[0193] In certain embodiments, the moiety -L 1 - is a group represented by the formula (IIc): [ka] where the dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond. and Said-L 1 -L 2 It is replaced by -Z, Said-L 1 - is optionally further substituted, provided that the hydrogen marked with an asterisk in formula (IIc) is -L 2 -Z or is not replaced by a substituent.

[0194] In certain embodiments, -L of formula (IIc) 1 - is one part - L 2 Replaced with -Z.

[0195] In certain embodiments, the moiety -L of formula (IIc) 1 - is not further substituted.

[0196] In certain embodiments, the moiety -L 1 - is a group represented by the formula (IIc-a): [ka] where the dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond. and Said-L 1 -L 2It is replaced by -Z, Said-L 1 - is optionally further substituted, provided that the hydrogen marked with an asterisk in formula (IIc-a) is -L 2 -Z or is not replaced by a substituent.

[0197] In certain embodiments, -L in formula (IIc-a) 1 - is one part - L 2 Replaced with -Z.

[0198] In certain embodiments, the moiety -L of formula (IIc-a) 1 - is not further substituted.

[0199] In certain embodiments, the moiety -L 1 - is a group represented by the formula (IIc-b): [ka] where the dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond. and Said-L 1 -L 2 It is replaced by -Z, Optionally, said -L 1 - is further substituted, provided that the hydrogen with an asterisk in formula (IIc-b) is -L 2 -Z or is not replaced by a substituent.

[0200] In certain embodiments, -L in formula (IIc-b) 1 - is one part - L 2 Replaced with -Z.

[0201] In certain embodiments, the moiety -L of formula (IIc-b) 1 - is not further substituted.

[0202] In certain embodiments, the moiety -L 1represents the formulae (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v): [ka] (In the formula, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; A dashed line with an asterisk indicates -L 2 -Z) is selected from the group consisting of Said-L 1 - is optionally further substituted, provided that the hydrogen marked with an asterisk in formulas (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v) is not replaced by a substituent.

[0203] In certain embodiments, the moiety -L of formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v) 1 - is not further substituted.

[0204] In certain embodiments, the moiety -L 1 - is a group represented by the formula (IIc-ii): [ka] (In the formula, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; A dashed line with an asterisk indicates -L 2 -Z) is.

[0205] In certain embodiments, -L in formula (IIc-ii) 1 - is one part - L 2 Replaced with -Z.

[0206] In certain embodiments, the moiety -L 1- represents the groups of the formulae (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv') and (IIc-v'): [ka] (In the formula, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; A dashed line with an asterisk indicates -L 2 -Z) is selected from the group consisting of Optionally, said -L 1 - is further substituted, with the proviso that the hydrogen marked with an asterisk in formulas (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv') and (IIc-v') is not replaced by a substituent.

[0207] In certain embodiments, the moiety -L of formula (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv') and (IIc-v') 1 - is not further substituted.

[0208] In certain embodiments, the moiety -L 1 - is a group represented by the formula (IIc-ii'): [ka] (In the formula, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; A dashed line with an asterisk indicates -L 2 -Z) is.

[0209] In certain embodiments, -L in formula (IIc-ii') 1 - is one part - L 2 Replaced with -Z.

[0210] In certain embodiments, the moiety -L 1- is a group of the formulae (IIc-i″), (IIc-ii″), (IIc-iii″) and (IIc-iv″): [ka] (In the formula, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; A dashed line with an asterisk indicates -L 2 -Z) is selected from the group consisting of Optionally, said -L 1 - is further substituted, with the proviso that the hydrogen marked with an asterisk in formulae (IIc-i''), (IIc-ii''), (IIc-iii'') and (IIc-iv'') is not replaced by a substituent.

[0211] In certain embodiments, the moiety -L of formula (IIc-i″), (IIc-ii″), (IIc-iii″) and (IIc-iv″) 1 - is not further substituted.

[0212] In certain embodiments, the moiety -L 1 - is a group represented by the formula (IIc-ii″): [ka] (In the formula, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; A dashed line with an asterisk indicates -L 2 -Z) is.

[0213] In certain embodiments, -L in formula (IIc-ii″) 1 - is one part - L 2 Replaced with -Z.

[0214] Formula (II), (IIa), (IIb), (IIb'), (IIc), (IIc-a), (IIc-b), (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv), (IIc-v), (I -L of Ic-i'), (IIc-ii'), (IIc-iii'), (IIc-iv'), (IIc-v'), (IIc-i''), (IIc-ii''), (IIc-iii'') and (IIc-iv'') 1 The optional further substituents of - are, in certain embodiments, as described above.

[0215] Another part - L 1 - is disclosed in WO 2016 / 020373 A1. Thus, in certain embodiments, the moiety -L 1 - is a group represented by the formula (III): [ka] (In the formula, The dashed lines indicate attachment by forming an amide or ester bond to a primary or secondary amine or hydroxyl of the CNP moiety, -D, respectively; -R 1 , -R 1a , -R 2 , -R 2a , -R 3 and -R 3a are, independently of each other, -H, -C(R 8 R 8a R 8b ), -C(=O)R 8 , -C≡N, -C(=NR 8 )R 8a , -CR 8 (=CR 8a R 8b ), -C≡CR 8 and -T, -R 4 , -R 5 and -R 5a are, independently of each other, -H, -C(R 9 R 9a R 9b ) and -T; a1 and a2 are independently 0 or 1; Each-R 6 , -R 6a , -R 7 , -R 7a , -R 8 , -R 8a , -R 8b , -R 9 , -R 9a , -R 9b are, independently of each other, -H, halogen, -CN, -COOR 10 , -OR 10 , -C(O)R 10 , -C(O)N(R 10 R 10a ), -S(O)2N(R 10 R 10a ), -S(O)N(R 10 R 10a ), -S(O)R 10 , -S(O)R 10 , -N(R 10 )S(O)2N(R 10a R 10b ), -SR 10 , -N(R 10 R 10a ), -NO2, -OC(O)R 10 , -N(R 10 )C(O)R 10a , -N(R 10 )S(O)2R 10a , -N(R 10 )S(O)R 10a , -N(R 10 )C(O)OR 10a , -N(R 10 )C(O)N(R 10a R 10b ), -OC(O)N(R 10 R 10a ), -T, C 1-20 Alkyl, C 2-20 Alkenyl, and C 2-20 alkynyl, wherein said -T, C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 Alkynyl may be one or more -R 11 optionally substituted with C 1-20Alkyl, C 2-20 Alkenyl and C 2-20 Alkynyl includes -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 12 )-, -S(O)2N(R 12 )-, -S(O)N(R 12 )-, -S(O)2-, -S(O)-, -N(R 12 )S(O)2N(R 12a )-, -S-, -N(R 12 )-, -OC(OR 12 )(R 12a )-, -N(R 12 )C(O)N(R 12a )- and -OC(O)N(R 12 )-, optionally interrupted by one or more groups selected from the group consisting of Each-R 10 , -R 10a , -R 10b are independently -H, -T, C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 alkynyl, wherein -T, C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 Alkynyl may be one or more -R 11 and wherein C is optionally substituted with 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 Alkynyl includes -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 12 )-, -S(O)2N(R 12 )-, -S(O)N(R 12 )-, -S(O)2-, -S(O)-, -N(R 12 )S(O)2N(R 12a )-, -S-, -N(R 12 )-, -OC(OR 12 )(R 12a )-, -N(R 12 )C(O)N(R 12a )- and -OC(O)N(R 12 )-, optionally interrupted by one or more groups selected from the group consisting of Each T is independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, wherein each T is independently one or more of the same or different —R 11 and optionally substituted with Each-R 11 are each independently a halogen, -CN, oxo (=O), -COOR 13 , -OR 13 , -C(O)R 13 , -C(O)N(R 13 R 13a ), -S(O)2N(R 13 R 13a ), -S(O)N(R 13 R 13a ), -S(O)R 13 , -S(O)R 13 , -N(R 13 )S(O)2N(R 13a R 13b ), -SR 13 , -N(R 13 R 13a ), -NO2, -OC(O)R 13 , -N(R 13 )C(O)R 13a , -N(R 13 )S(O)2R 13a , -N(R 13 )S(O)R 13a , -N(R 13 )C(O)OR 13a , -N(R 13 )C(O)N(R 13a R 13b ), -OC(O)N(R 13 R 13a ), and C 1-6 alkyl, wherein C 1-6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R 12 , -R 12a , -R 13 , -R 13a , -R 13b are independently -H and C 1-6alkyl, 1-6 the alkyl is optionally substituted with one or more halogens which may be the same or different; In some cases, Pair-R 1 / -R 1a , -R 2 / -R 2a , -R 3 / -R 3a , -R 6 / -R 6a , -R 7 / -R 7a One or more of the following, together with the atoms to which they are attached, form a C 3-10 forming a cycloalkyl or a 3- to 10-membered heterocyclyl; In some cases, Pair-R 1 / -R 2 , -R 1 / -R 3 , -R 1 / -R 4 , -R 1 / -R 5 , -R 1 / -R 6 , -R 1 / -R 7 , -R 2 / -R 3 , -R 2 / -R 4 , -R 2 / -R 5 , -R 2 / -R 6 , -R 2 / -R 7 , -R 3 / -R 4 , -R 3 / -R 5 , -R 3 / -R 6 , -R 3 / -R 7 , -R 4 / -R 5 , -R 4 / -R 6 , -R 4 / -R 7 , -R 5 / -R 6 , -R 5 / -R 7 , -R 6 / -R7 together with the atoms to which they are attached form ring A, A is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl and Said-L 1 -L 2 -Z, and optionally -L 1 - is further substituted, -L 2 - is a single chemical bond or spacer, -Z is a water-soluble polymer moiety.

[0216] -L in formula (III) 1 The optional further substituents of - are, in certain embodiments, as described above.

[0217] In certain embodiments, -L of formula (III) 1 - is one part - L 2 Replaced with -Z.

[0218] In certain embodiments, -L of formula (III) 1 - is not further substituted.

[0219] -L 1 Further embodiments of - are disclosed in EP 1536334 B1, WO 2009 / 009712 A1, WO 2008 / 034122 A1, WO 2009 / 143412 A2, WO 2011 / 082368 A2, and U.S. Pat. No. 8,618,124 B2, which are incorporated herein by reference in their entirety.

[0220] -L 1 Further embodiments of - are disclosed in U.S. Pat. No. 8,946,405 B2 and U.S. Pat. No. 8,754,190 B2, which are incorporated herein by reference in their entirety. Thus, the moiety -L1 - is a group represented by the formula (IV): [ka] (In the formula, The dashed line indicates a bond to the CNP moiety, -D, said bond being through a functional group of -D selected from the group consisting of -OH, -SH, and -NH; m is 0 or 1; -R 1 and -R 2 At least one or both of the following are, independently of each other, -CN, -NO2, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkenyl, optionally substituted alkynyl, -C(O)R 3 , -S(O)R 3 , -S(O)2R 3 , and -SR 4 is selected from the group consisting of -R 1 and -R 2 is selected from the group consisting of: -H, optionally substituted alkyl, optionally substituted arylalkyl, and optionally substituted heteroarylalkyl; -R 3 is -H, optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -OR 9 and -N(R 9 )2, -R 4 is selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Each-R 5is independently selected from the group consisting of: -H, optionally substituted alkyl, optionally substituted alkenylalkyl, optionally substituted alkynylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; -R 9 is selected from the group consisting of —H and optionally substituted alkyl; -Y- is absent and -X- is -O- or -S-; or -Y- is -N(Q)CH2- and -X- is -O-; Q is selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; In some cases, -R 1 and -R 2 may combine to form a 3- to 8-membered ring, and In some cases, both -R 9 together with the nitrogen to which they are attached form a heterocyclic ring) and Said-L 1 -L 2 -Z, and optionally -L 1 - is further substituted, -L 2 - is a single chemical bond or spacer, -Z is a water-soluble polymer moiety.

[0221] -L in formula (IV) 1 The optional further substituents of - are, in certain embodiments, as described above.

[0222] In certain embodiments, -L of formula (IV) 1 - is one part - L 2 Replaced with -Z.

[0223] In certain embodiments, -L of formula (IV) 1 - is not further substituted.

[0224] The terms used exclusively in connection with formula (IV) have the following meanings:

[0225] As used herein, the term "alkyl" includes straight-chain, branched, or cyclic saturated hydrocarbon groups of 1 to 8 carbons, or in some embodiments, 1 to 6 or 1 to 4 carbon atoms.

[0226] The term "alkoxy" includes an alkyl group attached to an oxygen, including methoxy, ethoxy, isopropoxy, cyclopropoxy, cyclobutoxy, and the like.

[0227] The term "alkenyl" includes non-aromatic unsaturated hydrocarbons containing a carbon-carbon double bond.

[0228] The term "alkynyl" includes non-aromatic unsaturated hydrocarbons containing a carbon-carbon triple bond.

[0229] The term "aryl" includes aromatic hydrocarbon groups of 6 to 18 carbons, and in certain embodiments, 6 to 10 carbons, including groups such as phenyl, naphthyl, and anthracenyl. The term "heteroaryl" includes aromatic rings containing 3 to 15 carbon atoms and at least one N, O, or S atom, and in certain embodiments, 3 to 7 carbon atoms, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl, and the like.

[0230] In certain embodiments, an alkenyl, alkynyl, aryl, or heteroaryl moiety may be coupled to the remainder of the molecule by an alkylene bond. The substituent in these situations will be referred to as alkenylalkyl, alkynylalkyl, arylalkyl, or heteroarylalkyl, indicating that an alkylene moiety is between the alkenyl, alkynyl, aryl, or heteroaryl moiety and the molecule to which the alkenyl, alkynyl, aryl, or heteroaryl is coupled.

[0231] The term "halogen" includes bromo, fluoro, chloro and iodo.

[0232] 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 exemplary groups provided above for the term "heteroaryl."

[0233] When the ring system is optionally substituted, suitable substituents are selected from the group consisting of alkyl, alkenyl, alkynyl, or additional rings, each of which is optionally further substituted. Optional substituents for any of the groups, including those described above, include halo, nitro, cyano, -OR, -SR, -NR, -OCOR, -NRCOR, -COOR, -CONR, -SOR, -SOR, -SONR, -SONR, where each R is independently alkyl, alkenyl, alkynyl, aryl, or heteroaryl, or two R groups, together with the atoms to which they are attached, form a ring.

[0234] -L 1 Further embodiments of - are disclosed in WO2013 / 036857A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 - is a compound of formula (V): [ka] (In the formula, The dashed line indicates the bond to the CNP moiety, -D, said bond being through the amine functional group of -D; -R 1 is an optionally substituted C1-C6 linear, branched, or cyclic alkyl, an optionally substituted aryl, an optionally substituted heteroaryl, an alkoxy, and -NR 5 2, -R 2 is selected from the group consisting of -H, optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl; -R 3 is selected from the group consisting of -H, optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl; -R 4 is selected from the group consisting of -H, optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl; Each-R 5 are each independently selected from the group consisting of: -H, optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl, or when taken together, two -R 5 can be cycloalkyl or cycloheteroalkyl) and Said-L 1 -L 2 -Z, and optionally -L 1 - is further substituted, -L 2 - is a single chemical bond or spacer, -Z is a water-soluble polymer moiety.

[0235] -L in formula (V) 1 The optional further substituents of - are, in certain embodiments, as described above.

[0236] In certain embodiments, -L in formula (V) 1 - is one part - L 2 Replaced with -Z.

[0237] In certain embodiments, -L in formula (V) 1 - is not further substituted.

[0238] The terms used exclusively in connection with formula (V) have the following meanings:

[0239] "Alkyl," "alkenyl," and "alkynyl" include straight-chain, branched, or cyclic hydrocarbon groups of 1 to 8, or 1 to 6, or 1 to 4 carbons, 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, they contain 1 to 6 C.

[0240] "Aryl" includes aromatic hydrocarbon groups of 6 to 18 carbons, and in certain embodiments, 6 to 10 carbons, including groups such as phenyl, naphthyl, and anthracene. "Heteroaryl" includes aromatic rings of 3 to 15 carbons and containing at least one N, O, or S atom, and in certain embodiments, 3 to 7 carbons and containing at least one N, O, or S atom, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl, and the like.

[0241] The term "substituted" refers to an alkyl, alkenyl, alkynyl, aryl, or heteroaryl group that contains 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, thio may be selected from carbamates, thioureas, ketones, sulfones, sulfonamides, aryls (including phenyl, naphthyl and anthracenyl), heteroaryls (including 5-membered heteroaryls such as pyrrole, imidazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, thiadiazole, triazole, oxadiazole and tetrazole, 6-membered heteroaryls such as pyridine, pyrimidine and pyrazine, and fused heteroaryls such as benzofuran, benzothiophene, benzoxazole, benzimidazole, indole, benzothiazole, benzisoxazole and benzisothiazole).

[0242] -L 1 Further embodiments of - are disclosed in U.S. Patent No. 7,585,837 B2, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 is a group represented by the formula (VI): [ka] (In the formula, The dashed line indicates the bond to the CNP moiety, -D, said bond being through the amine functional group of -D; R1 and R 2 are independently hydrogen, alkyl, alkoxy, alkoxyalkyl, aryl, alkaryl, aralkyl, halogen, nitro, -SO3H, -SO2NHR 5 , amino, ammonium, carboxyl, PO3H2, and OPO3H2; R 3 , R 4 and R 5 are independently selected from the group consisting of hydrogen, alkyl, and aryl; Said-L 1 -L 2 -Z, and optionally -L 1 - is further substituted, -L 2 - is a single chemical bond or spacer, -Z is a water-soluble polymer moiety.

[0243] Suitable substituents for formula (VI) include 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- to 7-membered heterocycle), or halogen moiety.

[0244] In certain embodiments, -L of formula (VI) 1 - is one part - L 2 Replaced with -Z.

[0245] -L in formula (VI) 1 The optional further substituents of - are, in certain embodiments, as described above.

[0246] In certain embodiments, -L of formula (VI) 1 - is not further substituted.

[0247] The terms used exclusively in connection with formula (VI) have the following meanings:

[0248] The terms "alkyl," "alkoxy," "alkoxyalkyl," "aryl," "alkaryl," and "aralkyl" refer to alkyl radicals of 1 to 8 carbon atoms, and in certain embodiments 1 to 4 carbon atoms, e.g., methyl, ethyl, propyl, isopropyl, and butyl, and aryl radicals of 6 to 10 carbon atoms, e.g., phenyl and naphthyl. The term "halogen" includes bromo, fluoro, chloro, and iodo.

[0249] -L 1 Further embodiments of - are disclosed in WO2002 / 089789A1, which is incorporated herein by reference in its entirety. Thus, the moiety -L 1 - is a group represented by the formula (VII): [ka] (In the formula, The dashed line indicates the bond to the CNP moiety, -D, said bond being through the amine functional group of -D; L1 is a bifunctional linking group, Y1 and Y2 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 3-8 Substituted cycloalkyl, aryl, substituted aryl, aralkyl, C 1-6 Heteroalkyl, substituted C 1-6 Heteroalkyl, C 1-6 Alkoxy, phenoxy and C 1-6 heteroalkoxy; Ar is a moiety that, when included in formula (VII), forms a polysubstituted aromatic hydrocarbon or polysubstituted heterocyclic group; X is a chemical bond, a moiety that is actively transported into a target cell, a hydrophobic moiety, or a combination thereof; y is 0 or 1) and Said-L 1 -L 2 -Z, and optionally -L 1 - is further substituted, -L 2 - is a single chemical bond or spacer, -Z is a water-soluble polymer moiety.

[0250] In certain embodiments, -L of formula (VII) 1 - is one part - L 2 Replaced with -Z.

[0251] -L in formula (VII) 1 The optional further substituents of - are, in certain embodiments, as described above.

[0252] In certain embodiments, -L of formula (VII) 1 - is not further substituted.

[0253] The terms used exclusively in connection with formula (VII) have the following meanings:

[0254] The term "alkyl" includes, for example, alkoxy, C 3-8 Linear, branched, substituted C, including cycloalkyl or substituted cycloalkyl 1-12 It shall be understood to include alkyl.

[0255] The term "substituted" shall be understood to include the addition and replacement of one or more atoms contained in a functional group or compound with one or more different atoms.

[0256] 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- shall be understood to include fluoro, chloro, iodo, and bromo.

[0257] In certain embodiments, -L 1 - is a substructure of formula (VIII) [ka] (In the formula, The dashed line with an asterisk indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; Unmarked dashed lines are -L 1 - indicates the bond to the rest of Including, Said-L 1 -L 2 -Z, and optionally -L 1 - is further substituted, where -L 2 - is a single chemical bond or spacer, -Z is a water-soluble polymer moiety.

[0258] In certain embodiments, -L of formula (VIII) 1 - is one part - L 2 Replaced with -Z.

[0259] -L in formula (VIII) 1 The optional further substituents of - are as described above.

[0260] In certain embodiments, -L of formula (VIII) 1 - is not further substituted.

[0261] In certain embodiments, -L 1 - is a substructure of formula (IX) [ka] (In the formula, The dashed line with an asterisk indicates attachment by forming a carbamate bond to the nitrogen of the CNP moiety, -D; Unmarked dashed lines are -L 1 - indicates the bond to the rest of Including, Said-L 1 -L 2 -Z, and optionally -L 1 - is further substituted, -L 2 - is a single chemical bond or spacer, -Z is a water-soluble polymer moiety.

[0262] -L in formula (IX) 1 The optional further substituents of - are as described above.

[0263] In certain embodiments, -L of formula (IX) 1 - is one part - L 2 Replaced with -Z.

[0264] In certain embodiments, -L of formula (IX) 1 - is not further substituted.

[0265] The moiety -D can be linked to -L via any functional group of the DH. 1 - may be linked to -L through the amine functional group of D-H 1 -, which may be the N-terminal amine functionality or the amine functionality provided by the lysine side chains, i.e., by the lysines at positions 9, 11, 15, 16, 20 and 26 when CNP has the sequence of SEQ ID NO: 24.

[0266] -L to the ring of the CNP moiety 1 Binding of - significantly reduces the affinity of the CNP conjugate for NPR-B compared to binding at the N-terminus or to the non-cyclic portion of CNP. This reduced affinity for NPR-B, in turn, reduces the risk of cardiovascular side effects such as hypotension.

[0267] Therefore, -L 1 In certain embodiments, - is attached to the side chain of an amino acid residue in the cyclic moiety of -D or to the backbone of the cyclic moiety of -D. 1 - is covalently and reversibly attached to the side chain of an amino acid residue in the ring portion of -D. When -D is a CNP moiety having the sequence of SEQ ID NO: 24, -L 1 - is, in certain embodiments, attached to the amine functionality provided by the lysine at position 26 of the corresponding drug DH.

[0268] Part-L 2 - is a chemical bond or a spacer moiety.

[0269] In certain embodiments, -L 2 - is a chemical bond.

[0270] In certain embodiments, -L 2 - is a spacer portion.

[0271] Part-L 2 - replaces any existing -H with -L, unless expressly excluded 1 - can be bonded to

[0272] -L 2 If - is other than a single chemical bond, -L 2 - stands for -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O)2N(R y1 )-, -S(O)N(R y1)-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(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 can have one or more identical or different -R y2 may be substituted with 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)2N(R y3 )-, -S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(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 )-, optionally interrupted by one or more groups selected from the group consisting of;

[0273] -R y1 and -R y1a are independent of each other: -H, -T, 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 can have one or more identical or different -R y2may be substituted with 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)2N(R y4 )-, -S(O)N(R y4 )-, -S(O)2-, -S(O)-, -N(R y4 )S(O)2N(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 )-, optionally interrupted by one or more groups selected from the group consisting of;

[0274] Each T independently represents phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 each T is independently selected from the group consisting of one or more of the same or different -R y2 may be substituted with;

[0275] Each-R y2 are independently halogen, -CN, oxo (=O), -COOR y5 , -OR y5 , -C(O)R y5 , -C(O)N(R y5 R y5a ), -S(O)2N(R y5 R y5a ), -S(O)N(R y5 R y5a ), -S(O)R y5 , -S(O)R y5 , -N(R y5 )S(O)2N(R y5a R y5b ), -SR y5 , -N(R y5 R y5a), -NO2, -OC(O)R y5 , -N(R y5 )C(O)R y5a , -N(R y5 )S(O)2R 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 alkyl; C 1-6 The 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 are independently -H and C 1-6 alkyl; C 1-6 The alkyl may be optionally substituted with one or more halogens which may be the same or different.

[0276] -L 2 If - is other than a single chemical bond, -L 2 - stands for -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O)2N(R y1 )-, -S(O)N(R y1 )-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(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 1-20 Alkyl, C 2-20 Alkenyl, and C 2-20alkynyl; -T-, C 1-20 Alkyl, C 2-20 Alkenyl, and C 2-20 Alkynyl can have one or more identical or different -R y2 may be substituted with C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-, -S(O)2N(R y3 )-, -S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(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 )-, optionally interrupted by one or more groups selected from the group consisting of;

[0277] -R y1 and -R y1a are independent of each other: -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 can have one or more identical or different -R y2 may be substituted with C 1-10 Alkyl, C 2-10 Alkenyl, and C 2-10 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 )-, -S(O)2N(R y4 )-, -S(O)N(R y4 )-, -S(O)2-, -S(O)-, -N(R y4 )S(O)2N(R y4a )-, -S-, -N(Ry4 )-, -OC(OR y4 )(R y4a )-, -N(R y4 )C(O)N(R y4a )- and -OC(O)N(R y4 )-, optionally interrupted by one or more groups selected from the group consisting of;

[0278] Each T independently represents phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 each T is independently selected from the group consisting of one or more of the same or different -R y2 may be substituted with;

[0279] -R y2 is halogen, -CN, oxo(=O), -COOR y5 , -OR y5 , -C(O)R y5 , -C(O)N(R y5 R y5a ), -S(O)2N(R y5 R y5a ), -S(O)N(R y5 R y5a ), -S(O)R y5 , -S(O)R y5 , -N(R y5 )S(O)2N(R y5a R y5b ), -SR y5 , -N(R y5 R y5a ), -NO2, -OC(O)R y5 , -N(R y5 )C(O)R y5a , -N(R y5 )S(O)2R 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 y5R y5a ), and C 1-6 alkyl; C 1-6 The alkyl may be optionally substituted with one or more identical or different halogens;

[0280] Each-R y3 , -R y3a , -R y4 , -R y4a , -R y5 , -R y5a and -R y5b are each independently -H and C 1-6 alkyl; C 1-6 The alkyl may be optionally substituted with one or more halogens which may be the same or different.

[0281] -L 2 If - is other than a single chemical bond, -L 2 - stands for -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O)2N(R y1 )-, -S(O)N(R y1 )-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(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 can have one or more identical or different -R y2 may be substituted with 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)2N(R y3 )-, -S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(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 )-, optionally interrupted by one or more groups selected from the group consisting of;

[0282] -R y1 and -R y1a are independently -H, -T, C 1-10 Alkyl, C 2-10 Alkenyl, and C 2-10 alkynyl;

[0283] Each T independently represents phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbocyclic ring, and 8- to 30-membered heterocyclic ring;

[0284] Each-R y2 is halogen and C 1-6 selected from the group consisting of alkyl;

[0285] Each-R y3 , -R y3a , -R y4 , -R y4a , -R y5 , -R y5a and -R y5b are each independently -H and C 1-6 alkyl; C 1-6 The alkyl may be optionally substituted with one or more halogens which may be the same or different.

[0286] In certain embodiments, -L 2 - is C1-20 alkyl chains, which are independently -O-, -T- and -C(O)N(R y1 )-, optionally interrupted by one or more groups selected from C 1-20 The alkyl chains are independently -OH, -T and -C(O)N(R y6 R y6a ) may be substituted with one or more groups selected from -R y1 , -R y6 , -R y6a are independently H and C 1-4 alkyl, T is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 It is selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycycle, and 8- to 30-membered heteropolycycle.

[0287] In certain embodiments, -L 2 - has a molecular weight in the range of 14 g / mol to 750 g / mol inclusive.

[0288] In certain embodiments, -L 2 - has a chain length of 1 to 20 atoms.

[0289] As used herein, the moiety -L 2 The term "chain length" in relation to -L 1 -L in the shortest link between - and -Z 2 -Refers to the number of atoms.

[0290] In certain embodiments, -L 2 - is of the following formula (i): [ka] During the ceremony, The dashed lines marked with asterisks indicate -L 1 - refers to the bond to; The unmarked dashed line indicates the bond to -Z; -R 1 -H, C 1-6Alkyl, 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.

[0291] In certain embodiments, -R of formula (i) 1 is selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. In certain embodiments, -R in formula (i) 1 is selected from the group consisting of -H, methyl, ethyl, and propyl. In certain embodiments, -R in formula (i) 1 is selected from the group consisting of —H and methyl. In certain embodiments, —R 1 is methyl.

[0292] In certain embodiments, n in formula (i) is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In certain embodiments, n in formula (i) is selected from the group consisting of 0, 1, 2, 3, 4, and 5. In certain embodiments, n in formula (i) is selected from the group consisting of 0, 1, 2, and 3. In certain embodiments, n in formula (i) is selected from the group consisting of 0 and 1. In certain embodiments, n in formula (i) is 0.

[0293] In certain embodiments, -L 2 -teeth, [ka] a moiety selected from the group consisting of: During the ceremony, The dashed lines marked with asterisks indicate -L 1 - indicates a bond to; The unmarked dashed line indicates the bond to -Z; Moieties (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi), (xii), (xiii), (xiv), (xv), (xvi) and (xvii) may further be optionally substituted.

[0294] In certain embodiments, -L 2 -teeth, [ka] selected from the group consisting of: The dashed lines marked with asterisks indicate -L 1 - indicates a bond to; The unmarked dashed line indicates the bond to -Z.

[0295] In certain embodiments, -L 2 -teeth, [ka] is selected from the group consisting of The dashed lines marked with asterisks indicate -L 1 - indicates a bond to; The unmarked dashed line indicates the bond to -Z.

[0296] In certain embodiments, -L 2 - is of the following formula (xvi): [ka] During the ceremony, The dashed lines marked with asterisks indicate -L 1 - indicates a bond to; The unmarked dashed line indicates the bond to -Z.

[0297] In certain embodiments, the moiety -L 1 -L 2 -teeth, [ka] selected from the group consisting of: During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0298] In certain embodiments, the moiety -L 1 -L 2 - is of the following formula (IId-ii): [ka] During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0299] In certain embodiments, the moiety -L 1 -L 2 - is of the following formula (IId-ii'): [ka] During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0300] In certain embodiments, the moiety -L 1 -L 2 -teeth, [ka] is selected from the group consisting of The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0301] In certain embodiments, the moiety -L 1-L 2 - is of the following formula (IId-iia): [ka] During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0302] In certain embodiments, the moiety -L 1 -L 2 - is of the following formula (IId-iia'): [ka] During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0303] In certain embodiments, the moiety -L 1 -L 2 -teeth, [ka] selected from the group consisting of: During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0304] In certain embodiments, the moiety -L 1 -L 2 - is of the following formula (IId-iib): [ka] During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0305] In certain embodiments, the moiety -L 1 -L 2 - is of the following formula (IId-iib'): [ka] During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk indicates the bond to -Z.

[0306] In certain embodiments, -Z of Formula (Ia) or (Ib) has a molecular weight in the range of 5 to 200 kDa. In certain embodiments, -Z of Formula (Ia) or (Ib) has a molecular weight in the range of 8 to 100 kDa. In certain embodiments, -Z of Formula (Ia) or (Ib) has a molecular weight in the range of 10 to 80 kDa. In certain embodiments, -Z of Formula (Ia) or (Ib) has a molecular weight in the range of 12 to 60 kDa. In certain embodiments, -Z of Formula (Ia) or (Ib) has a molecular weight in the range of 15 to 40 kDa. In certain embodiments, -Z of Formula (Ia) or (Ib) has a molecular weight of about 20 kDa. In certain embodiments, -Z of Formula (Ia) or (Ib) has a molecular weight of about 40 kDa.

[0307] The polymer moiety -Z of Formula (Ia) or (Ib) comprises a polymer. In certain embodiments, -Z of Formula (Ia) or (Ib) is 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(sialic acid), poly(hydroxybenzoates ... Poly(acrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyl oxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly Poly(hydroxypropyloxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinylamines), poly(vinyl methyl ethers), poly(vinyl The polymers include polymers selected from the group consisting of hydroxypropyl methylcelluloses, ...

[0308] In certain embodiments, -Z in formula (Ia) or (Ib) comprises a protein. Preferred proteins are selected from the group consisting of the carboxyl terminal peptide of chorionic gonadotropin described in US2012 / 0035101A1 (incorporated herein by reference); albumin; XTEN sequences described in WO2011123813A2 (incorporated herein by reference); proline / alanine random coil sequences described in WO2011 / 144756A1 (incorporated herein by reference); proline / alanine / serine random coil sequences described in WO2008 / 155134A1 and WO2013 / 024049A1 (incorporated herein by reference); and Fc fusion proteins.

[0309] In certain embodiments, -Z of formula (Ia) or (Ib) is polysarcosine. In certain embodiments, -Z of formula (Ia) or (Ib) comprises poly(N-methylglycine). In certain embodiments, -Z of formula (Ia) or (Ib) comprises a random coil protein moiety. In certain embodiments, -Z of formula (Ia) or (Ib) comprises one random coil protein moiety. In certain embodiments, -Z of formula (Ia) or (Ib) comprises two random coil protein moieties. In certain embodiments, -Z of formula (Ia) or (Ib) comprises three random coil protein moieties. In certain embodiments, -Z of formula (Ia) or (Ib) comprises four random coil protein moieties. In certain embodiments, -Z of formula (Ia) or (Ib) comprises five random coil protein moieties. In certain embodiments, -Z of formula (Ia) or (Ib) comprises six random coil protein moieties. In certain embodiments, -Z of formula (Ia) or (Ib) comprises 7 random coil protein moieties. In certain embodiments, -Z of formula (Ia) or (Ib) comprises 8 random coil protein moieties.

[0310] In certain embodiments, such random coil protein portions comprise at least 25 amino acid residues and at most 2000 amino acids. In certain embodiments, such random coil protein portions comprise at least 30 amino acid residues and at most 1500 amino acid residues. In certain embodiments, such random coil protein portions comprise at least 50 amino acid residues and at most 500 amino acid residues.

[0311] In certain embodiments, -Z in formula (Ia) or (Ib) comprises a random coil protein moiety in which at least 80%, and in certain embodiments, at least 85%, and in certain embodiments, at least 90%, and in certain embodiments, at least 95%, and in certain embodiments, at least 98%, and in certain embodiments, at least 99%, of the total number of amino acids forming said random coil protein moiety are selected from alanine and proline. In certain embodiments, at least 10%, but less than 75%, and in certain embodiments, less than 65%, of the total number of amino acid residues in such a random coil protein moiety are proline residues. In certain embodiments, such a random coil protein moiety is one described in WO2011 / 144756A1, the entire contents of which are incorporated herein by reference. In certain embodiments, -Z comprises at least one moiety 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 (hereby incorporated by reference). Moieties comprising such random coil proteins containing alanine and proline are referred to as "PA" or "PA moieties."

[0312] Thus, -Z in formula (Ia) or (Ib) comprises a PA moiety.

[0313] In certain embodiments, -Z of formula (Ia) or (Ib) comprises a random coil protein moiety in which at least 80%, in certain embodiments, at least 85%, in certain embodiments, at least 90%, in certain embodiments, at least 95%, in certain embodiments, at least 98%, and in certain embodiments, at least 99% of the total number of amino acids forming said random coil protein moiety are selected from alanine, serine, and proline. In certain embodiments, at least 4% but less than 40% of the total number of amino acid residues in such a random coil protein moiety are proline residues. In certain embodiments, such a random coil protein moiety is one described in WO2008 / 155134A1 (incorporated herein by reference in its entirety). In certain embodiments, -Z in Formula (Ia) or (Ib) comprises at least one moiety 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 (hereby incorporated by reference). Moieties comprising such random coil protein moieties containing alanine, serine, and proline are referred to as "PAS" or "PAS moieties."

[0314] Thus, -Z in formula (Ia) or (Ib) comprises a PAS moiety.

[0315] In certain embodiments, -Z of formula (Ia) or (Ib) comprises a random coil protein moiety in which at least 80%, in certain embodiments, at least 85%, in certain embodiments, at least 90%, in certain embodiments, at least 95%, in certain embodiments, at least 98%, and in certain embodiments, at least 99% of the total number of amino acids forming said random coil protein moiety are selected from alanine, glycine, and proline. A moiety comprising such a random coil protein moiety containing alanine, glycine, and proline is referred to as a "PAG" or a "PAG moiety."

[0316] Thus, -Z in formula (Ia) or (Ib) includes a PAG moiety.

[0317] In certain embodiments, -Z of formula (Ia) or (Ib) comprises a random coil protein moiety in which at least 80%, in certain embodiments, at least 85%, in certain embodiments, at least 90%, in certain embodiments, at least 95%, in certain embodiments, at least 98%, and in certain embodiments, at least 99% of the total number of amino acids forming said random coil protein moiety are selected from proline and glycine. Such a random coil protein moiety containing proline and glycine is referred to as "PG" or a "PG moiety."

[0318] In certain embodiments, such PG moieties include moieties of formula (a-0): [(Gly) p -Pro-(Gly) q ] r (a-0); During the ceremony, 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 ranging from 10 to 1000, inclusive; However, at least one of p and q is at least 1.

[0319] In certain embodiments, p in formula (a-0) is selected from the group consisting of 1, 2, and 3.

[0320] In certain embodiments, q in formula (a-0) is selected from 0, 1, and 2.

[0321] In certain embodiments, the PG moiety comprises the sequence of SEQ ID NO: 97: GGPGGPGPGGPGGPGPGGPG.

[0322] In certain embodiments, the PG moiety comprises a sequence of formula (a-0-a): (GGPGGPGPGGPGGPGPGGPG) v (a-0-a) where v is an integer ranging from 1 to 50, inclusive.

[0323] It is understood that the sequence of formula (a-0-a) comprises v repeats of the sequence of SEQ ID NO:97.

[0324] Thus, -Z in formula (Ia) or (Ib) contains a PG moiety.

[0325] In certain embodiments, -Z of formula (Ia) or (Ib) comprises a random coil protein moiety in which at least 80%, in certain embodiments, at least 85%, in certain embodiments, at least 90%, in certain embodiments, at least 95%, in certain embodiments, at least 98%, and in certain embodiments, at least 99% of the total number of amino acids forming said random coil protein moiety are selected from alanine, glycine, serine, threonine, glutamic acid, and proline. In certain embodiments, such a random coil protein moiety is one described in WO2010 / 091122A1 (incorporated herein by reference). In certain embodiments, -Z in Formula (Ia) or (Ib) is 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 disclosed in WO2010 / 091122A1 (hereby incorporated by reference). , 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.A moiety containing such a random coil protein moiety containing alanine, glycine, serine, threonine, glutamic acid, and proline is referred to as "XTEN" or "XTEN moiety," consistent with the designation in WO2010 / 091122A1.

[0326] Thus, -Z in formula (Ia) or (Ib) includes an XTEN moiety.

[0327] In certain embodiments, -Z of formula (Ia) or (Ib) is a hyaluronic acid-based polymer.

[0328] In certain embodiments, -Z of formula (Ia) or (Ib) is a polymer moiety disclosed in WO2013 / 024047A1 (hereby incorporated by reference).

[0329] In certain embodiments, -Z of formula (Ia) or (Ib) is a polymer moiety disclosed in WO2013 / 024048A1 (hereby incorporated by reference).

[0330] In certain embodiments, -Z in formula (Ia) or (Ib) is a PEG-based polymer. In certain embodiments, -Z is a branched or multi-branched PEG-based polymer.

[0331] In certain embodiments, -Z in Formula (Ia) or (Ib) is a branched polymer. In certain embodiments, -Z in Formula (Ia) or (Ib) is a branched polymer having 1, 2, 3, 4, 5, or 6 branch points. In certain embodiments, -Z in Formula (Ia) or (Ib) is a branched polymer having 1, 2, or 3 branch points. In certain embodiments, -Z in Formula (Ia) or (Ib) is a branched polymer having 1 branch point. In certain embodiments, -Z in Formula (Ia) or (Ib) is a branched polymer having 2 branch points. In certain embodiments, -Z in Formula (Ia) or (Ib) is a branched polymer having 3 branch points.

[0332] In certain embodiments, the branch points are selected from the group consisting of -N<, -CH< and >C<.

[0333] In certain embodiments, such branched moieties -Z of formula (Ia) or (Ib) are PEG-based.

[0334] In certain embodiments, such branching moieties-Z of Formula (Ia) or (Ib) have a molecular weight in the range of 5 kDa to 500 kDa, inclusive. In certain embodiments, such branching moieties-Z of Formula (Ia) or (Ib) have a molecular weight in the range of 10 kDa to 250 kDa, inclusive. In certain embodiments, such branching moieties-Z of Formula (Ia) or (Ib) have a molecular weight in the range of 10 kDa to 150 kDa, inclusive. In certain embodiments, such branching moieties-Z of Formula (Ia) or (Ib) have a molecular weight in the range of 12 kDa to 100 kDa, inclusive. In certain embodiments, such branching moieties-Z of Formula (Ia) or (Ib) have a molecular weight in the range of 15 kDa to 80 kDa, inclusive. In certain embodiments, such branching moieties-Z of Formula (Ia) or (Ib) have a molecular weight in the range of 10 kDa to 80 kDa, inclusive. In certain embodiments, the molecular weight is about 10 kDa. In certain embodiments, the molecular weight of such branching moiety-Z of Formula (Ia) or (Ib) is about 20 kDa. In certain embodiments, the molecular weight of such branching moiety-Z of Formula (Ia) or (Ib) is about 30 kDa. In certain embodiments, the molecular weight of such branching moiety-Z of Formula (Ia) or (Ib) is about 40 kDa. In certain embodiments, the molecular weight of such branching moiety-Z of Formula (Ia) or (Ib) is about 50 kDa. In certain embodiments, the molecular weight of such branching moiety-Z of Formula (Ia) or (Ib) is about 60 kDa. In certain embodiments, the molecular weight of such branching moiety-Z of Formula (Ia) or (Ib) is about 70 kDa. In certain embodiments, the molecular weight of such branching moiety-Z of Formula (Ia) or (Ib) is about 80 kDa. In certain embodiments, such branched moiety -Z of formula (Ia) or (Ib) has a molecular weight of about 40 kDa.

[0335] In certain embodiments, -Z comprises the following moiety: [ka]

[0336] In certain embodiments, -Z comprises an amide bond.

[0337] In certain embodiments, -Z of formula (Ia) or (Ib) comprises a moiety of formula (a): [ka] During the ceremony, The dashed line is -L 2 - or - represents the bond to the remainder of Z; BP a is a branch point selected from the group consisting of -N<, -CR< and >C<; -R is -H and C 1-6 selected from the group consisting of alkyl; a is BP a is 0 if -N< or -CR<, and a is BP a is 1 if >C<; -S a -, -S a′ -, -S a″ -and-S a″′ - are independently a chemical bond or 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 have one or more identical or different -R 1 may be substituted with 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)2N(R 2)-, -S(O)N(R 2 )-, -S(O)2-, -S(O)-, -N(R 2 )S(O)2N(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 )-, optionally interrupted by one or more groups selected from the group consisting of; Each -T- is independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 each -T- is independently selected from the group consisting of one or more of the same or different -R 1 may be substituted with; Each-R 1 are independently halogen, -CN, 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)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 3a , -N(R 3 )S(O)R 3a , -N(R 3 )C(O)OR 3a , -N(R 3 )C(O)N(R3a R 3b ), -OC(O)N(R 3 R 3a ) and C 1-6 alkyl; C 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; -P a′ , -P a″ and -P a″′ are independently polymer moieties.

[0338] Optionally, the moiety of formula (a) is substituted with one or more substituents.

[0339] In certain embodiments, BP of formula (a) a is -N<.

[0340] In certain embodiments, BP of formula (a) a is -CR<. In certain embodiments, -R is -H.

[0341] Thus, in certain embodiments, a in formula (a) is 0.

[0342] In certain embodiments, BP of formula (a) a is >C<.

[0343] In certain embodiments, -S of formula (a) a - is a chemical bond.

[0344] In certain embodiments, -S of formula (a) a -C 1-10 Alkyl, C 2-10 Alkenyl and C2-10 alkynyl, 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)2N(R 4 )-, -S(O)N(R 4 )-, -S(O)2-, -S(O)-, -N(R 4 )S(O)2N(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 )-, optionally interrupted by one or more chemical groups selected from the group consisting of -R 4 and -R 4a is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. In certain embodiments, -S in formula (a) a - is selected from the group consisting of methyl, ethyl, propyl, and butyl, which are -O-, -C(O)-, and -C(O)N(R 4 )-.

[0345] In certain embodiments, -S of formula (a) a′ is a chemical bond.

[0346] In certain embodiments, -S of formula (a) a′ -C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 alkynyl, 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)2N(R 4 )-, -S(O)N(R 4)-, -S(O)2-, -S(O)-, -N(R 4 )S(O)2N(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 )-, optionally interrupted by one or more chemical groups selected from the group consisting of -R 4 and -R 4a is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. In certain embodiments, -S in formula (a) a′ - is selected from the group consisting of methyl, ethyl, propyl, and butyl, which are -O-, -C(O)-, and -C(O)N(R 4 )-.

[0347] In certain embodiments, -S of formula (a) a″ - is a chemical bond.

[0348] In certain embodiments, -S of formula (a) a″ -C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 alkynyl, 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)2N(R 4 )-, -S(O)N(R 4 )-, -S(O)2-, -S(O)-, -N(R 4 )S(O)2N(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)-, optionally interrupted by one or more chemical groups selected from the group consisting of -R 4 and -R 4a is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. In certain embodiments, -S in formula (a) a″ - is selected from the group consisting of methyl, ethyl, propyl, and butyl, which are -O-, -C(O)-, and -C(O)N(R 4 )-.

[0349] In certain embodiments, -S of formula (a) a″′ - is a chemical bond.

[0350] In certain embodiments, -S of formula (a) a″′ -C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 alkynyl, 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)2N(R 4 )-, -S(O)N(R 4 )-, -S(O)2-, -S(O)-, -N(R 4 )S(O)2N(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 )-, optionally interrupted by one or more chemical groups selected from the group consisting of -R 4 and -R 4a is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. In certain embodiments, -S in formula (a) a″′- is selected from the group consisting of methyl, ethyl, propyl, and butyl, which are -O-, -C(O)-, and -C(O)N(R 4 )-.

[0351] In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′are independently 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(dimethyl acrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyl oxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyl octadecyl ... Poly(oxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinylamines), poly(vinyl methyl ethers), poly(vinylpyrrolidone) The polymers include polymers selected from the group consisting of: celluloses, silicones, celluloses, carbomethylcelluloses, hydroxypropylmethylcelluloses, 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.

[0352] In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′independently have a molecular weight in the range of 5 kDa to 50 kDa (inclusive), in certain embodiments in the range of 5 kDa to 40 kDa (inclusive), in certain embodiments in the range of 7.5 kDa to 35 kDa (inclusive), in certain embodiments in the range of 7.5 to 30 kDa (inclusive), and in certain embodiments in the range of 10 to 30 kDa (inclusive).

[0353] In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ In certain embodiments, -P of formula (a) has a molecular weight of about 5 kDa. a′ , -P a″ and -P a″′ has a molecular weight of about 7.5 kDa. In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of about 10 kDa. In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of about 12.5 kDa. In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of about 15 kDa. In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ has a molecular weight of approximately 20 kDa.

[0354] In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′independently comprise a PEG-based moiety comprising at least 20% PEG, in certain embodiments at least 30%, and in certain embodiments at least 40% PEG, in certain embodiments at least 50% PEG, in certain embodiments at least 60% PEG, in certain embodiments at least 70% PEG, in certain embodiments at least 80% PEG, and in certain embodiments at least 90% PEG.

[0355] In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ independently comprise a protein moiety, in certain embodiments a random coil protein moiety, in certain embodiments a random coil protein moiety selected from the group consisting of a PA, PAS, PAG, PG, and XTEN moiety.

[0356] In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ is a PA moiety. In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ is a PAS moiety. In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ is a PAG moiety. In certain embodiments, -P of formula (a) a′ , -P a″ and -P a″′ is a PG moiety. In certain embodiments, -P in formula (a) a′ , -P a″ and -P a″′ is the XTEN portion.

[0357] In certain embodiments, -Z comprises one moiety of formula (a). In certain embodiments, -Z comprises two moieties of formula (a). In other embodiments, -Z comprises three moieties of formula (a). In certain embodiments, -Z comprises four moieties of formula (a). In certain embodiments, -Z comprises five moieties of formula (a). In certain embodiments, -Z comprises six moieties of formula (a).

[0358] In certain embodiments, -Z comprises a moiety of formula (b): [ka] During the ceremony, The dashed line is -L 2 - represents the bond to or to the remainder of -Z; b1 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7 and 8; b2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7 and 8; b3 is an integer in the range of 150 to 1000, inclusive; in certain embodiments, in the range of 150 to 500, inclusive; and in certain embodiments, in the range of 200 to 460, inclusive; b4 is an integer in the range of 150 to 1000, inclusive; in certain embodiments, in the range of 150 to 500, inclusive; and in certain embodiments, in the range of 200 to 460, inclusive.

[0359] Optionally, the moiety of formula (b) is substituted with one or more substituents.

[0360] In certain embodiments, b3 and b4 in formula (b) are the same integer.

[0361] In certain embodiments, both b3 and b4 of formula (b) are integers in the range of 200 to 250, and in certain embodiments, b3 and b4 of formula (b) are about 225. In certain embodiments, both b3 and b4 of formula (b) are integers in the range of 400 to 500, and in certain embodiments, b3 and b4 of formula (b) are about 450.

[0362] In certain embodiments, b1 of formula (b) is selected from the group consisting of 0, 1, 2, 3, and 4. In certain embodiments, b1 of formula (b) is selected from the group consisting of 1, 2, and 3. In certain embodiments, b1 of formula (b) is 2.

[0363] In certain embodiments, b2 of formula (b) is selected from the group consisting of 1, 2, 3, 4, and 5. In certain embodiments, b2 of formula (b) is selected from the group consisting of 2, 3, and 4. In certain embodiments, b2 of formula (b) is 3.

[0364] In certain embodiments, b1 of Formula (b) is 2, b2 of Formula (b) is 3, and b3 and b4 are both about 450. In certain embodiments, b1 of Formula (b) is 2, b2 of Formula (b) is 3, and b3 and b4 are both about 225.

[0365] In certain embodiments, -Z comprises one moiety of formula (b). In certain embodiments, -Z comprises two moieties of formula (b). In certain embodiments, -Z comprises three moieties of formula (b). In certain embodiments, -Z comprises four moieties of formula (b). In certain embodiments, -Z comprises five moieties of formula (b). In certain embodiments, -Z comprises six moieties of formula (b).

[0366] In certain embodiments, -Z comprises a moiety of formula (c). [ka] During the ceremony, The dashed line is -L 2 - represents the bond to or to the remainder of -Z; c1 and c2 are independently integers in the range of 150 to 500, inclusive; in certain embodiments, in the range of 200 to 460, inclusive.

[0367] Optionally, the moiety of formula (c) is substituted with one or more substituents.

[0368] In certain embodiments, both c1 and c2 in formula (c) are the same integer.

[0369] In certain embodiments, c1 and c2 of formula (c) are in the range of 200 to 250, inclusive, and in certain embodiments, are about 225. In certain embodiments, c1 and c2 of formula (c) are in the range of 400 to 500, inclusive, and in certain embodiments, c1 and c2 of formula (c) are about 450.

[0370] In formula (c), c1 and c2 represent a branched PEG-based polymer, the moiety -Z containing at least 10% PEG, having one branch point and two PEG-based polymer arms, with a molecular weight of approximately 40 kDa. Therefore, each of the two PEG-based polymer arms has a molecular weight of approximately 20 kDa. In formula (c), c1 and c2 represent a branch point of -CH<.

[0371] In certain embodiments, -Z comprises one moiety of formula (c). In certain embodiments, -Z comprises two moieties of formula (c). In certain embodiments, -Z comprises three moieties of formula (c). In certain embodiments, -Z comprises four moieties of formula (c). In certain embodiments, -Z comprises five moieties of formula (c). In certain embodiments, -Z comprises six moieties of formula (c).

[0372] In certain embodiments, the moiety -Z is of formula (d): [ka] During the ceremony, The dashed line is -L 2 - indicates a bond to; -Z b -C 1-50 Alkyl, C 2-50 Alkenyl, and C 2-50 alkynyl; C 1-50Alkyl, C 2-50 Alkenyl, and C 2-50 Alkynyl may have one or more identical or different -R 1 may be substituted with 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)2N(R 2 )-, -S(O)N(R 2 )-, -S(O)2-, -S(O)-, -N(R 2 )S(O)2N(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 )-, optionally interrupted by one or more groups selected from the group consisting of; Each -T- is independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 each -T- is independently selected from the group consisting of one or more of the same or different -R 1 may be substituted with; Each-R 1 are independently halogen, -CN, 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)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a R 3b ), -SR 3 , -N(R 3R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 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; C 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).

[0373] Optionally, the moiety of formula (d) is substituted with one or more substituents.

[0374] In certain embodiments, BP of formula (d) a , -S a-, -S a′ -, -S a″ -, -S a″′ -, -P a′ , -P a″ , -P a″′ is as defined above for formula (a).

[0375] In certain embodiments, -Z in formula (d) a is of formula (b). In certain embodiments, b1, b2, b3, and b4 are as described for formula (b).

[0376] In certain embodiments, the moiety -Z of formula (Ia) or (Ib) is of formula (e): [ka] During the ceremony, The dashed line is -L 2 - indicates a bond to; e is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 and 15; -Z a is the following: [ka] and b1, b2, b3 and b4 are used as defined for formula (b).

[0377] Optionally, the moiety of formula (e) is substituted with one or more substituents.

[0378] In certain embodiments, b1, b2, b3, and b4 of formula (e) are as defined above for formula (b).

[0379] In certain embodiments, e in Formula (e) is 1. In certain embodiments, e in Formula (e) is 2. In certain embodiments, e in Formula (e) is 3. In certain embodiments, e in Formula (e) is 4. In certain embodiments, e in Formula (e) is 5. In certain embodiments, e in Formula (e) is 6. In certain embodiments, e in Formula (e) is 7. In certain embodiments, e in Formula (e) is 8. In certain embodiments, e in Formula (e) is 9. In certain embodiments, e in Formula (e) is 10. In certain embodiments, e in Formula (e) is 11. In certain embodiments, e in Formula (e) is 12. In certain embodiments, e in Formula (e) is 13. In certain embodiments, e in Formula (e) is 14. In certain embodiments, e in Formula (e) is 15.

[0380] In certain embodiments, e in formula (e) is selected from the group consisting of 2, 3, 4, 5, 6, 7, 8, and 9. In certain embodiments, e in formula (e) is selected from 3, 4, 5, and 6. In certain embodiments, e in formula (e) is 5.

[0381] In certain embodiments, e in formula (e) is 5, b1 in formula (e) is 2, b2 in formula (e) is 3, and b3 and b4 in formula (e) are both about 450.

[0382] In certain embodiments, the moiety -Z of formula (Ia) or (Ib) is of formula (ei) or (ei′): [ka] During the ceremony, The dashed line is -L 2 - indicates a bond to e is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 and 15; -Z a is the following: [ka] and b1, b2, b3 and b4 are used as defined for formula (b).

[0383] In certain embodiments, b1, b2, b3, and b4 of formulas (ei) and (ei′) are as defined above for formula (b).

[0384] In certain embodiments, e in formulas (ei) and (ei') is as described for formula (e).

[0385] In certain embodiments, b1 of formula (ei) and (ei') is 2, b2 of formula (ei) and (ei') is 3, and b3 and b4 of formula (ei) and (ei') are both about 450.

[0386] In certain embodiments, -Z of formula (Ia) or (Ib) is of formula (ei):

[0387] In certain embodiments, the moiety -Z is a branched PEG-based polymer comprising at least 10% PEG, has three branch points and four PEG-based polymer arms, and has a molecular weight of about 40 kDa. Thus, each of the four PEG-based polymer arms has a molecular weight of about 10 kDa. In certain embodiments, each of the three branch points is -CH<.

[0388] In certain embodiments, the moiety -Z is of formula (f): [ka] During the ceremony, The dashed line is -L 2 - indicates a bond to; BP f is a branch point selected from the group consisting of -N<, -CR< and >C<; -R is -H and C 1-6 selected from the group consisting of alkyl; f is BP f is 0 if -N< or -CR<, and f is BPf is 1 if >C<; -S f -, -S f′ -, -S f″ -and-S f″′ - are independently a chemical bond or independently 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 have one or more identical or different -R 1 may be substituted with 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)2N(R 2 )-, -S(O)N(R 2 )-, -S(O)2-, -S(O)-, -N(R 2 )S(O)2N(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 )-, optionally interrupted by one or more groups selected from the group consisting of; Each -T- is independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 each -T- is independently selected from the group consisting of one or more of the same or different -R 1 may be substituted with; Each R 1 are independently halogen, -CN, oxo (=O), -COOR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 R3a ), -S(O)2N(R 3 R 3a ), -S(O)N(R 3 R 3a ), -S(O)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 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; C 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′ , -Z a″ and -Z a″′ independently, [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).

[0389] Optionally, the moiety of formula (f) is substituted with one or more substituents.

[0390] In certain embodiments, BP of formula (f) a , -S a -, -S a′ -, -S a″ -, -S a″′ -, -P a′ , -P a″ and -P a″′ is as defined above for formula (a).

[0391] In certain embodiments, BP of formula (f) f is -CR< and r is 0. In certain embodiments, -R is -H.

[0392] In certain embodiments, -S of formula (f) f - is a chemical bond.

[0393] In certain embodiments, -Z in formula (f) a′ , -Z a″ and -Z a″′ have the same structure. In certain embodiments, -Z of formula (f) a′ , -Z a″ and -Z a″′ is that of equation (b).

[0394] In certain embodiments, b1, b2, b3, and b4 are as described for formula (b).

[0395] In certain embodiments, -S of formula (f) f - is a chemical bond, and BP in formula (f) a is -CR< and -R is -H. In certain embodiments, -S of formula (f) f- is a chemical bond, and BP in formula (f) a is -CR<, -R is -H, and -Z in formula (f) a′ , -Z a″ and -Z a″′ is that of equation (b).

[0396] In certain embodiments, -Z is of formula (g): [ka] During the ceremony, The dashed line is -L 2 - indicates a bond to; -S g -, -S g′ -and-S g″ - is independent, 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 have one or more identical or different -R 1 may be substituted with 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)2N(R 2 )-, -S(O)N(R 2 )-, -S(O)2-, -S(O)-, -N(R 2 )S(O)2N(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 )-, optionally interrupted by one or more groups selected from the group consisting of; Each -T- is independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10each -T- is independently selected from the group consisting of one or more of the same or different -R 1 may be substituted with; Each R 1 are independently halogen, -CN, 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)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 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; C 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 1-6The alkyl may be optionally substituted with one or more identical or different halogens; -Z a and -Z a′ independently, [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).

[0397] Optionally, the moiety of formula (g) is substituted with one or more substituents.

[0398] In certain embodiments, BP of formula (g) a , -S a -, -S a′ -, -S a″ -, -S a″′ -, -P a′ , -P a″ and -P a″′ is as defined above for formula (a).

[0399] In certain embodiments, -S in formula (g) g -C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 alkynyl, wherein one or more of the same or different -R 1 may be substituted with -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(R3 R 3a ), -S(O)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 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; C 1-6 The alkyl may be optionally 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.

[0400] In certain embodiments, -S in formula (g) g -C 1-6 alkyl.

[0401] In certain embodiments, -S in formula (g) g′ -C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 alkynyl, wherein one or more of the same or different -R 1 may be substituted with -R 1 is halogen, oxo (=O), -COOR 3 , -OR 3 , -C(O)R3 , -C(O)N(R 3 R 3a ), -S(O)2N(R 3 R 3a ), -S(O)N(R 3 R 3a ), -S(O)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 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; C 1-6 The alkyl may be optionally 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.

[0402] In certain embodiments, -S in formula (g) g′ -C 1-6 alkyl.

[0403] In certain embodiments, -S in formula (g) g″ -C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 alkynyl, which may be one or more of the same or different -R 1may be substituted with -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)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 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; C 1-6 The alkyl may be optionally 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.

[0404] In certain embodiments, -S in formula (g) g″ -C 1-6 alkyl.

[0405] In certain embodiments, -Z in formula (g) a and -Z a′have the same structure. In certain embodiments, -Z in formula (g) a and -Z a′ is that of equation (b).

[0406] In certain embodiments, -Z of formula (Ia) or (Ib) is of formula (gi): [ka] During the ceremony, The dashed line is -L 2 - indicates a bond to; -S g -, -S g′ -and-S g″ - is independent, 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 have one or more identical or different -R 1 may be substituted with 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)2N(R 2 )-, -S(O)N(R 2 )-, -S(O)2-, -S(O)-, -N(R 2 )S(O)2N(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 )-, optionally interrupted by one or more groups selected from the group consisting of; Each -T- is independently phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10each -T- is independently selected from the group consisting of one or more of the same or different -R 1 may be substituted with; Each R 1 are independently halogen, -CN, 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)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a R 3b ), -SR 3 , -N(R 3 R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 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; C 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 1-6The alkyl may be optionally substituted with one or more identical or different halogens; -Y a1 - and -Y a1′ -teeth, [ka] and; -Z a and -Z a′ independently, [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).

[0407] Optionally, the moiety of formula (gi) is substituted with one or more substituents.

[0408] In certain embodiments, -Y of formula (gi) a1 - and -Y a1′ - are both [ka] and During the ceremony, The dashed lines marked with asterisks are -Z a or -Z a′ is bonded to.

[0409] In certain embodiments, BP of formula (gi) a , -S a -, -S a′ -, -S a″ -, -S a″′ -, -P a′ , -P a″ and -P a″′is as defined above for formula (a).

[0410] In certain embodiments, -S of formula (gi) g -, -S g′ -and-S g″ - is as defined for formula (g). In certain embodiments, -Z in formula (gi) a and -Z a′ have the same structure. In certain embodiments, -Z of formula (gi) a and -Z a′ is of formula (b). In certain embodiments, b1, b2, b3, and b4 are as described for formula (b).

[0411] In certain embodiments, -Z is of formula (h): [ka] During the ceremony, The dashed line is -L 2 - indicates a bond to; Each-Z c is the following part: [ka] and During the ceremony, Each c1 is independently an integer in the range of 200-250.

[0412] Optionally, the moiety of formula (h) is substituted with one or more substituents.

[0413] In certain embodiments, both c1's of formula (h) are the same. In certain embodiments, both c1's of formula (h) are about 225.

[0414] In certain embodiments, -Z of formula (Ia) or (Ib) is of the following formula (ha): [ka] It is of During the ceremony, The dashed line is -L 2 - indicates a bond to; each k is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; -Y a1 - and -Y a1′ -teeth, [ka] and; Each-Z c The following part: [ka] and During the ceremony, Each c1 is independently an integer in the range of 200-250.

[0415] Optionally, the moiety of formula (ha) is substituted with one or more substituents.

[0416] In certain embodiments, each k in formula (ha) is independently selected from the group consisting of 2, 3, 4, 5, 6, and 7. In certain embodiments, both k in formula (ha) are the same.

[0417] In certain embodiments, both c1's of formula (ha) are the same.

[0418] In certain embodiments, both c1's of formula (ha) are about 225.

[0419] In certain embodiments, -Y of formula (ha) a1 - and -Y a1′ - are both [ka] and During the ceremony, The dashed line marked with an asterisk is -Z c is bonded to.

[0420] In certain embodiments, the moiety -Z is of the formula (hi): [ka] During the ceremony, The dashed line is -L 2 - indicates a bond to; Each-Z c The following part: [ka] and Each c1 is independently an integer in the range of 200-250.

[0421] Optionally, the moiety of formula (hi) is substituted with one or more substituents.

[0422] In certain embodiments, both c1's of formula (hi) are the same. In certain embodiments, both c1's of formula (hi) are about 225.

[0423] In certain embodiments, the moiety -Z of formula (Ia) or (Ib) is of formula (h-ia): [ka] During the ceremony, The dashed line is -L 2 - indicates a bond to; each k is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; -Y a1 - and -Y a1′ -teeth, [ka] and; Each-Z c The following part: [ka] and Each c1 is independently an integer in the range of 200-250.

[0424] In certain embodiments, each k in formula (h-ia) is independently selected from the group consisting of 2, 3, 4, 5, 6, and 7. In certain embodiments, both k in formula (h-ia) are the same.

[0425] In certain embodiments, both c1's of formula (h-ia) are the same. In certain embodiments, both c1's of formula (h-ia) are about 225.

[0426] In certain embodiments, -Y of formula (h-ia) a1 - and -Y a1′ - are both [ka] and The dashed line marked with an asterisk is -Z c is bonded to.

[0427] In certain embodiments, -Z of formula (Ia) or (Ib) comprises a moiety selected from the group consisting of: [ka] TIFF2025004178000081.tif239151TIFF2025004178000082.tif146159In the formula, The dashed line is -L 2 - indicates a bond to; s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14 and s15 are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10; -X d1 , -X d2 , -X d3 and -X d4 are each independently —OH, —SH, and —NR g1 R g2 is selected from the group consisting of: -X e1 , -X e2, -X e3 and -X e4 are independently -H, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 alkynyl; -R g1 and -R g2 are independently -H, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 alkynyl; -X 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 are independently -H, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 alkynyl; preferably -H; -Y d1 -, -Y d2 -, -Y d3 - and -Y d4 - are independent of each other, [ka] selected from the group consisting of: -Z d1 , -Z d2 , -Z d3 and -Z d4 are each independently a protein, in particular embodiments a random coil protein, in particular embodiments a random coil protein selected from the group consisting of PA, PAS, PAG, PG, and XTEN.

[0428] In certain embodiments, -Y in formula (j-iv), (jv) and (j-vi)d1 - and -Y d2 - and -Y of formula (j-vii) d1 -, -Y d2 -, -Y d3 - and -Y d4 -teeth, [ka] is.

[0429] In certain embodiments, -Y in formula (j-iv), (jv) and (j-vi) d1 - and -Y d2 - and -Y of formula (j-vii) d1 -, -Y d2 -, -Y d3 - and -Y d4 -teeth, [ka] and; During the ceremony, The dashed lines marked with asterisks are -Z d1 , -Z d2 , -Z d3 and -Z d4 The unmarked dashed line is pointing towards -L 2 -Heading towards.

[0430] In certain embodiments, -X in formula (ji) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 and -X f8 is -H; -X in formula (ji) d1 and -X d2 is -OH; -X in formula (ji) e1 and -X e2 is selected from the group consisting of —H and methyl; s1, s2, s3 and s4 in formula (ji) are selected from the group consisting of 2, 3, 4, 5 and 6.

[0431] In certain embodiments, -X in formula (ji) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 and -X f8 is -H; -X in formula (ji) d1 and -X d2 is -OH; -X in formula (ji) e1 and -X e2 is —H; s1, s2, s3 and s4 in formula (ji) are 4.

[0432] In certain embodiments, -Xf1, -Xf2, -Xf3, and -Xf4 of formula (j-ii) are -H; -Xd1, -Xd2, -Xd3, and -Xd2 of formula (j-ii) are -OH; -Xe1, -Xe2, -Xe3, and -Xe4 of formula (j-ii) are selected from the group consisting of -H and methyl; and s1, s2, s3, s4, and s5 of formula (j-ii) are selected from the group consisting of 1, 2, 3, 4, 5, and 6. In certain embodiments, -Xf1, -Xf2, -Xf3 and -Xf4 of formula (j-ii) are -H; -Xd1, -Xd2, -Xd3 and -Xd2 of formula (j-ii) are -OH; -Xe1, -Xe2, -Xe3 and -Xe4 of formula (j-ii) are -H; s1 is 4 of formula (j-ii), and s2, s3, s4 and s5 of formula (j-ii) are 1.

[0433] In certain embodiments, -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 of formula (j-iii) are selected from the group consisting of 2, 3, 4, 5 and 6. In certain embodiments, -X of 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.

[0434] In certain embodiments, -X in 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 in formula (j-iv) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; —Y d1 - and -Y d2 -teeth, [ka] is selected from the group consisting of:

[0435] In certain embodiments, -X in formula (j-iv) f1 , -X f2 , -X f3 , -X f4 , -X f5 and -X f6is -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, and s7 of formula (j-iv) is 4; -Y of formula (j-iv) d1 - and -Y d2 -teeth, [ka] is.

[0436] In certain embodiments, -X in 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, and s7 of formula (j-iv) is 4; -Y of formula (j-iv) d1 - and -Y d2 -teeth, [ka] and; In the formula, the dashed lines marked with asterisks represent -Z d1 , -Z d2 , -Z d3 and -Z d4 The unmarked dashed line is pointing towards -L 2 -Heading towards.

[0437] In certain embodiments, -X in formula (jv) f1 , -X f2 , -X f3 and -X f4 is -H; s1, s2, s3, s4 and s5 of formula (jv) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; -Y of formula (jv) d1 - and -Y d2 -teeth, [ka] is selected from the group consisting of:

[0438] In certain embodiments, -X in formula (jv) f1 , -X f2 , -X f3 and -X f4 is -H; s1 of formula (jv) is 3, s2 of formula (jv) is 2, s3 of formula (jv) is 1, s4 of formula (jv) is 2, s5 of formula (jv) is 1; -Y of formula (jv) d1 - and -Y d2 -teeth, [ka] is.

[0439] In certain embodiments, -X in formula (jv) f1 , -X f2 , -X f3 and -X f4 is -H; s1 of formula (jv) is 3, s2 of formula (jv) is 2, s3 of formula (jv) is 1, s4 of formula (jv) is 2, s5 of formula (jv) is 1; -Y of formula (jv) d1 - and -Y d2 -teeth, [ka] and; In the formula, the dashed lines marked with asterisks represent -Z d1 , -Z d2 , -Z d3 and -Z d4 The unmarked dashed line is pointing towards -L 2 -It is oriented towards.

[0440] In certain embodiments, -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 of formula (j-vi) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; -Y of formula (j-vi) d1 - and -Y d2 -teeth, [ka] is selected from the group consisting of:

[0441] In certain embodiments, -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 of formula (j-vi) is 4, s2 of formula (j-vi) is 5, s3 of formula (j-vi) is 2, s4 of formula (j-vi) is 4, s5 of formula (j-vi) is 4, s6 of formula (j-vi) is 5, s7 of formula (j-vi) is 2, s8 of formula (j-vi) is 4, s9 of formula (j-vi) is 4; -Y of formula (jv) d1 - and -Y d2 -teeth, [ka] is.

[0442] In certain embodiments, -X in formula (j-vi) f1 , -X f2 , -X f3 , -X f4 , -X f5 , -X f6 , -X f7 , -X f8 , -X f9 and -X f10is -H; s1 of formula (j-vi) is 4, s2 of formula (j-vi) is 5, s3 of formula (j-vi) is 2, s4 of formula (j-vi) is 4, s5 of formula (j-vi) is 4, s6 of formula (j-vi) is 5, s7 of formula (j-vi) is 2, s8 of formula (j-vi) is 4, s9 of formula (j-vi) is 4; -Y of formula (jv) d1 - and -Y d2 -teeth, [ka] and; In the formula, the dashed lines marked with asterisks represent -Z d1 , -Z d2 , -Z d3 and -Z d4 The unmarked dashed line is pointing towards -L 2 -It is oriented towards.

[0443] In certain embodiments, -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 d1 -, -Y d2 -, -Y d3 - and -Y d4 -teeth, [ka] is selected from the group consisting of:

[0444] In certain embodiments, -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; -H; s1 of formula (j-vii) is 4, s2 of formula (j-vii) is 4, s3 of formula (j-vii) is 5, s4 of formula (j-vii) is 2, s5 of formula (j-vii) is 4, s6 of formula (j-vii) is 5, s7 of formula (j-vii) is 2, s8 of formula (j-vii) is 4, s9 of formula (j-vii) is 4, s10 of formula (j-vii) is 5, s11 of formula (j-vii) is 2, s12 of formula (j-vii) is 4, s13 of formula (j-vii) is 5, s14 of formula (j-vii) is 2, s15 of formula (j-vii) is 4; -Y of formula (j-vii) d1 -, -Y d2 -, -Y d3 - and -Y d4 -teeth, [ka] is.

[0445] In certain embodiments, -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 f14is -H; -H; s1 of formula (j-vii) is 4, s2 of formula (j-vii) is 4, s3 of formula (j-vii) is 5, s4 of formula (j-vii) is 2, s5 of formula (j-vii) is 4, s6 of formula (j-vii) is 5, s7 of formula (j-vii) is 2, s8 of formula (j-vii) is 4, s9 of formula (j-vii) is 4, s10 of formula (j-vii) is 5, s11 of formula (j-vii) is 2, s12 of formula (j-vii) is 4, s13 of formula (j-vii) is 5, s14 of formula (j-vii) is 2, s15 of formula (j-vii) is 4; -Y of formula (j-vii) d1 -, -Y d2 -, -Y d3 - and -Y d4 -teeth, [ka] and; The dashed lines marked with asterisks are -Z d1 , -Z d2 , -Z d3 and -Z d4 The unmarked dashed line is pointing towards -L 2 -It is oriented towards.

[0446] In certain embodiments, -Z in formula (ji), (j-ii), (j-iii), (j-iv), (jv), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 have the same structure. In certain embodiments, -Z of formula (ji), (j-ii), (j-iii), (j-iv), (jv), (j-vi), and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 is a PA moiety. In certain embodiments, -Z in formulas (ji), (j-ii), (j-iii), (j-iv), (jv), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Zd4 is a PAS moiety. In certain embodiments, -Z in formulas (ji), (j-ii), (j-iii), (j-iv), (jv), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 is a PAG moiety. In certain embodiments, -Z in formulas (ji), (j-ii), (j-iii), (j-iv), (jv), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 is a PG moiety. In certain embodiments, -Z in formulas (ji), (j-ii), (j-iii), (j-iv), (jv), (j-vi) and (j-vii) d1 , -Z d2 , -Z d3 and -Z d4 is the XTEN portion.

[0447] In certain embodiments, the CNP conjugate has the formula (IIf): [ka] (In the formula, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed lines with asterisks indicate the structure: [ka] represents a bond to -Z having the formula Each-Z a teeth, [ka] where each c1 is independently an integer in the range of 200 to 250.

[0448] In certain embodiments, each c1 of Formula (IIf) is about 225.

[0449] In certain embodiments, the CNP conjugate has the formula (IIf-i): [ka] (In the formula, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed lines with asterisks indicate the structure: [ka] represents a bond to -Z having the formula Each-Z a teeth, [ka] wherein each c1 is independently an integer ranging from 200 to 250.

[0450] In certain embodiments, each c1 of Formula (IIf-i) is about 225.

[0451] In certain embodiments, the CNP conjugate has the formula (IIf-ii): [ka] (In the formula, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed lines with asterisks indicate the structure: [ka] represents a bond to -Z having the formula Each-Z a teeth, [ka] wherein each c1 is independently an integer ranging from 200 to 250.

[0452] In certain embodiments, each c1 in Formula (IIf-ii) is about 225.

[0453] In certain embodiments, -D of Formulas (IIf), (IIf-i), and (IIf-ii) is a CNP moiety, i.e., the conjugates of Formulas (IIf), (IIf-i), and (IIf-ii) are CNP conjugates. In certain embodiments, -D of Formulas (IIf), (IIf-i), and (IIf-ii) is a CNP moiety having the sequence of SEQ ID NO: 24, SEQ ID NO: 25, or SEQ ID NO: 30. In certain embodiments, -D of Formulas (IIf), (IIf-i), and (IIf-ii) is a CNP moiety having the sequence of SEQ ID NO: 24. In certain embodiments, the -D moiety of Formulas (IIf), (IIf-i), and (IIf-ii) is a CNP moiety having the sequence of SEQ ID NO: 20. In certain embodiments, the -D moiety of Formulas (IIf), (IIf-i), and (IIf-ii) is a CNP moiety having the sequence of SEQ ID NO: 21. In certain embodiments, the -D moiety of formula (IIf), (IIf-i), and (IIf-ii) is a CNP moiety having the sequence of SEQ ID NO: 22. In certain embodiments, the -D moiety of formula (IIf), (IIf-i), and (IIf-ii) is a CNP moiety having the sequence of SEQ ID NO: 23. In certain embodiments, the -D moiety of formula (IIf), (IIf-i), and (IIf-ii) is a CNP moiety having the sequence of SEQ ID NO: 30.

[0454] In certain embodiments, -D in formulas (IIf), (IIf-i) and (IIf-ii) is linked to -L through the nitrogen of the N-terminal amine functional group of CNP. 1 - is the CNP portion bound to

[0455] In certain embodiments, -D in formulas (IIf), (IIf-i) and (IIf-ii) is linked to -L through a nitrogen provided by the amine functionality of the lysine side chain of the CNP moiety. 1 - is the CNP portion bound to

[0456] In certain embodiments, when the CNP moiety is that of SEQ ID NO: 24, the lysine side chain is not part of the ring formed by the disulfide bridge between the cysteine ​​residues at positions 22 and 38.

[0457] Thus, in certain embodiments, when the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is connected to -L in the CNP conjugates of formulas (IIf), (IIf-i), and (IIf-ii) via the amine functionality provided by the side chain of lysine at position 9. 1 - is linked to.

[0458] In certain embodiments, when the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to -L in the CNP conjugates of formulas (IIf), (IIf-i), and (IIf-ii) via the amine functionality provided by the side chain of lysine at position 11. 1 - is linked to.

[0459] In certain embodiments, when the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to -L in the CNP conjugates of formulas (IIf), (IIf-i), and (IIf-ii) via the amine functionality provided by the side chain of lysine at position 15. 1 - is linked to.

[0460] In certain embodiments, when the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to -L in the CNP conjugates of formulas (IIf), (IIf-i), and (IIf-ii) via the amine functionality provided by the side chain of lysine at position 16. 1 - is linked to.

[0461] In certain embodiments, when the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to -L in the CNP conjugates of formulas (IIf), (IIf-i) and (IIf-ii) via the amine functionality provided by the side chain of lysine at position 20. 1 - is linked to.

[0462] In certain embodiments, when the CNP moiety is that of SEQ ID NO: 24, the lysine side chain is part of a ring formed by a disulfide bridge between the cysteine ​​residues at positions 22 and 38.

[0463] Thus, in certain embodiments, when the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to -L in the CNP conjugates of formulas (IIf), (IIf-i) and (IIf-ii) via the amine functionality provided by the side chain of lysine at position 26. 1 - is linked to.

[0464] In certain embodiments, the CNP conjugate has the formula (IIf-i), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 24, and is connected to -L through the amine functionality provided by the side chain of lysine at position 26. 1 - is bonded to

[0465] In certain embodiments, the CNP conjugate has the formula (IIf), where c1 is about 225, -D is a CNP moiety having the sequence of SEQ ID NO: 20, and -L is connected via the amine functionality provided by the side chain of lysine at position 30. 1 - is bonded to

[0466] In certain embodiments, the CNP conjugate has the formula (IIf-i), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 20, and is connected to -L through the amine functionality provided by the side chain of lysine at position 30. 1 - is bonded to

[0467] In certain embodiments, the CNP conjugate has the formula (IIf-ii), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 20, and is connected to -L through the amine functionality provided by the side chain of lysine at position 30. 1 - is bonded to

[0468] In certain embodiments, the CNP conjugate has the formula (IIf), where c1 is about 225, -D is a CNP moiety having the sequence of SEQ ID NO: 21, and -L is connected via the amine functionality provided by the side chain of lysine at position 29. 1 - is bonded to

[0469] In certain embodiments, the CNP conjugate has the formula (IIf-i), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 21, and is connected to -L through the amine functionality provided by the side chain of lysine at position 29. 1 - is bonded to

[0470] In certain embodiments, the CNP conjugate has the formula (IIf-ii), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 21, and is connected to -L through the amine functionality provided by the side chain of lysine at position 29. 1 - is bonded to

[0471] In certain embodiments, the CNP conjugate has the formula (IIf), where c1 is about 225, -D is a CNP moiety having the sequence of SEQ ID NO: 22, and -L is connected via the amine functionality provided by the side chain of lysine at position 28. 1 - is bonded to

[0472] In certain embodiments, the CNP conjugate has the formula (IIf-i), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 22, and is connected to -L through the amine functionality provided by the side chain of lysine at position 28. 1 - is bonded to

[0473] In certain embodiments, the CNP conjugate has the formula (IIf-ii), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 22, and is connected to -L through the amine functionality provided by the side chain of lysine at position 28. 1 - is bonded to

[0474] In certain embodiments, the CNP conjugate has the formula (IIf), where c1 is about 225, -D is a CNP moiety having the sequence of SEQ ID NO: 23, and -L is connected via the amine functionality provided by the side chain of lysine at position 27. 1 - is bonded to

[0475] In certain embodiments, the CNP conjugate has the formula (IIf-i), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 23, and is connected to -L through the amine functionality provided by the side chain of lysine at position 27. 1 - is bonded to

[0476] In certain embodiments, the CNP conjugate has the formula (IIf-ii), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 23, and is connected to -L through the amine functionality provided by the side chain of lysine at position 27. 1 - is bonded to

[0477] In certain embodiments, the CNP conjugate has the formula (IIf), where c1 is about 225, -D is a CNP moiety having the sequence of SEQ ID NO: 30, and -L is connected via the amine functionality provided by the side chain of lysine at position 27. 1 - is bonded to

[0478] In certain embodiments, the CNP conjugate has the formula (IIf-i), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 30, and is connected to -L through the amine functionality provided by the side chain of lysine at position 27. 1 - is bonded to

[0479] In certain embodiments, the CNP conjugate has the formula (IIf-ii), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 30, and is connected to -L through the amine functionality provided by the side chain of lysine at position 27. 1 - is bonded to

[0480] In certain embodiments, the CNP conjugate has the formula (IIf-ii), where c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 24, and is connected to -L through the amine functionality provided by the side chain of lysine at position 26. 1 - is bonded to

[0481] It is understood that the positions of the cysteines and lysines described above vary depending on the length of the CNP moiety, and that one skilled in the art would have no difficulty identifying corresponding cysteines and lysines in longer or shorter versions of the CNP moiety, and would also understand, for example, that some lysines may not be present in a shorter CNP moiety. It is further understood that, for example, as a result of site-directed mutagenesis, more lysine residues may be present in the non-cyclization and / or cyclization portions of the CNP moiety.

[0482] In certain embodiments, the CNP conjugate has the formula (IIf), where c1 is about 225, -D is a CNP moiety having the sequence of SEQ ID NO: 24, and -L is connected via the amine functionality provided by the side chain of lysine at position 26. 1 - is bonded to

[0483] In certain embodiments, the CNP conjugate of the invention has the formula (IIf'): [ka] (In the formula, The unmarked dashed line indicates attachment to the nitrogen provided by the side chain of lysine at position 26 of the CNP portion of SEQ ID NO: 24 by forming an amide bond; The dashed lines with asterisks indicate the structure: [ka] represents a bond to -Z having the formula each Z a teeth, [ka] where each c1 is independently an integer in the range of 200 to 250.

[0484] In certain embodiments, each c1 in Formula (IIf') is about 225.

[0485] In certain embodiments, the CNP conjugate has the formula (IIf-i′): [ka] (In the formula, The unmarked dashed line indicates attachment to the nitrogen provided by the side chain of lysine at position 26 of the CNP portion of SEQ ID NO: 24 by forming an amide bond; The dashed lines with asterisks indicate the structure: [ka] represents a bond to -Z having the formula each Z a teeth, [ka] wherein each c1 is independently an integer ranging from 200 to 250.

[0486] In certain embodiments, each c1 in Formula (IIf-i') is about 225.

[0487] In certain embodiments, the CNP conjugate has the formula (IIf-ii'): [ka] (In the formula, The unmarked dashed line indicates attachment to the nitrogen provided by the side chain of lysine at position 26 of the CNP portion of SEQ ID NO: 24 by forming an amide bond; The dashed lines with asterisks indicate the structure: [ka] represents a bond to -Z having the formula each Z a teeth, [ka] wherein each c1 is independently an integer ranging from 200 to 250.

[0488] In certain embodiments, each c1 in Formula (IIf-ii') is about 225.

[0489] In certain embodiments, the CNP conjugate is of formula (IIfa): [ka] During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk represents the structure: [ka] represents the bond to -Z, k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; Each-Z a teeth, [ka] and Each c1 is independently an integer in the range of 200-250.

[0490] In certain embodiments, k in Formula (IIfa) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[0491] In certain embodiments, each c1 of Formula (IIfa) is about 225.

[0492] In certain embodiments, the CNP conjugate is of formula (IIfa-i): [ka] During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk represents the structure: [ka] represents the bond to -Z, k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; Each-Z a teeth, [ka] and Each c1 is independently an integer in the range of 200-250.

[0493] In certain embodiments, k in formula (IIfa-i) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[0494] In certain embodiments, each c1 of Formula (IIfa-i) is about 225.

[0495] In certain embodiments, the CNP conjugate is of formula (IIfa-ii): [ka] During the ceremony, The unmarked dashed line indicates the attachment of the CNP moiety, -D, to the nitrogen by forming an amide bond; The dashed line marked with an asterisk represents the structure: [ka] represents the bond to -Z, k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; Each-Z a teeth, [ka] and Each c1 is independently an integer in the range of 200-250.

[0496] In certain embodiments, each c1 of Formula (IIfa-ii) is about 225.

[0497] In certain embodiments, the CNP portion of the CNP conjugates of formula (IIfa), (IIfa-i), and (IIfa-ii) has the sequence of SEQ ID NO:25.

[0498] In certain embodiments, the CNP portion of the CNP conjugates of formula (IIfa), (IIfa-i), and (IIfa-ii) has the sequence of SEQ ID NO:20.

[0499] In certain embodiments, the CNP portion of the CNP conjugates of formula (IIfa), (IIfa-i), and (IIfa-ii) has the sequence of SEQ ID NO:21.

[0500] In certain embodiments, the CNP portion of the CNP conjugates of formula (IIfa), (IIfa-i), and (IIfa-ii) has the sequence of SEQ ID NO:22.

[0501] In certain embodiments, the CNP portion of the CNP conjugates of formula (IIfa), (IIfa-i), and (IIfa-ii) has the sequence of SEQ ID NO:23.

[0502] In certain embodiments, the CNP portion of the CNP conjugates of formula (IIfa), (IIfa-i), and (IIfa-ii) has the sequence of SEQ ID NO:30.

[0503] In certain embodiments, the CNP portion of the CNP conjugates of formula (IIfa), (IIfa-i), and (IIfa-ii) has the sequence of SEQ ID NO:24.

[0504] In one embodiment, the CNP moiety is linked to -L in the CNP conjugates of formula (IIfa), (IIfa-i) and (IIfa-ii) via the nitrogen of the N-terminal amine functional group of CNP. 1 - is bonded to.

[0505] In certain embodiments, the CNP conjugate is of formula (IIfa'): [ka] During the ceremony, The unmarked dashed line indicates attachment to the nitrogen provided by the side chain of the 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: [ka] represents the bond to -Z, k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; each Z a teeth, [ka] and Each c1 is independently an integer in the range of 200-250.

[0506] In certain embodiments, k in formula (IIfa′) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[0507] In certain embodiments, each c1 of Formula (IIfa') is about 225.

[0508] In certain embodiments, the CNP conjugate is of the following formula (IIfa-i′): [ka] During the ceremony, The unmarked dashed line indicates attachment to the nitrogen provided by the side chain of the 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: [ka] represents the bond to -Z, k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; each Z a teeth, [ka] and Each c1 is independently an integer in the range of 200-250.

[0509] In certain embodiments, k in formula (IIfa-i′) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[0510] In certain embodiments, each c1 of Formula (IIfa-i') is about 225.

[0511] In certain embodiments, the CNP conjugate is of the following formula (IIfa-ii'): [ka] During the ceremony, The unmarked dashed line indicates attachment to the nitrogen provided by the side chain of the 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: [ka] represents the bond to -Z, k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; each Z a teeth, [ka] and Each c1 is independently an integer in the range of 200-250.

[0512] In certain embodiments, k in formula (IIfa-ii′) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[0513] In certain embodiments, each c1 of Formula (IIfa-ii') is about 225.

[0514] In certain embodiments, the dry formulations of the present invention may include one or more excipients, such as, for example, preservatives, stabilizers, anti-adsorption agents, cryoprotectants, antioxidants, and other adjuvants, etc. It is understood that a single excipient may have multiple functions, such as dual, triple, etc.

[0515] In certain embodiments, the dry pharmaceutical formulations of the present invention may include a preservative, such as a preservative selected from the group consisting of benzoic acid, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, potassium sorbate, chlorobutanol, benzyl alcohol, phenylmercuric nitrate thimerosal, sorbic acid, potassium sorbate, chlorocresol, benzalkonium chloride, 2-ethoxyethanol, chlorhexidine, chlorobutanol, phenylethyl alcohol, phenylmercuric acetate, m-cresol, and mixtures thereof.

[0516] In certain embodiments, the dry formulations of the present invention may comprise stabilizers such as stabilizers selected from the group consisting of alanine; arginine; aspartic acid; glycine; histidine; lysine; proline; sugars such as glucose, sucrose, trehalose, etc.; polyols such as glycerol, mannitol, sorbitol, etc.; salts such as potassium phosphate, sodium sulfate, etc.; chelating agents such as EDTA, hexaphosphate, etc.; ligands such as divalent metal ions; organic molecules such as other salts or phenol derivatives; and oligomers or polymers such as cyclodextrin, dextran, dendrimers, PEG, PVP, protamine, HSA, etc.

[0517] In certain embodiments, the dry pharmaceutical formulation of the present invention may contain an anti-adsorption agent, such as an anti-adsorption agent selected from the group consisting of ionic or non-ionic surfactants or other proteins or soluble polymers, used to competitively coat or adsorb to the inner surface of the formulation or formulation container, such as poloxamer (Pluronic F-68), PEG dodecyl ether (Brij35), dextran, polyethylene glycol, PEG-polyhistidine, BSA, HSA, gelatin, etc. The type and concentration of the excipient are selected depending on the effect to be avoided, but typically, a surfactant monolayer is formed at the interface at concentrations higher than the CMC.

[0518] In certain embodiments, the dry pharmaceutical formulation of the present invention can contain a cryoprotectant, such as a cryoprotectant selected from the group consisting of sugars, polyhydric alcohols, surfactants, amino acids, non-aqueous solvents, and peptides. During freeze-drying or spray-drying, the cryoprotectant can counteract the destabilizing effects caused by hydrogen bond disruption and water removal. Trehalose is particularly effective in reducing moisture-induced aggregation and also improves thermal stability, which may be caused by exposure of hydrophobic groups of compounds to water. Mannitol and sucrose can also be used as the sole lyoprotectant / cryoprotectant or in combination with each other, with a higher mannitol:sucrose ratio known to enhance the physical stability of compounds in dry pharmaceutical formulations. Mannitol can also be combined with trehalose. Trehalose can also be combined with sorbitol, or sorbitol can be used as the sole cryoprotectant. Starch or a starch derivative can also be used.

[0519] In certain embodiments, the dry pharmaceutical formulation of the present invention can comprise an antioxidant, such as an antioxidant selected from the group consisting of methionine, butylhydroxytoluene, butylhydroxyanisole, tocopherol, propyl gallate, ascorbic acid, sodium bisulfite, ethylenediaminetetraacetic acid (EDTA), cysteine, glutathione, monothioglycerol, poly(ethyleneimine), vitamin E, ectoine, morin, and mixtures thereof. In certain embodiments, the dry pharmaceutical formulation of the present invention can comprise an antioxidant, such as an antioxidant selected from the group consisting of methionine, butylhydroxytoluene, butylhydroxyanisole, tocopherol, propyl gallate, ascorbic acid, ethylenediaminetetraacetic acid (EDTA), poly(ethyleneimine), vitamin E, ectoine, morin, and mixtures thereof.

[0520] In certain embodiments, the dry pharmaceutical formulation of the present invention may comprise additional excipients selected from the group consisting of wetting agents, viscosity modifiers, and antibiotics.

[0521] In certain embodiments, the dry pharmaceutical formulation of the present invention comprises a CNP conjugate, succinic acid, trehalose, and Tris.

[0522] In certain embodiments, the dry pharmaceutical formulation according to the present invention comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 1.3 to 45.4 wt% Succinic acid: 0.2 to 3.2% by weight Trehalose dihydrate: 52.6-98.4% by weight Tris: 0.1~5.6% by weight Includes.

[0523] In certain embodiments, the dry pharmaceutical formulation according to the present invention comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 1.3 to 38.7 wt% Succinic acid: 0.2 to 3.2% by weight Trehalose dihydrate: 52.6-98.4% by weight Tris: 0.1~5.6% by weight Includes.

[0524] Due to the wide variation in patient weight ranges, e.g., 3-60 kg, in certain embodiments, it is advantageous to have dry pharmaceutical formulations with different strengths, i.e., different concentrations, of the CNP conjugate, e.g., high, medium, and low strengths. Thus, in one embodiment, the dry pharmaceutical formulation is provided in multiple concentrations, e.g., in 2 different concentrations, e.g., in 3 different concentrations, e.g., in 4 different concentrations, e.g., in 5 different concentrations, e.g., in 6 different concentrations, e.g., in 7 different concentrations, e.g., in 8 different concentrations, e.g., in 9 different concentrations, e.g., in 10 different concentrations, e.g., in 11 different concentrations, e.g., in 12 different concentrations.

[0525] In certain embodiments, the dry pharmaceutical formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 23.8-38.7 wt% Succinic acid: 0.2 to 3.1% by weight Trehalose dihydrate: 52.6-75.9% by weight Tris: 0.1~5.6% by weight Includes.

[0526] In certain embodiments, the dry pharmaceutical formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 5.8 to 12.4 wt% Succinic acid: 0.3 to 3.2% by weight Trehalose dihydrate: 78.8-93.8% by weight Tris: 0.1~5.6% by weight Includes.

[0527] In certain embodiments, the dry pharmaceutical formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 1.4 to 3.7 wt% Succinic acid: 0.3 to 3.2% by weight Trehalose dihydrate: 87.5-98.2% by weight Tris: 0.1~5.6% by weight Includes.

[0528] In certain embodiments, the dry pharmaceutical formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 28.8-33.2 wt% Succinic acid: 0.6 to 1.6% by weight Trehalose dihydrate: 62.4-70.4% by weight Tris: 0.2~2.8% by weight Includes.

[0529] In certain embodiments, the dry pharmaceutical formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 7.4-8.8 wt% Succinic acid: 0.8 to 2.0% by weight Trehalose dihydrate: 85.7-91.6% by weight Tris: 0.2~3.5% by weight Includes.

[0530] In certain embodiments, the dry pharmaceutical formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 2.0 to 2.5 wt% Succinic acid: 0.8 to 2.1% by weight Trehalose dihydrate: 91.7-97.0% by weight Tris: 0.2~3.8% by weight Includes.

[0531] In certain embodiments, the dry pharmaceutical formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 32.2 to 34.0 wt% Succinic acid: 0.9 to 1.0% by weight Trehalose dihydrate: 64.5-65.5% by weight Tris: 0.5~1.4% by weight Includes.

[0532] In certain embodiments, the dry pharmaceutical formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 8.3-8.9 wt% Succinic acid: 1.2 to 1.3% by weight Trehalose dihydrate: 88.6-89.1% by weight Tris: 0.7~1.8% by weight Includes.

[0533] In certain embodiments, the dry pharmaceutical formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 2.3-2.4 wt% Succinic acid: 1.3 to 1.4% by weight Trehalose dihydrate: 94.4-95.6% by weight Tris: 0.7~1.9% by weight Includes.

[0534] In certain embodiments, the dry pharmaceutical formulation comprises about 45.4 wt. % CNP conjugate, about 0.9 wt. % succinic acid, about 52.6 wt. % trehalose dihydrate, and about 1.1 wt. % Tris, based on the total weight of the dry pharmaceutical formulation.

[0535] In certain embodiments, the dry pharmaceutical formulation comprises 45.4 wt. % CNP conjugate, 0.9 wt. % succinic acid, 52.6 wt. % trehalose dihydrate, and 1.1 wt. % Tris, based on the total weight of the dry pharmaceutical formulation.

[0536] In certain embodiments, the dry pharmaceutical formulation comprises about 32.6 wt. % CNP conjugate, about 1.0 wt. % succinic acid, about 65.2 wt. % trehalose dihydrate, and about 1.2 wt. % Tris, based on the total weight of the dry pharmaceutical formulation.

[0537] In certain embodiments, the dry pharmaceutical formulation comprises 33 wt % CNP conjugate, 1 wt % succinic acid, 65 wt % trehalose dihydrate, and 1 wt % Tris, based on the total weight of the dry pharmaceutical formulation.

[0538] In certain embodiments, the dry pharmaceutical formulation comprises 32.6 wt % CNP conjugate, 1.0 wt % succinic acid, 65.2 wt % trehalose dihydrate, and 1.2 wt % Tris, based on the total weight of the dry pharmaceutical formulation.

[0539] In certain embodiments, the dry pharmaceutical formulation comprises about 8.2 wt. % CNP conjugate, about 1.2 wt. % succinic acid, about 89.1 wt. % trehalose dihydrate, and about 1.5 wt. % Tris, based on the total weight of the dry pharmaceutical formulation.

[0540] In certain embodiments, the dry pharmaceutical formulation comprises 8 wt % CNP conjugate, 1 wt % succinic acid, 89 wt % trehalose dihydrate, and 2 wt % Tris, based on the total weight of the dry pharmaceutical formulation.

[0541] In certain embodiments, the dry pharmaceutical formulation comprises 8.2 wt % CNP conjugate, 1.2 wt % succinic acid, 89.1 wt % trehalose dihydrate, and 1.5 wt % Tris, based on the total weight of the dry pharmaceutical formulation.

[0542] In certain embodiments, the dry pharmaceutical formulation comprises about 2.3 wt. % CNP conjugate, about 1.3 wt. % succinic acid, about 94.9 wt. % trehalose dihydrate, and about 1.6 wt. % Tris, based on the total weight of the dry pharmaceutical formulation.

[0543] In certain embodiments, the dry pharmaceutical formulation comprises 2 wt % CNP conjugate, 1 wt % succinic acid, 95 wt % trehalose dihydrate, and 2 wt % Tris, based on the total weight of the dry pharmaceutical formulation.

[0544] In certain embodiments, the dry pharmaceutical formulation comprises 2.3 wt % CNP conjugate, 1.3 wt % succinic acid, 94.9 wt % trehalose dihydrate, and 1.6 wt % Tris, based on the total weight of the dry pharmaceutical formulation.

[0545] The dry pharmaceutical formulation of the present invention is a dry pharmaceutical formulation that is stable for at least 6 months, such as at least 7 months, for example at least 8 months, such as at least 9 months, for example at least 10 months, for example at least 11 months, for example at least 12 months. In a particular embodiment, the dry pharmaceutical formulation is stable for at least 14 months, such as at least 16 months, for example at least 18 months, for example at least 20 months, for example at least 22 months, for example at least 24 months, such as at least 40 months, for example at least 60 months.

[0546] In certain embodiments, the dry pharmaceutical formulation of the present invention is stored at a temperature ranging from -80°C to 25°C, for example, from -20°C to 25°C, for example, from -15°C to 25°C, for example, from -10°C to 25°C, for example, from -5°C to 5°C, from -5°C to 25°C, for example, from 0°C to 5°C, for example, from 0°C to 25°C, for example, from 2°C to 10°C, or for example, from 4°C to 8°C. In certain embodiments, the dry pharmaceutical formulation is stored at 2°C. In certain embodiments, the dry pharmaceutical formulation is stored at 5°C. In certain embodiments, the dry pharmaceutical formulation is stored at 6°C. In certain embodiments, the dry pharmaceutical formulation is stored at 8°C. In certain embodiments, the dry pharmaceutical formulation is stored at 10°C. In certain embodiments, the dry pharmaceutical formulation is stored at 16°C. In certain embodiments, the dry pharmaceutical formulation is stored at 20°C. In certain embodiments, the dry pharmaceutical formulation is stored at 25°C. In certain embodiments, the dry pharmaceutical formulation is stored at 30°C. In certain embodiments, the dry pharmaceutical formulation is stored at 40°C.

[0547] In certain embodiments, the dried pharmaceutical formulation is stable for at least 12 months when stored at 0-10° C. In certain embodiments, the dried pharmaceutical formulation is stable for at least 12 months when stored at 2-10° C. In certain embodiments, the dried pharmaceutical formulation is stable for at least 12 months when stored at 4-8° C. In certain embodiments, the dried pharmaceutical formulation is stable for at least 12 months when stored at 5° C.

[0548] In certain embodiments, the dry pharmaceutical formulation is stable for at least 24 months when stored at 5° C. In certain embodiments, the dry pharmaceutical formulation is stable for at least 36 months when stored at 5° C. In certain embodiments, the dry pharmaceutical formulation is stable for at least 48 months when stored at 5° C. In certain embodiments, the dry pharmaceutical formulation is stable for at least 60 months when stored at 5° C.

[0549] In certain embodiments, the dry pharmaceutical formulation is stable for 6 months when stored at 25° C. In certain embodiments, the dry pharmaceutical formulation is stable for at least 6 months when stored at 25° C. In certain embodiments, the dry pharmaceutical formulation is stable for at least 12 months when stored at 25° C.

[0550] In certain embodiments, the dry pharmaceutical formulation is stable for 6 months when stored at 30° C. In certain embodiments, the dry pharmaceutical formulation is stable for at least 6 months when stored at 30° C. In certain embodiments, the dry pharmaceutical formulation is stable for at least 12 months when stored at 30° C.

[0551] In certain embodiments, the dry pharmaceutical formulation is stable for 6 months when stored at 40° C. In certain embodiments, the dry pharmaceutical formulation is stable for at least 6 months when stored at 40° C. In certain embodiments, the dry pharmaceutical formulation is stable for at least 12 months when stored at 40° C.

[0552] In certain embodiments, the dry pharmaceutical formulations of the present invention are provided as a single dose, ie, a solution comprising the dry pharmaceutical formulation of a CNP conjugate comprises one therapeutic dose.

[0553] A preferred method of drying is freeze-drying, ie the pharmaceutical formulation is a freeze-dried pharmaceutical formulation.

[0554] Another aspect of the invention is a method for producing a dry pharmaceutical formulation according to the invention, comprising the steps of: (i) mixing a CNP conjugate with at least a buffer and a bulking agent; (ii) adjusting the pH of the mixture of step (i); (iii) optionally filtering the mixture from step (ii); (iv) transferring the mixture from step (ii) or (iii) into a container in an amount corresponding to the desired number of doses; (v) drying the mixture; (vi) sealing the container Includes; The order of steps (ii) and (iii) may be reversed.

[0555] It will be appreciated that during step (v) of drying the mixture, the bulking agent may become a lyoproctant.

[0556] In certain embodiments, steps (ii) and (iii) are not reversed.

[0557] In certain embodiments, the CNP conjugate in step (i) is mixed with a buffer and a bulking agent.

[0558] The number of doses transferred into the container in step (iv) may be at least 1, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30, or may be greater than 30.

[0559] In certain embodiments, the method for producing a dry pharmaceutical formulation according to the present invention comprises: (i) mixing the CNP conjugate with at least succinic acid and trehalose dihydrate; CNP conjugate: 0.9 to 82.1 mg / mL Succinic acid: 1.3-57.6mM Trehalose dihydrate: 67-111.6 mg / mL obtaining a formulation comprising: (ii) adjusting the pH of the mixture of step (i) to a pH in the range of pH 4.0 to pH 6.0; (iii) optionally filtering the mixture from step (ii); (iv) transferring into a container an amount of the mixture from step (ii) or (iii) corresponding to the desired number of doses; (v) drying the mixture; (vi) sealing the container Includes; The order of steps (ii) and (iii) may be reversed.

[0560] In certain embodiments, steps (ii) and (iii) are not reversed.

[0561] In certain embodiments, in step (ii), the pH of the mixture of step (i) is adjusted with Tris.

[0562] In certain embodiments, the CNP conjugate in step (i) is mixed with succinic acid and trehalose dihydrate to form a CNP conjugate: 0.9 to 82.1 mg / mL Succinic acid: 1.3 to 57.6 mM Trehalose dihydrate: 67-111.6 mg / mL to obtain a formulation comprising:

[0563] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 19.8 to 73.6 mg / mL Succinic acid: 1.7 to 50 mM Trehalose dihydrate: 63-100 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.0 to pH 6.0.

[0564] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 19.8 to 73.6 mg / mL Succinic acid: 1.7 to 50 mM Trehalose dihydrate: 56-100 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.0 to pH 6.0.

[0565] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 4.2 to 16.5 mg / mL Succinic acid: 1.7 to 36.4 mM Trehalose dihydrate: 67-105 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.0 to pH 6.0.

[0566] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 1.0 to 4.4 mg / mL Succinic acid: 1.7 to 33 mM Trehalose dihydrate: 67-105 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.0 to pH 6.0.

[0567] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 27.5 to 50.5 mg / mL Succinic acid: 5.1 to 20.3 mM Trehalose dihydrate: 67-95 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.0 to pH 6.0.

[0568] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 5.8 to 10.8 mg / mL Succinic acid: 5.1 to 20.3 mM Trehalose dihydrate: 72-105 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.0 to pH 6.0.

[0569] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 1.5-2.9 mg / mL Succinic acid: 5.1 to 20.3 mM Trehalose dihydrate: 73-105 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.0 to pH 6.0.

[0570] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 37.4 to 42.9 mg / mL Succinic acid: 9.3-10.2mM Trehalose dihydrate: 71-87 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.5 to pH 5.5.

[0571] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 7.5-9.1 mg / mL Succinic acid: 9.3-11mM Trehalose dihydrate: 75-97 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.5 to pH 5.5.

[0572] In certain embodiments, the formulation in step (i) comprises: CNP conjugate: 1.9-2.4 mg / mL Succinic acid: 9.3-11mM Trehalose dihydrate: 77-97 mg / mL Including, In step (ii), the pH is adjusted to a range of pH 4.5 to pH 5.5.

[0573] In certain embodiments, the formulation in step (i) comprises about 60.4 mg / mL LCNP conjugate, about 10 mM succinic acid, and about 70 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0574] In a particular embodiment, the formulation in step (i) comprises 60.4 mg / mL LCNP conjugate, 10 mM succinic acid, 70 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0575] In certain embodiments, the formulation in step (i) comprises about 39.6 mg / mL LCNP conjugate, about 10 mM succinic acid, and about 79.0 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0576] In a particular embodiment, the formulation in step (i) comprises 40 mg / mL LCNP conjugate, 10 mM succinic acid, 79 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0577] In a particular embodiment, the formulation in step (i) comprises 39.6 mg / mL LCNP conjugate, 10 mM succinic acid, 79.0 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0578] In certain embodiments, the formulation in step (i) comprises about 8.2 mg / mL LCNP conjugate, about 10 mM succinic acid, and about 89.0 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0579] In a particular embodiment, the formulation in step (i) comprises 8 mg / mL LCNP conjugate, 10 mM succinic acid, and 89 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0580] In a particular embodiment, the formulation in step (i) comprises 8.2 mg / mL LCNP conjugate, 10 mM succinic acid, and 89.0 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0581] In certain embodiments, the formulation in step (i) comprises about 2.2 mg / mL LCNP conjugate, about 10 mM succinic acid, and about 89.5 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0582] In certain embodiments, the formulation in step (i) comprises 2 mg / mL LCNP conjugate, 10 mM succinic acid, and 90 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0583] In a particular embodiment, the formulation in step (i) comprises 2.2 mg / mL LCNP conjugate, 10 mM succinic acid, and 89.5 mg / mL trehalose dihydrate, and in step (ii) the pH is adjusted to pH 5.

[0584] Before administering the dry pharmaceutical formulation of the present invention to a patient in need thereof, the dry pharmaceutical formulation is reconstituted (reconstituted). The reconstitution of the dry pharmaceutical formulation into a reconstituted formulation is carried out by adding a predetermined amount of a reconstitution solution to the dry pharmaceutical formulation. Thus, a further aspect of the present invention is a method for reconstituting the dry pharmaceutical formulation of the present invention, comprising: (a) contacting a dry pharmaceutical formulation of the present invention with a reconstitution solution.

[0585] Another aspect of the present invention is a reconstituted pharmaceutical preparation obtainable from the method for reconstituting a dried pharmaceutical preparation of the present invention.

[0586] It is understood that the bulking agent of the dry pharmaceutical formulation of the present invention performs a bulking function only in the dry pharmaceutical formulation, i.e., not in the reconstituted pharmaceutical formulation, and when the dry pharmaceutical formulation is reconstituted, the bulking agent performs the function of an isotonic agent.

[0587] Reconstitution can be carried out in the container in which the dry pharmaceutical formulation comprising the CNP conjugate is provided, e.g., a vial; a syringe, e.g., a dual-chamber syringe; an ampoule; a cartridge, e.g., a dual-chamber cartridge; or the dry pharmaceutical formulation can be transferred to a different container, followed by reconstitution.

[0588] In certain embodiments, the container in which the dry pharmaceutical formulation is reconstituted is a vial.

[0589] In certain embodiments, the container for reconstitution of the dried pharmaceutical formulation is a syringe. In certain embodiments, the container for reconstitution of the dried pharmaceutical formulation is a dual-chamber syringe. In certain embodiments, the container for reconstitution of the dried pharmaceutical formulation is a cartridge. In certain embodiments, the container for reconstitution of the dried pharmaceutical formulation is a dual-chamber cartridge.

[0590] In certain embodiments, the dry pharmaceutical formulation according to the present invention is provided in a first chamber of a dual-chamber syringe and the reconstitution solution is provided in a second chamber of the dual-chamber syringe.

[0591] In certain embodiments, the dry pharmaceutical formulation according to the present invention is provided in a first chamber of a dual-chamber cartridge and the reconstitution solution is provided in a second chamber of the dual-chamber cartridge.

[0592] Another aspect of the present invention refers to a container containing the dried pharmaceutical formulation or reconstituted formulation of the present invention.

[0593] The reconstitution time of the dry pharmaceutical formulation has been found to be suitable for therapeutic settings. In certain embodiments, the addition of a surfactant, such as polysorbate 20 or polysorbate 80, to a formulation containing a CNP conjugate increases the reconstitution time of the dry pharmaceutical formulation. Thus, in certain embodiments, the dry pharmaceutical formulation according to the present invention does not contain a surfactant, such as polysorbate 20 or polysorbate 80.

[0594] The reconstitution solution is a sterile liquid, such as water or a buffer, which may contain further additives, such as preservatives and / or antimicrobial agents.

[0595] In certain embodiments, the reconstitution solution comprises one or more preservatives and / or antimicrobial agents and / or antioxidants.

[0596] In certain embodiments, the reconstitution solution includes one or more preservatives.

[0597] The preservative may be selected from the group consisting of m-cresol, benzoic acid, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, potassium sorbate, chlorobutanol, benzyl alcohol, phenylmercuric nitrate, thimerosal, sorbic acid, potassium sorbate, chlorocresol, benzalkonium chloride, 2-ethoxyethanol, chlorhexidine, chlorobutanol, phenylethyl alcohol, phenylmercuric acetate, and mixtures thereof.

[0598] In certain embodiments, the preservative is m-cresol. In certain embodiments, the preservative is benzyl alcohol. In certain embodiments, the preservative is benzoic acid. In certain embodiments, the preservative is phenol. In certain embodiments, the preservative is methylparaben. In certain embodiments, the preservative is ethylparaben. In certain embodiments, the preservative is propylparaben. In certain embodiments, the preservative is butylparaben. In certain embodiments, the preservative is potassium sorbate. In certain embodiments, the preservative is benzyl alcohol. In certain embodiments, the preservative is phenylmercuric nitrate. In certain embodiments, the preservative is thimerosal. In certain embodiments, the preservative is sorbic acid. In certain embodiments, the preservative is potassium sorbate. In certain embodiments, the preservative is chlorocresol. In certain embodiments, the preservative is benzalkonium chloride. In certain embodiments, the preservative is 2-ethoxyethanol. In certain embodiments, the preservative is chlorhexidine. In certain embodiments, the preservative is chlorobutanol. In certain embodiments, the preservative is phenylethyl alcohol. In certain embodiments, the preservative is phenylmercuric acetate.

[0599] In certain embodiments, the preservative has a concentration in the range of 1 to 10 mg / mL, 1.5 to 3.5 mg / mL, or 2 to 3 mg / mL.

[0600] The antioxidant may be selected from the group consisting of methionine, butylhydroxytoluene, butylhydroxyanisole, tocopherol, propyl gallate, ascorbic acid, ethylenediaminetetraacetic acid (EDTA), poly(ethyleneimine), vitamin E, and mixtures thereof.

[0601] In certain embodiments, the preservative is methionine. In certain embodiments, the preservative is butylhydroxytoluene. In certain embodiments, the preservative is butylhydroxyanisole. In certain embodiments, the preservative is tocopherol. In certain embodiments, the preservative is propyl gallate. In certain embodiments, the preservative is ethylenediaminetetraacetic acid. In certain embodiments, the preservative is poly(ethyleneimine). In certain embodiments, the preservative is vitamin E.

[0602] As defined herein, the term "methionine" is intended to encompass both D-methionine and L-methionine, as well as mixtures thereof. In certain embodiments, the term "methionine" refers to L-methionine. In certain embodiments, the term "methionine" refers to D-methionine. In certain embodiments, the term "methionine" refers to a mixture of D-methionine or L-methionine. In certain embodiments, the term "methionine" refers to L-methionine hydrochloride.

[0603] As defined herein, the term "EDTA" includes all forms of EDTA known in the art, e.g., EDTA salts, e.g., EDTA metal salts, e.g., disodium EDTA, dipotassium EDTA, calcium EDTA, dimagnesium EDTA, or mixtures thereof. In certain embodiments, EDTA refers to disodium EDTA. In certain embodiments, the term "EDTA" refers to calcium EDTA. In certain embodiments, the term "EDTA" refers to anhydrous EDTA.

[0604] In certain embodiments, the molar ratio of antioxidant to CNP moiety is about 0.1:1 to about 100:1. In certain embodiments, the molar ratio of antioxidant to CNP moiety is about 0.1:1 to about 70:1. In certain embodiments, the molar ratio of antioxidant to CNP moiety is about 0.1:1 to about 15:1. In certain embodiments, the molar ratio of antioxidant to CNP moiety is about 1:1 to about 10:1. In certain embodiments, the molar ratio of antioxidant to CNP moiety is about 3:1 to about 7:1.

[0605] In certain embodiments, the regeneration solution does not contain an antimicrobial agent.

[0606] In certain embodiments, the regeneration solution includes an antimicrobial agent.

[0607] In certain embodiments, the regeneration solution comprises one or more excipients.

[0608] In certain embodiments, the regeneration solution is sterile water.

[0609] In certain embodiments, the regeneration solution is sterile water containing 0.7-1.1% benzyl alcohol.

[0610] In certain embodiments, the regeneration solution is sterile water containing 0.9% benzyl alcohol.

[0611] In certain embodiments, the regeneration solution includes a pH adjuster.

[0612] As used herein, the term "pH adjuster" refers to a compound used to adjust the pH of the regeneration solution.

[0613] In certain embodiments, the pH adjuster can be an acid or an acid salt thereof, and the acid can be selected from the group consisting of acetic acid, citric acid, succinic acid, hydrochloric acid, phosphoric acid, carbonic acid, nitric acid, and mixtures thereof.

[0614] In certain embodiments, the pH adjuster can be a base or a basic salt thereof, and the base can be selected from the group consisting of Tris (tris(hydroxymethyl)aminomethane), sodium hydroxide, potassium hydroxide, lysine, and mixtures thereof.

[0615] After reconstitution, the single-dose formulation containing the CNP conjugate has a volume of 4 mL or less, e.g., about 0.03 to about 1.1 mL. In certain embodiments, the volume is about 0.03 mL. In certain embodiments, the volume is about 0.05 mL. In certain embodiments, the volume is about 0.1 mL. In certain embodiments, the volume is about 0.3 mL. In certain embodiments, the volume is about 0.8 mL. In certain embodiments, the volume is about 1 mL. In certain embodiments, the volume is about 2 mL. In certain embodiments, the volume is about 3 mL. In certain embodiments, the volume is about 4 mL.

[0616] In certain embodiments, the CNP conjugate is administered in a formulation in an amount sufficient to provide a therapeutically effective amount of CNP for at least 3 days, e.g., at least 4 days, e.g., at least 5 days, e.g., at least 6 days per administration.

[0617] In certain embodiments, the CNP conjugate is administered in a formulation in an amount sufficient to provide a therapeutically effective amount of CNP for one week.

[0618] The buffer maintains the pH of the reconstituted formulation within a desired range, in certain embodiments, the pH of the reconstituted formulation is 6 or less, since the reversible linkages within the CNP-conjugate may not be stable under basic conditions.

[0619] In certain embodiments, the pH of the reconstituted formulation is about pH 4 to about pH 6. In certain embodiments, the pH of the reconstituted formulation is about pH 4.5 to about pH 5.5. In certain embodiments, the pH of the reconstituted formulation is about 5. In certain embodiments, the pH of the reconstituted formulation is 5.

[0620] In certain embodiments, the buffering agent has a concentration in the reconstituted formulation ranging from 1.3 to 57.6 mM. In certain embodiments, the buffering agent has a concentration in the reconstituted formulation ranging from 1.7 to 33 mM. In certain embodiments, the buffering agent has a concentration in the reconstituted formulation ranging from 5.1 to 20.3 mM. In certain embodiments, the buffering agent has a concentration of about 10 mM in the reconstituted formulation.

[0621] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 0.9-82.1mg / mL Succinic acid: 1.3-57.6mM Trehalose dihydrate: 67-111.6 mg / mL Including, It has a pH range of pH 4.0 to pH 6.0.

[0622] The reconstituted formulations are also available in different strengths, i.e., different concentrations, of the CNP conjugate, for example, high, medium, and low strengths. Thus, it is another aspect of the present invention that the reconstituted formulations of the present invention are provided in different concentrations of the CNP conjugate, for example, in two different concentrations, for example, in three different concentrations, for example, in four different concentrations, for example, in five different concentrations, for example, in six different concentrations, for example, in seven different concentrations, for example, in eight different concentrations, for example, in nine different concentrations, for example, in ten different concentrations, for example, in eleven different concentrations, for example, in twelve different concentrations.

[0623] Those skilled in the art will recognize that there may be rounding errors in the concentrations of ingredients contained in the reconstituted formulations of the present invention, i.e., when starting from a particular dry formulation, the composition of the corresponding reconstituted formulation may vary to some extent due to necessary rounding.

[0624] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 19.8 to 73.6 mg / mL Succinic acid: 1.7 to 50 mM Trehalose dihydrate: 63-100 mg / mL and has a pH in the range of pH 4.0 to pH 6.0.

[0625] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 4.2 to 16.5 mg / mL Succinic acid: 1.7 to 36.4 mM Trehalose dihydrate: 67-105 mg / mL and has a pH in the range of pH 4.0 to pH 6.0.

[0626] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 1.0 to 4.4 mg / mL Succinic acid: 1.7 to 33 mM Trehalose dihydrate: 67-105 mg / mL and has a pH in the range of pH 4.0 to pH 6.0.

[0627] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 27.5 to 50.5 mg / mL Succinic acid: 5.1 to 20.3 mM Trehalose dihydrate: 67-95 mg / mL and has a pH in the range of pH 4.0 to pH 6.0.

[0628] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 5.8 to 10.8 mg / mL Succinic acid: 5.1 to 20.3 mM Trehalose dihydrate: 72-105 mg / mL and has a pH in the range of pH 4.0 to pH 6.0.

[0629] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 1.5-2.9 mg / mL Succinic acid: 5.1 to 20.3 mM Trehalose dihydrate: 73-105 mg / mL and has a pH in the range of pH 4.0 to pH 6.0.

[0630] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 37.4 to 42.9 mg / mL Succinic acid: 9.3-10.2mM Trehalose dihydrate: 71-87 mg / mL and has a pH in the range of pH 4.5 to pH 5.5.

[0631] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 7.5-9.1 mg / mL Succinic acid: 9.3-11mM Trehalose dihydrate: 75-97 mg / mL and has a pH in the range of pH 4.5 to pH 5.5.

[0632] In certain embodiments, the regenerating formulation comprises: CNP conjugate: 1.9-2.4 mg / mL Succinic acid: 9.3-11mM Trehalose dihydrate: 77-97 mg / mL and has a pH in the range of pH 4.5 to pH 5.5.

[0633] In a specific embodiment, the reconstituted formulation comprises about 60.4 mg / mL LCNP conjugate, about 10 mM succinic acid, about 70 mg / mL trehalose dihydrate, and has a pH of 5.

[0634] In a specific embodiment, the reconstituted formulation comprises 60.4 mg / mL LCNP conjugate, 10 mM succinic acid, 70 mg / mL trehalose dihydrate, and has a pH of 5.

[0635] In a specific embodiment, the reconstituted formulation comprises about 60.4 mg / mL LCNP conjugate, about 10 mM succinic acid, about 56 mg / mL trehalose dihydrate, and has a pH of 5.

[0636] In a specific embodiment, the reconstituted formulation comprises 60.4 mg / mL LCNP conjugate, 10 mM succinic acid, 56 mg / mL trehalose dihydrate, and has a pH of 5.

[0637] In certain embodiments, the reconstituted formulation comprises about 39.6 mg / mL LCNP conjugate, about 10 mM succinic acid, about 79.0 mg / mL trehalose dihydrate, and has a pH of 5.

[0638] In a specific embodiment, the reconstituted formulation comprises 40 mg / mL LCNP conjugate, 10 mM succinic acid, 79 mg / mL trehalose dihydrate, and has a pH of 5.

[0639] In a specific embodiment, the reconstituted formulation comprises 39.6 mg / mL LCNP conjugate, 10 mM succinic acid, 79.0 mg / mL trehalose dihydrate, and has a pH of 5.

[0640] In certain embodiments, the reconstituted formulation comprises about 8.2 mg / mL LCNP conjugate, about 10 mM succinic acid, about 89.0 mg / mL trehalose dihydrate, and has a pH of 5.

[0641] In a specific embodiment, the reconstituted formulation comprises 8 mg / mL LCNP conjugate, approximately 10 mM succinic acid, 89 mg / mL trehalose dihydrate, and has a pH of 5.

[0642] In a specific embodiment, the reconstituted formulation comprises 8.2 mg / mL LCNP conjugate, 10 mM succinic acid, 89.0 mg / mL trehalose dihydrate, and has a pH of 5.

[0643] In certain embodiments, the reconstituted formulation comprises about 2.2 mg / mL LCNP conjugate, about 10 mM succinic acid, about 89.5 mg / mL trehalose dihydrate, and has a pH of 5.

[0644] In a specific embodiment, the reconstituted formulation comprises 2 mg / mL LCNP conjugate, 10 mM succinic acid, 90 mg / mL trehalose dihydrate, and has a pH of 5.

[0645] In a specific embodiment, the reconstituted formulation comprises 2.2 mg / mL LCNP conjugate, 10 mM succinic acid, 89.5 mg / mL trehalose dihydrate, and has a pH of 5.

[0646] The regenerative formulations can be administered by injection, infusion, intradermal, subcutaneous, intramuscular, intravenous, intraosseous, and intraperitoneal.

[0647] In certain embodiments, the regenerative formulation comprising the CNP conjugate is administered by subcutaneous injection.

[0648] In certain embodiments, the regenerative formulation comprising the CNP conjugate is administered by subcutaneous injection using a syringe, needle, pen injector, or autoinjector.

[0649] In certain embodiments, the regenerative formulation comprising the CNP conjugate is administered by subcutaneous injection using a syringe.

[0650] In certain embodiments, the regenerative formulation comprising the CNP conjugate is administered by subcutaneous injection using a pen syringe.

[0651] In certain embodiments, the regenerative formulation containing the CNP conjugate is administered by subcutaneous injection using an auto-injector.

[0652] Another aspect of the present invention is the dried or reconstituted pharmaceutical formulation of the present invention for use as a medicament.

[0653] In another aspect, the invention relates to a dried or reconstituted pharmaceutical formulation of the invention for use in the treatment, suppression, delay or prevention of one or more diseases treatable, suppressed, delayed or preventable by CNP.

[0654] In certain embodiments, the present invention relates to a dried or reconstituted pharmaceutical formulation of the present invention for use in the treatment of one or more diseases treatable by CNP.

[0655] A further aspect of the present invention is a method for treating, suppressing, delaying or preventing one or more diseases treatable by CNP in a patient, comprising administering to the patient a therapeutically effective amount of a regenerative pharmaceutical formulation of the present invention.

[0656] In certain embodiments, the patient is an adult. In certain embodiments, the patient is a pediatric patient.

[0657] In certain embodiments, the one or more diseases that can be treated, inhibited, delayed, or prevented by CNP are selected from the group consisting of bone-related disorders, e.g., bone-related disorders such as skeletal dysplasia; cancer; autoimmune diseases; fibrosis; inflammatory diseases; central nervous system diseases, e.g., neurodegenerative diseases; infectious diseases; pulmonary diseases; heart and vascular diseases; metabolic diseases; and eye diseases.

[0658] In certain embodiments, the one or more disorders that can be treated, inhibited, delayed, or prevented by CNP are achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasia, lethal dysplasia, osteogenesis imperfecta, achondrogenesis imperfecta, chondrodysplasia punctata, homozygous achondroplasia, chiroptera dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly syndrome, rhizometaphyseal dysplasia, Jansen metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, osteogenesis imperfecta, osseous dysplasia varus, femoral brachiosteogenesis imperfecta, Langer metapodial dysplasia, Niebergelt metapodial dysplasia, Robinow syndrome, Reinhardt syndrome, acroostosis imperfecta, peripheral osteogenesis imperfecta, deficiency, Kneist dysplasia, fibrochondrogenesis, Roberts syndrome, distal intermedius dysplasia, micromelia, Morquio syndrome, Kneist syndrome, metamorphic dysplasia, spondyloepiphyseal dysplasia, neurofibromatosis, Legius syndrome, Leopard syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, cardio-facio-cutaneous 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 Crouzondermoskeletal syndrome) syndrome), dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, segmental dysplasia, enchondromatosis, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis, bone poikilosis, hemorrhagic shock, hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, renal edema, hepatic edema, acute renal failure, chronic renal failure, glaucoma, ocular hypertension, multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, breast cancer, growth retardation, cranial deformities, orthodontic defects defects), cervical spinal cord compression, spinal cord stenosis, hydrocephalus, hearing loss due to chronic otitis, cardiovascular disease, neurological disease and obesity.

[0659] In certain embodiments, the one or more disorders that can be treated, inhibited, delayed, or prevented by CNP include achondroplasia, e.g., homozygous achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasia, lethal dysplasia, osteogenesis imperfecta, achondroplasia, chondrodysplasia punctata, chiropteran dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly syndrome, rhizal chondrodysplasia punctata (proximal short chondrodysplasia punctata), Jansen metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, osteogenesis imperfecta, osseous dysplasia volvulus, femoral brachiosteogenesis imperfecta, Langer metapodial dysplasia, Niebergelt metapodial dysplasia, Robinow syndrome, Reinhardt syndrome, acromioclavicular dysplasia, Hypoplasia, peripheral osteogenesis imperfecta, Kneist dysplasia, fibrochondrogenesis, Roberts syndrome, distal intermedius dysplasia, micromelia, Morquio syndrome, Kneist syndrome, metamorphic dysplasia, spondyloepiphyseal dysplasia, neurofibromatosis, Legius syndrome, Leopard syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, cardio-facio-cutaneous syndrome, Costello syndrome, SHOX deficiency, idiopathic dwarfism, growth hormone deficiency, osteoarthritis, cleidocranial dysplasia, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzondermoskeletal syndrome) syndrome), dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, segmental dysplasia, enchondromatosis, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis, bone poikilosis, hemorrhagic shock, hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, renal edema, hepatic edema, acute renal failure, chronic renal failure, glaucoma, ocular hypertension, multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, breast cancer, growth retardation, cranial deformities, orthodontic defectsdefects), cervical spinal cord compression, spinal cord stenosis, hydrocephalus, hearing loss due to chronic otitis, obesity, disorders involving aberrant RAS-mitogen activated protein kinase signaling, pulmonary hypertension, vascular disorders, endothelial dysfunction, cirrhosis, hepatic ascites, hepatic fibrosis, hepatorenal syndrome, asthma, pulmonary fibrosis, chronic kidney disease, cardiorenal syndrome, dyspnea and lysosomal storage diseases, e.g., mucopolysaccharidosis.

[0660] In certain embodiments, the one or more disorders that can be treated, inhibited, delayed, or prevented by CNP include arrhythmias, such as cardiac arrhythmias or sinus arrhythmia; atrial fibrillation; atrial flutter; bradycardia; Brugada syndrome; premature beats; commotio cordis; heart block; long QT syndrome; parasystoles; preexcitation syndrome; tachycardia; ventricular fibrillation; ventricular flutter; cardiac conduction disorders; low cardiac output; cardiac hypertrophy; dilated cardiomyopathy; hypertrophy, such as left ventricular hypertrophy or right ventricular hypertrophy; cardiomyopathies, such as alcoholic, dilated, hypertrophic, restrictive, diabetic, or Chagas cardiomyopathy; arrhythmogenic right ventricular dysplasia; endocardial fibroelastosis; endomyocardial fibrosis; glycogen storage disease type IIb; Kearns-Sayre syndrome; myocardial reperfusion injury; myocarditis; sarcoglycanopathies; endocarditis, such as bacterial or non-infective endocarditis; cardiac arrest; sudden cardiac arrest; The cardiovascular disease is one or more selected from the group consisting of cardiac death; out-of-hospital cardiac arrest; cardio-renal syndrome; paroxysmal dyspnea; cardiac edema, heart failure such as diastolic or systolic heart failure; valvular heart disease; aortic regurgitation; aortic stenosis; heart valve prolapse; mitral regurgitation; mitral stenosis; pulmonary atresia; pulmonary regurgitation; pulmonary stenosis; tricuspid atresia; tricuspid regurgitation; tricuspid stenosis; myocardial ischemia; acute coronary syndrome; angina pectoris; coronary artery disease; Kounis syndrome; myocardial infarction; pulmonary heart disease; ventricular dysfunction such as left or right ventricular dysfunction; ventricular outflow tract obstruction; aortic stenosis, pulmonary stenosis; hypertension; atherosclerosis; restenosis; critical limb ischemia; peripheral arterial disease; ischemia such as ischemia-reperfusion injury or ischemic injury; cardiac edema and abnormal fluid retention in myocardial edema.

[0661] In certain embodiments, the one or more diseases that can be treated, inhibited, delayed, or prevented by CNP is selected from the group consisting of ischemic heart disease, such as myocardial infarction; congestive heart failure; arrhythmia, and atherosclerosis.

[0662] In certain embodiments, the one or more diseases that can be treated, inhibited, delayed, or prevented by CNP are one or more central nervous system diseases selected from the group consisting of cerebral ischemia, such as ischemic hypoxia; cerebral infarction; transient ischemic attack; vertebrobasilar insufficiency; cerebrovascular disease; stroke; intracranial hemorrhage; corneal neovascularization; corneal transplantation; graft-versus-host disease; graft rejection; glaucoma, such as angle-closure, neovascular, open-angle, or low-tension glaucoma; ischemic optic neuropathy; central serous chorioretinopathy; retinopathies, such as diabetic or hypertensive retinopathy; retinal degeneration; macular degeneration; geographic atrophy; macular edema; Stargardt's disease; vitreomacular dystrophy; wet macular degeneration; retinoschisis; retinal detachment; retinal perforation; retinal hemorrhage; retinal neovascularization; retinal vein occlusion; retinal artery occlusion; retinopathy of prematurity; and proliferative vitreoretinopathy.

[0663] In certain embodiments, the one or more disorders are selected from the group consisting of hypophosphatasia, hypochondroplasia, Muenke syndrome, hypertension, osteogenesis imperfecta, and achondroplasia.

[0664] In certain embodiments, the one or more diseases treatable with the CNP invention are hypophosphatasia. In certain embodiments, the one or more diseases treatable with CNP are achondroplasia. In certain embodiments, the one or more diseases treatable with CNP are Muenke syndrome. In certain embodiments, the one or more diseases treatable with CNP are hypertension. In certain embodiments, the one or more diseases treatable with CNP are osteogenesis imperfecta. In certain embodiments, the one or more diseases treatable with CNP are achondroplasia. [Example]

[0665] Materials and Methods Unless otherwise stated, all materials were commercially available.

[0666] RP-HPLC was used to determine the content and purity of compound (1) and to detect free CNP-38. Mobile phase A consisted of 0.05% TFA in water, and mobile phase B consisted of 0.04% TFA in acetonitrile. A Waters Acquity CSH C18, 130 Å, 1.7 μm, 2.1 × 100 mm column was used. The flow rate was set at 0.3 mL / min, detection was performed at a wavelength of 215 nm, and the column operating temperature was 60 °C (±1 °C). Samples were diluted with formulation buffer containing 0.5% Tween®-20. Content was determined by peak area comparison to a reference solution.

[0667] The purity of compound (1) was determined using SE-HPLC. The mobile phase consisted of 15 mM sodium phosphate pH 7.40, 135 mM sodium chloride, and 0.2% Pluronic F-68 in water. A GE Superdex 200 Increase 10 / 300 GL column was used. The flow rate was set at 0.75 mL / min, detection was performed at 215 nm, and the column operating temperature was room temperature. Samples were diluted with formulation buffer containing 0.05% Pluronic F-68.

[0668] Peptide mapping of compound 1 was used to evaluate the conversion of aspartate 28 to isoaspartate 28 and the oxidation of methionine 33 to methionine oxide 33 (Met(O)). Thermolysin digestion of 1 was performed at a thermolysin / CNP-38 ratio of 1:20 (by weight) at 37°C for 7 h at pH 7.5. The resulting peptide mixture was separated by RP-HPLC on a Waters Acquity UPLC HSST3, 100 Å, 1.8 μm, 2.1 × 150 mm column using 0.10% (v / v) TFA in water as mobile phase A and 0.09% TFA in acetonitrile as mobile phase B, with detection at 210 nm. The flow rate was set at 0.28 mL / min, and the column operating temperature was 45°C (±1°C). Thermolysin fragments containing aspartate (28) and methionine (33) were characterized via LC-MS. In a typical analysis, fragments are compared to a reference mixture containing these fragment peptides. Fragments were quantified relative to their individual peak areas relative to the peak area of ​​the corresponding unmodified fragment. Under the conditions of thermolysin digestion and subsequent RP-HPLC, the formation of the isoaspartate product of aspartate (28) and the oxidation of methionine (33) could be quantified for (1).

[0669] Biopotency analysis was performed by in vitro release of CNP-38 from (1) followed by biopotency analysis of the released CNP-38 in a cellular assay.

[0670] In vitro release and RP-HPLC quantification of CNP-38 from (1): (1) was incubated at pH 10.0 and 15°C for 24 hours. The pH was adjusted by 1:3.5 (volume ratio) dilution of the reconstituted formulation with release buffer (0.5 M boric acid, 10 mM methionine, 2.383 g / L Pluronic F-68; pH adjusted to 10.0 with 4 M aqueous NaOH). (For example, 50 μL of a 3.6 mg CNP-38 equivalent / mL solution of (1) was diluted with 175 μL of release buffer.) After incubation, release was stopped by 1:1 (volume ratio) dilution with 5% (volume ratio) acetic acid. The amount of released CNP-38 was determined by peak area comparison to a standard solution of known CNP-38 content.

[0671] Measurement of the biopotency of functional cGMP-stimulated release of CNP-38 in HEK293 cells: An NPR-B-overexpressing Hek293 cell line was generated as follows. The coding region for the NPR-B reference sequence was cloned into a lentiviral vector under a CMV promoter for constitutive receptor expression. A bicistronic element located on the vector for puromycin resistance was used as a eukaryotic cell selectable marker. After transduction, stably growing cell pools were subjected to qRT-PCR to confirm receptor mRNA expression compared to parental Hek293 cells. Stimulation of the NPR-B receptor by CNP-38 results in intracellular production of the second messenger cGMP, which is detected by a commercially available cGMP assay.

[0672] Cells were routinely cultured in DMEM / Glutamax / HEPES medium containing 10% FBS and 1% puromycin solution (370 μg / mL) at 37°C and 5% CO2. For each assay, cells were suspended in DMEM + 2% BSA and incubated at 37°C and 5% CO2 for 3 hours. Serial dilutions of the released CNP mixture in stimulation buffer (DMEM + 2% BSA + 0.1 mM IBMX + 0.1% Tween®-20) were prepared and added to the cells (further diluted 1:2 with the CNP-38 serial dilutions). After 60 minutes of incubation at 37°C and 5% CO2, cells were lysed and cGMP levels were determined using a commercially available cGMP TR-FRET assay (Cisbio, cGMP kit, catalog number 62GM2PEB). Potency was determined using a four-parameter logistic curve fit in PLA software validated by parallel line analysis, compared to a CNP-38 standard curve.

[0673] The amounts of degraded CNP-38 (deimidated and aspartimide variants) were determined using RP-HPLC. Mobile phase A consisted of 0.05% TFA in water, and mobile phase B consisted of 0.04% TFA in acetonitrile. A Waters Acquity HSS T3 C18, 100 Å, 1.8 μm, 2.1 × 100 mm column was used. The flow rate was set at 0.5 mL / min, detection was performed at 215 nm, and the column operating temperature was set at 30 °C. The deimidated and aspartimide variants were quantified relative to the peak area of ​​unmodified CNP-38.

[0674] [Example 1] Synthesis of compound (1) [ka] Compound (1) was synthesized according to the method described in WO2017 / 118693 for conjugate 11i.

[0675] [Example 2] Stability test of freeze-dried formulation containing compound (1) Eight different formulations (F1–F8) containing compound (1) were prepared as shown in Table 1. 200 μL of each formulation was filled into vials and lyophilized. The water content of the lyophilized material was determined by Karl Fischer titration to be 0.4% or less for all formulations. Each formulation contained an appropriate amount of compound (1) to achieve a CNP-38 concentration of 3.6 mg / mL after reconstitution with 200 μL of water. The formulations were placed in an incubator set to maintain temperatures of 25°C or 40°C. After 3 months (T3M), the formulations were reconstituted with water and analyzed. The results indicate a favorable purity profile of compound (1) in the trehalose-containing formulation compared to the mannitol-containing formulation. Furthermore, it was observed that no additives were required to prevent methionine oxidation.

[0676] [Table 1]

[0677] [Table 2]

[0678] [Example 3] Stability testing of liquid and freeze-dried formulations containing compound (1) Formulation (F9) (1) was prepared, and 200 μL of the formulation was filled into a vial and lyophilized. The lyophilized formulation contained an appropriate amount of CNP-38 to achieve a concentration of 3.0 mg / mL after reconstitution with 200 μL of water. The lyophilized product and the liquid formulation used to prepare it were placed in an incubator set to maintain a temperature of 40°C.

[0679] After 21 days (T21D), the lyophilisate was reconstituted with water and the formulation was analysed. The results indicate that lyophilisation protects 1 from degradation such as methionine oxidation and isoaspartate formation.

[0680] [Table 3]

[0681] [Table 4]

[0682] [Example 4] (1) Stability test of formulations containing surfactants Three different formulations of (1) (F10-F12) were prepared, and 1080 μL of each was filled into vials and lyophilized. Each formulation contained an appropriate amount of (1) to achieve a CNP-38 concentration of 3.5 mg / mL after reconstitution with 1000 μL of water. Some formulations contained polysorbate 20 (PS20) or polysorbate 80 (PS80) as a surfactant. The lyophilized formulations were placed in an incubator set to maintain 40°C / 75% RH. After 1 month (T1M) and 3 months (T3M), the formulations were reconstituted with 1 mL of water and analyzed.

[0683] Formulations F11 and F12 containing surfactants show a slight increase in methionine oxidation of (1) under stress conditions and a clear effect on the regeneration time (without shaking) after the addition of 1 mL of water compared to F10.

[0684] [Table 5]

[0685] [Table 6]

[0686] [Example 5] Long-term stability test of lyophilized formulations containing compound (1) Lyophilized formulations of compound (1) (F13) were prepared by lyophilizing 1080 μL in vials. The formulations contained an appropriate amount of (1) to achieve nominal concentrations of 3.9 mg CNP-38 / vial and 3.6 mg CNP-38 / mL after reconstitution with 1000 μL of water. The formulations were placed in incubators set to maintain 5°C, 25°C / 60% RH, 30°C / 65% RH, and 40°C / 75% RH, respectively. After 3 months (T3M), 4 months (T4M), 6 months (T6M), 9 months (T9M), 12 months (T12M), 18 months (T18M), and 24 months (T24M), the formulations were reconstituted with 1000 μL of water and analyzed.

[0687] The results demonstrate high stability of Compound (1) in lyophilized formulation F13, as shown herein in terms of content, purity, and biopotency. Furthermore, the formulation exhibited good stability when tested in the relevant pharmaceutical methods. The lyophilized formulation was characterized by a white cake, and no change in cake appearance was detected regardless of storage. The residual moisture content was very low (0.03%) after lyophilization and increased only slightly during the test period. There was no significant change in reconstitution time during the storage period. Visual inspection revealed that the samples were substantially free of visible particles throughout the test for all samples; low amounts of subvisible particles, ≥ 25 μm and ≥ 10 μm, were observed (measured by flow-through microscopy) regardless of storage temperature and storage time. A slight change in color from B9 to >B8 was observed upon storage at 5°C for 12 months, but no color increase was observed upon storage at 25°C and 30°C for 6 months or at 40°C for 3 months. (Color was assessed using a spectral colorimeter, and absolute color assessment was performed according to the European Pharmacopoeia, 8th Edition, Monograph 2.2.2.) There was no change in turbidity (measured using a turbidimeter) of lyophilized samples stored for up to 12 months at 2-8°C, 25°C / 60% relative humidity, and 30°C / 65% relative humidity. A negligible increase in turbidity was measured in samples stored for 3 months at 40°C / 75% relative humidity. No change in appearance was observed upon storage at 5°C for up to 12 months, with values ​​of less than 1 NTU (nephelometric turbidity unit). Values ​​remained below 1 NTU for 6 months at 25°C, 30°C, and 40°C. pH was unaffected by storage. Finally, osmolality at TO was within the physiological range and no changes were detected during the study.

[0688] [Table 7]

[0689] [Table 8]

[0690] [Example 6] Stability Test of a Low-Dose Formulation Containing Compound (1) In a vial, a lyophilized formulation (F"14") of (1) was prepared by lyophilizing 1060 μL. The formulation was assumed to contain an amount of (1) sufficient to obtain a nominal concentration of CNP-38 0.80 mg / vial and CNP-38 0.75 mg / mL after reconstitution. The formulation was placed in an incubator set to maintain 5°C, 25°C / 60% RH, 30°C / 65% RH, and 40°C / 75% RH. After 1 month (T1M), 3 months (T3M), 6 months (T6M), and 12 months (T12M), the formulation was reconstituted with 1.0 mL of water and analyzed.

[0691] The results show a high stability of Compound (1) in the lyophilized formulation F14, as shown herein in terms of content, purity, and biological activity. Furthermore, the formulation showed good stability when tested by relevant pharmaceutical methods. The lyophilized formulation was characterized by a white cake, and no change in cake appearance was detected regardless of storage. The residual moisture content was very low after lyophilization (0.31%) and increased slightly during the test period. No significant change in reconstitution time was observed during the storage period. Visual observation showed that the reconstituted solution essentially contained no visible particles at the available time points (evaluated according to Ph.Eur. 2.9.20 and USP <790>). Furthermore, for all samples, a low amount of invisible particles, i.e., particles of 0 ≥ 25 μm and 5 ≥ 10 μm, were observed regardless of storage temperature and storage time (measured by flow-through microscopy). A slight increase in color tone from =WFI to <B9 was observed after 3 months of storage at 30°C and 40°C (determined according to Ph.Eur. 2.2.2 using the b-scale). During the 3-month storage period, the transparency was not affected, and the reconstituted solution was found to be =WFI (determined according to Ph.Eur. 2.2.1). The pH was not affected by storage. Finally, the osmotic pressure at T0 was within the physiological range, and no change was detected over 3 months.

[0692] [Table 9]

[0693] [Table 10]

[0694] [Example 7] Stability of CNP-38 at different pH values Preformulation studies were conducted to evaluate the stability of CNP-38 at different pH values. CNP-38 (synthesized according to the method described in WO2017 / 118693) was incubated at approximately 1 mg / mL in solution in succinate buffer at pH 4.0, 4.5, 5.0, 5.5, and 6.0 at 37°C. The amount of degraded CNP-38 (the sum of the deamidated and aspartimide variants of CNP-38) was assessed by RP-HPLC after 7 days. The amount of degraded CNP-38 was 2.7% at pH 4.0, 1.6% at pH 4.5, 1.0% at pH 5.0, 2.9% at pH 5.5, and 3.3% at pH 6.0, indicating that pH 5.0 is preferred for CNP-38 formulations.

[0695] [Example 8] Stability testing of compound (1)-containing formulations reconstituted with water containing antioxidants and / or preservatives Lyophilized formulations of compound (1) (F13) were prepared by lyophilizing 1080 μL in vials. The formulations contained an amount of (1) appropriate to achieve a nominal concentration of 3.9 mg CNP-38 / vial and 3.6 mg CNP-38 / mL. The water contained antioxidants and / or preservatives (R1-R5, Table 11). Vials containing the reconstituted formulations were placed in an incubator set to maintain 5°C. After 4 weeks (T4W), samples were analyzed.

[0696] The results show that compound (1) is stable after regeneration under the test conditions and is not affected by the presence of antioxidants and / or preservatives (Table 12). The pH of the regeneration solution was not affected by antioxidants and / or preservatives after regeneration.

[0697] [Table 11]

[0698] [Table 12]

[0699] Abbreviation BSA - Bovine serum albumin cGMP - cyclic guanosine monophosphate CI - confidence interval DMEM - Dulbecco's Modified Eagle's Medium FBS - Fetal bovine serum HEPES-4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid IBMX-3-isobutyl-1-methylxanthine Isoasp - Isoaspartate LC-MS - Liquid chromatography coupled to mass spectrometry Met(O)-Methionine sulfoxide PLA-Parallel Line Analysis PS20 - Polysorbate 20 PS80 - Polysorbate 80 RH - Relative Humidity RP-HPLC - Reversed-Phase High-Performance Liquid Chromatography SE-HPLC - Size Exclusion High Performance Liquid Chromatography TFA - Trifluoroacetic acid TR-FRET - Time-resolved fluorescence energy transfer UPLC - Ultra High Performance Liquid Chromatography.

[0700] The present specification includes the following sequence information: SEQUENCE LISTING <110> Ascendis Pharma Growth Disorders A / S <120> Dry Pharmaceutical Formulations of CNP Conjugates <130> PA24-566 <141> 2020-02-10 <150> EP 19156488.9 <151> 2019-02-11 <160> 97 <170> PatentIn version 3.5 <210> 1 <211> 22 <212> PRT <213> Homo sapiens <220> <221> DISULFID <222> (6)..(22) <400> 1 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 1 5 10 15 Met Ser Gly Leu Gly Cys 20 <210> 2 <211> 53 <212> PRT <213> Homo sapiens <220> <221> DISULFID <222> (37)..(53) <400> 2 Asp Leu Arg Val Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu Leu 1 5 10 15 Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly 20 25 30 Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met 35 40 45 Ser Gly Leu Gly Cys 50 <210> 3 <211> 54 <212> PRT <213> Artificial Sequence <220> <223> G-CNP-53 <220> <221> DISULFID <222> (38)..(54) <400> 3 Gly Asp Leu Arg Val Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu 1 5 10 15 Leu Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys 20 25 30 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 35 40 45 Met Ser Gly Leu Gly Cys 50 <210> 4 <211> 54 <212> PRT <213> Artificial Sequence <220> <223> M-CNP-53 <220> <221> DISULFID <222> (38)..(54) <400> 4 Met Asp Leu Arg Val Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu 1 5 10 15 Leu Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys 20 25 30 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 35 40 45 Met Ser Gly Leu Gly Cys 50 <210> 5 <211> 54 <212> PRT <213> Artificial Sequence <220> <223> P-CNP-53 <220> <221> DISULFID <222> (38)..(54) <400> 5 Pro Asp Leu Arg Val Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu 1 5 10 15 Leu Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys 20 25 30 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 35 40 45 Met Ser Gly Leu Gly Cys 50 <210> 6 <211> 53 <212> PRT <213> Artificial Sequence <220> <223> CNP-53 M48N <220> <221> DISULFID <222> (37)..(53) <400> 6 Asp Leu Arg Val Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu Leu 1 5 10 15 Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly 20 25 30 Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Asn 35 40 45 Ser Gly Leu Gly Cys 50 <210> 7 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> CNP-53 with deletion of amino acids 15-31 <220> <221> DISULFID <222> (20)..(36) <400> 7 Asp Leu Arg Val Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Gly Leu 1 5 10 15 Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser 20 25 30 Gly Leu Gly Cys 35 <210> 8 <211> 52 <212> PRT <213> Artificial Sequence <220> <223> CNP-52 <220> <221> DISULFID <222> (36)..(52) <400> 8 Leu Arg Val Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu Leu Gln 1 5 10 15 Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu 20 25 30 Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser 35 40 45 Gly Leu Gly Cys 50 <210> 9 <211> 51 <212> PRT <213> Artificial Sequence <220> <223> CNP-51 <220> <221> DISULFID <222> (35)..(51) <400> 9 Arg Val Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu Leu Gln Glu 1 5 10 15 His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser 20 25 30 Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly 35 40 45 Leu Gly Cys 50 <210> 10 <211> 50 <212> PRT <213> Artificial Sequence <220> <223> CNP-50 <220> <221> DISULFID <222> (34)..(50) <400> 10 Val Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu Leu Gln Glu His 1 5 10 15 Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys 20 25 30 Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu 35 40 45 Gly Cys 50 <210> 11 <211> 49 <212> PRT <213> Artificial Sequence <220> <223> CNP-49 <220> <221> DISULFID <222> (33)..(49) <400> 11 Asp Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu Leu Gln Glu His Pro 1 5 10 15 Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly 20 25 30 Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly 35 40 45 Cys <210> 12 <211> 48 <212> PRT <213> Artificial Sequence <220> <223> CNP-48 <220> <221> DISULFID <222> (32)..(48) <400> 12 Thr Lys Ser Arg Ala Ala Trp Ala Arg Leu Leu Gln Glu His Pro Asn 1 5 10 15 Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys 20 25 30 Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 35 40 45 <210> 13 <211> 47 <212> PRT <213> Artificial Sequence <220> <223> CNP-47 <220> <221> DISULFID <222> (31)..(47) <400> 13 Lys Ser Arg Ala Ala Trp Ala Arg Leu Leu Gln Glu His Pro Asn Ala 1 5 10 15 Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe 20 25 30 Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 35 40 45 <210> 14 <211> 46 <212> PRT <213> Artificial Sequence <220> <223> CNP-46 <220> <221> DISULFID <222> (30)..(46) <400> 14 Ser Arg Ala Ala Trp Ala Arg Leu Leu Gln Glu His Pro Asn Ala Arg 1 5 10 15 Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly 20 25 30 Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 35 40 45 <210> 15 <211> 45 <212> PRT <213> Artificial Sequence <220> <223> CNP-45 <220> <221> DISULFID <222> (29)..(45) <400> 15 Arg Ala Ala Trp Ala Arg Leu Leu Gln Glu His Pro Asn Ala Arg Lys 1 5 10 15 Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu 20 25 30 Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 35 40 45 <210> 16 <211> 44 <212> PRT <213> Artificial Sequence <220> <223> CNP-44 <220> <221> DISULFID <222> (28)..(44) <400> 16 Ala Ala Trp Ala Arg Leu Leu Gln Glu His Pro Asn Ala Arg Lys Tyr 1 5 10 15 Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys 20 25 30 Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 35 40 <210> 17 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> CNP-44 with a deletion of amino acids 14-22 <220> <221> DISULFID <222> (19)..(35) <400> 17 Ala Ala Trp Ala Arg Leu Leu Gln Glu His Pro Asn Ala Gly Leu Ser 1 5 10 15 Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly 20 25 30 Leu Gly Cys 35 <210> 18 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> CNP-44 with a deletion of amino acids 15-22 <220> <221> DISULFID <222> (20)..(36) <400> 18 Ala Ala Trp Ala Arg Leu Leu Gln Glu His Pro Asn Ala Arg Gly Leu 1 5 10 15 Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser 20 25 30 Gly Leu Gly Cys 35 <210> 19 <211> 43 <212> PRT <213> Artificial Sequence <220> <223> CNP-43 <220> <221> DISULFID <222> (27)..(43) <400> 19 Ala Trp Ala Arg Leu Leu Gln Glu His Pro Asn Ala Arg Lys Tyr Lys 1 5 10 15 Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu 20 25 30 Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 35 40 <210> 20 <211> 42 <212> PRT <213> Artificial Sequence <220> <223> CNP-42 <220> <221> DISULFID <222> (26)..(42) <400> 20 Trp Ala Arg Leu Leu Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly 1 5 10 15 Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp 20 25 30 Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 35 40 <210> 21 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> CNP-41 <220> <221> DISULFID <222> (25)..(41) <400> 21 Ala Arg Leu Leu Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala 1 5 10 15 Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg 20 25 30 Ile Gly Ser Met Ser Gly Leu Gly Cys 35 40 <210> 22 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> CNP-40 <220> <221> DISULFID <222> (24)..(40) <400> 22 Arg Leu Leu Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn 1 5 10 15 Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile 20 25 30 Gly Ser Met Ser Gly Leu Gly Cys 35 40 <210> 23 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> CNP-39 <220> <221> DISULFID <222> (23)..(39) <400> 23 Leu Leu Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys 1 5 10 15 Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly 20 25 30 Ser Met Ser Gly Leu Gly Cys 35 <210> 24 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> CNP-38 <220> <221> DISULFID <222> (22)..(38) <400> 24 Leu Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys 1 5 10 15 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 20 25 30 Met Ser Gly Leu Gly Cys 35 <210> 25 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> CNP-37 <220> <221> DISULFID <222> (21)..(37) <400> 25 Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly 1 5 10 15 Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met 20 25 30 Ser Gly Leu Gly Cys 35 <210> 26 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> CNP-37 mit Q1pQ (pQ = pyroglutamate) <220> <221> MISC_FEATURE <222> (1)..(1) <223> X = pyroglutamate <220> <221> DISULFID <222> (21)..(37) <400> 26 Xaa Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly 1 5 10 15 Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met 20 25 30 Ser Gly Leu Gly Cys 35 <210> 27 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> G-CNP-37 <220> <221> DISULFID <222> (22)..(38) <400> 27 Gly Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys 1 5 10 15 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 20 25 30 Met Ser Gly Leu Gly Cys 35 <210> 28 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> P-CNP-37 <220> <221> DISULFID <222> (22)..(38) <400> 28 Pro Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys 1 5 10 15 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 20 25 30 Met Ser Gly Leu Gly Cys 35 <210> 29 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> M-CNP-37 <220> <221> DISULFID <222> (22)..(38) <400> 29 Met Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys 1 5 10 15 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 20 25 30 Met Ser Gly Leu Gly Cys 35 <210> 30 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> PG-CNP-37 <220> <221> DISULFID <222> (23)..(39) <400> 30 Pro Gly Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys 1 5 10 15 Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly 20 25 30 Ser Met Ser Gly Leu Gly Cys 35 <210> 31 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> MG-CNP-37 <220> <221> DISULFID <222> (23)..(39) <400> 31 Met Gly Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys 1 5 10 15 Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly 20 25 30 Ser Met Ser Gly Leu Gly Cys 35 <210> 32 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> CNP-37 M32N <220> <221> DISULFID <222> (21)..(37) <400> 32 Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly 1 5 10 15 Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Asn 20 25 30 Ser Gly Leu Gly Cys 35 <210> 33 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> G-CNP-37 M32N <220> <221> DISULFID <222> (22)..(38) <400> 33 Gly Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys 1 5 10 15 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 20 25 30 Asn Ser Gly Leu Gly Cys 35 <210> 34 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> G-CNP-37 K14Q <220> <221> DISULFID <222> (22)..(38) <400> 34 Gly Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Gln Lys 1 5 10 15 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 20 25 30 Met Ser Gly Leu Gly Cys 35 <210> 35 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> G-CNP-37 K14P <220> <221> DISULFID <222> (22)..(38) <400> 35 Gly Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Pro Lys 1 5 10 15 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 20 25 30 Met Ser Gly Leu Gly Cys 35 <210> 36 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> G-CNP-37 K14Q, deletion of amino acid 15 <220> <221> DISULFID <222> (21)..(37) <400> 36 Gly Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Gln Gly 1 5 10 15 Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met 20 25 30 Ser Gly Leu Gly Cys 35 <210> 37 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> G-CNP-37 K14Q, K15Q <220> <221> DISULFID <222> (22)..(38) <400> 37 Gly Gln Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Gln Gln 1 5 10 15 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 20 25 30 Met Ser Gly Leu Gly Cys 35 <210> 38 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> CNP-36 <220> <221> DISULFID <222> (20)..(36) <400> 38 Glu His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu 1 5 10 15 Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser 20 25 30 Gly Leu Gly Cys 35 <210> 39 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> CNP-35 <220> <221> DISULFID <222> (19)..(35) <400> 39 His Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser 1 5 10 15 Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly 20 25 30 Leu Gly Cys 35 <210> 40 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> CNP-34 <220> <221> DISULFID <222> (18)..(34) <400> 40 Pro Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys 1 5 10 15 Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu 20 25 30 Gly Cys <210> 41 <211> 33 <212> PRT <213> Artificial Sequence <220> <223> CNP-33 <220> <221> DISULFID <222> (17)..(33) <400> 41 Asn Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly 1 5 10 15 Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly 20 25 30 Cys <210> 42 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> CNP-32 <220> <221> DISULFID <222> (16)..(32) <400> 42 Ala Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys 1 5 10 15 Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 20 25 30 <210> 43 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> CNP-31 <220> <221> DISULFID <222> (15)..(31) <400> 43 Arg Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe 1 5 10 15 Gly Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 20 25 30 <210> 44 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> CNP-30 <220> <221> DISULFID <222> (14)..(30) <400> 44 Lys Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly 1 5 10 15 Leu Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 20 25 30 <210> 45 <211> 29 <212> PRT <213> Artificial Sequence <220> <223> CNP-29 <220> <221> DISULFID <222> (13)..(29) <400> 45 Tyr Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu 1 5 10 15 Lys Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 20 25 <210> 46 <211> 28 <212> PRT <213> Artificial Sequence <220> <223> CNP-28 <220> <221> DISULFID <222> (12)..(28) <400> 46 Lys Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys 1 5 10 15 Leu Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 20 25 <210> 47 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> GHKSEVAHRF-CNP-28 <220> <221> DISULFID <222> (22)..(38) <400> 47 Gly His Lys Ser Glu Val Ala His Arg Phe Lys Gly Ala Asn Lys Lys 1 5 10 15 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 20 25 30 Met Ser Gly Leu Gly Cys 35 <210> 48 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> CNP-27 <220> <221> DISULFID <222> (11)..(27) <400> 48 Gly Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu 1 5 10 15 Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 20 25 <210> 49 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> CNP-27 K4Q, K5Q <220> <221> DISULFID <222> (11)..(27) <400> 49 Gly Ala Asn Gln Gln Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu 1 5 10 15 Asp Arg Ile Gly Ser Met Ser Gly Leu Gly Cys 20 25 <210> ...

Claims

1. A dry pharmaceutical formulation, the pharmaceutical formulation comprising a CNP conjugate, a buffer, and a bulking agent, the CNP conjugate comprising a CNP moiety covalently and reversibly bound to a polymer moiety.

2. 2. The dry pharmaceutical formulation of claim 1, wherein the buffering agent is selected from the group consisting of succinic acid, citric acid, lactic acid, acetic acid, glutamic acid, fumaric acid, aspartic acid, glutaric acid, phosphoric acid, histidine, gluconic acid, tartaric acid, malic acid, and mixtures thereof.

3. 3. The dry pharmaceutical formulation of claim 1 or 2, wherein the buffer is succinic acid.

4. 4. The dry pharmaceutical formulation according to any one of claims 1 to 3, wherein the bulking agent is selected from the group consisting of trehalose, mannitol, sucrose, raffinose, gelatin, lactose, dipotassium hydrogen phosphate, sorbitol, xylitol, glycine, histidine, hydroxyethyl starch, dextrose, dextran, propylene glycol and mixtures thereof.

5. The dry pharmaceutical formulation according to any one of claims 1 to 4, wherein the bulking agent is selected from the group consisting of trehalose, sucrose and glycine.

6. The dry pharmaceutical formulation according to any one of claims 1 to 5, wherein the bulking agent is trehalose.

7. The dry pharmaceutical formulation of the invention according to any one of claims 1 to 6, wherein the formulation comprises one or more further excipients.

8. 8. The dry pharmaceutical formulation of claim 7, wherein the one or more additional excipients are selected from the group consisting of preservatives, stabilizers, anti-adsorption agents, cryoprotectants, antioxidants and other auxiliary agents.

9. The dry pharmaceutical formulation of the present invention according to any one of claims 1 to 8, wherein the dry pharmaceutical formulation comprises a stabilizer.

10. The dry pharmaceutical formulation of the present invention according to any one of claims 1 to 9, wherein the dry pharmaceutical formulation comprises a preservative.

11. The dry pharmaceutical formulation of the present invention according to any one of claims 1 to 10, wherein the dry pharmaceutical formulation comprises an adsorption inhibitor.

12. The dry pharmaceutical formulation of the present invention according to any one of claims 1 to 11, wherein the formulation comprises a cryoprotectant.

13. The dry pharmaceutical formulation of the present invention according to any one of claims 1 to 12, wherein the formulation comprises an antioxidant.

14. The dry pharmaceutical formulation of the present invention according to any one of claims 1 to 13, wherein the formulation comprises further excipients selected from the group consisting of wetting agents, viscosity modifiers and antibiotics.

15. The dry pharmaceutical formulation of any one of claims 1 to 14, wherein the pharmaceutical formulation comprises a pH adjusting agent.

16. 16. The dry pharmaceutical formulation of claim 15, wherein the pH adjusting agent is selected from the group consisting of Tris, sodium hydroxide, potassium hydroxide, lysine, and mixtures thereof.

17. 17. The dry pharmaceutical formulation of claim 15 or 16, wherein the pH adjusting agent is Tris.

18. The dry pharmaceutical formulation of any one of claims 1 to 17, wherein the pharmaceutical formulation comprises a CNP conjugate, succinic acid, trehalose and Tris.

19. 19. The dry pharmaceutical formulation of any one of claims 1 to 18, wherein the CNP portion has the sequence of 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:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, or SEQ ID NO:

30.

20. The dry pharmaceutical formulation of any one of claims 1 to 19, wherein the CNP portion has the sequence of SEQ ID NO:

24.

21. The polymer moiety may be 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(ethyl oxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyl oxazolines), poly(hydroxypropyl methacrylates ... (iminocarbonates), poly(lactic acids), poly(lactic acid-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinyl methyl ethers), poly(vinylpyrrolidones), silicones, celluloses, 21. The dry pharmaceutical formulation according to any one of claims 1 to 20, comprising a polymer selected from the group consisting of carbomethylcelluloses, hydroxypropylmethylcelluloses, 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.

22. The formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 1.3 to 45.4% by weight Succinic acid: 0.2 to 3.2% by weight Trehalose dihydrate: 52.6 to 98.4% by weight Tris: 0.1 to 5.6% by weight 22. The dry pharmaceutical formulation according to any one of claims 1 to 21, comprising:

23. The formulation comprises, based on the total weight of the dry pharmaceutical formulation: CNP conjugate: 1.3 to 38.7% by weight Succinic acid: 0.2 to 3.2% by weight Trehalose dihydrate: 52.6 to 98.4% by weight Tris: 0.1 to 5.6% by weight 23. The dry pharmaceutical formulation according to any one of claims 1 to 22, comprising:

24. The dry pharmaceutical formulation according to any one of claims 1 to 23, wherein the CNP conjugate is of the following formula (Ia) or (Ib): 【Chemistry 1】 [In the formula, -D is a CNP moiety; -L １ - is a reversible linker moiety; -L ２ - is a single chemical bond or a spacer moiety; -Z is a polymer moiety; x is an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, and 16; and y is an integer selected from the group consisting of 1, 2, 3, 4, and 5.

25. 25. The dry pharmaceutical formulation of claim 24, wherein x in formula (Ia) is an integer selected from the group consisting of 1, 2, 3, 4, 6 and 8.

26. 25. The dry pharmaceutical formulation of claim 24, wherein y in formula (Ib) is an integer selected from the group consisting of 2, 3, 4 and 5.

27. 26. The dry pharmaceutical formulation of claim 24 or 25, wherein the CNP conjugate is of formula (Ia) and x is 1.

28. -L １ The dry pharmaceutical formulation of any one of claims 24 to 27, wherein - is linked to -D via an amide linkage.

29. -Z and -L ２ The dry pharmaceutical formulation of any one of claims 24 to 28, wherein the linkage between is a stable linkage.

30. -L ２ The dry pharmaceutical formulation of any one of claims 24 to 29, wherein - is a spacer moiety.

31. -L ２ The dry pharmaceutical formulation according to any one of claims 24 to 30, wherein - has a molecular weight in the range of 14 g / mol to 750 g / mol.

32. -L ２ 32. The dry pharmaceutical formulation according to any one of claims 24 to 31, wherein - has a chain length of 1 to 20 atoms.

33. -L ２ The dry pharmaceutical formulation according to any one of claims 24 to 32, wherein - is of formula (i): 【Chemistry 2】 [In the formula, The dotted line marked with an asterisk is -L １ - indicates a bond to; The unmarked dotted line indicates the bond to -Z; -R １ is -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.

34. 34. The dry pharmaceutical formulation of claim 33, wherein n in formula (i) is selected from the group consisting of 3, 4, 5, 6, 7, 8 and 9.

35. 35. The dry pharmaceutical formulation of any one of claims 24 to 34, wherein -Z has a molecular weight in the range of 5 to 200 kDa, inclusive.

36. -Z is 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(ethyl oxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl oxazolines), poly(hydroxy methacrylates), poly(hydroxypropyl methacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyl oxazolines), poly(imide poly(lactic acids), poly(lactic acid-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinyl methyl ethers), poly(vinylpyrrolidones), silicones, cellulose, carbamide, cellulose acetate, cellulose acetates ...

36. The dry pharmaceutical formulation according to any one of claims 24 to 35, comprising a polymer selected from the group consisting of polymethylcelluloses, hydroxypropylmethylcelluloses, 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.

37. A method for producing a dry pharmaceutical formulation according to any one of claims 1 to 36, comprising the steps of: The method further comprising: (i) mixing the CNP conjugate with at least a buffer and a bulking agent; (ii) adjusting the pH of the mixture of step (i); (iii) optionally filtering the mixture from step (ii); (iv) transferring an amount of the mixture from step (ii) or (iii) into a container corresponding to a desired number of doses; (v) drying the mixture; (vi) sealing the container. Including; The process wherein the order of steps (ii) and (iii) is reversed.

38. A method for reconstituting a dry pharmaceutical formulation according to any one of claims 1 to 36, comprising the steps of: The method further comprising: (a) contacting a dry pharmaceutical formulation according to any one of claims 1 to 36 with a reconstitution solution, A method comprising:

39. A regenerated pharmaceutical preparation obtainable by the regeneration method of claim 38.

40. The dried pharmaceutical preparation according to any one of claims 1 to 36 or the regenerated pharmaceutical preparation according to claim 39, for use as a medicine.

41. The dry pharmaceutical formulation according to any one of claims 1 to 36 or the regenerative pharmaceutical formulation according to claim 39, for use in the treatment, suppression, delay or prevention of one or more diseases that can be treated, suppressed, delayed or prevented by CNP.

42. 42. The dry pharmaceutical formulation for use according to claim 41, wherein the disease is selected from the group consisting of bone-related disorders, such as skeletal dysplasia; cancer; autoimmune diseases; fibrosis; inflammatory diseases; central nervous system diseases, such as neurodegenerative diseases; infectious diseases; lung diseases; heart and vascular diseases; metabolic diseases; and eye diseases.

43. 43. A dry pharmaceutical formulation for use according to claim 41 or 42, wherein the disease is achondroplasia.

44. A method for treating, inhibiting, delaying or preventing one or more diseases treatable by CNP in a patient, comprising administering to the patient a therapeutically effective amount of the regenerative pharmaceutical formulation described in claim 39.

45. 45. The method of claim 44, wherein the disease is selected from the group consisting of bone-related disorders, e.g. skeletal dysplasia; cancer; autoimmune diseases; fibrosis; inflammatory diseases; central nervous system diseases, e.g. neurodegenerative diseases; infectious diseases; pulmonary diseases; heart and vascular diseases; metabolic diseases and eye diseases.

46. 46. ​​The method of claim 44 or 45, wherein the disease is achondroplasia.

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