Liquid pharmaceutical formulations of CNP compounds

Abstract

Liquid pharmaceutical formulations comprising a CNP compound, a buffer, an isotonic agent, a preservative, and optionally an antioxidant, and methods of making and using such formulations.

Description

[0001] The present application relates to liquid pharmaceutical formulations comprising a CNP compound, a buffer, an isotonicity agent, a preservative, and optionally an antioxidant, and methods of making and using such formulations.

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

[0003] The human pre-pro-CNP (prepro-CNP) that comprises 126 amino acid whose, further by furin protease cleavage to produce CNP-53.CNP-53 has biological activity, but it is usually processed into 22 amino acid forms with biological activity in circulation, i.e. CNP-22, by unknown mechanism.The biological activity of CNP is strictly regulated, and its removal from blood plasma is very fast.Therefore, in view of the short half-life in vivo of CNP (after intravenous administration 2 minutes), its application as therapeutic agent in children's colony is challenging, because it can need continuous infusion.

[0004] A variety of methods for increasing the in vivo half-life of CNP have been explored. For example, Lorget et al. (Am. J. Hum. Genet. 91, 1108-1114, 2012) disclosed a recombinant CNP-39 (also known as BMN111), consisting of the 37 C-terminal amino acids of human CNP-53 plus glycine and proline added to the N-terminus of the peptide, which mimics the pharmacological activity of human CNP.

[0005] In WO 2016 / 110577 A1, WO 2017 / 118693 A1, WO 2017 / 118698 A1, WO 2017 / 118700A1, WO 2017 / 118703A1, WO 2017 / 118704A1 and WO2017 / 118707A1, the expansion of the half-life scope in vivo of CNP based on being puted together with water-soluble carrier part (such as PEG) via reversible prodrug linker has been explored.The pharmaceutical preparation of such reversible CNP conjugates must provide enough CNP conjugate stability, to avoid premature release of CNP during storage, wherein carrier is connected to CNP via reversible bonding.In the case that the reversible bonding between carrier and CNP degrades during storage, the concentration of fast-acting drug (readily available drug) can increase, and this can cause excessive risk when using, and then cause hypotension. Furthermore, any drug released during storage is subject to rapid renal clearance upon administration to a patient, so the time over which the long-acting composition provides therapeutically relevant amounts of drug is reduced. This poses a risk of unmet medical need.

[0006] Since degradation products that may form during storage can impair the biological activity of the CNP moiety, their formation needs to be minimized. Furthermore, in reversible CNP conjugates, the reversible linkage between the CNP moiety and the carrier moiety makes storage of pharmaceutical formulations containing the CNP conjugates challenging.

[0007] WO 2020 / 165081 A1 teaches dry drug formulations comprising reversible CNP conjugates, which require reconstitution in an aqueous solution to provide an injectable drug.

[0008] It is also important to identify suitable liquid pharmaceutical formulations of CNP compounds, particularly reversible CNP conjugates, which comprise CNP covalently linked to a carrier moiety via a reversible linker, where the peptide exhibits an acceptable impurity profile. Furthermore, it is particularly important to identify suitable liquid formulations, particularly for reversible CNP conjugates, that exhibit limited premature release of CNP even after extended storage.

[0009] Therefore, there remains a need in the art for ready-to-use liquid formulations of CNP compounds, particularly reversible CNP conjugates, which exhibit storage stability and provide ease of use.

[0010] It is therefore an object of the present application to at least partially overcome the above-mentioned disadvantages.

[0011] This object is achieved using a liquid pharmaceutical formulation comprising a CNP compound, a buffer, an isotonicity agent, a preservative and optionally an antioxidant.

[0012] Surprisingly, it was found that the liquid pharmaceutical formulation of the present application allows stable long-term storage. The liquid pharmaceutical formulation of the present application shows a substantial degree of peptide stability after storage for several months and does not require freezing it, such as storing it in a refrigerator.

[0013] Within the meaning of this application, the terms are used as follows.

[0014] As used herein, the term "about" in combination with a numerical value is used to indicate a range of values, which is plus or minus no more than 20% of the numerical value, including the endpoint value; in certain embodiments, no more than 10% of the numerical value; in certain embodiments, no more than 8% of the numerical value; in certain embodiments, no more than 5% of the numerical value; and in certain embodiments, no more than 2% of the numerical value. For example, the phrase "about 200" is used to mean a range of 200 + / - 10% and including the endpoint value, i.e., a range of 180 to 220 and including the endpoint value; in certain embodiments, 200 + / - 8%, i.e., a range of 184 to 216 and including the endpoint value; in certain embodiments, a range of 200 + / - 5% and including the endpoint value, i.e., a range of 190 to 210 and including the endpoint value; and in certain embodiments, 200 + / - 2%, i.e., a range of 196 to 204 and including the endpoint value. It should be understood that a percentage given as "about 20%" does not mean "20% + / - 10%", i.e. a range of 10% to 30% and including the endpoint values, but "about 20%" means a range of 18 to 22% and including the endpoint values, i.e. plus and minus 10% of the value 20.

[0015] As used herein, the term "antimicrobial agent" refers to a chemical substance, such as a chemical substance that kills or inhibits the growth of microorganisms (eg, bacteria, fungi, yeasts, protozoa, molds) and / or destroys viruses.

[0016] As used herein, the term "anti-adsorption agent" primarily refers to an ionic or nonionic surfactant, protein, or soluble polymer that is used to competitively coat or adsorb to the inner surface of the container containing the formulation. The excipient concentration and type selected depends on the effect to be avoided, but generally a monolayer of surfactant is formed at the interface above the critical micelle concentration (CMC).

[0017] As used herein, the term "buffer" or "buffering agent" refers to a compound that maintains the pH within a desired range. Physiologically tolerable buffers include, for example, sodium phosphate, histidine, bicarbonate, citrate, acetate, sulfate, nitrate, chloride, and pyruvate. Antacids such as Mg(OH)2 or ZnCO3 may also be used.

[0018] As used herein, the term "CNP" refers to all CNP polypeptides, in certain embodiments, from mammalian species, such as from humans and mammalian species, particularly from humans and murine species, as well as variants, analogs, orthologs, homologs, derivatives, and fragments thereof, characterized in that they regulate 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 WO 2009 / 067639 A2 and WO 2010 / 135541 A2 are hereby incorporated by reference into this document.

[0019] As used herein, the term "CNP polypeptide variant" refers to a polypeptide from the same species that differs from a reference CNP polypeptide. Generally, the 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 has at least 70%, 80%, 90%, or 95% identity to a reference CNP polypeptide. A polypeptide having an amino acid sequence that is at least, for example, 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 sequence may include up to 5 amino acid changes per 100 amino acids of the query amino acid sequence. These changes in the reference sequence may occur at the amino (N-terminal) or carboxyl (C-terminal) terminal position of the reference amino acid sequence, or anywhere between these terminal positions, and may be interspersed individually between residues in the reference sequence or in one or more contiguous groups within the reference sequence. The query sequence may be the entire amino acid sequence of a reference sequence or any fragment thereof as specified herein. Such CNP polypeptide variants may be naturally occurring variants, such as naturally occurring allelic variants encoded by one of several alternative forms of CNP occupying a given locus on a chromosome or organism, or isomers encoded by naturally occurring splice variants derived from a single primary transcript. Alternatively, CNP polypeptide variants may be known non-naturally occurring variants, and they may be made by mutagenesis techniques known in the art. It is known in the art that one or more amino acids can be deleted at the N-terminus or C-terminus of a biologically active peptide or protein without substantial loss of biological function. Such N- and / or C-terminal deletions are also encompassed by the term CNP polypeptide variant.

[0020] Those skilled in the art will also recognize that some amino acid sequences of CNP polypeptides can be altered without significantly affecting the structure or function of the peptide. Such mutants include deletions, insertions, inversions, duplications, and substitutions selected according to general rules known in the art, resulting in little or no 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 note that there are two main approaches for studying the tolerance of amino acid sequences to variation.

[0021] As used herein, the term "CNP analogs" refers to CNPs from different and unrelated organisms that perform the same function in each organism but are not derived from an ancestral structure shared by the organism's ancestors. Instead, similar CNPs arose independently and later evolved to perform the same or similar functions. In other words, similar CNP polypeptides are polypeptides with completely different amino acid sequences that perform the same biological activity, namely, regulating the growth, proliferation, and differentiation of cartilage growth plate chondrocytes.

[0022] As used herein, the term "CNP orthologs" refers to CNPs from two different species whose sequences are related to each other via a common homologous CNP in an ancestral species, but have evolved to become divergent from each other.

[0023] As used herein, the term "CNP homolog" refers to CNPs from different organisms that perform the same function in each organism and are derived from an ancestral structure shared by the ancestors of the organisms. In other words, homologous CNP polypeptides are polypeptides that have very similar amino acid sequences and possess the same biological activity, namely, 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 has at least 40%, 50%, 60%, 70%, 80%, 90%, or 95% identity to a reference CNP polypeptide.

[0024] Thus, a CNP polypeptide can be, for example: (i) a polypeptide in which at least one amino acid residue is substituted with a conservative or non-conservative amino acid residue, in certain embodiments, a conservative amino acid residue, and such substituted amino acid residue may or may not be an amino acid residue encoded by the genetic code; and / or (ii) a polypeptide in which at least one amino acid residue includes a substituent group; and / or (iii) a polypeptide in which a 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 polypeptide in which other amino acids are fused to the CNP polypeptide, such as an IgG Fc fusion region peptide, or a leader or secretory sequence, or a polypeptide or precursor protein sequence useful for purification of the above forms.

[0025] As used herein, the term "CNP polypeptide fragment" refers to any peptide that comprises a contiguous span of a portion of the amino acid sequence of a CNP polypeptide.

[0026] More specifically, a CNP polypeptide fragment comprises at least 6, such as at least 8, at least 10, or at least 17, consecutive amino acids of a CNP polypeptide. A CNP polypeptide fragment can also be described as a subclass of CNP polypeptides comprising 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. Further included are classes of CNP polypeptide fragments having a length of at least 6 amino acids, which, as described above, are further specified by their N-terminal and C-terminal positions. The term "CNP polypeptide fragment" as an individual species also encompasses all CNP polypeptide fragments having a length of at least 6 amino acids, as described above, which can be specifically specified by their N-terminal and C-terminal positions. That is, on any given CNP polypeptide amino acid sequence, a fragment having a length of at least 6 consecutive amino acid residues can occupy every combination of N-terminal and C-terminal positions.

[0027] Since the term CNP includes variants, analogs, orthologs, homologs, derivatives and fragments of the above-mentioned CNPs, all references to a particular position within a reference sequence also include the equivalent position in variants, analogs, orthologs, homologs, derivatives and fragments of the CNP portion, even if not explicitly mentioned.

[0028] Naturally occurring CNP-22 (SEQ ID NO: 1) has the following sequence:

[0029] GLSKGCFGLKLDRIGSMSGLGC,

[0030] The cysteines at positions 6 and 22 are linked by a disulfide bond.

[0031] In certain embodiments, the term "CNP" also refers to the following peptide sequence:

[0032] SEQ ID NO: 2 (CNP-53):

[0033] DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0034] SEQ ID NO: 3 (G-CNP-53):

[0035] GDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0036] SEQ ID NO:4(M-CNP-53):

[0037] MDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0038] SEQ ID NO:5(P-CNP-53):

[0039] PDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0040] SEQ ID NO:6(CNP-53M48N):

[0041] DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC;

[0042] SEQ ID NO:7(CNP-53Δ15-31):

[0043] DLRVDTKSRAAWARGLSKGCFGLKLDRIGSMSGLGC;

[0044] SEQ ID NO:8(CNP-52):

[0045] LRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0046] SEQ ID NO:9(CNP-51):

[0047] RVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0048] SEQ ID NO:10(CNP-50):

[0049] VDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0050] SEQ ID NO:11(CNP-49):

[0051] DTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0052] SEQ ID NO:12(CNP-48):

[0053] TKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0054] SEQ ID NO:13(CNP-47):

[0055] KSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0056] SEQ ID NO:14(CNP-46):

[0057] SRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0058] SEQ ID NO:15(CNP-45):

[0059] RAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0060] SEQ ID NO:16(CNP-44):

[0061] AAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0062] SEQ ID NO:17(CNP-44Δ14-22):

[0063] AAWARLLQEHPNAGLSKGCFGLKLDRIGSMSGLGC;

[0064] SEQ ID NO:18(CNP-44Δ15-22):

[0065] AAWARLLQEHPNARGLSKGCFGLKLDRIGSMSGLGC;

[0066] SEQ ID NO:19(CNP-43):

[0067] AWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0068] SEQ ID NO:20(CNP-42):

[0069] WARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0070] SEQ ID NO: 21 (CNP-41):

[0071] ARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0072] SEQ ID NO: 22 (CNP-40):

[0073] RLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0074] SEQ ID NO: 23 (CNP-39):

[0075] LLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0076] SEQ ID NO: 24 (CNP-38):

[0077] LQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC,

[0078] The cysteines at positions 22 and 38 are connected by a disulfide bond;

[0079] SEQ ID NO: 25 (CNP-37):

[0080] QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0081] SEQ ID NO: 26 (CNP-37Q1pQ, wherein pQ = pyroglutamic acid):

[0082] pQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0083] SEQ ID NO: 27 (G-CNP-37):

[0084] GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0085] SEQ ID NO: 28 (P-CNP-37):

[0086] PQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0087] SEQ ID NO:29(M-CNP-37):

[0088] MQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0089] SEQ ID NO:30(PG-CNP-37):

[0090] PGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0091] SEQ ID NO:31(MG-CNP-37):

[0092] MGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0093] SEQ ID NO:32(CNP-37M32N):

[0094] QEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC;

[0095] SEQ ID NO:33(G-CNP-37 M32N):

[0096] GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC;

[0097] SEQ ID NO:34(G-CNP-37 K14Q):

[0098] GQEHPNARKYKGANQKGLSKGCFGLKLDRIGSMSGLGC;

[0099] SEQ ID NO:35(G-CNP-37 K14P):

[0100] GQEHPNARKYKGANPKGLSKGCFGLKLDRIGSMSGLGC;

[0101] SEQ ID NO:36(G-CNP-37 K14Q,Δ15):

[0102] GQEHPNARKYKGANQGLSKGCFGLKLDRIGSMSGLGC;

[0103] SEQ ID NO:37(G-CNP-37 K14Q,K15Q):

[0104] GQEHPNARKYKGANQQGLSKGCFGLKLDRIGSMSGLGC;

[0105] SEQ ID NO:38(CNP-36):

[0106] EHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0107] SEQ ID NO:39(CNP-35):

[0108] HPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0109] SEQ ID NO:40(CNP-34):

[0110] PNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0111] SEQ ID NO:41(CNP-33):

[0112] NARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0113] SEQ ID NO:42(CNP-32):

[0114] ARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0115] SEQ ID NO:43(CNP-31):

[0116] RKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0117] SEQ ID NO:44(CNP-30):

[0118] KYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0119] SEQ ID NO:45(CNP-29):

[0120] YKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0121] SEQ ID NO:46(CNP-28):

[0122] KGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0123] SEQ ID NO:47(GHKSEVAHRF-CNP-28):

[0124] GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0125] SEQ ID NO:48(CNP-27):

[0126] GANKKGLSKGCFGLKLDRIGSMSGLGC;

[0127] SEQ ID NO:49(CNP-27K4Q,K5Q):

[0128] GANQQGLSKGCFGLKLDRIGSMSGLGC;

[0129] SEQ ID NO:50(CNP-27K4R,K5R):

[0130] GANRRGLSKGCFGLKLDRIGSMSGLGC;

[0131] SEQ ID NO:51(CNP-27K4P,K5R):

[0132] GANPRGLSKGCFGLKLDRIGSMSGLGC;

[0133] SEQ ID NO:52(CNP-27K4S,K5S):

[0134] GANSSGLSKGCFGLKLDRIGSMSGLGC;

[0135] SEQ ID NO:53(CNP-27K4P,K5R):

[0136] GANGANPRGLSRGCFGLKLDRIGSMSGLGC;

[0137] SEQ ID NO:54(CNP-27K4R,K5R,K9R):

[0138] GANRRGLSRGCFGLKLDRIGSMSGLGC;

[0139] SEQ ID NO:55(CNP-27K4R,K5R,K9R,M22N):

[0140] GANRRGLSRGCFGLKLDRIGSNSGLGC;

[0141] SEQ ID NO:56(P-CNP-27K4R,K5R,K9R):

[0142] PGANRRGLSRGCFGLKLDRIGSMSGLGC;

[0143] SEQ ID NO:57(M-CNP-27K4R,K5R,K9R):

[0144] MGANRRGLSRGCFGLKLDRIGSMSGLGC;

[0145] SEQ ID NO:58(HSA fragment-CNP-27):

[0146] GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLG;

[0147] SEQ ID NO:59(HSA fragment-CNP-27M22N):

[0148] GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSNSGLGC;

[0149] SEQ ID NO:60(M-HSA fragment-CNP-27):

[0150] MGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0151] SEQ ID NO:61(P-HSA fragment-CNP-27):

[0152] PGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0153] SEQ ID NO:62(CNP-26):

[0154] ANKKGLSKGCFGLKLDRIGSMSGLGC;

[0155] SEQ ID NO:63(CNP-25):

[0156] NKKGLSKGCFGLKLDRIGSMSGLGC;

[0157] SEQ ID NO:64 (CNP-24):

[0158] KKGLSKGCFGLKLDRIGSMSGLGC;

[0159] SEQ ID NO:65 (CNP-23):

[0160] KGLSKGCFGLKLDRIGSMSGLGC;

[0161] SEQ ID NO:66 (R-CNP-22):

[0162] RGLSKGCFGLKLDRIGSMSGLGC;

[0163] SEQ ID NO:67 (ER-CNP-22):

[0164] ERGLSKGCFGLKLDRIGSMSGLGC;

[0165] SEQ ID NO:68 (R-CNP-22K4R):

[0166] RGLSRGCFGLKLDRIGSMSGLGC;

[0167] SEQ ID NO:69 (ER-CNP-22 4KR):

[0168] ERGLSRGCFGLKLDRIGSMSGLGC; [[ID=​​​​​​​​​​​​​​​​​​​

[0176] GHHSHEQHPHGANPRGLSKGCFGLKLDRIGSMSGLGC;

[0177] SEQ ID NO:74 (IgG1 (F c ) fragment-CNP-22):

[0178] GQPREPQVYTLPPSGLSKGCFGLKLDRIGSMSGLGC;

[0179] SEQ ID NO: 75 (HSA fragment-CNP-22):

[0180] GQHKDDNPNLPRGANPRGLSKGCFGLKLDRIGSMSGLGC;

[0181] SEQ ID NO: 76 (HSA fragment-CNP-22):

[0182] GERAFKAWAVARLSQGLSKGCFGLKLDRIGSMSGLGC;

[0183] SEQ ID NO: 77 (Osteoporin NPR C inhibitor fragment-CNP22):

[0184] FGIPMDRIGRNPRGLSKGCFGLKLDRIGSMSGLGC;

[0185] SEQ ID NO: 78 (FGF2 heparin binding domain fragment-CNP22):

[0186] GKRTGQYKLGSKTGPGPKGLSKGCFGLKLDRIGSMSGLGC;

[0187] SEQ ID NO:79(IgG1(F c ) fragment-CNP-22K4R):

[0188] GQPREPQVYTGANQQGLSRGCFGLKLDRIGSMSGLGC;

[0189] SEQ ID NO: 80 (HSA fragment-CNP-22K4R):

[0190] GVPQVSTSTGANQQGLSRGCFGLKLDRIGSMSGLGC;

[0191] SEQ ID NO: 81 (fibronectin fragment-CNP-22K4R):

[0192] GQPSSSSQSTGANQQGLSRGCFGLKLDRIGSMSGLGC;

[0193] SEQ ID NO: 82 (fibronectin fragment-CNP-22K4R):

[0194] GQTHSSGTQSGANQQGLSRGCFGLKLDRIGSMSGLGC;

[0195] SEQ ID NO: 83 (fibronectin fragment-CNP-22K4R):

[0196] GSTGQWHSESGANQQGLSRGCFGLKLDRIGSMSGLGC;

[0197] SEQ ID NO: 84 (zinc finger fragment-CNP-22K4R):

[0198] GSSSSSSSSGANQQGLSRGCFGLKLDRIGSMSGLGC;

[0199] SEQ ID NO:85 (CNP-21):

[0200] LSKGCFGLKLDRIGSMSGLGC;

[0201] SEQ ID NO:86 (CNP-20):

[0202] SKGCFGLKLDRIGSMSGLGC;

[0203] SEQ ID NO:87 (CNP-19):

[0204] KGCFGLKLDRIGSMSGLGC;

[0205] SEQ ID NO:88 (CNP-18):

[0206] GCFGLKLDRIGSMSGLGC;

[0207] SEQ ID NO:89 (CNP-17):

[0208] CFGLKLDRIGSMSGLGC;

[0209] SEQ ID NO:90 (BNP fragment-CNP-17-BNP fragment):

[0210] SPKMVQGSGCFGLKLDRIGSMSGLGCKVLRRH;

[0211] SEQ ID NO:91 (CNP-38L1G):

[0212] GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0213] SEQ ID NO:92 (Ac-CNP-37; wherein Ac = acetyl):

[0214] Ac-QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;

[0215] SEQ ID NO:93:

[0216] QEHPNARX1YX2GANX3X4GLSX5GCFGLX6LDRIGSMSGLGC,

[0217] wherein X1, X2, X3, X4, X5 and X6 are independently selected from the group consisting of K, R, P, S and Q, provided 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, provided that at least one of X1, X2, X3, X4, X5 and X6 is R;

[0218] SEQ ID NO:94:

[0219] QEHPNARKYKGANX1X2GLSX3GCFGLX4LDRIGSMSGLGC,

[0220] wherein X1, X2, X3 and X4 are independently selected from the group consisting of K, R, P, S and Q, provided 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 the group consisting of K and R, provided that at least one of X1, X2, X3 and X4 is R;

[0221] SEQ ID NO:95:

[0222] QEHPNARKYKGANX1X2GLSKGCFGLKLDRIGSMSGLGC,

[0223] wherein X1X2 is selected from the group consisting of KR, RK, KP, PK, SS, RS, SR, QK, QR, KQ, RQ, RR, and QQ.

[0224] It will be understood that in SEQ ID NOs: 2 to 95, the equivalents of cysteines at positions 22 and 38 in SEQ ID NO: 24 are also linked by a disulfide bond.

[0225] The term "CNP" also includes poly (amino acid) conjugates having a sequence as described above, but having a backbone comprising amide and non-amide bonds, such as ester bonds, for example ester peptides. A ester peptide is a chain of amino acid residues wherein the backbone comprises an amide (peptide) and an ester bond. Thus, as used herein, the term "side chain" refers to a portion of the α-carbon of the amino acid moiety that is connected to the amino acid moiety if the amino acid moiety is connected via an amine bond (as in a polypeptide), or to any portion comprising a carbon atom that is connected to the poly (amino acid) conjugate backbone, as in the case of ester peptides. In certain embodiments, the term "CNP" refers to a polypeptide having a backbone formed by an amide (peptide) bond.

[0226] As used herein, the term "ring portion" refers to an extension of the continuous amino acid residues of CNP, which is located between two cysteine ​​residues forming an intramolecular disulfide bridge, or between the homologous amino acid residues connected by a chemical linker. Preferably, the ring portion is located between two cysteine ​​residues forming an intramolecular disulfide bridge. These two cysteines correspond to the 22nd and 38th cysteine ​​in the CNP-38 (SEQ ID NO: 24) sequence. Therefore, if CNP has the sequence of CNP-38, the 23rd to 37th amino acids are located in the ring portion.

[0227] Regardless of the length of the CNP, the sequence of the loop portion of the wild-type CNP is FGLKLDRIGSMSG LG (SEQ ID NO: 96). In certain embodiments, the loop portion of the CNP is FGLKLDRIGSN SGLG (SEQ ID NO: 97).

[0228] Since the term CNP includes variants, analogs, orthologs, homologs, derivatives and fragments of the above-mentioned CNP, the term "loop portion" also includes corresponding variants, analogs, orthologs, homologs, derivatives and fragments of the sequence of SEQ ID NO: 96. Therefore, all references to specific positions within the reference sequence include equivalent positions in variants, analogs, orthologs, homologs, derivatives and fragments of the CNP portion, even if not explicitly mentioned.

[0229] As used herein, the term "CNP agonist" refers to any molecule that activates natriuretic peptide receptor B (NPR-B). In certain embodiments, the EC of the CNP agonist is 50 The NPR-B activity is up to 50-fold higher than that of CNP-22 (SEQ ID NO: 1).

[0230] As used herein, the term "CNP conjugate" refers to any compound, crystal, or mixture comprising at least one CNP moiety conjugated to at least one other moiety.

[0231] As used herein, the term "reversible CNP conjugate" refers to a CNP conjugate comprising at least one CNP moiety that is covalently and reversibly conjugated to a carrier moiety and from which the at least one CNP moiety is released with a release half-life of at least 6 hours under physiological conditions.

[0232] As used herein, the term "stable CNP conjugate" refers to a CNP conjugate comprising at least one CNP moiety covalently and stably conjugated to a carrier moiety, ie via a stable linkage having a half-life of more than 6 months under physiological conditions.

[0233] As used herein, the phrase "CNP conjugate, wherein the CNP portion" is followed by an amount in mg / ml, meaning that the liquid formulation contains the CNP conjugate, but only the CNP portion is taken into account for the corresponding amount, without taking into account the complete CNP conjugate, i.e., without taking into account other moieties in the CNP conjugate other than the CNP portion, such as the carrier portion. The amount of the CNP portion in the CNP conjugate can be determined by quantitative amino acid analysis or by any known analytical method that allows for quantification of an unknown sample compared to a CNP conjugate having a known content of the CNP portion.

[0234] As used herein, the term "cryoprotectant" refers to a compound that is added to a formulation to protect the CNP compound during the freezing stage.

[0235] As used herein, the term "C 1-4 "Alkyl" alone or in combination means a straight or branched chain alkyl moiety having 1 to 4 carbon atoms. If present at the end of a molecule, a straight or branched chain C 1-4 Examples of alkyl groups are: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. 1-4 When the alkyl group is connected, then the 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 atom of an alkyl carbon may be optionally substituted by a substituent as defined above. 1-4 An alkyl group may be interrupted by one or more moieties as defined below.

[0236] As used herein, the term "C 1-6"Alkyl" alone or in combination means a straight chain or branched chain alkyl moiety having 1 to 6 carbon atoms. If present at the end of a molecule, straight chain 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 the alkyl group is connected, then the 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 carbon may optionally be substituted by a substituent as defined above. 1-6 An alkyl group may be interrupted by one or more moieties as defined below.

[0237] 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, wherein C 1-10 、C 1-20 or C 1-50 Each hydrogen atom of a carbon may optionally be substituted by a substituent as defined above. Optionally, C 1-10 、C 1-20 Alkyl, or C 1-50 An alkyl group may be interrupted by one or more moieties as defined below.

[0238] As used herein, the term "C 2-6 "Alkenyl" alone or in combination means a straight or branched hydrocarbon moiety having 2 to 6 carbon atoms and containing at least one carbon-carbon double bond. Examples, if present at the end of a molecule, are: -CH=CH2, -CH=CH-CH3, -CH2-CH=CH2, -CH=CHCH2-CH3 and -CH=CH-CH=CH2. When the two parts of the molecule are connected by C 2-6 When alkenyl is connected, then such C 2-6 An example of an alkenyl group is -CH=CH-. 2-6 Each hydrogen atom of the alkenyl moiety may be optionally substituted by a substituent as defined above. 2-6 An alkenyl group may be interrupted by one or more moieties as defined below.

[0239] Therefore, the term "C 2-10 Alkenyl", "C 2-20 Alkenyl" or "C2-50 "Alkenyl" alone or in combination means a straight or branched chain hydrocarbon moiety having 2 to 10, 2 to 20 or 2 to 50 carbon atoms containing at least one carbon-carbon double bond. 2-10 Alkenyl, C 2-20 Alkenyl or C 2-50 Each hydrogen atom of the alkenyl group may be optionally substituted by a substituent as defined above. 2-10 Alkenyl, C 2-20 Alkenyl or C 2-50 An alkenyl group may be interrupted by one or more moieties as defined below.

[0240] As used herein, the term "C 2-6 "Alkynyl" alone or in combination means a straight or branched hydrocarbon moiety having 2 to 6 carbon atoms and containing at least one carbon-carbon triple bond. If present at the end of a molecule, examples are: -C≡CH, -CH2-C≡CH, CH2-CH2-C≡CH and CH2-C≡C-CH3. When two parts of a molecule are linked through an alkynyl group, an example is -C≡C-. C 2-6 Each hydrogen atom of the alkynyl group may be optionally substituted by a substituent as defined above. Optionally, one or more double bonds may be present. Optionally, C 2-6 An alkynyl group may be interrupted by one or more moieties as defined below.

[0241] Therefore, as used herein, the term "C 2-10 Alkynyl", "C 2-20 Alkynyl" and "C 2-50 "Alkynyl" alone or in combination means a straight or branched chain hydrocarbon moiety having 2 to 10, 2 to 20 or 2 to 50 carbon atoms, containing at least one carbon-carbon triple bond. 2-10 Alkynyl, C 2-20 Alkynyl or C 2-50 Each hydrogen atom of the alkynyl group may be optionally substituted 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 An alkynyl group may be interrupted by one or more moieties as defined below.

[0242] 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 2-10Alkynyl, C 2-20 Alkenyl or C 2-50 Alkynyl groups may be optionally interrupted by one or more moieties, which in certain embodiments are selected from the group consisting of:

[0243]

[0244] in,

[0245] Dashed lines indicate attachment to the remainder of a moiety or reagent; and

[0246] -R and -R a are independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

[0247] 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 substituted by a substituent as defined above. 3-10 "Cycloalkyl" also includes bridged bicyclic rings such as norbornane or norbornene.

[0248] As used herein, the term "8- to 30-membered carbon polycyclyl" or "8- to 30-membered carbon polycyclic ring" refers to a cyclic moiety of two or more rings having 8 to 30 ring atoms, wherein two adjacent rings share at least one ring atom and may contain up to the maximum number of double bonds (fully, partially or unsaturated aromatic or non-aromatic). In certain embodiments, the 8- to 30-membered carbon polycyclyl refers to a cyclic moiety of two, three, four or five rings, in certain embodiments two, three or four rings.

[0249] As used herein, the term "3- to 10-membered heterocyclyl" or "3- to 10-membered heterocycle" means a ring having 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms, which may contain up to the maximum number of double bonds (fully, partially or unsaturated aromatic or non-aromatic rings), wherein at least one ring atom, up to 4 ring atoms, are substituted with a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)2-), oxygen and nitrogen (including =N(O)-), and wherein the ring is attached to the rest of the molecule via a carbon or nitrogen atom. Examples of 3- to 10-membered heterocycles include, but are not limited to, aziridine, oxirane, thiirane, aziridine, oxirane, thiirene, azetidine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazole, isothiazolin, 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 heterocyclic group or the 3- to 10-membered heterocyclic group may be substituted with a substituent as defined below.

[0250] As used herein, the term "8- to 11-membered heterobicyclyl" or "8- to 11-membered heterobicycle" means a heterocyclic moiety having two rings of 8 to 11 ring atoms, wherein at least one ring atom is shared by both rings, and may contain up to the maximum number of double bonds (fully, partially or unsaturated aromatic or non-aromatic rings), wherein at least one ring atom, up to 6 ring atoms, are substituted with a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)2-), oxygen and nitrogen (including =N(O)-), and wherein the ring is attached to the rest of the molecule via a carbon or nitrogen atom. Examples of 8- to 11-membered heterobicyclic rings are: indole, indoline, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzisothiazole, benzimidazole, benzimidazolinone, quinoline, quinazoline, dihydroquinazoline, quinoline, dihydroquinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, decahydroisoquinoline, tetrahydroisoquinoline, dihydroisoquinoline, benzazepine, purine and pteridine. The term 8- to 11-membered heterobicyclic ring also includes spirocyclic structures of two rings, such as 1,4-dioxa-8-azaspiro[4.5]decane, or bridged heterocycles such as 8-azabicyclo[3.2.1]octane. Each hydrogen atom of an 8- to 11-membered heterobicyclic ring or an 8- to 11-membered heterobicyclic ring carbon can be substituted by a substituent as defined below.

[0251] Similarly, the term "8- to 30-membered heteropolycyclyl" or "8- to 30-membered heteropolycycle" refers to a heterocyclic moiety having two or more rings, in certain embodiments three, four, or five rings, of 8 to 30 ring atoms, wherein two adjacent rings share at least one ring atom, and may contain up to the maximum number of double bonds (fully, partially, or unsaturated aromatic or non-aromatic rings), wherein at least one ring atom, up to 10 ring atoms, are substituted by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)2-), oxygen, and nitrogen (including =N(O)-), and wherein the ring is attached to the rest of the molecule via a carbon or nitrogen atom.

[0252] It is to be understood that the phrase "-R x / -R y The pairs are joined together with the atoms to which they are attached to form C 3-10 Cycloalkyl or 3- to 10-membered heterocyclic group":

[0253]

[0254] Meaning-R x and -R y The following structure is formed:

[0255]

[0256] Where R is C 3-10 a cycloalkyl group or a 3- to 10-membered heterocyclic group.

[0257] It is also to be understood that the phrase "-R x / -R y The pairs are joined together with the atoms to which they are attached to form ring A":

[0258]

[0259] Meaning R x and R y The following structure is formed:

[0260]

[0261] As used herein, the term "dry pharmaceutical formulation" means providing a pharmaceutical formulation that can be stored in dry form before administration, such as at least 1 week, such as at least 2 weeks, such as at least 1 month, such as at least 6 months, such as at least 12 months, such as at least 18 months, such as at least 24 months, such as at least 36 months, or such as at least 48 months. Suitable methods for drying are spray drying and lyophilization, i.e., freeze drying. The residual water content of such a dry pharmaceutical formulation comprising a CNP compound (particularly a CNP conjugate) is up to 5%, in certain embodiments less than 2%, and in certain embodiments less than 1%, as determined by Karl Fischer. In certain embodiments, the dry pharmaceutical formulation is dried by lyophilization.

[0262] As used herein, the term "drug" refers to a substance used to treat, cure, prevent, or diagnose disease or to otherwise enhance physical or mental health. If a drug, such as CNP, is conjugated to another moiety, the portion of the resulting product derived from the drug is referred to as the "drug moiety."

[0263] As used herein, the term "EC 50 ” refers to the concentration of CNP agonist or CNP conjugate that elicits half-maximal cGMP production. The EC values ​​of CNP agonists, CNP conjugates, and CNP-22 were measured by 50 NPR-B activity in the form of: NIH-3T3 (mouse embryonic fibroblast cell line) cells expressing NPR-B on the cell surface were cultured and incubated with CNP agonist, CNP conjugate, corresponding released CNP or CNP-22, and the production of the intracellular second messenger cGMP was determined using a standard cGMP assay. Specifically, the assay was performed as follows:

[0264] (1) Mouse NIH-3T3 cells expressing endogenous NPR-B were cultured in DMEM F-12 medium containing 5% fetal bovine serum (FBS) and 5 mM glutamine at 37°C and 5% CO2.

[0265] (2) For each assay, 50,000 cells were resuspended in Dulbecco's PBS containing IBMX and incubated with different concentrations of CNP agonist, CNP conjugate, corresponding released CNP, or CNP-22;

[0266] (3) After incubation at 37°C and 5% CO2 for 30 min, cells were lysed and cGMP levels were measured;

[0267] (4) Generate EC from defined cGMP levels 50 value.

[0268] Preferably, in step (2), the concentration of IBMX is 0.5 mM.

[0269] Step (3) can be performed using any assay for measuring cGMP, which is a standard procedure known to those skilled in the art. Preferably, step (3) is performed using a cGMP time-resolved fluorescence resonance energy transfer (cGMP TR-FRET) assay, more preferably using a cGMP TR-FRET assay from Cisbio (Cat. No. 62GM2PEB). Since in such experiments, reversible CNP conjugates release a certain amount of CNP, which can distort the results, the measurement of NPR-B activity of the CNP conjugate is preferably made in the form of a stable CNP conjugate that does not release CNP.

[0270] As used herein, the term "excipient" refers to a compound used together with a CNP compound, such as a buffer, isotonicity regulator, preservative, stabilizer, anti-adsorbent, oxidation protectant or other adjuvant. However, in some cases, an excipient can have dual or triple functions. The term "excipient" can also refer to a diluent, adjuvant or vehicle used together with a CNP compound. Such pharmaceutical excipients can be sterile liquids, such as water and oil, including oils from petroleum, animal, plant or synthetic sources, including but not limited to: peanut oil, soybean oil, mineral oil, sesame oil, etc. When the pharmaceutical preparation is administered orally, water is a preferred excipient. When the pharmaceutical preparation is administered intravenously or subcutaneously, saline and dextrose aqueous solution are preferred excipients. In certain embodiments, saline solution, dextrose aqueous solution and glycerol solution are used as liquid excipients for injection solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, mannitol, trehalose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skimmed milk powder, glycerol, propylene, ethylene glycol, water, ethanol, etc. The pharmaceutical preparation may also contain small amounts of wetting agents or emulsifiers, pH buffers such as, for example, acetates, succinates, Tris (tris(hydroxymethyl)aminomethane), carbonates, phosphates, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), MES (2-(N-morpholino)ethanesulfonic acid), or may contain detergents such as Poloxamers, poloxamines, CHAPS, Or amino acid, such as glycine, lysine or histidine. These pharmaceutical preparations can be in the form of solution, suspension, emulsion, tablet, pill, capsule, powder, sustained release agent. Pharmaceutical preparation can be mixed with suppository together with traditional adhesive and excipient (such as triglyceride). Oral preparation can comprise the excipient of standard, such as pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate etc. Such preparation can contain the CNP compound of therapeutically effective amount, and the excipient of appropriate amount, so that suitable administration form is provided for patient. Preparation should be applicable to the mode of administration.

[0271] As used herein, the term "preparation" or "pharmaceutical preparation" refers to a preparation containing one or more CNP compounds and one or more excipients, as well as any product formed directly or indirectly by the combination, complexation or aggregation of any two or more components in the composition, or by the dissociation of one or more components, or by other types of reactions or interactions of one or more components. Therefore, the pharmaceutical preparation of the present application encompasses any preparation or composition made by mixing one or more CNP compounds with pharmaceutically acceptable excipients (such as buffers and isotonic agents).

[0272] As used herein, the term "free form" of a drug refers to the unmodified, pharmacologically fully active form of the drug, eg, after release from a reversible CNP conjugate.

[0273] As used herein, the term "functional group" refers to a group of atoms that can react with other groups of atoms. Functional groups include, but are not limited to, the following groups: carboxylic acid (-(C=O)OH), primary or secondary amine (-NH2, -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, phosphoric acid (-O(P=O)OHOH), phosphonic acid (-O(P=O)OHH), haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine, disulfide, sulfonamide, sulfuric acid, vinyl sulfone, vinyl ketone, diazoalkane, ethylene oxide, and aziridine.

[0274] As used herein, the term "halogen" means fluorine, chlorine, bromine or iodine. It is generally preferred that the halogen is fluorine or chlorine.

[0275] As used herein, the term "interrupted" means that the moiety is inserted between two carbon atoms, or (if the insertion is at one of the moiety's termini) between a carbon or heteroatom and a hydrogen atom, in certain embodiments between a carbon and a hydrogen atom.

[0276] As used herein, the term "isotonic agent" refers to a compound that minimizes pain, irritation, and tissue damage caused by cell damage due to differences in osmotic pressure between the injected solution and plasma.

[0277] As used herein, the term "liquid pharmaceutical formulation" means a pharmaceutical formulation that is provided and can be stored in liquid form for at least 1 week, such as at least 2 weeks, such as at least 1 month, such as at least 6 months, such as at least 12 months, such as at least 18 months, such as at least 24 months, such as at least 36 months, or such as at least 48 months prior to administration.

[0278] As used herein, the term "lyophilized formulation" means that the formulation containing the CNP compound is first frozen and then dehydrated by reducing pressure. This term does not exclude additional drying steps that occur during the manufacturing process before the formulation is filled into the final container.

[0279] As used herein, the terms "lyophilization" or "freeze drying" are interchangeable and refer to a dehydration process characterized by freezing the formulation, then reducing the surrounding pressure, and optionally heating to allow the frozen water in the formulation to sublime directly from the solid phase to a gas. Generally, the sublimated water is collected by desublimation.

[0280] As used herein, the term "moiety" refers to a portion of a molecule that lacks one or more atoms compared to the corresponding reagent. For example, if a reagent of the formula "HXH" reacts with another reagent and becomes part of the reaction product, the structure of the corresponding moiety of the reaction product is "HX-" or "-X-", with each "-" indicating attachment to another moiety. Thus, the drug moiety (e.g., CNP moiety) is released from the reversible CNP conjugate as a drug, such as CNP.

[0281] It should be understood that if a sequence or chemical structure of a group of atoms is provided that connects two moieties or interrupts a moiety, the sequence or chemical structure may be connected to the two moieties in either direction unless otherwise expressly stated. For example, the moiety "-C(O)N(R 1 )-” can be used as “-C(O)N(R 1 )-" or as "-N(R 1 )C(O)-" connects two parts, or interrupts a part. Similarly, part:

[0282]

[0283] It can be connected to two parts or interrupted to one part, as:

[0284] or as

[0285] In the case that the CNP part comprises one or more acidic or basic groups, pharmaceutical preparation also comprises its corresponding pharmaceutically or toxicologically acceptable salt, is specifically its pharmaceutically available salt.Therefore, can exist or use the CNP part that comprises one or more acidic groups, for example as alkali metal salt, alkaline earth metal salt or as ammonium salt.The more accurate example of such salt comprises sodium salt, potassium salt, calcium salt, magnesium salt or the salt containing ammonia or organic amine, as for example ethamine, ethanolamine, triethanolamine or amino acid, and other salt and amine well known by persons skilled in the art.Comprise the CNP part of one or more basic groups and promptly be protonated group, can exist, and can use with the form of it and inorganic or organic acid addition salt. The example of suitable acid comprises: hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, tosic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethyl acetic 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 acid well known by persons skilled in the art.For those skilled in the art, known other methods that basic group is converted into cationic, such as the alkylation of amido, obtain the suitable counterion of positively charged ammonium group and salt.If the CNP part comprises acidic and basic group simultaneously, according to the application's pharmaceutical preparation, except mentioned salt form, also comprise inner salt or betaine (zwitterion).Each salt can be by customary method well known by persons skilled in the art, for example, by these conjugates are contacted in solvent or dispersant with organic or inorganic acid or alkali, or by obtaining with the anion exchange or cation exchange of other salt.

[0286] As used herein, the term "natural product" refers to a purified organic compound isolated from a natural source and produced by a primary or secondary metabolic pathway.

[0287] As used herein, the term "oligonucleotide" refers to double-stranded or single-stranded RNA and DNA, preferably having 2 to 1000 nucleotides, and any modifications thereof. Modifications include, for example, modifications that provide other chemical groups that incorporate additional charge, polarizability, hydrogen bonding, electrostatic interactions, and mobility into the nucleic acid ligand base or the entire nucleic acid ligand. Such modifications include, for example: 2'-sugar modification, 5-pyrimidine modification, 8-purine modification, modification at the exocyclic amine, substitution of 4-thiouridine, substitution of 5-bromo- or 5-iodo-uracil; backbone modification, methylation, unconventional base pairing combinations such as the isobases isocytidine and isoguanidine. Modifications can also include 3' and 5' modifications, such as capping and stereochemical changes. The term also includes aptamers.

[0288] As used herein, the term "oxidation protectant" or "antioxidant" refers to a compound that inhibits the oxidation of other compounds, such as peptides.

[0289] As used herein, the term "pH adjusting agent" refers to a compound used to adjust the pH of a solution.

[0290] As used herein, the term "pharmaceutically acceptable" means a substance that does not cause harm when administered to a patient, and preferably means a substance approved by regulatory agencies such as EMA (Europe) and / or FDA (USA) and / or any other national regulatory agencies for use in animals, preferably humans.

[0291] As used herein, the term "physiological conditions" refers to an aqueous buffer at pH 7.4 and 37°C.

[0292] As used herein, the term "polypeptide" refers to a chain of at least 2 and at most 50 amino acid monomeric moieties linked by peptide (amide) bonds. For simplicity, for CNPs only, sequences with more than 50 amino acids will also be referred to as "polypeptides."

[0293] As used herein, the term "preservative" refers to a chemical substance that has an antimicrobial effect and / or prevents chemical degradation.

[0294] As used herein, the term "protein" refers to a chain of more than 50 amino acid monomer moieties linked by peptide bonds, wherein preferably no more than 12,000 amino acid monomers are linked by peptide bonds, such as no more than 10,000 amino acid monomer moieties, no more than 8,000 amino acid monomer moieties, no more than 5,000 amino acid monomer moieties, or no more than 2,000 amino acid monomer moieties.

[0295] As used herein, the term "polymer" means a molecule comprising a repeating structural unit (i.e., a monomer), which is connected by a chemical bond in a side chain, cyclic, branched, cross-linked or dendritic manner or a combination thereof, which can be synthetic or biologically derived or a combination of the two. It should be understood that polymers may also include one or more other chemical groups and / or moieties, such as one or more functional groups. In certain embodiments, soluble polymers have a molecular weight of at least 0.5 kDa, such as a molecular weight of at least 1 kDa, a molecular weight of at least 2 kDa, a molecular weight of at least 3 kDa, or a molecular weight of at least 5 kDa. If polymer is soluble, in certain embodiments, it has a molecular weight of at most 1000 kDa, such as at most 750 kDa, such as at most 500 kDa, such as at most 300 kDa, such as at most 200 kDa, such as at most 100 kDa.

[0296] It will be understood that even though the side chain of each amino acid may be different, a protein or polypeptide is a polymer in which amino acids are repeating structural units.

[0297] As used herein, the term "polymeric" or "polymeric moiety" means an agent or moiety that comprises one or more polymers or polymeric moieties. A polymeric agent or moiety may also optionally comprise one or more other moieties, which in certain embodiments are selected from the group consisting of:

[0298] ·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 tetrahydronaphthyl; and

[0299] A key selected from the group consisting of:

[0300]

[0301] in,

[0302] A dashed line indicates attachment to the remainder of the moiety or agent, and

[0303] -R and -R a are independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

[0304] It will be understood by those skilled in the art that the polymer products obtained from a polymerization reaction do not all have the same molecular weight, but rather exhibit a distribution of molecular weights. Therefore, as used herein, the terms molecular weight range, molecular weight, range of the number of monomers in a polymer, and number of monomers in a polymer refer to the number average molecular weight and average number of monomers, i.e., the arithmetic mean of the molecular weight of a polymer or polymeric portion and the arithmetic mean of the number of monomers in a polymer or polymeric portion.

[0305] Thus, in a polymeric moiety comprising "x" monomer units, any integer given for "x" corresponds to the arithmetic mean number of monomers. Any integer range given for "x" provides a range of integers within which the arithmetic mean number of monomers lies. An integer for "x" given as "about x" means that the arithmetic mean number of monomers lies within an integer range of x + / - 10%, in certain embodiments within an integer range of x + / - 8%, in certain embodiments within an integer range of x + / - 5%, and in certain embodiments within an integer range of x + / - 2%.

[0306] As used herein, with respect to a moiety or agent, the term "PEG-based" means that the moiety or agent comprises PEG. In certain embodiments, the PEG-based moiety or agent comprises at least 10% (w / w) PEG, such as at least 20% (w / w) PEG, such as at least 30% (w / w) PEG, such as at least 40% (w / w) PEG, such as at least 50% (w / w), such as at least 60% (w / w) PEG, such as at least 70% (w / w) PEG, such as at least 80% (w / w) PEG, such as at least 90% (w / w) PEG, such as at least 95% (w / w) PEG. The remaining weight percentage of the PEG-based moiety or agent is other moieties selected from the following moieties and bonds:

[0307] C1 -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 tetrahydronaphthyl; and

[0308] A key selected from the group consisting of:

[0309]

[0310] in,

[0311] A dashed line indicates attachment to the remainder of the moiety or agent, and

[0312] -R and -R a are independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

[0313] As used herein, with respect to a moiety or agent, the term "PEG-based comprising at least X% PEG" means that the moiety or agent comprises at least X% (w / w) of ethylene glycol units (-CH2CHO-), wherein the ethylene glycol units may be arranged in blocks, alternating arrangements, or may be randomly distributed in the moiety or agent, and in certain embodiments, all of the ethylene glycol units of the moiety or agent are present in one block; the remaining weight percentage of the PEG-based moiety or agent, in certain embodiments, is other moieties selected from the following moieties and linkages:

[0314] C1 -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 tetrahydronaphthyl; and

[0315] A key selected from the group consisting of:

[0316]

[0317] in,

[0318] A dashed line indicates attachment to the remainder of the moiety or agent, and

[0319] -R and -R a are independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

[0320] As used herein, the term "hyaluronic acid-based comprising at least X% hyaluronic acid" is used accordingly.

[0321] Those of ordinary skill in the art also recognize that the reversible CNP conjugates comprised in the liquid preparation of the present application are prodrugs.As used herein, term " prodrug " refers to that drug moiety (such as CNP moiety) is reversibly and covalently conjugated to carrier moiety (such as-Z) by reversible linker moiety.Prodrug releases reversible and covalently bound drug moiety in the form of its corresponding drug.In other words, prodrug is the reversible conjugate comprising drug moiety (such as CNP moiety, it is covalently and reversibly conjugated via reversible linker moiety and carrier moiety, wherein carrier moiety and reversible conjugation of reversible linker moiety are direct or by spacer.Such prodrug or conjugate release the drug moiety preceded by conjugation in the form of free drug.

[0322] As used herein, the term "random coil" refers to a peptide or protein that adopts / has / forms, in certain embodiments, a conformation that substantially lacks defined secondary and tertiary structure, as determined by circular dichroism spectroscopy in an aqueous buffer at ambient temperature and pH 7.4. In certain embodiments, the ambient temperature is about 20° C. (i.e., 18° C. to 22° C.), and in certain embodiments, the ambient temperature is 20° C.

[0323] As used herein, the term "reversible linkage" is a linkage that is cleavable in the absence of an enzyme under physiological conditions (aqueous buffer at pH 7.4, 37° C.) with a half-life ranging from one hour to six months, such as one hour to four months, such as one hour to three months, one hour to two months, or one hour to one month. Thus, a stable linkage is one that has a half-life of more than six months under physiological conditions (aqueous buffer at pH 7.4, 37° C.).

[0324] As used herein, the term "agent" means a compound comprising at least one functional group for reacting with a functional group of another compound or drug. It should be understood that drugs comprising functional groups such as primary or secondary amines or hydroxyl functional groups are also reagents.

[0325] As used herein, the term "reversible linker moiety" is a moiety that is covalently conjugated to a drug moiety (e.g., a CNP moiety) through a reversible bond and is also covalently conjugated to a carrier moiety, such as -Z, wherein the covalent conjugation to the carrier moiety is direct or through a spacer moiety (e.g., -L). 2 In certain embodiments, -Z and -L 2 -The bond between them is a stable bond.

[0326] As used herein, the term "reconstitution" means adding a liquid to a dry pharmaceutical formulation to restore the formulation's original form, such as a solution.

[0327] As used herein, the term "small molecule" or "small drug molecule" refers to an organic compound or drug having a molecular weight of less than 1000 Da, such as less than 900 Da or less than 800 Da.

[0328] As used herein, the term "sealed container" means a container that is closed in a manner that is airtight, does not allow exchange of gases between the outside and the inside, and maintains the sterility of the contents.

[0329] As used herein, the term "spacer" or "spacer moiety" refers to a moiety suitable for connecting two moieties. Suitable spacers may be selected from the group consisting of: C 1-50 Alkyl, C 2-50 Alkenyl or C 2-50 Alkynyl, where C 1-50 Alkyl, C 2-50 Alkenyl or C 2-50 The alkynyl group may be optionally interrupted by one or more groups selected from the group consisting of: -NH-, -N(C 1-4 alkyl)-、-O-、-S-、-C(O)-、-C(O)NH-、-C(O)N(C 1-4 alkyl)-, -OC(O)-, -S(O)-, -S(O)2-, 4- to 7-membered heterocyclyl, phenyl and naphthyl.

[0330] As used herein, the term "substituted" means that one or more -H atoms of a molecule or moiety are replaced with a different atom or group of atoms, which atom or group of atoms is referred to as a "substituent."

[0331] In certain embodiments, such one or more substituents are independently selected from the group consisting of 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 x1R x1a )、-S(O)2R 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; wherein -T 0 、C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl is optionally substituted by one or more identical or different -R x2 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-、-S(O)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 )-;

[0332] -Rx1 、-R x1a 、-R x1b are independently selected from the following groups: -H, -T 0 、C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl; wherein -T 0 、C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl is optionally substituted by one or more identical or different -R x2 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-、-S(O)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 )-;

[0333] Each T 0 Independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; wherein each T 0 are optionally replaced by one or more identical or different -R x2 replace;

[0334] Each-R x2 Independently selected from the group consisting of 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)2Rx4 、-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; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[0335] Each-R x3 、-R x3a 、-R x4 、-R x4a 、-R x4b Independently selected from the group consisting of: -H and C 1-6 Alkyl; among which C 1-6 The alkyl group is optionally substituted with one or more same or different halogens.

[0336] In certain embodiments, one or more substituents are independently selected from the group consisting of 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)2R x1 、-S(O)R x1 、-N(R x1 )S(O)2N(R x1a R x1b ),-SR x1 、-N(R x1 Rx1a )、-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 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl; wherein -T 0 、C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl is optionally substituted by one or more identical or different -R x2 substituted, and wherein C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-、-S(O)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 )-;

[0337] Each-R x1 、-R x1a 、-R x1b 、-R x3 、-R x3a Independently selected from the group consisting of: -H, halogen, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl;

[0338] Each-T 0 Independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; wherein each T 0 independently optionally with one or more identical or different -R x2 replace;

[0339] Each-R x2 Independently selected from the group consisting of 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)2R 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; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[0340] Each-R x4 、-R x4a 、-R x4b Independently selected from the group consisting of: -H, halogen, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl.

[0341] In certain embodiments, one or more substituents are independently selected from the group consisting of 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)2R 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; wherein -T 0 、C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl is optionally substituted by one or more identical or different -R x2 substituted, and wherein C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-、-S(O)2N(R x3 )-、-S(O)N(R x3 )-、-S(O)2-、-S(O)-、-N(Rx3 )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 )-;

[0342] Each-R x1 、-R x1a 、-R x1b 、-R x3 、-R x3a Independently selected from the group consisting of: -H, halogen, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl;

[0343] Each-T 0 Independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; wherein each T 0 are optionally replaced by one or more identical or different -R x2 replace;

[0344] In certain embodiments, up to 6 -H atoms of the optionally substituted molecule are independently substituted with substituents, e.g., 5 -H atoms are independently substituted with substituents, 4 -H atoms are independently substituted with substituents, 3 -H atoms are independently substituted with substituents, 2 -H atoms are independently substituted with substituents, or 1 -H atom is substituted with a substituent.

[0345] As used herein, with respect to pharmaceutical formulations comprising a reversible CNP conjugate, the terms "stable" and "stability" mean that after storage for a period of time, such as one month, two months, four months, six months, eight months, twelve months, eighteen months, twenty-four months, thirty-six months, forty-eight months, sixty months, in particular, after a specified storage time, the pharmaceutical formulation contains less than 5% of free CNP and less than 20% (such as less than 10% or such as less than 5%) of impurities, such as impurities resulting from methionine oxidation. Such storage is preferably carried out at 2 to 8° C., i.e., the formulation is in liquid form. Impurities can be quantified by RP-HPLC or SEC based on their respective peak areas relative to the total peak area of ​​all CNP compound-related peaks, in particular, the peak area of ​​the CNP conjugate-related peak in the chromatogram. Impurities in the CNP portion and the CNP portion of the reversible CNP conjugate can be determined by proteolytic digestion or peptide mapping and quantified based on their respective peak areas relative to the peak area of ​​the corresponding unmodified proteolytic peptide. Thus, with respect to a pharmaceutical formulation comprising a CNP agonist or a stabilized CNP conjugate, the terms "stable" and "stability" mean that after storage for a period of time, such as one month, two months, four months, six months, eight months, twelve months, eighteen months, twenty-four months, thirty-six months, forty-eight months, sixty months, in particular after a specified storage time, the pharmaceutical formulation contains less than 20% (such as less than 10% or such as less than 5%) of impurities, such as impurities resulting from methionine oxidation. Impurities can be quantified by RP-HPLC or SEC based on their respective peak areas relative to the total peak areas of all CNP agonist or stabilized CNP conjugate-related peaks, in particular the peak areas of the CNP conjugate-related peaks in the chromatogram. Impurities in the CNP agonist can be determined by proteolytic digestion or peptide mapping analysis and quantified based on their respective peak areas relative to the peak areas of the corresponding unmodified proteolytic peptides.

[0346] As used herein, the term "stabilizer" refers to a compound used to stabilize a CNP compound. Stabilization can be achieved by enhancing the peptide's stabilizing power or by direct binding of the excipient to the CNP compound.

[0347] As used herein, the term "surfactant" refers to a wetting agent that lowers the surface tension of a liquid.

[0348] As used herein, the term "therapeutically effective amount" means an amount that is enough to cure, alleviate or partially stop the clinical manifestations of a given disease and its complication. The effective amount of each purpose will depend on the severity of the disease or injury and the weight and overall state of the experimenter. It should be understood that conventional experiments can be used, by constructing a matrix of values ​​and testing the different points in the matrix to determine appropriate dosage, which is all within the scope of the common skills of trained physicians. Within the scope of the application, the therapeutically effective amount relates to the dosage that is intended to achieve the therapeutic effect that continues for a period of time, i.e. at least one day, such as two days, such as three days, such as four days, such as five days, such as six days, such as one week or such as two weeks.

[0349] As used herein, the term "traceless linker" means a reversible linker that releases the drug in its free form upon cleavage.

[0350] As used herein, the term "unit dose" means the amount of drug administered to a patient as a single dose.

[0351] As used herein, with respect to carrier moieties such as polymeric moieties, the term "water-soluble" means that when such moieties are part of a CNP conjugate, at least 1 g of the CNP conjugate comprising such water-soluble carrier moiety can be dissolved in 1 liter of water at 20°C to form a homogeneous solution.

[0352] In general, the terms “comprise” and “comprising” also cover “consist of” and “consist of” or “consist essentially of”.

[0353] When the present disclosure refers to a range of parameter values, it should be understood that the disclosure also discloses the integers within the range and the sub-ranges formed between the integers in the range.

[0354] In certain embodiments, the CNP compound is a CNP conjugate.

[0355] In certain embodiments, the CNP compound is a reversible CNP conjugate, wherein the CNP conjugate comprises at least one CNP moiety covalently and reversibly conjugated to a carrier moiety, and under physiological conditions, the at least one CNP moiety is released therefrom with a release half-life of at least 6 hours.

[0356] In certain embodiments, the CNP compound is a stable CNP conjugate, wherein the CNP conjugate comprises at least one CNP moiety covalently and stably conjugated to a carrier moiety, ie, via a stable linkage with a half-life of more than 6 months under physiological conditions.

[0357] In certain embodiments, the CNP compound is a stable CNP conjugate, wherein the CNP conjugate comprises a CNP moiety covalently and stably conjugated to a fatty acid derivative.

[0358] In certain embodiments, the CNP portion has a sequence 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: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, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ IDNO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94 and SEQ ID NO:95.

[0359] In certain embodiments, 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. 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. In certain embodiments, the CNP portion has the sequence of SEQ ID NO: 30.

[0360] In certain embodiments, the ring portion of the CNP moiety has SEQ ID NO: 96 or SEQ ID NO: 97. In certain embodiments, the ring portion of the CNP moiety has SEQ ID NO: 96. In certain embodiments, the ring portion of the CNP moiety has SEQ ID NO: 97.

[0361] In certain embodiments, the carrier moiety is defined as the variable -Z, which is described in more detail elsewhere herein.

[0362] In order to maintain a certain pH or pH range, liquid pharmaceutical formulations contain buffers. Buffers maintain the pH of liquid pharmaceutical formulations within the desired range.

[0363] The buffer can be selected from the group consisting of acetic acid, succinic acid, citric acid, lactic acid, glutamic acid, fumaric acid, aspartic acid, glutaric acid, phosphoric acid, histidine, gluconic acid, tartaric acid, malic acid, Tris (tris (hydroxymethyl) aminomethane) and mixtures thereof. It will be apparent to those skilled in the art that corresponding conjugate bases or salts of the buffer, such as acetate, succinate, citrate, lactate, glutamate, fumarate, aspartate, glutarate, phosphate, gluconate, tartrate, malate and mixtures thereof, are also encompassed.

[0364] In certain embodiments, the buffer is selected from the group consisting of acetic acid, succinic acid, citric acid, lactic acid, glutamic acid, fumaric acid, aspartic acid, glutaric acid, phosphoric acid, histidine, gluconic acid, tartaric acid, and malic acid.

[0365] In certain embodiments, the buffer is acetic acid. In certain embodiments, the buffer is succinic acid. In certain embodiments, the buffer is citric acid. In certain embodiments, the buffer is lactic acid. In certain embodiments, the buffer is glutamic acid. In certain embodiments, the buffer is fumaric acid. In certain embodiments, the buffer is aspartic acid. In certain embodiments, the buffer is glutaric acid. In certain embodiments, the buffer is phosphoric acid. In certain embodiments, the buffer is histidine. In certain embodiments, the buffer is gluconic acid. In certain embodiments, the buffer is tartaric acid. In certain embodiments, the buffer is malic acid.

[0366] Acetic acid was found to be a particularly useful buffering agent because its use did not result in the formation of acetylated adduct impurities under long-term storage conditions, as detected by RP-HPLC and / or RP-HPLC-MS.

[0367] In certain embodiments, the buffering agent is not citric acid.

[0368] In certain embodiments, the concentration of the buffer ranges from 5 to 50 mM. In certain embodiments, the concentration of the buffer ranges from 10 to 50 mM. In certain embodiments, the concentration of the buffer is about 10 mM.

[0369] In certain embodiments, the pH of the liquid pharmaceutical formulation is no higher than 6 because the reversible bonds within the reversible CNP conjugate may be unstable under alkaline conditions.

[0370] In certain embodiments, the pH of the liquid pharmaceutical formulation is from about pH 3.5 to about pH 5.5. In certain embodiments, the pH of the liquid pharmaceutical formulation is from about pH 3.5 to about pH 5.0. In certain embodiments, the pH of the liquid pharmaceutical formulation is from about pH 4.5 to about pH 5.0. In certain embodiments, the pH of the liquid pharmaceutical formulation is from about pH 4.0 to about pH 5.0. In certain embodiments, the pH of the liquid pharmaceutical formulation is about 4.5. In certain embodiments, the pH of the liquid pharmaceutical formulation is about 4.0. In certain embodiments, the pH of the liquid pharmaceutical formulation is 4.5. In certain embodiments, the pH of the liquid pharmaceutical formulation is 4.0.

[0371] The liquid pharmaceutical formulation according to the present application comprises an isotonic agent. The isotonic agent can be selected from the group consisting of mannitol, trehalose, sucrose, raffinose, gelatin, lactose, calcium hydrogen phosphate, sorbitol, xylitol, glycine, histidine, ethanol, hydroxyethyl starch, potassium chloride, sodium chloride, glucose, dextran, polysucrose, Propylene glycol, glycerin and mixtures thereof.

[0372] In certain embodiments, the isotonic agent is selected from the group consisting of mannitol, trehalose, sucrose, raffinose, gelatin, lactose, dibasic calcium phosphate, sorbitol, xylitol, glycine, histidine, ethanol, hydroxyethyl starch, potassium chloride, sodium chloride, glucose, dextran, propylene glycol, glycerol, and mixtures thereof.

[0373] In certain embodiments, the isotonic agent is selected from the group consisting of mannitol, trehalose, sucrose, raffinose, gelatin, lactose, dibasic calcium phosphate, sorbitol, xylitol, glycine, histidine, ethanol, hydroxyethyl starch, potassium chloride, sodium chloride, glucose, dextran, glycerol, and propylene glycol.

[0374] In certain embodiments, the isotonic agent is mannitol. In certain embodiments, the isotonic agent is trehalose, such as trehalose dihydrate. In certain embodiments, the isotonic agent is sucrose. In certain embodiments, the isotonic agent is raffinose. In certain embodiments, the isotonic agent is gelatin. In certain embodiments, the isotonic agent is lactose. In certain embodiments, the isotonic agent is calcium hydrogen phosphate. In certain embodiments, the isotonic agent is sorbitol. In certain embodiments, the isotonic agent is xylitol. In certain embodiments, the isotonic agent is glycine. In certain embodiments, the isotonic agent is histidine. In certain embodiments, the isotonic agent is ethanol. In certain embodiments, the isotonic agent is hydroxyethyl starch. In certain embodiments, the isotonic agent is potassium chloride. In certain embodiments, the isotonic agent is sodium chloride. In certain embodiments, the isotonic agent is glucose. In certain embodiments, the isotonic agent is dextran. In certain embodiments, the isotonic agent is polysucrose In certain embodiments, the isotonic agent is propylene glycol. In certain embodiments, the isotonic agent is glycerol.

[0375] In certain embodiments, the isotonicity agent is selected from the group consisting of mannitol and trehalose.

[0376] In certain embodiments, the isotonic agent is a mixture of mannitol and trehalose.

[0377] As defined herein, the term "trehalose" is intended to encompass all salts and hydration states of trehalose, such as anhydrous trehalose or trehalose dihydrate. In certain embodiments, the term "trehalose" refers to anhydrous trehalose. In certain embodiments, the term "trehalose" refers to trehalose dihydrate.

[0378] As defined herein, the term "mannitol" is intended to encompass D-mannitol and L-mannitol and mixtures thereof. In certain embodiments, the term "mannitol" refers to L-mannitol. In certain embodiments, the term "mannitol" refers to D-mannitol. In certain embodiments, the term "mannitol" refers to a mixture of L-mannitol and D-mannitol.

[0379] In certain embodiments, the concentration range of the isotonic agent is 10 to 200 mg / ml. In certain embodiments, the concentration of the isotonic agent is about 100 mg / ml. In certain embodiments, the concentration of the isotonic agent is about 84 mg / ml. In certain embodiments, the concentration of the isotonic agent is about 80 mg / ml. In certain embodiments, the concentration of the isotonic agent is about 50 mg / ml. In certain embodiments, the concentration of the isotonic agent is about 41 mg / ml.

[0380] In certain embodiments, the liquid formulation is substantially isotonic, implying that its osmolality is about 250-350 mOsm / kg water. The liquid formulation can also be hypertonic (>350 mOsm / kg water) or hypotonic (<250 mOsm / kg water).

[0381] In certain embodiments, reducing sugars should be avoided because they can react with the CNP moiety. Thus, in certain embodiments, the pharmaceutical formulation does not contain reducing sugars.

[0382] The liquid pharmaceutical preparation according to the present application contains a preservative. The preservative can be selected from the group consisting of m-cresol, benzyl alcohol, benzoic acid, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, potassium sorbate, chlorobutanol, benzyl alcohol, phenylmercuric nitrate, thimerosal, sorbic acid, potassium sorbate, chlorocresol, benzalkonium chloride, ethylene glycol monoethyl ether, chlorhexidine, chlorobutanol, phenylethyl alcohol, phenylmercuric acetate, and mixtures thereof.

[0383] 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 chlorocresol. 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 benzyl alcohol. In certain embodiments, the preservative is benzyl mercuric 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 ethylene glycol monoethyl ether. 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.

[0384] In certain embodiments, the concentration of the preservative ranges from 1 to 6 mg / ml. In certain embodiments, the concentration of the preservative ranges from 3 to 6 mg / ml. In certain embodiments, the concentration of the preservative ranges from 2 to 3 mg / ml. In certain embodiments, the concentration of the preservative is about 3 mg / ml. In certain embodiments, the concentration of the preservative is 3 mg / ml.

[0385] The liquid pharmaceutical formulation according to the present application may further comprise a pH adjusting agent. In certain embodiments, the pH adjusting agent is a base. Examples of bases may be selected from the group consisting of sodium hydroxide, Tris (tris(hydroxymethyl)aminomethane), potassium hydroxide, lysine, and mixtures thereof.

[0386] In certain embodiments, the pH adjusting agent is a base selected from the group consisting of sodium hydroxide, sodium acetate, Tris (tris(hydroxymethyl)aminomethane), potassium hydroxide, and lysine.

[0387] In certain embodiments, the pH adjusting agent is selected from the group consisting of sodium hydroxide and Tris (tris(hydroxymethyl)aminomethane).

[0388] In certain embodiments, the pH adjuster is sodium hydroxide. In certain embodiments, the pH adjuster is sodium acetate. In certain embodiments, the pH adjuster is Tris. In certain embodiments, the pH adjuster is potassium hydroxide. In certain embodiments, the pH adjuster is lysine.

[0389] In certain embodiments, the pH adjusting agent is an acid. Examples of acids may be selected from the group consisting of hydrochloric acid, phosphoric acid, carbonic acid, nitric acid, and mixtures thereof.

[0390] In certain embodiments, the pH adjuster is an acid selected from the group consisting of hydrochloric acid, phosphoric acid, carbonic acid, and nitric acid.

[0391] In certain embodiments, the pH adjusting agent is hydrochloric acid. In certain embodiments, the pH adjusting agent is phosphoric acid. In certain embodiments, the pH adjusting agent is carbonic acid. In certain embodiments, the pH adjusting agent is nitric acid.

[0392] In certain embodiments, the pH adjusting agent is a mixture of at least one base and at least one acid. In certain embodiments, the pH adjusting agent is a mixture of a base and an acid. In certain embodiments, the pH adjusting agent is a mixture of sodium hydroxide and hydrochloric acid.

[0393] In certain embodiments, the concentration range of the pH adjusting agent or the mixture of pH adjusting agents is 0.01 to 5 mg / ml. In certain embodiments, the concentration range of the pH adjusting agent or the mixture of pH adjusting agents is 0.04 to 2.5 mg / ml. In certain embodiments, the concentration range of the pH adjusting agent or the mixture of pH adjusting agents is 0.08 to 1.25 mg / ml. In certain embodiments, the concentration of the pH adjusting agent or the mixture of pH adjusting agents is about 0.4 mg / ml. It should be understood that in the case of a mixture of pH adjusting agents, the concentration provided refers to the total concentration of all pH adjusting agents.

[0394] Liquid pharmaceutical preparations according to the application optionally include antioxidants. The example of antioxidant can be selected from the group consisting of methionine, butylated hydroxytoluene, butylated hydroxyanisole, tocopherol, propyl gallate, ascorbic acid, sodium bisulfite, ethylenediaminetetraacetic acid (EDTA), cysteine, glutathione, monothioglycerol, poly(ethyleneimine), vitamin E, ectoine, morin and mixtures thereof.

[0395] In certain embodiments, the antioxidant is selected from the group consisting of methionine, butylated hydroxytoluene, butylated hydroxyanisole, tocopherol, propyl gallate, ascorbic acid, sodium bisulfite, ethylenediaminetetraacetic acid (EDTA), cysteine, glutathione, monothioglycerol, poly(ethyleneimine), vitamin E, ectoine, and morin.

[0396] It has been observed that methionine is particularly useful in preventing oxidation of residues, such as methionine residues in CNP compounds, to, for example, methionine sulfoxide or methionine sulfone.

[0397] In certain embodiments, the antioxidant is methionine. In certain embodiments, the antioxidant is ascorbic acid. In certain embodiments, the antioxidant is butylated hydroxytoluene. In certain embodiments, the antioxidant is butylated hydroxyanisole. In certain embodiments, the antioxidant is tocopherol. In certain embodiments, the antioxidant is propyl gallate. In certain embodiments, the antioxidant is sodium bisulfite. In certain embodiments, the antioxidant is monothioglycerol. In certain embodiments, the antioxidant is EDTA. In certain embodiments, the antioxidant is cysteine. In certain embodiments, the antioxidant is glutathione. In certain embodiments, the antioxidant is polyethyleneimine. In certain embodiments, the antioxidant is vitamin E. In certain embodiments, the antioxidant is ectoine. In certain embodiments, the antioxidant is morin.

[0398] As defined herein, the term "methionine" is intended to encompass D-methionine and L-methionine, and 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.

[0399] As defined herein, the term "EDTA" is intended to encompass all forms of EDTA known in the art, such as EDTA salts, including EDTA metal salts, such as EDTA disodium salt, EDTA dipotassium salt, EDTA calcium salt, EDTA dimagnesium salt, or mixtures thereof. In certain embodiments, EDTA refers to EDTA disodium salt. In certain embodiments, the term "EDTA" refers to EDTA dicalcium salt. In certain embodiments, the term "EDTA" refers to anhydrous EDTA.

[0400] In certain embodiments, the liquid pharmaceutical formulation does not contain sodium bisulfite. In certain embodiments, the liquid pharmaceutical formulation does not contain cysteine. In certain embodiments, the liquid pharmaceutical formulation does not contain glutathione. In certain embodiments, the liquid pharmaceutical formulation does not contain monothioglycerol. In certain embodiments, the liquid pharmaceutical formulation does not contain ascorbic acid.

[0401] In certain embodiments, the concentration of the antioxidant ranges from about 0.1 mg / ml to about 1.5 mg / ml. In certain embodiments, the concentration of the antioxidant ranges from about 0.5 mg / ml to about 1.0 mg / ml.

[0402] In certain embodiments, the molar ratio of the antioxidant to the CNP or CNP agonist is from about 0.1:1 to about 100:1. In certain embodiments, the molar ratio of the antioxidant to the CNP or CNP agonist is from about 0.1:1 to about 70:1. In certain embodiments, the molar ratio of the antioxidant to the CNP or CNP agonist is from about 0.1:1 to about 15:1. In certain embodiments, the molar ratio of the antioxidant to the CNP or CNP agonist is from about 1:1 to about 10:1. In certain embodiments, the molar ratio of the antioxidant to the CNP or CNP agonist is from about 3:1 to about 7:1.

[0403] In certain embodiments, the liquid pharmaceutical formulations of the present application do not contain an antioxidant.

[0404] The liquid pharmaceutical formulation according to the present application optionally contains a detergent. Examples of detergents can be selected from the group consisting of octyl sucrose, Triton X-100, Triton X-114, polysorbate 20, polysorbate 80, NP-40, CA-630, Brij-35, Brij-58, n-dodecyl-β-maltoside, octyl-β-glucoside, octylthioglucosidoside, sodium lauryl sulfate, CHAPS, F-127 and F-68.

[0405] In certain embodiments, the liquid pharmaceutical formulations of the present application do not contain detergents.

[0406] Formulations intended for human administration are preferably prepared under Good Manufacturing Practice (GMP), which is approved or approvable for the preparation of drugs for human administration by the FDA or regulatory agencies in countries outside the U.S. (e.g., the European Medicines Agency). Generally, the formulation is sterile, for example, by sterile filtration using a 0.2 μm or 0.22 μm filter.

[0407] In certain embodiments, the liquid pharmaceutical formulation of the present application comprises a CNP agonist at a concentration of about 0.1 to about 20 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 0.5 to about 15 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 0.75 to about 10 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 1 to about 8 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 2 to about 6 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 10 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 8 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 6 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 4 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 2 mg / ml. In certain embodiments, the concentration of the CNP agonist is about 1 mg / ml.

[0408] In certain embodiments, the liquid pharmaceutical formulation of the present application comprises a CNP agonist at a concentration of 0.1 to 20 mg / ml. In certain embodiments, the concentration of the CNP agonist is 0.5 to 15 mg / ml. In certain embodiments, the concentration of the CNP agonist is 0.75 to 10 mg / ml. In certain embodiments, the concentration of the CNP agonist is 1 to 8 mg / ml. In certain embodiments, the concentration of the CNP agonist is 2 to 6 mg / ml. In certain embodiments, the concentration of the CNP agonist is 10 mg / ml. In certain embodiments, the concentration of the CNP agonist is 8 mg / ml. In certain embodiments, the concentration of the CNP agonist is 6 mg / ml. In certain embodiments, the concentration of the CNP agonist is 4 mg / ml. In certain embodiments, the concentration of the CNP agonist is 2 mg / ml. In certain embodiments, the concentration of the CNP agonist is 1 mg / ml.

[0409] In certain embodiments, the liquid pharmaceutical preparation of the present application comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 0.1 to about 20 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 0.5 to about 15 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 0.75 to about 10 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 1 to about 8 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 2 to about 6 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 10 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 8 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 6 mg / ml. In certain embodiments, the liquid pharmaceutical formulation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 4 mg / ml. In certain embodiments, the liquid pharmaceutical formulation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 2 mg / ml. In certain embodiments, the liquid pharmaceutical formulation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of about 1 mg / ml. It should be understood that the above-mentioned concentrations provided refer to the amount of the CNP portion, rather than the entire CNP conjugate.

[0410] In certain embodiments, the liquid pharmaceutical preparation of the present application comprises a CNP conjugate, wherein the CNP portion is present at a concentration of 0.1 to 20 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of 0.5 to 15 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of 0.75 to 10 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of 1 to 8 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of 2 to 6 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of 10 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of 8 mg / ml. In certain embodiments, the liquid pharmaceutical preparation comprises a CNP conjugate, wherein the CNP portion is present at a concentration of 6 mg / ml. In certain embodiments, the liquid pharmaceutical formulation comprises a CNP conjugate wherein the CNP portion is present at a concentration of 4 mg / ml. In certain embodiments, the liquid pharmaceutical formulation comprises a CNP conjugate wherein the CNP portion is present at a concentration of 2 mg / ml. In certain embodiments, the liquid pharmaceutical formulation comprises a CNP conjugate wherein the CNP portion is present at a concentration of 1 mg / ml. It should be understood that the concentrations provided above refer to the amount of the CNP portion, not the entire CNP conjugate.

[0411] In certain embodiments, the reversible CNP conjugate has the structure of Formula (Ia) or (Ib):

[0412]

[0413] in,

[0414] -D is the CNP part;

[0415] -L 1 - is a reversible joint portion;

[0416] -L 2 - is a single chemical bond or spacer moiety;

[0417] -Z is a carrier moiety;

[0418] 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

[0419] y is an integer selected from the group consisting of 2, 3, 4 and 5.

[0420] -D of formula (Ia) or (Ib) is covalently and reversibly conjugated to -L 1 -.

[0421] In certain embodiments, the liquid pharmaceutical formulation of the present application comprises:

[0422]

[0423] In certain embodiments, the liquid pharmaceutical formulation of the present application comprises:

[0424]

[0425] In certain embodiments, -Z of formula (Ia) or (Ib) is a polymeric moiety. In certain embodiments, -Z of formula (Ia) or (Ib) is a water-soluble polymeric moiety.

[0426] In certain embodiments, x of Formula (Ia) is an integer selected from the group consisting of 1, 2, 3, 4, 6, and 8. In certain embodiments, x of Formula (Ia) is an integer selected from the group consisting of 1, 2, 4, and 6. In certain embodiments, x of Formula (Ia) is an integer selected from the group consisting of 1, 4, and 6, and in certain embodiments, x of Formula (Ia) is 1.

[0427] In certain embodiments, y of Formula (Ib) is an integer selected from the group consisting of 3, 4, and 5. In certain embodiments, y of Formula (Ib) is an integer selected from the group consisting of 4 and 5. In certain embodiments, y of Formula (Ib) is an integer selected from the group consisting of 2 and 3. In certain embodiments, y of Formula (Ib) is 2. In certain embodiments, y of Formula (Ib) is 3. In certain embodiments, y of Formula (Ib) is 4. In certain embodiments, y of Formula (Ib) is 5.

[0428] In certain embodiments, the reversible CNP conjugate has formula (Ia) where x=1.

[0429] 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.

[0430] In certain embodiments, -D of Formula (Ia) or (Ib) has a sequence of SEQ ID NO: 20. In certain embodiments, -D of Formula (Ia) or (Ib) has a sequence of SEQ ID NO: 21. In certain embodiments, -D of Formula (Ia) or (Ib) has a sequence of SEQ ID NO: 22. In certain embodiments, -D of Formula (Ia) or (Ib) has a sequence of SEQ ID NO: 23. In certain embodiments, -D of Formula (Ia) or (Ib) has a sequence of SEQ ID NO: 24. In certain embodiments, -D of Formula (Ia) or (Ib) has a sequence of SEQ ID NO: 25.

[0431] The moiety -L of formula (Ia) or (Ib) 1 - is conjugated to a functional group of a side chain of an amino acid residue of -D, to an N-terminal amine functional group or a C-terminal carboxyl functional group of -D, or to a nitrogen atom in the main polypeptide chain of -D. The connection to the N-terminus or C-terminus can be made directly through the corresponding amine or carboxyl functional group, respectively, or indirectly, wherein the spacer part is first conjugated to the amine or carboxyl functional group, wherein the spacer part -L 1 - is conjugated.

[0432] The moiety -L of formula (Ia) or (Ib) 1 - is a reversible linker from which the drug, i.e., DH, is released in its free or unmodified form, i.e., -L 1- is a scarless linker. Suitable reversible linkers are known in the art, such as the reversible linker moieties disclosed in, for example, WO2005 / 099768A2, WO2006 / 136586A2, WO2011 / 089216A1 and WO2013 / 024053A1, which are incorporated herein by reference.

[0433] In certain embodiments, -L 1 - is a reversible linker as described in WO2011 / 012722A1, WO2011 / 089214A1, WO2011 / 089215A1, WO2013 / 024052A1 and WO2013 / 160340A1, which are incorporated herein by reference.

[0434] Part-L 1 - can be connected to -D by any type of bond, provided that it is reversible. In certain embodiments, -L 1 - is linked to -D via a bond selected from the group consisting of an amide, an ester, a carbamate, an acetal, an aminal, an imine, an oxime, a hydrazone, a disulfide, and an acylguanidine. In certain embodiments, -L 1 - is linked to -D via a bond selected from the group consisting of amide, ester, carbamate and acylguanidine. It will be appreciated that these bonds themselves may not be reversible, but are included in -L 1 The adjacent group in - can make the bond reversible.

[0435] In certain embodiments, the moiety -L 1 - is linked to -D via an amide bond.

[0436] Part-L 1 - disclosed in WO2009 / 095479A2. Thus, in certain embodiments, the moiety -L 1 -is formula (II):

[0437]

[0438] The dashed line indicates the connection to the nitrogen of -D by forming an amide bond, where -D is a CNP moiety;

[0439] -X-

[0440] Yes-C(R 4 R 4a )-、-N(R 4 )-、-O-、-C(R 4 R 4a )-C(R 5 R 5a )-、-C(R 5 R 5a )-C(R 4R 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 )-;

[0441] X 1 is C or S(O);

[0442] -X 2 Yes-C(R 8 R 8a )-or-C(R 8 R 8a )-C(R 9 R 9a )-;

[0443] =X 3 is =O, =S or =N-CN;

[0444] -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 Independently selected from the group consisting of: -H and C 1-6 alkyl;

[0445] -R 3 、-R 3a Independently selected from the group consisting of: -H and C 1-6 Alkyl, provided that -R 3 、-R 3a If one or both are not -H, they are passed to sp 3 - a hybridized carbon atom connected to the N atom to which it is attached;

[0446] -R 7 Yes-N(R 10 R 10a ) or -NR 10-(C=O)-R 11 ;

[0447] R 7a 、-R 10 、-R 10a 、-R 11 Independently of each other, -H or C 1-6 alkyl;

[0448] Optionally, -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 pairs form a chemical bond;

[0449] Optionally, -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 pairs are linked to the atoms to which they are attached to form C 3-10 cycloalkyl or 3- to 10-membered heterocyclic group;

[0450] Optionally, -R 1 / -R 4 、-R 1 / -R 5 、-R 1 / -R 6 、-R 1 / -R 7a 、-R 4 / -R 5 、-R 4 / -R 6 、-R 8 / -R 9 、-R 2 / -R 3 One or more of the pairs are linked to the atoms to which they are attached to form ring A;

[0451] Optionally, -R 3 / -R 3aare linked together with the nitrogen atom to which they are attached to form a 3- to 10-membered heterocyclic ring;

[0452] A is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; and

[0453] Where -L 1 -Use -L 2 -Z replaces, and -L 1 - is optionally further substituted, provided that the hydrogens marked with an asterisk in formula (II) are not replaced by -L 2 -Z or a substituent;

[0454] in,

[0455] -L 2 - is a single chemical bond or spacer; and

[0456] -Z is a carrier moiety, preferably a water-soluble polymeric moiety.

[0457] In certain embodiments, -L of formula (II) 1 -Being a part-L 2 -Z replaces.

[0458] In certain embodiments, -L of formula (II) 1 -Not to be further superseded.

[0459] It should be understood that if R 3 / -R 3a Together with the nitrogen atom to which they are attached to form a 3- to 10-membered heterocyclic ring, only such 3- to 10-membered heterocyclic rings can be formed in which the atom directly attached to the nitrogen is sp 3 -hybridized carbon atoms. In other words, R 3 / -R 3a Such 3- to 10-membered heterocyclic rings formed together with the nitrogen atom to which they are attached have the following structure:

[0460]

[0461] in,

[0462] The dotted line indicates the difference with -L 1 -The rest of the connection;

[0463] The ring contains 3 to 10 atoms, including at least one nitrogen; and

[0464] R # and R ## Represents sp 3-Hybridized carbon atoms.

[0465] It is also to be understood that the 3- to 10-membered heterocyclic ring may be further substituted.

[0466] -R of formula (II) 3 / -R 3a Illustrative embodiments of suitable 3- to 10-membered heterocyclic rings formed together with the nitrogen atom to which they are attached are as follows:

[0467]

[0468] in,

[0469] Dashed lines indicate connections to the rest of the molecule; and

[0470] -R is selected from the group consisting of -H and C 1-6 alkyl.

[0471] -L of formula (II) 1 - may be optionally further substituted. Generally speaking, any substituent may be used as long as it does not affect the fragmentation principle, i.e. the hydrogens marked with an asterisk in formula (II) are not replaced and the part of formula (II)

[0472]

[0473] The nitrogen is still part of the primary, secondary or tertiary amine, i.e. -R 3 and -R 3a Independently of each other -H or through sp 3 The -hybridized carbon atom is connected to -N<.

[0474] In certain embodiments, -R 1 or -R 1a Use -L 2 In certain embodiments, -R 2 or -R 2a Use -L 2 In certain embodiments, -R 3 or -R 3a Use -L 2 In certain embodiments, -R 4 Use -L 2 In certain embodiments, -R 5 or -R 5a Use -L 2 In certain embodiments, -R 6 Use -L 2 In certain embodiments, -R7 or -R 7a Use -L 2 In certain embodiments, -R 8 or -R 8a Use -L 2 In certain embodiments, -R 9 or -R 9a Use -L 2 -Z replaces.

[0475] In certain embodiments, -R 4 Use -L 2 -Z replaces.

[0476] In certain embodiments, -X- of formula (II) is -C(R 4 R 4a )-or-N(R 4 In certain embodiments, -X- of formula (II) is -C(R 4 R 4a )-.

[0477] In certain embodiments, -X of formula (II) 1 - is C.

[0478] In certain embodiments, =X of formula (II) 3 =O.

[0479] In certain embodiments, -X of formula (II) 2 -for-C(R 8 R 8a )-.

[0480] In certain embodiments, -R 8 and -R 8a In certain embodiments, -R 8 and -R 8a In certain embodiments, at least one of -R 8 and -R 8a All are -H.

[0481] In certain embodiments, -R 1 and -R 1a In certain embodiments, -R 1 and -R 1a In certain embodiments, at least one of -R 1 and -R1a All are -H.

[0482] In certain embodiments, -R 2 and -R 2a In certain embodiments, -R 2 and -R 2a In certain embodiments, at least one of -R 2 and -R 2a All are -H.

[0483] In certain embodiments, -R 3 and -R 3a In certain embodiments, -R 3 and -R 3a In certain embodiments, at least one of -R in formula (II) is methyl. 3 and -R 3a In certain embodiments, -R 3 and -R 3a In certain embodiments, -R 3 is -H, and -R 3a It is a methyl group.

[0484] In certain embodiments, -R 4 and -R 4a In certain embodiments, -R 4 and -R 4a In certain embodiments, at least one of -R 4 and -R 4a All are -H.

[0485] In certain embodiments, the moiety -L 1 -is formula (IIa):

[0486]

[0487] The dotted line indicates the connection to the nitrogen of -D through the formation of an amide bond, wherein -D is a CNP moiety;

[0488] -R 1 、-R 1a 、-R 2 、-R 2a 、-R 3 、-R 3a, -R 4 , -R 4a and -X 2 - used as defined in formula (II);

[0489] and wherein -L 1 - is replaced by -L 2 -Z and wherein -L 1 - is optionally further substituted, provided that the hydrogen marked with an asterisk in formula (IIa)

[0490] is not substituted by -L 2 -Z or a substituent.

[0491] In certain embodiments, -L of formula (IIa) 1 - is replaced with a moiety -L 2 -Z.

[0492] In certain embodiments, the moiety -L of formula (IIa) 1 - is not further substituted.

[0493] In certain embodiments, -R of formula (IIa) 1 and -R 1a are independently selected from the group consisting of: -H, methyl, and ethyl. In certain embodiments, -R of formula (IIa) 1 and -R 1a at least one of which is -H. In certain embodiments, -R of formula (IIa) 1 and -R[[ID=4!6]] 1a are both -H. In certain embodiments, -R of formula (IIa) M 4 and -R 4a are independently selected from the group consisting of: -H, methyl, and ethyl. In certain embodiments, -R of formula (IIa) 4 and -R<000!0650>at least one of which is -H. In certain embodiments, -R of formula (IIa) 4 and -R 4a M are both -H.

[0494] In certain embodiments, -X of formula (IIa) 2 - is -C(R 8 R 8a ).

[0495] In certain embodiments, -R of formula (IIa) 8 and -R 8a are independently selected from the group consisting of: -H, methyl, and ethyl. In certain embodiments, -R of formula (IIa) 8 and -R 8aIn certain embodiments, at least one of -R 8 and -R 8a All are -H.

[0496] In certain embodiments, -R 2 and -R 2a In certain embodiments, -R 2 and -R 2a In certain embodiments, at least one of -R 2 and -R 2a All are -H.

[0497] In certain embodiments, -R 3 and -R 3a In certain embodiments, -R of formula (IIa) is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. 3 and -R 3a In certain embodiments, at least one of -R in formula (IIa) is methyl. 3 and -R 3a In certain embodiments, -R 3 and -R 3a In certain embodiments, -R 3 is -H, and -R 3a It is a methyl group.

[0498] In certain embodiments, the moiety -L 1 -is formula (IIb):

[0499]

[0500] The dashed line indicates the connection to the nitrogen of -D via formation of an amide bond, where -D is a CNP moiety;

[0501] -R 2 、-R 2a 、-R 3 、-R 3a and -X 2 - used as defined in formula (II); and

[0502] Where -L 1 -Use -L 2 -Z replaces and where -L 1 - is optionally further substituted, provided that the hydrogens marked with an asterisk in formula (IIb) are not replaced by -L 2 -Z or a substituent.

[0503] In certain embodiments, -L of formula (IIb) 1 -Use a part-L 2 In certain embodiments, the moiety -L of formula (IIb) is substituted with -Z. 1 -Not to be further superseded.

[0504] In certain embodiments, -X of Formula (IIb) 2 -for-C(R 8 R 8a )-.

[0505] In certain embodiments, -R 8 and -R 8a In certain embodiments, -R of formula (IIb) is independently selected from the group consisting of -H, methyl, and ethyl. 8 and -R 8a In certain embodiments, at least one of -R 8 and -R 8a All are -H.

[0506] In certain embodiments, -R 2 and -R 2a In certain embodiments, -R of formula (IIb) is independently selected from the group consisting of -H, methyl, and ethyl. 2 and -R 2a In certain embodiments, at least one of -R 2 and -R 2a All are -H.

[0507] In certain embodiments, -R 3 and -R 3a In certain embodiments, -R of formula (IIb) is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. 3 and -R 3a In certain embodiments, at least one of -R in formula (IIb) is methyl. 3 and -R 3a In certain embodiments, -R 3 and -R 3a In certain embodiments, -R 3 is -H, and -R 3a It is a methyl group.

[0508] In certain embodiments, the moiety -L 1 -is formula (IIb'):

[0509]

[0510] in,

[0511] The dashed line indicates the connection to the nitrogen of -D via formation of an amide bond, where -D is the CNP moiety;

[0512] The dashed line marked with an asterisk indicates the connection to -L 2 -;

[0513] -R 2 、-R 2a 、-R 3 、-R 3a and -X 2 - used as defined in formula (II); and

[0514] Where -L 1 - is optionally further substituted, provided that the hydrogens marked with an asterisk in formula (IIb') are not replaced by a substituent.

[0515] In certain embodiments, the moiety -L of formula (IIb') 1 -Not to be further superseded.

[0516] In certain embodiments, -X of Formula (IIb') 2 -for-C(R 8 R 8a )-.

[0517] In certain embodiments, -R 8 and -R 8a In certain embodiments, -R of Formula (IIb') 8 and -R 8a In certain embodiments, at least one of -R 8 and -R 8a All are -H.

[0518] In certain embodiments, -R 2 and -R 2a In certain embodiments, -R of Formula (IIb') 2 and -R 2a In certain embodiments, at least one of -R 2 and -R 2a All are -H.

[0519] In certain embodiments, -R3 and -R 3a In certain embodiments, -R 3 and -R 3a In certain embodiments, at least one of -R 3 and -R 3a In certain embodiments, -R 3 and -R 3a In certain embodiments, -R 3 is -H, and -R 3a It is a methyl group.

[0520] In certain embodiments, the moiety -L 1 -is formula (IIc):

[0521]

[0522] The dashed line indicates the connection to the nitrogen of -D by forming an amide bond, where -D is a CNP moiety;

[0523] and

[0524] Where -L 1 -Use -L 2 -Z replaces and where -L 1 - is optionally further substituted, provided that the hydrogens marked with an asterisk in formula (IIc) are not replaced by -L 2 -Z or a substituent.

[0525] In certain embodiments, -L of formula (IIc) 1 -Use a part-L 2 -Z replaces.

[0526] In certain embodiments, the moiety -L of formula (IIc) 1 -Not to be further superseded.

[0527] In certain embodiments, the moiety -L 1 -is formula (IIc-a):

[0528]

[0529] The dashed line indicates the connection to the nitrogen of -D by forming an amide bond, where -D is a CNP moiety;

[0530] and

[0531] Where -L 1 -Use -L 2-Z replaces and where -L 1 - is optionally further substituted, provided that the formula (IIc-a)

[0532] The hydrogens marked with an asterisk in the -L 2 -Z or a substituent.

[0533] In certain embodiments, -L of formula (IIc-a) 1 -Use a part-L 2 -Z replaces.

[0534] In certain embodiments, the moiety -L of formula (IIc-a) 1 -Not to be further superseded.

[0535] In certain embodiments, the moiety -L 1 -is formula (IIc-b):

[0536]

[0537] The dashed line indicates the connection to the nitrogen of -D via formation of an amide bond, where -D is a CNP moiety;

[0538] and

[0539] Where -L 1 -Use -L 2 -Z replaces and where -L 1 - is optionally further substituted, provided that the formula (IIc-b)

[0540] The hydrogens marked with an asterisk in the -L 2 -Z or a substituent.

[0541] In certain embodiments, -L of formula (IIc-b) 1 -Use a part-L 2 -Z replaces.

[0542] In certain embodiments, the moiety -L of formula (IIc-b) 1 -Not to be further superseded.

[0543] In certain embodiments, the moiety -L 1 - is selected from the group consisting of formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v):

[0544]

[0545]

[0546] in,

[0547] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0548] and

[0549] The dashed line marked with an asterisk indicates the connection to -L 2 -Z; and

[0550] -L 1 - is optionally further substituted, provided that the hydrogens marked with an asterisk in formulae (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v) are not replaced by a substituent.

[0551] In certain embodiments, the moiety -L of formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv), and (IIc-v) 1 -Not to be further superseded.

[0552] In certain embodiments, the moiety -L 1 -is formula (IIc-ii):

[0553]

[0554] in,

[0555] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0556] and

[0557] The dashed line marked with an asterisk indicates the connection to -L 2 -Z.

[0558] In certain embodiments, -L of formula (IIc-ii) 1 -Being a part-L 2 -Z replaces.

[0559] In certain embodiments, the moiety -L 1 - is selected from the group consisting of formula (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv') and (IIc-v'):

[0560]

[0561]

[0562] in,

[0563] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0564] The dashed line marked with an asterisk indicates the connection to -L 2 -Z; and

[0565] -L 1 - is optionally further substituted, provided that it is of the formula (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv')

[0566] The hydrogen atoms marked with an asterisk in (IIc-v') and (IIc-v') are not replaced by substituents.

[0567] In certain embodiments, the moiety -L of formula (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv'), and (IIc-v') 1 -Not to be further superseded.

[0568] In certain embodiments, the moiety -L 1 -is the formula (IIc-ii'):

[0569]

[0570] in,

[0571] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0572] The dashed line marked with an asterisk indicates the connection to -L 2 -Z.

[0573] In certain embodiments, -L of formula (IIc-ii') 1 -Use a part-L 2 -Z replaces.

[0574] In certain embodiments, the moiety -L 1 - is selected from the group consisting of formula (IIc-i"), (IIc-ii"), (IIc-iii") and (IIc-iv"):

[0575]

[0576] in,

[0577] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0578] The dashed line marked with an asterisk indicates the connection to -L 2 -Z; and

[0579] -L 1- is optionally further substituted, provided that it is of the formula (IIc-i"), (IIc-ii"), (IIc-iii") and (IIc-iv")

[0580] The hydrogen atoms marked with an asterisk are not replaced by substituents.

[0581] In certain embodiments, the moiety -L of formula (IIc-i"), (IIc-ii"), (IIc-iii"), and (IIc-iv") 1 -Not to be further superseded.

[0582] In certain embodiments, the moiety -L 1 -is the formula (IIc-ii"):

[0583]

[0584] in,

[0585] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0586] and

[0587] The dashed line marked with an asterisk indicates the connection to -L 2 -Z.

[0588] In certain embodiments, -L of formula (IIc-ii") 1 -Being a part-L 2 -Z replaces.

[0589] In certain embodiments, Formula (II), (IIa), (IIb), (IIb'), (IIc), (IIc-a), (IIc-b), (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv), (II -L of c-v), (IIc-i'), (IIc-ii'), (IIc-iii'), (IIc-iv'), (IIc-v'), (IIc-i”), (IIc-ii”), (IIc-iii”) and (IIc-iv”) 1 The other optional substituents of - are as described above.

[0590] WO 2016 / 020373 A1 discloses another part - L 1 -. Thus, in certain embodiments, the moiety -L 1 -is formula (III):

[0591]

[0592] in,

[0593] The dotted line indicates the connection with the primary amine, secondary amine, or hydroxyl group of -D by forming an amide or ester bond, respectively.

[0594] Where -D is the CNP moiety;

[0595] -R 1 、-R 1 、-R 2 、-R 2 a、-R 3 and -R 3 a are independently selected from the following groups: -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;

[0596] -R 4 、-R 5 and -R 5a are independently selected from the group consisting of: -H, -C(R 9 R 9a R 9b ) and -T;

[0597] a1 and a2 are independently 0 or 1;

[0598] Each-R 6 、-R 6a 、-R 7 、-R 7a 、-R 8 、-R 8a 、-R 8b 、-R 9 、-R 9a 、-R 9b are independently selected from the group consisting of: -H, halogen, -CN, -COOR 10 、-OR 10 、-C(O)R 10 、-C(O)N(R 10 R 10 a)、-S(O)2N(R 10 R 10a )、-S(O)N(R 10 R 10a )、-S(O)2R 10 、-S(O)R 10 、-N(R10 )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 -T, C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 Alkynyl groups are optionally substituted with one or more identical or different -R 11 substituted, and wherein C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -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 )-;

[0599] Each-R 10 、-R 10a 、-R 10b Independently selected from the group consisting of -H, -T, C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20Alkynyl; wherein -T, C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 Alkynyl groups are optionally substituted with one or more identical or different -R 11 substituted, and wherein C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -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 )-;

[0600] Each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclyl and 8- to 11-membered heterobicyclyl; wherein each T is independently optionally replaced by one or more identical or different -R 11 replace;

[0601] Each-R 11 are independently selected from: 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)2R 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; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[0602] Each-R 12 、-R 12a 、-R 13 、-R 13a 、-R 13b Independently selected from the group consisting of: -H and C 1-6 Alkyl; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[0603] Optionally, -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 pairs are linked to the atoms to which they are attached to form C 3-10 cycloalkyl or 3- to 10-membered heterocyclic group;

[0604] Optionally, -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 / -R5 、-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 / -R 7 One or more of the pairs are linked to the atoms to which they are attached to form ring A;

[0605] A is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl;

[0606] Where -L 1 -Use -L 2 -Z replaces and where -L 1 - is optionally further substituted;

[0607] in,

[0608] -L 2 - is a single chemical bond or spacer; and

[0609] -Z is a carrier moiety, preferably a water-soluble polymeric moiety.

[0610] In certain embodiments, -L of formula (III) 1 The other optional substituents of - are as described above.

[0611] In certain embodiments, -L of formula (III) 1 -Use a part-L 2 -Z replaces.

[0612] In certain embodiments, -L in formula (III) 1 -Not to be further superseded.

[0613] -L is disclosed in EP1536334B1, WO 2009 / 009712 A1, WO 2008 / 034122 A1, WO 2009 / 143412 A2, WO 2011 / 082368 A2 and US 8618124 B2. 1 -other embodiments, which are incorporated herein by reference in their entirety.

[0614] US8946405B2 and US8754190B2 disclose -L 1 -other embodiments, which are incorporated herein by reference in their entirety. Thus, part -L 1 -is formula (IV):

[0615]

[0616] in,

[0617] The dotted line represents the connection to -D (which is a CNP moiety), and wherein the connection is via a

[0618] -D functional groups: -OH, -SH and -NH2;

[0619] m is 0 or 1;

[0620] -R 1 and -R 2 At least one or two of them are independently selected from the following groups: -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 ,

[0621] -R 1 and -R 2 One and only one of the following is selected from the group consisting of: -H, optionally substituted alkyl, optionally substituted arylalkyl, and optionally substituted heteroarylalkyl;

[0622] -R 3 Selected from the group consisting of -H, optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -OR 9 and -N(R 9 )2;

[0623] -R 4 selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;

[0624] Each -R 5 is 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;

[0625] -R 9 is selected from the group consisting of: -H and optionally substituted alkyl;

[0626] -Y- is absent and -X- is -O- or -S-; or

[0627] -Y- is -N(Q)CH2- and -X- is -O-;

[0628] Q is selected from the group consisting of: optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;

[0629] Optionally, -R 1 and -R 2 may be linked to form a 3- to 8-membered ring; and

[0630] Optionally, -R 9 together with the nitrogen to which it is attached forms a heterocycle;

[0631] wherein -L 1 - is replaced by -L 2 -Z and wherein -L 1 - is optionally further substituted;

[0632] wherein,

[0633] -L 2 - is a single bond or a spacer; and

[0634] -Z is a carrier moiety, preferably a water-soluble polymeric moiety.

[0635] In certain embodiments, the optional further substituents of -L 1 - of formula (IV) are as described above.

[0636] In certain embodiments, -L 1 - of formula (IV) is replaced with a moiety -L 2 -Z. In certain embodiments, -L 1 - of formula (IV) is not further substituted.

[0637] Only in the context of formula (IV), the terms used have the following meanings:

[0638] As used herein, the term "alkyl" includes straight chain, branched chain, or cyclic saturated hydrocarbon groups of 1 to 8 carbon atoms, or in some embodiments, 1 to 6 or 1 to 4 carbon atoms.

[0639] The term "alkoxy" includes alkyl groups bonded to oxygen, including methoxy, ethoxy, isopropoxy, cyclopropyloxy, cyclobutyloxy, and the like.

[0640] The term "alkenyl" includes non-aromatic unsaturated hydrocarbons having carbon-carbon double bonds.

[0641] The term "alkynyl" includes non-aromatic unsaturated hydrocarbons having a carbon-carbon triple bond.

[0642] The term "aryl" includes aromatic hydrocarbon groups of 6 to 18 carbon atoms, and in certain embodiments, 6 to 10 carbon atoms, including groups such as phenyl, naphthyl, and anthracenyl. The term "heteroaryl" includes aromatic rings containing 3 to 15 carbon atoms, and in certain embodiments, 3 to 7 carbon atoms, 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.

[0643] In certain embodiments, the alkenyl, alkynyl, aryl, or heteroaryl moiety can be coupled to the remainder of the molecule through an alkylene linkage. In these cases, the substituent will be referred to as an alkenylalkyl, alkynylalkyl, arylalkyl, or heteroarylalkyl group, indicating that the alkylene moiety is located between the alkenyl, alkynyl, aryl, or heteroaryl moiety and the molecule to which the alkenyl, alkynyl, aryl, or heteroaryl group is coupled.

[0644] The term "halogen" includes bromine, fluorine, chlorine and iodine.

[0645] The term "heterocycle" refers to a 4- to 8-membered aromatic or non-aromatic ring containing 3 to 7 carbon atoms and at least one atom of N, O, or S. Examples are piperidinyl, piperazinyl, tetrahydropyranyl, pyrrolidinyl, and tetrahydrofuranyl, as well as the exemplary groups provided above for the term "heteroaryl".

[0646] When the ring system is optionally substituted, suitable substituents are selected from the group consisting of alkyl, alkenyl, alkynyl, or another ring, each optionally further substituted. Optional substituents on any group, including the above, include halogen, nitro, cyano, -OR, -SR, -NR2, -OCOR, -NRCOR, -COOR, -CONR2, -SOR, -S02R, -SONR2, -SONR2, wherein 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.

[0647] WO 2013 / 036857 A1 discloses -L1 -another embodiment of the present invention, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 -is formula (V):

[0648]

[0649] in,

[0650] The dashed line indicates the connection to -D (which is a CNP moiety), and wherein the connection is through the amine functionality of -D;

[0651] -R 1 Selected from the group consisting of: optionally substituted C1-C6 linear, branched or cyclic alkyl; optionally substituted aryl; optionally substituted heteroaryl; alkoxy; and -NR 5 2;

[0652] -R 2 is selected from the group consisting of: -H; optionally substituted C1-C6 alkyl; optionally substituted aryl; and optionally substituted heteroaryl;

[0653] -R 3 is selected from the group consisting of: -H; optionally substituted C1-C6 alkyl; optionally substituted aryl; and optionally substituted heteroaryl;

[0654] -R 4 is selected from the group consisting of: -H; optionally substituted C1-C6 alkyl; optionally substituted aryl; and optionally substituted heteroaryl;

[0655] Each-R 5 are independently selected from the following groups: -H; optionally substituted C1-C6 alkyl; optionally substituted aryl; and optionally substituted heteroaryl; or when two -R 5 When taken together, they may be cycloalkyl or cycloheteroalkyl;

[0656] Where -L 1 -Use -L 2 -Z replaces and where -L 1 - is optionally further substituted;

[0657] in,

[0658] -L 2 - is a single chemical bond or spacer; and

[0659] -Z is a carrier moiety, preferably a water-soluble polymeric moiety.

[0660] In certain embodiments, -L of formula (V) 1 The other optional substituents of - are as described above.

[0661] In certain embodiments, -L of formula (V) 1 -Use a part-L 2 In certain embodiments, -L of formula (V) 1 -Not to be further superseded.

[0662] In the context of formula (V) only, the terms used have the following meanings:

[0663] "Alkyl," "alkenyl," and "alkynyl" include straight, branched, or cyclic hydrocarbon groups of 1-8 carbons, 1-6 carbons, or 1-4 carbons, wherein alkyl is a saturated hydrocarbon, alkenyl includes one or more carbon-carbon double bonds, and alkynyl includes one or more carbon-carbon triple bonds. Unless otherwise specified, these contain 1-6 carbons.

[0664] "Aryl" includes aromatic hydrocarbon groups of 6-18 carbon atoms, and in certain embodiments, 6-10 carbon atoms, including groups such as phenyl, naphthyl, and anthracenyl. "Heteroaryl" includes aromatic rings containing 3-15 carbon atoms, and in certain embodiments, 3-7 carbon atoms, containing at least one N, O, or S atom, including groups such as pyrrolyl, pyridinyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl, and the like.

[0665] The term "substituted" means an alkyl, alkenyl, alkynyl, aryl or heteroaryl group containing one or more substituents replacing one or more hydrogen atoms. The substituents can generally be selected from: halogen, including F, Cl, Br and I; lower alkyl, including linear, branched and cyclic; lower haloalkyl, including fluoroalkyl, chloroalkyl, bromoalkyl and iodoalkyl; OH; lower alkoxy, including linear, branched and cyclic; SH; lower alkylthio, including linear, branched and cyclic; amino, alkylamino, dialkylamino, silyl (including alkylsilyl, alkoxysilyl and arylsilyl); nitro; cyano; carbonyl; carboxylic acid, carboxylic ester, carboxylic acid ... Amine, aminocarbonyl; aminoacyl; carbamate; urea; thiocarbamate; thiourea; ketone; sulfone; sulfonamide; aryl, including phenyl, naphthyl and anthracenyl; heteroaryl, including 5-membered heteroaryl (including pyrrole, imidazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, thiadiazole, triazole, oxadiazole and tetrazole), 6-membered heteroaryl (including pyridine, pyrimidine, pyrazine, and fused heteroaryl, including benzofuran, benzothiophene, benzoxazole, benzimidazole, indole, benzothiazole, benzisoxazole and benzisothiazole).

[0666] WO 2022 / 115563 A1 discloses -L 1 -another embodiment of the present invention, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1-is formula (Va):

[0667]

[0668] The dashed line marked with an asterisk indicates the 2 -Z linkage, and the unlabeled dashed line indicates linkage with -D.

[0669] In certain embodiments, -L 1 - is formula (Va), and the dashed line marked with an asterisk represents -L 2 -Z linkage, and the unlabeled dashed line indicates linkage with -D, where -D is the CNP portion of the following amino acid sequence:

[0670] SEQ ID NO: 106:

[0671] LQEHPQARKYKGAQKKGLSKGCFGLKLDRIGSMSGLGC,

[0672] wherein the cysteines at positions 22 and 38 are linked by a disulfide bond; and

[0673] Which with -L 1 -linkage occurs at the N-terminus or a loop of the peptide.

[0674] US 7585837 B2 discloses -L 1 -another embodiment of the present invention, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 -is formula (VI):

[0675]

[0676] in,

[0677] The dotted line represents the connection to -D (which is the CNP moiety), and wherein the connection is through the amine function of -D

[0678] conducted by the group;

[0679] R 1 and R 2 independently selected from the group consisting of hydrogen, alkyl, alkoxy, alkoxyalkyl, aryl, alkaryl, aralkyl, halogen, nitro, -SO3H, -SO2NHR 5 , amino, ammonium, carboxyl, PO3H2 and OPO3H2;

[0680] R 3 、R 4 and R 5 independently selected from the group consisting of hydrogen, alkyl, and aryl;

[0681] Where -L1 - substituted by -L 2 - substituted by -Z and -L 1 - optionally further substituted;

[0682] wherein,

[0683] -L 2 - is a single chemical bond or a spacer; and

[0684] -Z is a carrier moiety, preferably a water-soluble polymeric moiety.

[0685] Suitable substituents of formula (VI) are alkyl (such as C 1-6 alkyl), alkenyl (such as C 2-6 alkenyl), alkynyl (such as C 2-6 alkynyl), aryl (such as phenyl), heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl (such as an aromatic 4- to 7-membered heterocycle) or a halogen moiety.

[0686] In certain embodiments, -L of formula (VI) 1 - is substituted with a moiety -L 2 -Z.

[0687] In certain embodiments, the optional further substituents of -L of formula (VI) are as described above. 1 - are as described above.

[0688] In certain embodiments, -L of formula (VI) 1 - is not further substituted.

[0689] Only in the context of formula (VI), the terms used have the following meanings:

[0690] The terms "alkyl", "alkoxy", "alkoxyalkyl", "aryl", "alkaryl" and "aralkyl" mean an alkyl radical having 1-8 carbon atoms (in certain embodiments 1-4 carbon atoms), such as methyl, ethyl, propyl, isopropyl and butyl; and an aryl radical having 6-10 carbon atoms, such as phenyl and naphthyl. The term "halogen" includes bromine, fluorine, chlorine and iodine.

[0691] Another embodiment of -L is disclosed in WO 2002 / 089789 A1, which is hereby incorporated by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 - is of formula (VII): 1 - is of formula (VII):

[0692]

[0693] wherein,

[0694] The dotted line represents the connection to -D (which is the CNP moiety), and wherein the connection is through the amine function of -D

[0695] conducted by the group;

[0696] L1 is a bifunctional linking group;

[0697] Y1 and Y2 are independently O, S or NR 7 ;

[0698] R 2 、R 3 、R 4 、R 5 、R 6 and R 7 Independently selected from the group consisting of 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;

[0699] Ar is a moiety that forms a polysubstituted aromatic hydrocarbon or a polysubstituted heterocyclic group when included in formula (VII);

[0700] X is a chemical bond or a moiety that is actively transported to the target cell, a hydrophobic moiety, or a combination thereof,

[0701] y is 0 or 1;

[0702] Where -L 1 -Use -L 2 -Z replaces and -L 1 - is optionally further substituted;

[0703] in,

[0704] -L 2 - is a single chemical bond or spacer; and

[0705] -Z is a carrier moiety, preferably a water-soluble polymeric moiety.

[0706] In certain embodiments, -L of formula (VII) 1 -Being a part-L 2 -Z replaces.

[0707] In certain embodiments, -L of formula (VII) 1 The other optional substituents of - are as described above.

[0708] In certain embodiments, -L of formula (VII) 1 -Not to be further superseded.

[0709] In the context of formula (VII) only, the terms used have the following meanings:

[0710] The term "alkyl" is understood to include, for example, straight chain, branched, substituted C 1-12 Alkyl, including alkoxy, C 3-8 Cycloalkyl or substituted cycloalkyl.

[0711] The term "substituted" should be understood to include adding or replacing one or more atoms contained in a functional group or compound with one or more different atoms.

[0712] Substituted alkyl groups include carboxyalkyl, aminoalkyl, dialkylamino, hydroxyalkyl, and mercaptoalkyl; substituted cycloalkyl groups include moieties such as 4-chlorocyclohexyl; aryl groups include moieties such as naphthyl; substituted aryl groups include moieties such as 3-bromophenyl; aralkyl groups include moieties such as toluoyl; heteroalkyl groups include moieties such as ethylthiophene; substituted heteroalkyl groups include moieties such as 3-methoxythiophene; alkoxy groups include moieties such as methoxy; and phenoxy groups include moieties such as 3-nitrophenoxy. Halogen is understood to include fluorine, chlorine, iodine, and bromine.

[0713] In certain embodiments, -L 1 - comprising a substructure of formula (VIII):

[0714]

[0715] in,

[0716] The dashed line marked with an asterisk indicates attachment to the nitrogen of -D via formation of an amide bond, where -D is a CNP moiety;

[0717] Unmarked dashed lines indicate the same 1 - the rest of the connection; and

[0718] Where -L 1 -Use -L 2 -Z replaces and where -L 1 - is optionally further substituted;

[0719] in,

[0720] -L 2 - is a single chemical bond or spacer; and

[0721] -Z is a carrier moiety, preferably a water-soluble polymeric moiety.

[0722] In certain embodiments, -L of formula (VIII) 1 -Use a part-L 2 -Z replaces.

[0723] In certain embodiments, -L of formula (VIII) 1 The other optional substituents of - are as described above.

[0724] In certain embodiments, -L of formula (VIII) 1 -Not to be further superseded.

[0725] In certain embodiments, -L 1 - comprising a substructure of formula (IX):

[0726]

[0727] in,

[0728] The dashed line marked with an asterisk indicates attachment to the nitrogen of -D via formation of an amide bond, where -D is a CNP moiety;

[0729] Unmarked dashed lines indicate the same 1 - the rest of the connection; and

[0730] Where -L 1 -Use -L 2 -Z replaces and -L 1 - is optionally further substituted;

[0731] in,

[0732] -L 2 - is a single chemical bond or spacer; and

[0733] -Z is a carrier moiety, preferably a water-soluble polymeric moiety.

[0734] -L of formula (IX) 1 The other optional substituents of - are as described above.

[0735] In certain embodiments, -L of formula (IX) 1 -Use a part-L 2 -Z replaces.

[0736] In certain embodiments, -L of formula (IX) 1 -Not to be further superseded.

[0737] The -D moiety can be linked to -L via any functional group of DH 1 -, and connected to -L through the amine functional group of DH 1If the CNP has the sequence of SEQ ID NO: 24, this may be the N-terminal amine function or an amine function provided by a lysine side chain (ie by lysine at positions 9, 11, 15, 16, 20 and 26).

[0738] Compared with the attachment at the N-terminus or the non-ring portion of the CNP, -L 1 - Linkage to the ring of the CNP moiety significantly reduces the affinity of the CNP conjugate for NPR-B, and this reduction in affinity for NPR-B in turn reduces the risk of cardiovascular side effects, such as hypotension.

[0739] Thus, in certain embodiments, -L 1 - is conjugated to the side chain of an amino acid residue of the ring portion of -D or to the main chain of the ring portion of -D. In certain embodiments, -L 1 - is covalently and reversibly conjugated to the side chain of an amino acid residue of the ring portion of -D. In certain embodiments, if -D is a CNP moiety having the sequence of SEQ ID NO: 24, then -L 1 - Conjugation to the amine function provided by lysine at position 26 of the corresponding drug DH.

[0740] Part-L 2 - is a chemical bond or a spacer moiety. In certain embodiments, -L 2 - is a chemical bond. In certain embodiments, -L 2 - is the spacer portion.

[0741] Part-L 2 - can be linked to -L by replacing any existing -H 1 -, unless expressly excluded.

[0742] When -L 2 -When it is not a single chemical bond, -L 2 - is selected from the group consisting of -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 1-50Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl; wherein -T-, C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl groups are optionally substituted with one or more identical or different -R y2 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-、-S(O)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 )-;

[0743] -R y1 and -R y1a are independently selected from the following groups: -H, -T, C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl; wherein -T, C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl groups are optionally substituted with one or more identical or different -R y2 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -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 )-;

[0744] Each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each T is independently optionally replaced by one or more identical or different -R y2 replace;

[0745] Each-R y2 Independently selected from the group consisting of 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)2R 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; among which C 1-6 Alkyl is optionally substituted with one or more halogens which may be the same or different; and each -R y3 、-R y3a 、-R y4 、-R y4a 、-R y5 、-Ry5a and -R y5b Independently selected from the group consisting of: -H and C 1-6 Alkyl, where C 1-6 The alkyl group is optionally substituted with one or more same or different halogens.

[0746] When -L 2 -When it is not a single chemical bond, -L 2 - is selected from the group consisting of -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 2-20 Alkynyl; wherein -T-, C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 Alkynyl groups are optionally substituted with one or more identical or different -R y2 substituted, and wherein C 1-20 Alkyl, C 2-20 Alkenyl and C 2-20 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-、-S(O)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 )-;

[0747] -R y1 and -R y1aare independently selected from the following groups: -H, -T, C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl; wherein -T, C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl groups are optionally substituted with one or more identical or different -R y2 substituted, and wherein C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -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 )-;

[0748] Each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each T is independently optionally replaced by one or more identical or different -R y2 replace;

[0749] -R y2 Selected from the group consisting of 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)2R y5 、-S(O)R y5 、-N(R y5 )S(O)2N(R y5a R y5b ),-SRy5 、-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; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different; and

[0750] Each-R y3 、-R y3a 、-R y4 、-R y4a 、-R y5 、-R y5a and -R y5b are independently selected from the group consisting of: -H and C 1-6 Alkyl, where C 1-6 The alkyl group is optionally substituted with one or more same or different halogens.

[0751] When -L 2 -When it is not a single chemical bond, -L 2 - is selected from the group consisting of -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; wherein -T-, C 1-50Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl groups are optionally substituted with one or more identical or different -R y2 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-、-S(O)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 )-;

[0752] -R y1 and -R y1a Independently selected from the group consisting of -H, -T, C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl;

[0753] Each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbon polycyclyl and 8- to 30-membered heteropolycyclyl; each -R y2 Independently selected from the group consisting of halogen and C 1-6 alkyl; and each -R y3 、-R y3a 、-R y4 、-R y4a 、-R y5 、-R y5a and -R y5b are independently selected from the group consisting of: -H and C 1-6 Alkyl, where C 1-6 The alkyl group is optionally substituted with one or more same or different halogens.

[0754] In certain embodiments, -L 2 -It's C 1-20 The alkyl chain is optionally interrupted by one or more groups independently selected from the group consisting of -O-, -T-, and -C(O)N(Ry1 )-; and the C 1-20 The alkyl chain is optionally substituted with one or more groups independently selected from the group consisting of: -OH, -T, and -C(O)N(R y6 R y6a ); where -R y1 、-R y6 、-R y6a Independently selected from the group consisting of H and C 1-4 Alkyl, and wherein T is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclic group, 8- to 11-membered heterobicyclic group, 8- to 30-membered carbopolycyclic group and 8- to 30-membered heteropolycyclic group.

[0755] In certain embodiments, -L 2 - having a molecular weight of 14 to 750 g / mol.

[0756] In certain embodiments, -L 2 - has a chain length of 1 to 20 atoms.

[0757] As used herein, with respect to part-L 2 -, the term "chain length" refers to the 1 -L is the shortest connection between -Z and -L 2 -The atomic number of .

[0758] In certain embodiments, -L 2 -is formula (i):

[0759]

[0760] in,

[0761] The dashed line marked with an asterisk indicates the connection to -L 1 -;

[0762] Unlabeled dashed lines indicate connections to -Z;

[0763] -R 1 Selected from the group consisting of: -H, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl;

[0764] n is selected from the following group: 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,

[0765] 15, 16, 17 and 18; and

[0766] wherein the moiety of formula (i) is optionally further substituted.

[0767] In certain embodiments, -R 1 is selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. 1 is selected from the group consisting of -H, methyl, ethyl, and propyl. In certain embodiments, -R 1 In certain embodiments, -R 1 It's methyl.

[0768] 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.

[0769] In certain embodiments, -L 2 - is a portion selected from the group consisting of:

[0770]

[0771]

[0772] in,

[0773] The dashed line marked with an asterisk indicates the connection to -L 1 -;

[0774] The unlabeled dashed line indicates connection to -Z; and

[0775] wherein moieties (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi), (xii), (xiii), (xiv), (xv), (xvi) and (xvii) are optionally further substituted.

[0776] In certain embodiments, -L 2 -Selected from the group consisting of:

[0777]

[0778] in,

[0779] The dashed line marked with an asterisk indicates the connection to -L 1 -;and

[0780] Unlabeled dashed lines indicate connections to -Z.

[0781] In certain embodiments, -L 2 -Selected from the group consisting of:

[0782]

[0783] in,

[0784] The dashed line marked with an asterisk indicates the connection to -L 1 -;and

[0785] Unlabeled dashed lines indicate connections to -Z.

[0786] In certain embodiments, -L 2 -is formula (xvi):

[0787]

[0788] in,

[0789] The dashed line marked with an asterisk indicates the connection to -L 1 -;and

[0790] Unlabeled dashed lines indicate connections to -Z.

[0791] In certain embodiments, the moiety -L 1 -L 2 -Selected from the group consisting of:

[0792]

[0793] in,

[0794] The unlabeled dashed line indicates attachment to the nitrogen of -D via formation of an amide bond, wherein -D is a CNP moiety; and

[0795] The dashed line marked with an asterisk indicates the connection to -Z.

[0796] In certain embodiments, the moiety -L 1 -L 2 -is the formula (IId-ii):

[0797]

[0798] in,

[0799] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0800] and

[0801] The dashed line marked with an asterisk indicates the connection to -Z.

[0802] In certain embodiments, the moiety -L1 -L 2 -is the formula (IId-ii'):

[0803]

[0804] in,

[0805] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0806] and

[0807] The dashed line marked with an asterisk indicates the connection to -Z.

[0808] In certain embodiments, the moiety -L 1 -L 2 -Selected from the group consisting of:

[0809]

[0810] in,

[0811] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0812] and

[0813] The dashed line marked with an asterisk indicates the connection to -Z.

[0814] In certain embodiments, the moiety -L 1 -L 2 -is the formula (IId-iia):

[0815]

[0816] in,

[0817] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0818] and

[0819] The dashed line marked with an asterisk indicates the connection to -Z.

[0820] In certain embodiments, the moiety -L 1 -L 2 -is the formula (IId-iia'):

[0821]

[0822] in,

[0823] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0824] and

[0825] The dashed line marked with an asterisk indicates the connection to -Z.

[0826] In certain embodiments, the moiety -L 1 -L 2 -Selected from the group consisting of:

[0827]

[0828] in,

[0829] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0830] and

[0831] The dashed line marked with an asterisk indicates the connection to -Z.

[0832] In certain embodiments, the moiety -L 1 -L 2 -is the formula (IId-iib):

[0833]

[0834] in,

[0835] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0836] and

[0837] The dashed line marked with an asterisk indicates the connection to -Z.

[0838] In certain embodiments, the moiety -L 1 -L 2 -is the formula (IId-iib'):

[0839]

[0840] in,

[0841] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[0842] and

[0843] The dashed line marked with an asterisk indicates the connection to -Z.

[0844] In certain embodiments, -Z of formula (Ia) or (Ib) has a molecular weight of 5 to 200 kDa. In certain embodiments, -Z of formula (Ia) or (Ib) has a molecular weight of 8 to 100 kDa. In certain embodiments, -Z of formula (Ia) or (Ib) has a molecular weight of 10 to 80 kDa. In certain embodiments, -Z of formula (Ia) or (Ib) has a molecular weight of 12 to 60 kDa. In certain embodiments, -Z of formula (Ia) or (Ib) has a molecular weight 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.

[0845] The polymeric moiety -Z of formula (Ia) or (Ib) comprises a polymer. In certain embodiments, -Z of formula (Ia) or (Ib) comprises a polymer selected from the group consisting of poly(2-methacryloyl-ethoxyphosphorylcholine), poly(acrylic acid), poly(acrylate), poly(acrylamide), poly(alkoxy) polymers, poly(amide), poly(amidoamine), poly(amino acid), poly(anhydride), poly(asparagine), poly(butyric acid), poly(glycolic acid), polybutylene terephthalate, poly(caprolactone), poly(carbonate), poly(cyanoacrylate), poly(dimethylacrylamide), poly(ester), poly(ethylene), poly(ethylene glycol), poly(ethylene oxide), poly(ethyl phosphate), poly(ethyl oxazoline), poly(glycolic acid), poly(hydroxyethyl acrylate), poly(hydroxyethyl-oxazoline), poly(hydroxymethyl acrylate), poly(hydroxypropyl methacrylamide), poly(hydroxypropyl methacrylate),

[0014] Examples of the present invention include, but are not limited to, oligosaccharides, oligosaccharides, oligosaccharides (e.g., oligosaccharides), ...

[0846] In certain embodiments, -Z of Formula (Ia) or (Ib) comprises a protein. Preferred proteins are selected from the group consisting of: the carboxyl-terminal peptide of chorionic gonadotropin as described in US2012 / 0035101 A1, which is incorporated herein by reference; albumin; XTEN as described in WO 2011123813 A2 TM sequences, which are incorporated herein by reference; proline / alanine random coil sequences as described in WO 2011 / 144756 A1, which are incorporated herein by reference; proline / alanine / serine random coil sequences as described in WO 2008 / 155134 A1 and WO 2013 / 024049 A1, which are incorporated herein by reference; and Fc-fusion proteins.

[0847] 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 portion. In certain embodiments, -Z of formula (Ia) or (Ib) comprises one random coil protein portion. In certain embodiments, -Z of formula (Ia) or (Ib) comprises two random coil protein portions. In certain embodiments, -Z of formula (Ia) or (Ib) comprises three random coil protein portions. In certain embodiments, -Z of formula (Ia) or (Ib) comprises four random coil protein portions. In certain embodiments, -Z of formula (Ia) or (Ib) comprises five random coil protein portions. In certain embodiments, -Z of formula (Ia) or (Ib) comprises six random coil protein portions. In certain embodiments, -Z of formula (Ia) or (Ib) comprises seven random coil protein moieties. In certain embodiments, -Z of formula (Ia) or (Ib) comprises eight random coil protein moieties.

[0848] In certain embodiments, such random coil protein portion comprises at least 25 amino acid residues and at most 2000 amino acids. In certain embodiments, such random coil protein portion comprises at least 30 amino acid residues and at most 1500 amino acid residues. In certain embodiments, such random coil protein portion comprises at least 50 amino acid residues and at most 500 amino acid residues.

[0849] In certain embodiments, -Z of formula (Ia) or (Ib) comprises a fatty acid derivative. In certain embodiments, -Z of formula (Ia) or (Ib) is a fatty acid derivative. In certain embodiments, -Z of formula (Ia) is a fatty acid derivative, and x is 1.

[0850] In certain embodiments, -Z of formula (Ia) or (Ib) is a fatty acid derivative as disclosed in WO 2006 / 097537 A2, which is incorporated herein by reference.

[0851] In certain embodiments, -Z of formula (Ia) or (Ib) comprises a fatty acid derivative as disclosed in WO 2021 / 055497 A1, which is incorporated herein by reference. Thus, in certain embodiments, -Z of formula (Ia) or (Ib) has the following structure (w):

[0852]

[0853] The dotted line represents the -L of formula (Ia) or (Ib). 2 -or-L 1 -connect.

[0854] In certain embodiments, -Z is of formula (w), and -L 1 - is formula (V).

[0855] In certain embodiments, -ZL 2 -L 1 - is the formula (wa):

[0856]

[0857] The dotted line represents the connection with -D of formula (Ia) or (Ib).

[0858] In certain embodiments, the CNP has a sequence selected from the group consisting of:

[0859] PGQEHPQARRYRGAQRRGLSRGCFGLKLDRIGSMSGLGC (SEQ ID NO:98);

[0860] PGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC(SEQ ID NO:30);

[0861] PGQEHPNARRYRGANRRGLSRGCFGLKLDRIGSMSGLGC (SEQ ID NO:99); and

[0862] PGQEHPQARKYKGAQKKGLSKGCFGLKLDRIGSMSGLGC (SEQ ID NO: 100).

[0863] In certain embodiments, the CNP has a sequence selected from the group consisting of SEQ ID NO: 98, SEQ ID NO: 30, SEQ ID NO: 99, and SEQ ID NO: 100, Z is of formula (w) and -L 1 - is a reversible linker moiety. In certain embodiments, CNP has a sequence selected from the group consisting of SEQ ID NO: 98, SEQ ID NO: 30, SEQ ID NO: 99, and SEQ ID NO: 100, Z is of formula (w) and -L 1 - is formula (v). In certain embodiments, the CNP has a sequence selected from the group consisting of SEQ ID NO: 98, SEQ ID NO: 30, SEQ ID NO: 99, and SEQ ID NO: 100, -ZL 2 -L 1 - is the formula (wa). 1 - may be linked to the CNP via a lysine other than the lysine within the ring structure, or may be linked to the N-terminus.

[0864] In certain embodiments, the CNPs of SEQ ID NO: 98, SEQ ID NO: 30, SEQ ID NO: 99 and SEQ ID NO: 100 further comprise an acetyl group, such as an acetyl group at the N-terminus of the peptide. In certain embodiments, the CNPs of SEQ ID NO: 98, SEQ ID NO: 30, SEQ ID NO: 99 and SEQ ID NO: 100 further comprise an -OH or -NH2 group at the C-terminus. In certain embodiments, the CNPs of SEQ ID NO: 98, SEQ ID NO: 30, SEQ ID NO: 99 and SEQ ID NO: 100 and -L 1 - attached to a residue in the CNP loop portion or to a site outside the CNP portion.

[0865] In certain embodiments, -L 1 - linked to a lysine residue, such as the lysine residues indicated in bold in SEQ ID NO: 98, SEQ ID NO: 30, SEQ ID NO: 99 and SEQ ID NO: 100:

[0866] PGQEHPQARRYRGAQRRGLSRGCFGLKLDRIGSMSGLGC (SEQ ID NO:98);

[0867] PGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC(SEQ ID NO:30);

[0868] PGQEHPNARRYRGANRRGLSRGCFGLKLDRIGSMSGLGC (SEQ ID NO:99); and

[0869] PGQEHPQARKYKGAQKKGLSKGCFGLKLDRIGSMSGLGC (SEQ ID NO: 100).

[0870] In certain embodiments, the CNP is selected from the group consisting of:

[0871] Ac-PGQEHPQARRYRGAQRRGLSRGCFGLKLDRIGSMSGLGC (SEQ ID NO: 101);

[0872] Ac-PGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC-NH2 (SEQ ID NO: 102);

[0873] Ac-PGQEHPNARRYRGANRRGLSRGCFGLKLDRIGSMSGLGC (SEQ ID NO: 103);

[0874] Ac-PGQEHPNARRYRGANRRGLSRGCFGLKLDRIGSMSGLGC-NH2 (SEQ ID NO: 104); and

[0875] Ac-PGQEHPQARRYRGAQRRGLSRGCFGLKLDRIGSMSGLGC-NH2 (SEQ ID NO: 105).

[0876] In certain embodiments, -Z of formula (Ia) or (Ib) is a hyaluronic acid-based polymer. In certain embodiments, -Z of formula (Ia) or (Ib) is a polymeric moiety as disclosed in WO 2013 / 024047 A1, which is incorporated herein by reference.

[0877] In certain embodiments, -Z of Formula (Ia) or (Ib) is a polymeric moiety as disclosed in WO 2013 / 024048 A1, which is incorporated herein by reference.

[0878] In certain embodiments, -Z of Formula (Ia) or (Ib) is a PEG-based polymer. In certain embodiments, -Z is a branched or multi-armed PEG-based polymer.

[0879] In certain embodiments,-Z of formula (Ia) or (Ib) is a branched polymer. In certain embodiments,-Z of formula (Ia) or (Ib) is a branched polymer with one, two, three, four, five or six branch points. In certain embodiments,-Z of formula (Ia) or (Ib) is a branched polymer with one, two or three branch points. In certain embodiments,-Z of formula (Ia) or (Ib) is a branched polymer with one branch point. In certain embodiments,-Z of formula (Ia) or (Ib) is a branched polymer with two branch points. In certain embodiments,-Z of formula (Ia) or (Ib) is a branched polymer with three branch points.

[0880] In certain embodiments, the branch point is selected from the group consisting of: -N<, -CH<, and >C<.

[0881] In certain embodiments, such branched moiety -Z of Formula (Ia) or (Ib) is PEG-based.

[0882] In certain embodiments, such side chain moiety-Z of formula (Ia) or (Ib) has a molecular weight of 5kDa to 500kDa, and includes 5kDa and 500kDa. In certain embodiments, such side chain moiety-Z of formula (Ia) or (Ib) has a molecular weight of 10kDa to 250kDa, and includes 10kDa and 250kDa. In certain embodiments, such side chain moiety-Z of formula (Ia) or (Ib) has a molecular weight of 10kDa to 150kDa, and includes 10kDa and 150kDa. In certain embodiments, such side chain moiety-Z of formula (Ia) or (Ib) has a molecular weight of 12kDa to 100kDa, and includes 12kDa and 100kDa. In certain embodiments, such side chain moiety-Z of formula (Ia) or (Ib) has a molecular weight of 15kDa to 80kDa, and includes 15kDa and 80kDa. In certain embodiments, such branched chain portion-Z of formula (Ia) or (Ib) has a molecular weight of 10kDa to 80kDa, including 10kDa and 80kDa. In certain embodiments, the molecular weight is about 10kDa. In certain embodiments, the molecular weight of such branched chain portion-Z of formula (Ia) or (Ib) is about 20kDa. In certain embodiments, the molecular weight of such branched chain portion-Z of formula (Ia) or (Ib) is about 30kDa. In certain embodiments, the molecular weight of such branched chain portion-Z of formula (Ia) or (Ib) is about 40kDa. In certain embodiments, the molecular weight of such branched chain portion-Z of formula (Ia) or (Ib) is about 50kDa. In certain embodiments, the molecular weight of such branched chain portion-Z of formula (Ia) or (Ib) is about 60kDa. In certain embodiments, the molecular weight of such branched chain portion-Z of formula (Ia) or (Ib) is about 70kDa. In certain embodiments, the molecular weight of such branched moiety-Z of formula (Ia) or (Ib) is about 80 kDa. In certain embodiments, the molecular weight of such branched moiety-Z of formula (Ia) or (Ib) is about 40 kDa.

[0883] In certain embodiments, -Z comprises the moiety:

[0884]

[0885] In certain embodiments, -Z comprises an amide bond.

[0886] In certain embodiments, -Z of formula (Ia) or (Ib) comprises a moiety of formula (a)

[0887]

[0888] in,

[0889] The dotted line indicates the difference with -L 2 - or - the rest of the Z connection;

[0890] BP a is a branch point selected from the group consisting of: -N<, -CR<, and >C<;

[0891] -R is selected from the group consisting of -H and C 1-6 alkyl;

[0892] If BP a If -N< or -CR<, then a is 0; and if BP a is >C<, then a is 1;

[0893] -S a -、-S a’ -、-S a” -and-S a”’ - are independently a chemical bond or selected from the group consisting of: C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl; wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl groups are optionally substituted with one or more identical or different -R 1 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 )-、-S(O)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 )-;

[0894] Each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbon polycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally replaced by one or more identical or different -R 1 replace;

[0895] Each-R 1 Independently selected from the group consisting of 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)2R 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;

[0896] Among them C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[0897] Each-R 2 、-R 2a 、-R 3 、-R 3a and -R 3b Independently selected from the group consisting of: -H and C 1-6 Alkyl, where C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different; and

[0898] -P a’ 、-P a” and -P a”’ are independently polymeric moieties.

[0899] Optionally, the moiety of formula (a) is substituted with one or more substituents.

[0900] In certain embodiments, BP of formula (a) a In certain embodiments, BP of formula (a) a In certain embodiments, -R is -H. Thus, in certain embodiments, a of formula (a) is 0. In certain embodiments, BP of formula (a) is a is >C<.

[0901] In certain embodiments, -S of formula (a) a - is a chemical bond.

[0902] In certain embodiments, -S of formula (a) a -Selected from the following group: C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl, where C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 The alkynyl group is optionally interrupted by one or more chemical groups selected from the group consisting of: -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 )-; where -R 4 and -R 4a In certain embodiments, -S of formula (a) is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. a - is selected from the group consisting of methyl, ethyl, propyl, butyl, optionally interrupted by one or more chemical groups selected from the group consisting of -O-, -C(O)- and -C(O)N(R 4 )-.

[0903] In certain embodiments, -S of formula (a) a’ - is a chemical bond.

[0904] In certain embodiments, -S of formula (a) a’-Selected from the following group: C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl, where C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 The alkynyl group is optionally interrupted by one or more chemical groups selected from the group consisting of: -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 )-; where -R 4 and -R 4a In certain embodiments, -S of formula (a) is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. a’ - is selected from the group consisting of methyl, ethyl, propyl, butyl, optionally interrupted by one or more chemical groups selected from the group consisting of -O-, -C(O)- and -C(O)N(R 4 )-.

[0905] In certain embodiments, -S of formula (a) a” - is a chemical bond.

[0906] In certain embodiments, -S of formula (a) a” -Selected from the following group: C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl, where C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 The alkynyl group is optionally interrupted by one or more chemical groups selected from the group consisting of: -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(OR4 )(R 4a )-、-N(R 4 )C(O)N(R 4a )-and-OC(O)N(R 4 )-; where -R 4 and -R 4a In certain embodiments, -S of formula (a) is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. a” - is selected from the group consisting of methyl, ethyl, propyl, butyl, optionally interrupted by one or more chemical groups selected from the group consisting of -O-, -C(O)- and -C(O)N(R 4 )-.

[0907] In certain embodiments, -S of formula (a) a”’ - is a chemical bond.

[0908] In certain embodiments, -S of formula (a) a”’ -Selected from the following group: C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl, where C 1-10 Alkyl, C 2-10 Alkenyl and C 2-10 The alkynyl group is optionally interrupted by one or more chemical groups selected from the group consisting of: -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 )-; where -R 4 and -R 4a In certain embodiments, -S of formula (a) is independently selected from the group consisting of -H, methyl, ethyl, propyl, and butyl. a”’ - is selected from the group consisting of methyl, ethyl, propyl, butyl, optionally interrupted by one or more chemical groups selected from the group consisting of -O-, -C(O)- and -C(O)N(R 4 )-.

[0909] In certain embodiments, -P of formula (a) a’ 、-Pa” and -P a”’ independently comprising a polymer selected from the group consisting of poly(2-methacryloyl-ethoxyphosphorylcholine), poly(acrylic acid), poly(acrylate), poly(acrylamide), poly(alkoxy) polymers, poly(amide), poly(amidoamine), poly(amino acid), poly(anhydride), poly(asparagine), poly(butyric acid), poly(glycolic acid), polybutylene terephthalate, poly(caprolactone), poly(carbonate), poly(cyanoacrylate), poly(dimethylacrylamide), poly(ester), poly(ethylene), poly(ethylene glycol), poly(ethylene oxide), poly(ethyl phosphate), poly(ethyl oxazoline), poly(glycolic acid), poly(hydroxyethyl acrylate), poly(hydroxyethyl-oxazoline), poly(hydroxymethyl acrylate), poly(hydroxypropyl methacrylamide), poly(hydroxypropyl methacrylate), poly(hydroxypropyl oxazoline),

[0014] The present invention also includes but is not limited to the following: polyols, emulsifiers, and oligomers ...

[0910] In certain embodiments, -P of formula (a) a’ 、-P a” and -P a”’ independently having a molecular weight of 5 kDa to 50 kDa, inclusive of 5 kDa and 500 kDa; in certain embodiments, having a molecular weight of 5 kDa to 40 kDa, inclusive of 5 kDa and 40 kDa; in certain embodiments, having a molecular weight of 7.5 kDa to 35 kDa, inclusive of 7.5 kDa and 35 kDa; in certain embodiments, having a molecular weight of 7.5 to 30 kDa, inclusive of 7.5 and 30 kDa; in certain embodiments, having a molecular weight of 10 to 30 kDa, inclusive of 10 and 30 kDa.

[0911] In certain embodiments, -P of formula (a) a’ 、-P a” and -P a”’ Has a molecular weight of about 5 kDa. In certain embodiments, -P of formula (a) 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”’ It has a molecular weight of approximately 20 kDa.

[0912] 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 comprising a PEG-based portion comprising at least 20% PEG, in certain embodiments at least 30% PEG, 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, in certain embodiments at least 90% PEG.

[0913] In certain embodiments, -P of formula (a) a’ 、-P a” and -P a”’ independently comprises a protein portion; in certain embodiments comprises a random coil protein portion; and in certain embodiments comprises a random coil protein portion selected from the group consisting of PA, PAS, PAG, PG, and XTEN TM part.

[0914] 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”’ 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”’ In certain embodiments, -P of formula (a) a’ 、-P a” and -P a”’ It is XTEN TM part.

[0915] In certain embodiments, -Z comprises one moiety of formula (a). In certain embodiments, -Z comprises two moieties of formula (a). In another embodiment, -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).

[0916] In certain embodiments, -Z comprises a moiety of formula (b):

[0917]

[0918] in,

[0919] The dotted line indicates the difference with -L 2 - or - the rest of the Z connection;

[0920] b1 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8;

[0921] b2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8;

[0922] b3 is an integer from 150 to 1000, inclusive; in certain embodiments, ranges from 150 to 500, inclusive; and in certain embodiments, ranges from 200 to 460, inclusive; and

[0923] b4 is an integer from 150 to 1000, inclusive; in certain embodiments, ranges from 150 to 500, inclusive; and in certain embodiments, ranges from 200 to 460, inclusive.

[0924] Optionally, the moiety of formula (b) is substituted with one or more substituents.

[0925] In certain embodiments, b3 and b4 of formula (b) are the same integer. In certain embodiments, b3 and b4 of formula (b) are both integers from 200 to 250, and in certain embodiments, b3 and b4 of formula (b) are about 225. In certain embodiments, b3 and b4 of formula (b) are both integers from 400 to 500, and in certain embodiments, b3 and b4 of formula (b) are about 450.

[0926] In certain embodiments, b1 in formula (b) is selected from the group consisting of 0, 1, 2, 3, and 4. In certain embodiments, b1 in formula (b) is selected from the group consisting of 1, 2, and 3. In certain embodiments, b1 in formula (b) is 2.

[0927] 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.

[0928] 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.

[0929] 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).

[0930] In certain embodiments, -Z comprises a moiety of formula (c):

[0931]

[0932] in,

[0933] The dotted line indicates the difference with -L 2 - or - the rest of the Z connection;

[0934] c1 and c2 are independently integers from 150 to 500, inclusive; in certain embodiments,

[0935] The integers are 200 to 460, inclusive.

[0936] Optionally, the moiety of formula (c) is substituted with one or more substituents.

[0937] In certain embodiments, c1 and c2 in formula (c) are both the same integer.

[0938] In certain embodiments, c1 and c2 of formula (c) range from 200 to 250, inclusive, and in certain embodiments, are about 225. In certain embodiments, c1 and c2 of formula (c) range from 400 to 500, inclusive, and in certain embodiments, are about 450.

[0939] In certain embodiments, the moiety -Z is a branched PEG-based polymer comprising at least 10% PEG, having one branch point and two PEG-based polymer arms, and having a molecular weight of about 40 kDa. Thus, each of the two PEG-based polymer arms has a molecular weight of about 20 kDa. In certain embodiments, the branch point is -CH<.

[0940] 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).

[0941] In certain embodiments, the moiety -Z is of formula (d):

[0942]

[0943] in,

[0944] The dotted line indicates the connection to -L 2 -;

[0945] -Z b -Selected from the following group: C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl; wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl groups are optionally substituted with one or more identical or different -R 1 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 )-、-S(O)2N(R 2 )-、-S(O)N(R 2)-、-S(O)2-、

[0946] -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 )-;

[0947] Each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbon polycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally replaced by one or more identical or different -R 1 replace;

[0948] Each-R 1 Independently selected from the group consisting of 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)2R 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(R3 R 3a ) and C 1-6 Alkyl; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[0949] Each-R 2 、-R 2a 、-R 3 、-R 3a and -R 3b Independently selected from the group consisting of: -H and C 1-6 Alkyl, where C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[0950] And -Z a for

[0951]

[0952] in,

[0953] BP a 、-S a -、-S a’ -、-S a” -、-S a”’ -、-P a’ 、-P a” 、-P a”’ and a are used as defined in formula (a).

[0954] Optionally, the moiety of formula (d) is substituted with one or more substituents.

[0955] 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).

[0956] In certain embodiments, -Z of formula (d) a In certain embodiments, b1, b2, b3 and b4 are as described in formula (b).

[0957] In certain embodiments, the moiety -Z of formula (Ia) or (Ib) is of formula (e):

[0958]

[0959] in,

[0960] The dotted line indicates the connection to -L 2 -;

[0961] e is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, and 15;

[0962] and

[0963] -Z a for

[0964]

[0965] in,

[0966] b1, b2, b3 and b4 are used as defined in formula (b).

[0967] Optionally, the moiety of formula (e) is substituted with one or more substituents.

[0968] In certain embodiments, b1, b2, b3, and b4 for formula (e) are as defined above for formula (b).

[0969] In certain embodiments, e of formula (e) is 1. In certain embodiments, e of formula (e) is 2. In certain embodiments, e of formula (e) is 3. In certain embodiments, e of formula (e) is 4. In certain embodiments, e of formula (e) is 5. In certain embodiments, e of formula (e) is 6. In certain embodiments, e of formula (e) is 7. In certain embodiments, e of formula (e) is 8. In certain embodiments, e of formula (e) is 9. In certain embodiments, e of formula (e) is 10. In certain embodiments, e of formula (e) is 11. In certain embodiments, e of formula (e) is 12. In certain embodiments, e of formula (e) is 13. In certain embodiments, e of formula (e) is 14. In certain embodiments, e of formula (e) is 15.

[0970] In certain embodiments, e of formula (e) is selected from the group consisting of 2, 3, 4, 5, 6, 7, 8, and 9. In certain embodiments, e of formula (e) is selected from the group consisting of 3, 4, 5, and 6. In certain embodiments, e in formula (e) is 5.

[0971] In certain embodiments, e of formula (e) is 5, b1 of formula (e) is 2, b2 of formula (e) is 3, and b3 and b4 of formula (e) are both about 450.

[0972] In certain embodiments, the moiety -Z of formula (Ia) or (Ib) is of formula (ei) or (e-i'):

[0973]

[0974] in,

[0975] The dotted line indicates the connection to -L 2 -;

[0976] e is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, and 15;

[0977] and

[0978] -Z a for

[0979]

[0980] in,

[0981] b1, b2, b3 and b4 are used as defined in formula (b).

[0982] In certain embodiments, b1, b2, b3, and b4 for formula (ei) and (ei') are as defined above for formula (b).

[0983] In certain embodiments, e of formula (ei) and (e-i') is as described for formula (e).

[0984] In certain embodiments, b1 of formula (ei) and (e-i') is 2, b2 of formula (ei) and (e-i') is 3, and b3 and b4 of formula (ei) and (e-i') are both about 450.

[0985] In certain embodiments, -Z in formula (Ia) or (Ib) is of formula (ei).

[0986] In certain embodiments, the moiety -Z is a PEG-based polymer comprising at least 10% PEG branches, having three branch points and four PEG-based polymer arms, and having 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<.

[0987] In certain embodiments, the moiety -Z is of formula (f):

[0988]

[0989] in,

[0990] The dotted line indicates the connection to -L 2 -;

[0991] BP fis a branch point selected from the group consisting of: -N<, -CR<, and >C<;

[0992] -R is selected from the group consisting of -H and C 1-6 alkyl;

[0993] If BP f If -N< or -CR<, then f is 0; and if BP f If >C<, then f is 1; -S f -、-S f’ -、-S f” -and-S f”’ - is independently a chemical bond or is independently selected from the group consisting of: C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl; wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl groups are optionally substituted with one or more identical or different -R 1 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 )-、-S(O)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 )-;

[0994] Each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbon polycyclyl and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally replaced by one or more identical or different -R 1 replace;

[0995] Each-R 1 Independently selected from the group consisting of 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)2R 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; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[0996] Each-R 2 、-R 2a 、-R 3 、-R 3a and -R 3b Independently selected from the group consisting of: -H and C 1-6 Alkyl, where C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[0997] And -Z a’ 、-Z a” and -Z a”’ Independently

[0998]

[0999] in,

[1000] BP a 、-S a -、-S a’ -、-Sa” -、-S a”’ -、-P a’ 、-P a” 、-P a”’ and a are used as defined in formula (a).

[1001] Optionally, the moiety of formula (f) is substituted with one or more substituents.

[1002] 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).

[1003] In certain embodiments, BP of formula (f) f is -CR<, and r is 0. In certain embodiments, -R is -H.

[1004] In certain embodiments, -S of formula (f) f - is a chemical bond.

[1005] In certain embodiments, -Z of formula (f) a’ 、-Z a” and -Z a”’ In certain embodiments, -Z of formula (f) a’ 、-Z a” and -Z a”’ It is formula (b).

[1006] In certain embodiments, b1, b2, b3, and b4 are as described in formula (b).

[1007] In certain embodiments, -S of formula (f) f - is a chemical bond, BP of formula (f) a In certain embodiments, -S of formula (f) is -CR<, wherein -R is -H. f - is a chemical bond, BP of formula (f) a is -CR<, -R is -H, and -Z of formula (f) a’ 、-Z a” and -Z a”’ It is formula (b).

[1008] In certain embodiments, -Z is of formula (g):

[1009]

[1010] in,

[1011] The dotted line indicates the connection to -L 2 -;

[1012] -S g -、-S g’ -and-S g” - independently selected from the group consisting of: C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl; wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl groups are optionally substituted with one or more identical or different -R 1 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 )-、-S(O)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 )-;

[1013] Each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbon polycyclyl and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally replaced by one or more identical or different -R 1 replace;

[1014] Each-R 1 Independently selected from the group consisting of 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)2R 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; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[1015] Each-R 2 、-R 2a 、-R 3 、-R 3a and -R 3b Independently selected from the group consisting of: -H and C 1-6 Alkyl, where C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[1016] And -Z a and -Z a’ Independently

[1017]

[1018] in,

[1019] 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).

[1020] Optionally, the moiety of formula (g) is substituted with one or more substituents.

[1021] 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).

[1022] In certain embodiments, -S of formula (g) g -Selected from the following group: C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl, optionally substituted with one or more identical or different -R 1 replace;

[1023] Where -R 1 Independently selected from the group consisting of 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)2R 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 R3a ) and C 1-6 Alkyl; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different; and

[1024] -R 3 、-R 3a and -R 3b Independently selected from the group consisting of -H, methyl, ethyl, propyl and butyl.

[1025] In certain embodiments, -S of formula (g) g -Selected from C 1-6 alkyl.

[1026] In certain embodiments, -S in formula (g) g’ -Selected from the following group: C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl, optionally substituted with one or more identical or different -R 1 replace,

[1027] Where -R 1 Selected from the group consisting of 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)2R 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(R3 R 3a ) and C 1-6 alkyl;

[1028] Among them C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different; and

[1029] -R 3 、-R 3a and -R 3b Independently selected from the group consisting of -H, methyl, ethyl, propyl and butyl.

[1030] In certain embodiments, -S of formula (g) g’ -Selected from C 1-6 alkyl.

[1031] In certain embodiments, -S in formula (g) g” -Selected from the following group: C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl, optionally substituted with one or more identical or different -R 1 replace,

[1032] Where -R 1 Selected from the group consisting of 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)2R 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;

[1033] Among them C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different; and

[1034] -R 3 、-R 3a and -R 3b Independently selected from the group consisting of -H, methyl, ethyl, propyl and butyl.

[1035] In certain embodiments, -S of formula (g) g” -Selected from C 1-6 alkyl.

[1036] In certain embodiments, -Z of formula (g) a and -Z a’ In certain embodiments, -Z of formula (g) a and -Z a’ It is formula (b).

[1037] In certain embodiments, -Z of formula (Ia) or (Ib) is of formula (gi):

[1038]

[1039] in,

[1040] The dotted line indicates the connection to -L 2 -;

[1041] -S g -、-S g’ -and-S g” - independently selected from the group consisting of: C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl; wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 Alkynyl groups are optionally substituted with one or more identical or different -R 1 substituted, and wherein C 1-50 Alkyl, C 2-50 Alkenyl and C 2-50 The alkynyl group is optionally interrupted by one or more groups selected from the group consisting of: -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 2 )-、-S(O)2N(R 2 )-、-S(O)N(R2 )-、-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 )-;

[1042] Each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbon polycyclyl and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally replaced by one or more identical or different -R 1 replace;

[1043] Each-R 1 Independently selected from the group consisting of 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)2R 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 R3a ) and C 1-6 Alkyl; among which C 1-6 The alkyl group is optionally substituted with one or more halogen groups which may be the same or different;

[1044] Each-R 2 、-R 2a 、-R 3 、-R 3a and -R 3b Independently selected from the group consisting of: -H and C 1-6 Alkyl, where C 1-6 Alkyl is optionally substituted with one or more halogens which may be the same or different; -Y a1 -and-Y a1‘ for

[1045]

[1046] And -Z a and -Z a’ Independently

[1047]

[1048] in,

[1049] 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).

[1050] Optionally, the moiety of formula (gi) is substituted with one or more substituents.

[1051] In certain embodiments, -Y of formula (gi) a1 -and-Y a1‘ Both The dashed lines marked with asterisks represent connections to -Z a or -Z a’ .

[1052] 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).

[1053] In certain embodiments, -S of formula (gi) g -、-S g’ -and-S g” - as defined for formula (g).

[1054] In certain embodiments, -Z of formula (gi) a and -Z a’ In certain embodiments, -Z of formula (gi) a and -Z a’ In certain embodiments, b1, b2, b3, and b4 are as described for formula (b).

[1055] In certain embodiments, -Z is of formula (h):

[1056]

[1057] in,

[1058] The dotted line indicates the connection to -L 2 -;and

[1059] Each-Z c For part

[1060]

[1061] in,

[1062] Each c1 is independently an integer from about 200 to 250;

[1063] Optionally, the moiety of formula (h) is substituted with one or more substituents.

[1064] In certain embodiments, both c1 of formula (h) are the same. In certain embodiments, both c1 of formula (h) are about 225.

[1065] In certain embodiments, -Z of formula (Ia) or (Ib) is of formula (ha):

[1066]

[1067] in,

[1068] The dotted line indicates the connection to -L 2 -;

[1069] Each k is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12;

[1070] -Y a1 -and-Ya1‘ -for

[1071]

[1072] and

[1073] Each-Z c For part

[1074]

[1075] in,

[1076] Each c1 is independently an integer from about 200 to 250.

[1077] Optionally, the moiety of formula (ha) is substituted with one or more substituents.

[1078] In certain embodiments, each k of Formula (ha) is independently selected from the group consisting of 2, 3, 4, 5, 6, and 7. In certain embodiments, two k of Formula (ha) are the same.

[1079] In certain embodiments, two c1 of formula (ha) are the same.

[1080] In certain embodiments, both c1 of formula (ha) are about 225.

[1081] In certain embodiments, -Y of formula (ha) a1 -and-Y a1‘ Both The dashed line marked with an asterisk indicates the connection to -Z c .

[1082] In certain embodiments, the moiety -Z is of formula (hi):

[1083]

[1084] in,

[1085] The dotted line indicates the connection to -L 2 -;and

[1086] Each-Z c For part

[1087]

[1088] wherein each c1 is independently an integer from 200 to 250.

[1089] Optionally, the moiety of formula (hi) is substituted with one or more substituents.

[1090] In certain embodiments, both c1 of formula (hi) are the same. In certain embodiments, both c1 of formula (hi) are about 225.

[1091] In certain embodiments, the moiety -Z of formula (Ia) or (Ib) is of formula (h-ia):

[1092]

[1093] in,

[1094] The dotted line indicates the connection to -L 2 -;

[1095] Each k is independently selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12;

[1096] -Y a1 -and-Y a1‘ -for

[1097]

[1098] and

[1099] Each-Z c For part

[1100]

[1101] wherein each c1 is independently an integer from 200 to 250.

[1102] In certain embodiments, each k of Formula (h-ia) is independently selected from the group consisting of 2, 3, 4, 5, 6, and 7. In certain embodiments, two k of Formula (h-ia) are the same.

[1103] In certain embodiments, both c1 of formula (h-ia) are the same. In certain embodiments, both c1 of formula (h-ia) are about 225.

[1104] In certain embodiments, -Y of formula (h-ia) a1 -and-Y a1‘ -Both The dashed line marked with an asterisk indicates the connection to -Z c .

[1105] In certain embodiments, moiety -Z of formula (Ia) or (Ib) comprises a moiety selected from the group consisting of:

[1106]

[1107]

[1108]

[1109] in,

[1110] The dotted line indicates the connection to -L 2 -;

[1111] s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14 and s15 are independently selected from the following group: 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10;

[1112] -X d1 、-X d2 、-X d3 and -X d4 are independently selected from the group consisting of: -OH, -SH and -NR g1 R g2 ; preferably -OH;

[1113] -X e1 、-X e2 、-X e3 and -X e4 are independently selected from the following groups: -H, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl;

[1114] -R g1 and -R g2 are independently selected from the following groups: -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 selected from the following groups: -H, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl; preferably -H;

[1115] -Y d1 -、-Y d2 -、-Y d3 -and-Y d4- independently selected from the group consisting of:

[1116] and

[1117] -Z d1 、-Z d2 、-Z d3 and -Z d4 are independently proteins, in certain embodiments random coil proteins, and in certain embodiments random coil proteins selected from the group consisting of PA, PAS, PAG, PG, and XTEN TM .

[1118] In certain embodiments, -Y of 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 for:

[1119]

[1120] In certain embodiments, -Y of 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 for:

[1121]

[1122] The dashed lines marked with asterisks are towards -Z d1 、-Z d2 、-Z d3 and -Z d4 , and the unmarked dotted line goes towards -L 2 -.

[1123] In certain embodiments, -X of formula (ji) f1 、-X f2 、-X f3 、-X f4 、-X f5 、-X f6 、-X f7 and -X f8 -H; -X of 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; and s1, s2, s3 and s4 of formula (ji) are selected from the group consisting of 2, 3, 4, 5 and 6.

[1124] In certain embodiments, -X of formula (ji) f1 、-X f2 、-X f3 、-X f4 、-X f5 、-X f6 、-X f7 and -X f8 -H; -X of formula (ji) d1 and -X d2 is -OH; -X in formula (ji) e1 and -X e2 is -H; and s1, s2 and s3 of formula (ji) are 4.

[1125] In certain embodiments, -X of formula (j-ii) f1 、-X f2 、-X f3 and -X f4 is -H; and -X of formula (j-ii) d1 、-X d2 、-X d3 and -X d4 is -OH; -X in formula (j-ii) e1 、-X e2 、-X e3 and -X e4 is selected from the group consisting of -H and methyl; s1, s2, s3, s4 and s5 of formula (j-ii) are selected from the group consisting of 2, 3, 4, 5 and 6. In certain embodiments, -X of formula (j-ii) f1 、-X f2 、-X f3 and -X f4 -H; -X of formula (j-ii) d1 、-X d2 、-X d3 and -X d4 is -OH; -X in formula (j-ii) e1 、-X e2 、-X e3 and -X e4 is -H; s1 of formula (j-ii) is 4 and s2, s3, s4 and s5 of formula (j-ii) are 1.

[1126] 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 -H; -X of 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; and 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 -H; -X of 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; and s1, s2 and s3 of formula (j-iii) are 4.

[1127] In certain embodiments, -X of formula (j-iv) f1 、-X f2 、-X f3 、-X f4 、-X f5 and -X f6 -H; s1, s2, s3, s4, s5, s6 and s7 of formula (j-iv) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; -Y d1 -and-Y d2 -Selected from the group consisting of:

[1128]

[1129] In certain embodiments, -X of formula (j-iv)f1 、-X f2 、-X f3 、-X f4 、-X f5 and -X f6 is -H; s1 of formula (j-iv) is 3, s2 of formula (j-iv) is 5, s3 of formula (j-iv) is 2, s4 of formula (j-iv) is 4, s5 of formula (j-iv) is 5, s6 of formula (j-iv) is 2, and s7 of formula (j-iv) is 4; and Y of formula (j-iv) d1 -and-Y d2 -for

[1130]

[1131] In certain embodiments, -X of formula (j-iv) f1 、-X f2 、-X f3 、-X f4 、-X f5 and -X f6 is -H; s1 of formula (j-iv) is 3, s2 of formula (j-iv) is 5, s3 of formula (j-iv) is 2, s4 of formula (j-iv) is 4, s5 of formula (j-iv) is 5, s6 of formula (j-iv) is 2, and s7 of formula (j-iv) is 4; and Y of formula (j-iv) d1 -and-Y d2 -for

[1132]

[1133] The dashed lines marked with asterisks are towards -Z d1 、-Z d2 、-Z d3 and -Z d4 , and the unmarked dotted line goes towards -L 2 -.

[1134] In certain embodiments, -X of formula (jv) f1 、-X f2 、-X f3 and -X f4 -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 -Selected from the group consisting of:

[1135]

[1136] In certain embodiments, -X of 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, and s5 of formula (jv) is 1; and Y of formula (jv) d1 -and-Y d2 -for

[1137]

[1138] In certain embodiments, -X of 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, and s5 of formula (jv) is 1; and Y of formula (jv) d1 -and-Y d2 -for

[1139]

[1140] The dashed lines marked with asterisks are towards -Z d1 、-Z d2 、-Z d3 and -Z d4 , and the unmarked dotted line goes towards -L 2 -.

[1141] In certain embodiments, -X of formula (j-vi) f1 、-X f2 、-X f3 、-X f4 、-X f5 、-X f6 、-X f7 、-X f8 、-X f9 and -X f10 -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 -Selected from the group consisting of:

[1142]

[1143] In certain embodiments, -X of formula (j-vi) f1 、-X f2 、-Xf3 、-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, and s9 of formula (j-vi) is 4; and Y of formula (j-vi) d1 -and-Y d2 -for

[1144]

[1145] In certain embodiments, -X of 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, and s9 of formula (j-vi) is 4; and Y of formula (j-vi) d1 -and-Y d2 -for

[1146]

[1147] The dashed lines marked with asterisks are towards -Z d1 、-Z d2 、-Z d3 and -Z d4 , and the unmarked dotted line goes towards -L 2 -.

[1148] In certain embodiments, -X of formula (j-vii) f1 、-X f2 、-X f3 、-X f4 、-X f5 、-X f6、-X f7 、-X f8 、-X f9 、-X f10 、-X f11 、-X f12 、-X f13 and -X f14 is -H; s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14 and s15 of formula (j-vii) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; -Y of formula (j-vii) d1 -、-Y d2 、-Y d3 -and-Y d4 -Selected from the group consisting of:

[1149]

[1150] In certain embodiments, -X of formula (j-vii) f1 、-X f2 、-X f3 、-X f4 、-X f5 、-X f6 、-X f7 、-X f8 、-X f9 、-X f10 、-X f11 、-X f12 、-X f13 and -X f14 is -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, and s15 of formula (j-vii) is 4; and -Y of formula (j-vii) d1 -、-Y d2 、-Y d3 -and-Y d4 -for

[1151] In certain embodiments, -X of formula (j-vii) f1 、-X f2 、-X f3、-X f4 、-X f5 、-X f6 、-X f7 、-X f8 、-X f9 、-X f10 、-X f11 、-X f12 、-X f13 and -X f14 is -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, and s15 of formula (j-vii) is 4; and -Y of formula (j-vii) d1 -、-Y d2 、-Y d3 -and-Y d4 -for

[1152]

[1153] The dashed lines marked with asterisks are towards -Z d1 、-Z d2 、-Z d3 and -Z d4 , and the unmarked dotted line goes towards -L 2 -.

[1154] 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 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 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 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 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 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 XTEN TM part.

[1155] In certain embodiments, the reversible CNP conjugate is of formula (Iif)

[1156]

[1157] in,

[1158] The unlabeled dashed line indicates attachment to the nitrogen of -D via formation of an amide bond, wherein -D is a CNP moiety; and

[1159] The dashed line marked with an asterisk indicates the connection to -Z with the following structure:

[1160]

[1161] in,

[1162] Each-Z a for

[1163]

[1164] in,

[1165] Each c1 is independently an integer from 200 to 250.

[1166] In certain embodiments, each c1 of Formula (Iif) is about 225.

[1167] In certain embodiments, the reversible CNP conjugate is of formula (Iif-i):

[1168]

[1169] in,

[1170] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[1171] and

[1172] The dashed line marked with an asterisk indicates the connection to -Z with the following structure:

[1173]

[1174] in,

[1175] Each-Z a for

[1176]

[1177] in,

[1178] Each c1 is independently an integer from 200 to 250.

[1179] In certain embodiments, each c1 of Formula (Iif-i) is about 225.

[1180] In certain embodiments, the CNP conjugate is of formula (Iif-ii):

[1181]

[1182] in,

[1183] The unlabeled dashed line indicates attachment to the nitrogen of -D via amide bond formation, where -D is the CNP moiety;

[1184] and

[1185] The dashed line marked with an asterisk indicates the connection to -Z with the following structure:

[1186]

[1187] in,

[1188] Each-Z a for

[1189]

[1190] in,

[1191] Each c1 is independently an integer from 200 to 250.

[1192] In certain embodiments, each c1 of Formula (Iif-ii) is about 225.

[1193] In certain embodiments, -D of formula (Iif), (Iif-i) and (Iif-ii) is a CNP moiety, i.e., the reversible conjugates of formula (Iif), (Iif-i) and (Iif-ii) are CNP conjugates. In certain embodiments, -D of formula (Iif), (Iif-i) and (Iif-ii) is a CNP moiety having a sequence of SEQ ID NO: 24, SEQ ID NO: 25 or SEQ ID NO: 30. In certain embodiments, -D of formula (Iif), (Iif-i) and (Iif-ii) is a CNP moiety having a sequence of SEQ ID NO: 24. In certain embodiments, -D of formula (Iif), (Iif-i) and (Iif-ii) is a CNP moiety having a sequence of SEQ ID NO: 20. In certain embodiments, -D of formula (Iif), (Iif-i) and (Iif-ii) is a CNP moiety having a sequence of SEQ ID NO: 21. In certain embodiments, -D of formula (Iif), (Iif-i) and (Iif-ii) is a CNP moiety having the sequence of SEQ ID NO: 22. -D of formula (Iif), (Iif-i) and (Iif-ii) is a CNP moiety having the sequence of SEQ ID NO: 23. In certain embodiments, -D of formula (Iif), (Iif-i) and (Iif-ii) is a CNP moiety having the sequence of SEQ ID NO: 30.

[1194] In certain embodiments, -D of formula (Iif), (Iif-i), and (Iif-ii) is a CNP moiety linked to -L via the nitrogen of the N-terminal amine functional group of the CNP. 1 -.

[1195] In certain embodiments, -D of Formula (Iif), (Iif-i) and (Iif-ii) is a CNP moiety linked to -L via the nitrogen provided by the amine functionality of the lysine side chain of the CNP moiety. 1 -.

[1196] In certain embodiments, if the CNP moiety is SEQ ID NO: 24, the lysine side chain is not part of the loop formed by the disulfide bridge between the cysteine ​​residues at positions 22 and 38.

[1197] Thus, in certain embodiments, if the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to the -L in the reversible CNP conjugates of formula (Iif), (Iif-i) and (Iif-ii) via the amine functionality provided by the lysine side chain at position 9. 1 -.

[1198] In certain embodiments, if the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to the -L in the reversible CNP conjugates of formula (Iif), (Iif-i) and (Iif-ii) via the amine functionality provided by the lysine side chain at position 11. 1 -.

[1199] In certain embodiments, if the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to the -L in the reversible CNP conjugates of formula (Iif), (Iif-i) and (Iif-ii) via the amine functionality provided by the lysine side chain at position 15. 1 -.

[1200] In certain embodiments, if the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to the -L in the reversible CNP conjugates of formula (Iif), (Iif-i) and (Iif-ii) via the amine functionality provided by the lysine side chain at position 16. 1 -.

[1201] In certain embodiments, if the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to the -L in the reversible CNP conjugates of formula (Iif), (Iif-i) and (Iif-ii) via the amine functionality provided by the lysine side chain at position 20. 1 -.

[1202] In certain embodiments, if the CNP moiety is SEQ ID NO: 24, the lysine side chain is part of a loop formed by a disulfide bridge between the cysteine ​​residues at positions 22 and 38.

[1203] Thus, in certain embodiments, if the CNP has the sequence of SEQ ID NO: 24, the CNP moiety is linked to the -L in the CNP conjugate of formula (Iif) via the amine functionality provided by the lysine side chain at position 26. 1 -.

[1204] In certain embodiments, the reversible CNP conjugate is of formula (Iif-i), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 24 and is linked to -L via the amine functional group provided by the lysine side chain at position 26. 1-.

[1205] In certain embodiments, the reversible CNP conjugate is of formula (Iif), wherein c1 is about 225, -D is a CNP moiety having a sequence of SEQ ID NO: 20 and is linked to -L via an amine functional group provided by the lysine side chain at position 30. 1 -.

[1206] In certain embodiments, the reversible CNP conjugate is of formula (Iif-i), wherein c1 is about 225, -D is a CNP moiety having the sequence of SEQ ID NO: 20 and is linked to -L via the amine functional group provided by the lysine side chain at position 30. 1 -.

[1207] In certain embodiments, the reversible CNP conjugate is of formula (Iif-ii), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 20 and is linked to -L via the amine functional group provided by the lysine side chain at position 30. 1 -.

[1208] In certain embodiments, the reversible CNP conjugate is of formula (Iif), wherein c1 is about 225, -D is a CNP moiety having a sequence of SEQ ID NO: 21 and is linked to -L via the amine functional group provided by the lysine side chain at position 29. 1 -.

[1209] In certain embodiments, the reversible CNP conjugate is of formula (Iif-i), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 21 and is linked to -L via the amine functional group provided by the lysine side chain at position 29. 1 -.

[1210] In certain embodiments, the reversible CNP conjugate is of formula (Iif-ii), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 21 and is linked to -L via the amine functional group provided by the lysine side chain at position 29. 1 -.

[1211] In certain embodiments, the reversible CNP conjugate is of formula (Iif), wherein c1 is about 225, -D is a CNP moiety having a sequence of SEQ ID NO: 22 and is linked to -L via the amine functional group provided by the lysine side chain at position 28. 1 -.

[1212] In certain embodiments, the reversible CNP conjugate is of formula (Iif-i), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 22 and is linked to -L via the amine functional group provided by the lysine side chain at position 28.1 -.

[1213] In certain embodiments, the reversible CNP conjugate is of formula (Iif-ii), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 22 and is linked to -L via the amine functional group provided by the lysine side chain at position 28. 1 -.

[1214] In certain embodiments, the reversible CNP conjugate is of formula (Iif), wherein c1 is about 225, -D is a CNP moiety having a sequence of SEQ ID NO: 23 and is linked to -L via the amine functional group provided by the lysine side chain at position 27. 1 -.

[1215] In certain embodiments, the reversible CNP conjugate is of formula (Iif-i), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 23 and is linked to -L via the amine functional group provided by the lysine side chain at position 27. 1 -.

[1216] In certain embodiments, the reversible CNP conjugate is of formula (Iif-ii), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 23 and is linked to -L via the amine functional group provided by the lysine side chain at position 27. 1 -.

[1217] In certain embodiments, the reversible CNP conjugate is of formula (Iif), wherein c1 is about 225, -D is a CNP moiety having the sequence of SEQ ID NO: 30 and is linked to -L via the amine functional group provided by the lysine side chain at position 27. 1 -.

[1218] In certain embodiments, the reversible CNP conjugate is of formula (Iif-i), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO: 30 and is linked to -L via the amine functional group provided by the lysine side chain at position 27. 1 -.

[1219] In certain embodiments, the reversible CNP conjugate is of formula (Iif-ii), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 30 and is linked to -L via the amine functional group provided by the lysine side chain at position 27. 1 -.

[1220] In certain embodiments, the reversible CNP conjugate is of formula (Iif-ii), wherein c1 is about 225, the CNP portion has the sequence of SEQ ID NO: 24 and is linked to -L via the amine functional group provided by the lysine side chain at position 26. 1 -.

[1221] It will be appreciated that the positions of the aforementioned cysteines and lysines will vary depending on the length of the CNP portion, and that one skilled in the art will readily identify the corresponding cysteines and lysines in longer or shorter versions of the CNP portion. It will also be appreciated that, for example, some lysine residues may not be present in a shorter CNP portion. It will be further appreciated that, for example, as a result of site-directed mutagenesis, there may be more lysine residues in the non-ring-forming portion and / or the ring-forming portion of the CNP portion.

[1222] In certain embodiments, the reversible CNP conjugate is of formula (Iif), wherein c1 is about 225, -D is a CNP moiety having a sequence of SEQ ID NO: 24 and is linked to -L via the amine functional group provided by the lysine side chain at position 26. 1 -.

[1223] In certain embodiments, the reversible CNP conjugate is of formula (Iif'):

[1224]

[1225] in,

[1226] The unlabeled dashed line represents the connection to the nitrogen provided by the lysine side chain at position 26 of the CNP portion of SEQ ID NO: 24 via formation of an amide bond; and

[1227] The dashed line marked with an asterisk indicates the connection to -Z with the following structure

[1228]

[1229] in,

[1230] Each-Z a for

[1231]

[1232] in,

[1233] Each c1 is independently an integer from 200 to 250.

[1234] In certain embodiments, each c1 of Formula (Iif') is about 225.

[1235] In certain embodiments, the reversible CNP conjugate is of formula (Iif-i'):

[1236]

[1237] in,

[1238] The unlabeled dashed line represents the connection to the nitrogen provided by the lysine side chain at position 26 of the CNP portion of SEQ ID NO: 24 via formation of an amide bond; and

[1239] The dashed line marked with an asterisk indicates the connection to -Z with the following structure

[1240]

[1241] in,

[1242] Each-Z a for

[1243]

[1244] in,

[1245] Each c1 is independently an integer from 200 to 250.

[1246] In certain embodiments, each c1 of Formula (Iif-i') is about 225.

[1247] In certain embodiments, the reversible CNP conjugate is of formula (Iif-ii'):

[1248]

[1249] in,

[1250] The unlabeled dashed line represents the connection to the nitrogen provided by the lysine side chain at position 26 of the CNP portion of SEQ ID NO: 24 via formation of an amide bond; and

[1251] The dashed line marked with an asterisk indicates the connection to -Z with the following structure

[1252]

[1253] in,

[1254] Each-Z a for

[1255]

[1256] in,

[1257] Each c1 is independently an integer from 200 to 250.

[1258] In certain embodiments, each c1 of Formula (Iif-ii') is about 225.

[1259] In certain embodiments, the reversible CNP conjugate is of formula (Iifa):

[1260]

[1261] in,

[1262] The unlabeled dashed line indicates attachment to the nitrogen of -D via formation of an amide bond, wherein -D is a CNP moiety; and

[1263] The dashed line marked with an asterisk indicates the connection to -Z with the following structure

[1264]

[1265] in,

[1266] k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12;

[1267] Each Z a for

[1268]

[1269] in,

[1270] Each c1 is independently an integer from 200 to 250.

[1271] In certain embodiments, k of Formula (Iifa) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[1272] In certain embodiments, each c1 of Formula (Iifa) is about 225.

[1273] In certain embodiments, the reversible CNP conjugate is of formula (lifa-i):

[1274]

[1275] in,

[1276] The unlabeled dashed line indicates attachment to the nitrogen of -D via formation of an amide bond, wherein -D is a CNP moiety; and

[1277] The dashed line marked with an asterisk indicates the connection to -Z with the following structure

[1278]

[1279] in,

[1280] k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12;

[1281] Each Za for

[1282]

[1283] in,

[1284] Each c1 is independently an integer from 200 to 250.

[1285] In certain embodiments, k of Formula (Iifa-i) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[1286] In certain embodiments, each c1 of Formula (Iifa-i) is about 225.

[1287] In certain embodiments, the reversible CNP conjugate is of formula (lifa-ii):

[1288]

[1289] in,

[1290] The unlabeled dashed line indicates attachment to the nitrogen of -D via formation of an amide bond, wherein -D is a CNP moiety; and

[1291] The dashed line marked with an asterisk indicates the connection to -Z with the following structure

[1292]

[1293] in,

[1294] k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12;

[1295] Each Z a for

[1296]

[1297] in,

[1298] Each c1 is independently an integer from 200 to 250.

[1299] In certain embodiments, each c1 of Formula (Iifa-ii) is about 225.

[1300] In certain embodiments, the CNP portion of the reversible CNP conjugates of Formula (Iifa), (Iifa-i), and (Iifa-ii) has the sequence of SEQ ID NO:25.

[1301] In certain embodiments, the CNP portion of the reversible CNP conjugates of Formula (Iifa), (Iifa-i), and (Iifa-ii) has the sequence of SEQ ID NO:20.

[1302] In certain embodiments, the CNP portion of the reversible CNP conjugates of Formula (Iifa), (Iifa-i), and (Iifa-ii) has the sequence of SEQ ID NO: 21.

[1303] In certain embodiments, the CNP portion of the reversible CNP conjugates of Formula (Iifa), (Iifa-i), and (Iifa-ii) has the sequence of SEQ ID NO:22.

[1304] In certain embodiments, the CNP portion of the reversible CNP conjugates of Formula (Iifa), (Iifa-i), and (Iifa-ii) has the sequence of SEQ ID NO:23.

[1305] In certain embodiments, the CNP portion of the reversible CNP conjugates of Formula (Iifa), (Iifa-i), and (Iifa-ii) has the sequence of SEQ ID NO:30.

[1306] In certain embodiments, the CNP portion of the reversible CNP conjugates of Formula (Iifa), (Iifa-i), and (Iifa-ii) has the sequence of SEQ ID NO:24.

[1307] In certain embodiments, the CNP moiety is linked to the -L in the reversible CNP conjugates of formula (Iifa), (Iifa-i) and (Iifa-ii) via the nitrogen of the N-terminal amine functional group of the CNP. 1 -.

[1308] In certain embodiments, the reversible CNP conjugate is of formula (Iifa'):

[1309]

[1310] in,

[1311] The unlabeled dashed line represents the connection to the nitrogen provided by the lysine side chain at position 26 of the CNP portion of SEQ ID NO: 24 via formation of an amide bond; and

[1312] The dashed line marked with an asterisk indicates the connection to -Z with the following structure

[1313]

[1314] in,

[1315] k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12;

[1316] Each Z a for

[1317]

[1318] in,

[1319] Each c1 is independently an integer from 200 to 250.

[1320] In certain embodiments, k of Formula (Iifa') is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[1321] In certain embodiments, each c1 of Formula (Iifa') is about 225.

[1322] In certain embodiments, the reversible CNP conjugate is of formula (lifa-i'):

[1323]

[1324] in,

[1325] The unlabeled dashed line represents the connection to the nitrogen provided by the lysine side chain at position 26 of the CNP portion of SEQ ID NO: 24 via formation of an amide bond; and

[1326] The dashed line marked with an asterisk indicates the connection to -Z with the following structure

[1327]

[1328] in,

[1329] k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12;

[1330] Each Z a for

[1331]

[1332] in,

[1333] Each c1 is independently an integer from 200 to 250.

[1334] In certain embodiments, k of Formula (Iifa-i′) is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[1335] In certain embodiments, each c1 of Formula (Iifa-i') is about 225.

[1336] In certain embodiments, the reversible CNP conjugate is of formula (lifa-ii'):

[1337]

[1338] in,

[1339] The unlabeled dashed line represents the connection to the nitrogen provided by the lysine side chain at position 26 of the CNP portion of SEQ ID NO: 24 via formation of an amide bond; and

[1340] The dashed line marked with an asterisk indicates the connection to -Z with the following structure:

[1341]

[1342] in,

[1343] k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12;

[1344] Each Z a for

[1345]

[1346] in,

[1347] Each c1 is independently an integer from 200 to 250.

[1348] In certain embodiments, k of Formula (Iifa-ii') is selected from the group consisting of 2, 3, 4, 5, 6, and 7.

[1349] In certain embodiments, each c1 of Formula (Iifa-ii') is about 225.

[1350] In certain embodiments, the CNP compound is a CNP agonist selected from the group consisting of a small molecule, a natural product, an oligonucleotide, a polypeptide, and a protein.

[1351] In certain embodiments, the CNP agonist comprises a small molecule. In certain embodiments, the CNP agonist is a small molecule.

[1352] In certain embodiments, the CNP agonist comprises a natural product. In certain embodiments, the CNP agonist is a natural product.

[1353] In certain embodiments, the CNP agonist comprises an oligonucleotide. In certain embodiments, such an oligonucleotide is selected from the group consisting of an antisense oligonucleotide, an aptamer, RNAi, and siRNA. In certain embodiments, the CNP agonist is an oligonucleotide selected from the group consisting of an antisense oligonucleotide, an aptamer, RNAi, and siRNA.

[1354] In certain embodiments, the CNP agonist comprises a protein. In certain embodiments, the CNP agonist is a protein.

[1355] In certain embodiments, the CNP agonist comprises a polypeptide. In certain embodiments, the CNP agonist is a polypeptide.

[1356] In certain embodiments, the CNP agonist comprises a CNP molecule or portion. In certain embodiments, the CNP agonist is CNP.

[1357] In certain embodiments, the CNP agonist comprises a CNP molecule or portion having the sequence of SEQ ID NO:24, SEQ ID NO:25, or SEQ ID NO:30.

[1358] In certain embodiments, the CNP agonist is a CNP having the sequence of SEQ ID NO: 24, SEQ ID NO: 25, or SEQ ID NO: 30. In certain embodiments, the CNP agonist is a CNP having the sequence of SEQ ID NO: 24. In certain embodiments, the CNP agonist is a CNP having the sequence of SEQ ID NO: 25. In certain embodiments, the CNP agonist is a CNP having the sequence of SEQ ID NO: 30.

[1359] In certain embodiments, the liquid pharmaceutical formulations of the present application may contain one or more other excipients, such as, for example, stabilizers, anti-adsorption agents, cryoprotectants and other adjuvants. It should be understood that an excipient may have multiple functions, such as dual or triple functions.

[1360] In certain embodiments, the liquid pharmaceutical formulation of the present application may contain a stabilizer, such as a stabilizer selected from the group consisting of alanine; arginine; aspartic acid; glycine; histidine; lysine; proline; sugars such as glucose, sucrose, trehalose; polyols such as glycerol, mannitol, sorbitol; salts such as potassium phosphate, sodium sulfate; chelating agents such as EDTA, hexaphosphate; ligands such as divalent metal ions; other salts or organic molecules such as phenol derivatives; oligomers or polymers such as cyclodextrins, dextran, dendrimers, PEG, PVP, protamine, and HAS.

[1361] In certain embodiments, the liquid pharmaceutical preparation of the present application may comprise 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, which are used to competitively coat or adsorb to the inner surface of the preparation or preparation container, such as pluronic acid. TM F-68), PEG lauryl ether ( 35), dextran, polyethylene glycol, PEG-polyhistidine, BSA, HSA, and gelatin. The excipient concentration and type chosen depends on the effect to be avoided, but generally a monolayer of surfactant is formed at the interface just above the CMC.

[1362] In certain embodiments, the liquid pharmaceutical preparation of the present application may include a cryoprotectant, such as a cryoprotectant selected from the group consisting of sugars, polyols, surfactants, amino acids, anhydrous solvents, and peptides. During freezing or spray drying, cryoprotectants can counteract the unstable effects caused by hydrogen bond rupture and water removal. Trehalose is particularly effective in reducing the aggregation caused by moisture, and also improves the thermal stability that the hydrophobic groups of the compound may cause when exposed to water. Mannitol and sucrose can also be used as independent freeze-drying / cryoprotectants, or are used in combination with each other, wherein a higher ratio of mannitol: sucrose is known to enhance the physical stability of the compound in the liquid pharmaceutical preparation. Mannitol can also be combined with trehalose. Trehalose can also be combined with sorbitol, or sorbitol can be used as an independent protective agent. Starch or starch derivatives can also be used.

[1363] In certain embodiments, the liquid pharmaceutical formulations of the present application may comprise additional excipients selected from the group consisting of a wetting agent, a viscosity modifier, and an antibiotic.

[1364] The applicant unexpectedly discovered that in the liquid pharmaceutical formulations of the present application, the reversible bond between the CNP and the carrier is stable, eliminating the need for freeze-drying and reconstitution from the freeze-dried material. However, if desired, the liquid pharmaceutical formulations of the present application can be dried, such as by freeze-drying, to form a dry, such as freeze-dried, liquid pharmaceutical formulation.

[1365] In certain embodiments, the liquid pharmaceutical formulation of the present application comprises a CNP conjugate, acetic acid, mannitol, m-cresol, and methionine.

[1366] In certain embodiments, the liquid pharmaceutical formulation of the present application comprises a CNP conjugate, acetic acid, trehalose, m-cresol, and methionine.

[1367] Due to the huge difference in patient weight ranges, such as, for example, 3-80 kg, in certain embodiments, it is beneficial to have liquid pharmaceutical formulations of CNP conjugates of different strengths (meaning different concentrations) (such as high, medium and low strengths). Therefore, in one embodiment, the liquid pharmaceutical formulation is provided in more than one concentration, such as at two different concentrations, such as at three different concentrations, such as at four different concentrations, such as at five different concentrations, such as at six different concentrations, such as at seven different concentrations, such as at eight different concentrations, such as at nine different concentrations, such as at ten different concentrations, such as at eleven different concentrations, such as at twelve different concentrations.

[1368] In certain embodiments, the present application provides liquid pharmaceutical formulations for storage at 2 to 8°C for at least 6 months.

[1369] In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 2 to 8°C for at least 12 months. In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 2 to 8°C for at least 24 months. In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 2 to 8°C for at least 36 months. In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 2 to 8°C for at least 48 months.

[1370] In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 2 to 8°C for 6 to 48 months. In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 2 to 8°C for 6 to 36 months. In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 2 to 8°C for 6 to 24 months. In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 2 to 8°C for 6 to 12 months.

[1371] In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 25° C. for one month. In certain embodiments, the present application provides a liquid pharmaceutical formulation for storage at 30° C. for one month.

[1372] The liquid pharmaceutical formulation as described above is a liquid pharmaceutical formulation that is stable for at least 6 months, such as at least 7 months, such as at least 8 months, such as at least 9 months, such as at least 10 months, such as at least 11 months, such as at least 12 months. In certain embodiments, the liquid pharmaceutical formulation is stable for at least 14 months, such as at least 16 months, such as at least 18 months, such as at least 20 months, such as at least 22 months, such as at least 24 months, such as at least 36 months, such as at least 48 months.

[1373] In certain embodiments, the liquid pharmaceutical formulation is stable for at least 36 months when stored at 2° C. to 8° C. In certain embodiments, the liquid pharmaceutical formulation is stable for at least 36 months when stored at 2° C. In certain embodiments, the liquid pharmaceutical formulation is stable for at least 6 months when stored at 5° C. In certain embodiments, the liquid pharmaceutical formulation is stable for at least 2 weeks when stored at 30° C.

[1374] In certain embodiments, the above-mentioned liquid pharmaceutical preparations can be stored at a temperature range of -80°C to 30°C, such as -20°C to 25°C, such as -15°C to 25°C, such as -10°C to 25°C, such as -5°C to 25°C, such as -0°C to 25°C, such as 2°C to 8°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 2°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 4°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 5°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 8°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 10°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 16°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 20°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 25°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 30°C. In certain embodiments, the liquid pharmaceutical preparation is stored at 40°C.

[1375] The liquid pharmaceutical preparation of the present application comprises at least about 80%, such as at least 85%, 90%, 95%, 96%, 97%, 98% or 99% of the native form of the peptide (CNP or CNP portion) after storage for at least 3 months, 6 months, 12 months or at least 24 months. Storage can be carried out at 5°C, 8°C, 15°C, 20°C or 25°C, and purity can be determined by any method known to those skilled in the art, such as by SE-HPLC or RP-HPLC.

[1376] In certain embodiments, the liquid pharmaceutical formulation comprises about 90%, 95%, 96%, 97% or 99% of the native form of the peptide (CNP or CNP portion) after storage at 5°C for 3 months or 6 months, as determined, for example, by SE-HPLC or RP-HPLC.

[1377] In certain embodiments, the liquid pharmaceutical formulation comprises about 80%, such as at least 85%, 90%, 95%, 96%, 97%, 98% or 99% of the native form of the peptide (CNP or CNP portion) after storage at 5°C for 24 months as determined by SE-HPLC or RP-HPLC.

[1378] In certain embodiments, the liquid pharmaceutical formulation comprises about 80%, 90%, or 95% of the native form of the peptide (CNP or CNP portion) after storage at 25°C for 6 months as determined by SE-HPLC or RP-HPLC.

[1379] The present application includes a method for preparing a liquid pharmaceutical formulation according to the present application, comprising the following steps:

[1380] (i) mixing a CNP compound with at least a buffer, an isotonicity agent, a preservative, and optionally an antioxidant;

[1381] (ii) adjusting the pH of the mixture of step (i);

[1382] (iii) optionally, filtering the mixture of step (ii);

[1383] (iv) transferring an amount of the mixture from step (ii) or (iii) equal to the desired number of doses into a container;

[1384] (v) a sealed container; and

[1385] The order of steps (ii) and (iii) can optionally be reversed.

[1386] In certain embodiments, steps (ii) and (iii) are not reversed.

[1387] In certain embodiments, the CNP compound in step (i) is mixed with a buffer, an isotonicity agent, a preservative, and optionally an antioxidant.

[1388] In certain embodiments, the method for preparing the liquid pharmaceutical formulation according to the present application comprises the following steps:

[1389] (i) mixing a CNP compound with at least acetic acid, mannitol, m-cresol and optionally an antioxidant;

[1390] (ii) adjusting the pH of the mixture of step (i);

[1391] (iii) optionally, filtering the mixture of step (ii);

[1392] (iv) transferring an amount of the mixture from step (ii) or (iii) equal to the desired number of doses into a container;

[1393] (v) a sealed container; and

[1394] The order of steps (ii) and (iii) can optionally be reversed.

[1395] In certain embodiments, steps (ii) and (iii) are not reversed.

[1396] In certain embodiments, the CNP compound in step (i) is mixed with acetic acid, mannitol, m-cresol, and optionally an antioxidant.

[1397] In certain embodiments, the method for preparing the liquid pharmaceutical formulation according to the present application comprises the following steps:

[1398] (i) mixing the CNP conjugate with at least acetic acid, mannitol, m-cresol and optionally methionine to produce a formulation comprising:

[1399]

[1400] (ii) adjusting the pH of the mixture of step (i) to pH 3.0 to pH 5.5 with NaOH and HCl;

[1401] (iii) optionally, filtering the mixture of step (ii);

[1402] (iv) transferring an amount of the mixture from step (ii) or (iii) equal to the desired number of doses into a container;

[1403] (v) a sealed container; and

[1404] The order of steps (ii) and (iii) can optionally be reversed.

[1405] In certain embodiments, the method for preparing the liquid pharmaceutical formulation according to the present application comprises the following steps:

[1406] (i) mixing the CNP conjugate with at least acetic acid, trehalose, m-cresol and optionally methionine to produce a formulation comprising:

[1407]

[1408] (ii) adjusting the pH of the mixture of step (i) to pH 3.0 to pH 5.5 with NaOH and HCl;

[1409] (iii) optionally, filtering the mixture of step (ii);

[1410] (iv) transferring an amount of the mixture from step (ii) or (iii) equal to the desired number of doses into a container;

[1411] (vi) a sealed container; and

[1412] The order of steps (ii) and (iii) can optionally be reversed.

[1413] In certain embodiments, steps (ii) and (iii) are not reversed.

[1414] Another aspect of the present application relates to a container for the liquid pharmaceutical preparation of the present application. The container may be suitable for storing and dispensing the preparation. In certain embodiments, the container comprises the liquid pharmaceutical preparation of the present application. When such a container is used, the kit may additionally comprise a dispenser, such as a syringe or a reservoir with capacity markings.

[1415] In certain embodiments, the container is made of glass. In certain embodiments, the inner surface of the container is coated with an inert material. In certain embodiments, the inert material is silicon.

[1416] In certain embodiments, the container may be selected from the group consisting of: a vial; a syringe, such as a dual chamber syringe; an ampoule, and a cartridge, such as a dual chamber cartridge.

[1417] In certain embodiments, the cartridge is used with a pen injector, such as an autoinjector.

[1418] In certain embodiments, the liquid pharmaceutical formulation of the present application is provided as a single dose, meaning that the container containing the liquid pharmaceutical formulation contains one therapeutic dose. It is clear to those skilled in the art that when the liquid pharmaceutical formulation is provided as a single dose, no preservative is present in the formulation.

[1419] In certain embodiments, the liquid drug formulation comprises multiple doses, meaning that the container containing the liquid drug formulation contains more than one therapeutic dose.

[1420] In certain embodiments, the multi-dose liquid pharmaceutical formulation comprises at least 2 doses, such as at least 4 doses, such as at least 6 doses, such as at least 8 doses, such as at least 10 doses, such as at least 12 doses of a CNP agonist or CNP; and in certain embodiments such as at least 14 doses.

[1421] In certain embodiments, the multi-dose liquid pharmaceutical formulation comprises at least 2, 4, 6, 8, 10, 12, or 14 doses of a CNP agonist or CNP.

[1422] Thus, in another aspect of the present application, a liquid pharmaceutical formulation is provided as a multi-dose formulation.

[1423] A single dose formulation comprising a CNP compound, particularly a CNP conjugate, has a volume of no more than 4 ml, such as from 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 0.5 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 1.5 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.

[1424] In certain embodiments, the CNP compound, particularly a CNP conjugate, is in a formulation with sufficient dosage to provide a therapeutically effective amount of CNP for at least three days, such as at least four days, such as at least five days, such as at least six days, in one application.

[1425] In certain embodiments, the CNP compound, particularly a CNP conjugate, is in a formulation at a sufficient dosage to provide a therapeutically effective amount of CNP for one week.

[1426] The liquid pharmaceutical formulations of the present application can be administered, for example, via topical, enteral or parenteral administration, as well as by methods of external application, injection or infusion, including intra-articular, peri-articular, intradermal, subcutaneous, intramuscular, intravenous, intraosseous, intraperitoneal, intrathecal, intracapsular, intraorbital, intravitreal, intratympanic, intravesical, intracardiac, transtracheal, subcutaneous, subcapsular, subarachnoid, intraspinal, intraventricular, intrasternal injection, infusion, intranasal, oral, transpulmonary and transdermal administration, direct delivery to the brain via an implanted device that allows delivery of the present application or similar formulations to brain tissue or brain fluid (e.g., Ommaya reservoir), direct intracerebroventricular injection or infusion, injection or infusion into the brain or an area associated with the brain, injection into the subchoroidal space, retro-orbital injection and intraocular instillation, preferably via subcutaneous injection.

[1427] In certain embodiments, the liquid pharmaceutical formulations of the present application are administered via subcutaneous injection.

[1428] In certain embodiments, the liquid drug formulations of the present application are administered via subcutaneous injection using a syringe and needle or a pen injector (such as an autoinjector).

[1429] In certain embodiments, the liquid pharmaceutical formulations of the present application are administered via subcutaneous injection using a syringe and needle.

[1430] In certain embodiments, the liquid drug formulations of the present application are administered via subcutaneous injection using a pen injector.

[1431] In certain embodiments, the liquid pharmaceutical formulations of the present application are administered via subcutaneous injection using an automatic injector.

[1432] In certain embodiments, the period of time between two consecutive subcutaneous administrations, i.e., the administration interval, is at least every 12 hours, 24 hours, 36 hours, 48 ​​hours, 60 hours, 72 hours, 84 hours, 96 hours, 108 hours, 120 hours, 132 hours, 144 hours, 156 hours, one week, two weeks, three weeks, or four weeks.

[1433] In certain embodiments, the time period between two consecutive subcutaneous administrations is 12 hours. In certain embodiments, the time period between two consecutive subcutaneous administrations is 24 hours. In certain embodiments, the time period between two consecutive subcutaneous administrations is 48 hours. In certain embodiments, the time period between two consecutive subcutaneous administrations is 72 hours. In certain embodiments, the time period between two consecutive subcutaneous administrations is 96 hours. In certain embodiments, the time period between two consecutive subcutaneous administrations is 120 hours. In certain embodiments, the time period between two consecutive subcutaneous administrations is 144 hours. In certain embodiments, the time period between two consecutive subcutaneous administrations is one week.

[1434] Another aspect of the present application is the use of the liquid pharmaceutical formulation of the present application as a medicament.

[1435] In another aspect, the present application relates to the use of the liquid pharmaceutical formulation of the present application in treating, controlling, delaying or preventing one or more diseases that can be treated, controlled, delayed or prevented with a CNP agonist (particularly CNP).

[1436] In certain embodiments, the present application relates to the use of the liquid pharmaceutical formulation of the present application in treating one or more diseases that can be treated with a CNP agonist (particularly CNP).

[1437] Another aspect of the present application is a method for treating, controlling, delaying or preventing one or more diseases treatable with a CNP agonist (particularly CNP) in a patient, comprising administering to the patient a therapeutically effective amount of the liquid pharmaceutical formulation of the present application.

[1438] In certain embodiments, the patient is an adult. In certain embodiments, the patient is a pediatric patient.

[1439] The liquid pharmaceutical formulation of the present application can be administered for at least six months, one year, five years, ten years, until the patient is 18 years old, until the patient's epiphysis is closed, or indefinitely. In certain embodiments, the liquid pharmaceutical formulation can be administered until the patient reaches the age of 18. In certain embodiments, the liquid pharmaceutical formulation can be administered until the patient's epiphysis is closed.

[1440] In certain embodiments, the one or more diseases that can be treated, controlled, delayed or prevented with a CNP agonist (particularly CNP) are selected from the group consisting of bone-related disorders, such as skeletal dysplasia; cancer; autoimmune diseases; fibrotic diseases; inflammatory diseases; central nervous system diseases, such as neurodegenerative diseases; infectious diseases; lung diseases; cardiac and vascular diseases; metabolic diseases and ophthalmic diseases.

[1441] In certain embodiments, the one or more diseases that can be treated, controlled, delayed or prevented with a CNP agonist (particularly CNP) are selected from the group consisting of achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasias, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, chondrodysplasia punctata, homozygous achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly, proximal achondrodysplasia punctata, Jansen type metaphyseal dysplasia, congenital spondyloepiphyseal dysplasia, atelosteogenesis, diastrophic dysplasia, spondyloepiphyseal ... dysplasia), congenital short femur, Langer mesomelic dysplasia, Nievergelt mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acromegaly, peripheral dysostosis, Kniest dysplasia, fibrochondrogenesis, Roberts syndrome, acromesomelic dysplasia, micromelia, Morquio syndrome, Kniest syndrome, metatrophic dysplasia, spondyloepimetaphyseal dysplasia, neurofibromatosis, LEOPARD syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome syndrome), cardiofaciocutaneous syndrome, Costello syndrome, SHOX gene deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysplasia, craniosynostosis (eg, Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome),syndrome or Crouzonodermoskeletal syndrome), dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, segmental dysplasia, enchondroma, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome syndrome), osteosclerosis, osteopoikilosis, hemorrhagic shock, hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, hydronephrosis, edema due to hepatopathy, acute renal insufficiency, chronic renal insufficiency, glaucoma, increased intraocular pressure, multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, breast cancer, growth retardation, skull deformity, orthodontic defects, cervical spinal cord compression, spinal stenosis, hydrocephalus, hearing loss due to chronic otitis media, cardiovascular disease, neurologic disease, and obesity.

[1442] In certain embodiments, the one or more diseases that can be treated, controlled, delayed or prevented with a CNP agonist (particularly CNP) are selected from the group consisting of: achondroplasia (e.g., homozygous achondroplasia), hypochondroplasia, short stature, dwarfism, osteochondrodysplasia, lethal dysplasia, osteogenesis imperfecta, achondrodysplasia punctata, flexor dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly, proximal limb punctate chondrodysplasia, Jansen type metaphyseal dysplasia, congenital spondyloepiphyseal dysplasia, osteodysplasia, malformations, congenital short femur, Langer type limb dysplasia, Niever Gelt-type mediomelia, Robinow syndrome, Reinhardt syndrome, acromegaly, peripheral skeletal dysplasia, Kniest dysplasia, fibrochondrogenesis, Roberts syndrome, acromegaly, phocomelia, Morquio syndrome, Kniest syndrome, variant spondyloepiphyseal dysplasia, spondyloepiphyseal dysplasia, neurofibromatosis, Rex syndrome, leopard skin syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Rex syndrome, cardiofaciocutaneous syndrome, Costello syndrome, SHOX gene defects Syndrome, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysplasia, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzonodermoskeletal syndrome), dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, segmental dysplasia, enchondroma, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, and rickets. Levy syndrome, Marfan syndrome, McCune-Albright syndrome, osteosclerosis, systemic brittle bone sclerosis, hemorrhagic shock, hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, hydronephrosis, hepatic edema, acute renal insufficiency, chronic renal insufficiency, glaucoma, increased intraocular pressure, multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, breast cancer, growth retardation, skull deformity, orthodontic defects, cervical spinal cord compression, spinal stenosis, hydrocephalus, hearing loss caused by chronic otitis media, obesity, diseases involving abnormal RAS-mitogen-activated protein kinase signaling, pulmonary hypertension, vascular disease,Endothelial dysfunction, cirrhosis, ascites, hepatic fibrosis, hepatorenal syndrome, asthma, pulmonary fibrosis, chronic kidney disease, cardiorenal syndrome, dyspnea, and lysosomal storage disorders (e.g., mucopolysaccharidoses).

[1443] In certain embodiments, the one or more diseases that can be treated, controlled, delayed or prevented with a CNP agonist (particularly CNP) are one or more cardiovascular diseases selected from the group consisting of arrhythmias (e.g., atrial or sinus arrhythmias), atrial fibrillation, atrial flutter, bradycardia, Brugada syndrome, premature heart complexes, commotio cordis, heart block, long QT syndrome, parasystole, preexcitation syndrome, tachycardia, ventricular fibrillation, ventricular flutter, cardiac conduction system disease, low cardiac output, cardiomegaly, dilated cardiomyopathy, hypertrophy (e.g., left ventricular hypertrophy or right ventricular hypertrophy), cardiomyopathy (e.g., alcoholic, dilated, hypertrophic, restrictive, diabetic or Chagas cardiomyopathy), arrhythmic right ventricular dysplasia, endocardial fibroelastosis, endomyocardial fibrosis, glycogen storage disease type IIb, Kearns-Sayre syndrome, syndrome), myocardial reperfusion injury, myocarditis, sarcoglycanosis, endocarditis (e.g., bacterial or noninfective endocarditis), cardiac arrest, sudden cardiac death, out-of-hospital cardiac arrest, cardiorenal syndrome, paroxysmal dyspnea, cardiac edema, heart failure (e.g., diastolic or systolic), valvular heart disease, aortic insufficiency; aortic stenosis, heart valve prolapse, mitral insufficiency, mitral stenosis, pulmonary atresia, pulmonary insufficiency, pulmonary stenosis, tricuspid atresia, tricuspid insufficiency, tricuspid stenosis, myocardial ischemia, acute coronary syndrome, angina pectoris, coronary artery disease, Kounis syndrome syndrome), myocardial infarction, cor pulmonale, 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), abnormal intracardiac fluid accumulation, and myocardial edema.

[1444] In certain embodiments, the one or more diseases that can be treated, controlled, delayed or prevented with a CNP agonist (particularly CNP) are selected from the group consisting of ischemic heart disease (such as myocardial infarction), congestive heart failure, arrhythmia and atherosclerosis.

[1445] In certain embodiments, the one or more diseases that can be treated, controlled, delayed or prevented with a CNP agonist (particularly CNP) are one or more central nervous system diseases selected from the group consisting of cerebral ischemia (e.g., ischemic hypoxia), cerebral infarction, transient ischemic attack, vertebrobasilar insufficiency, cerebrovascular disorder, stroke, intracranial hemorrhage, corneal neovascularization, corneal transplantation, graft-versus-host disease, graft rejection, glaucoma (e.g., angle-closure, neovascular, open-angle or low-tension glaucoma), ischemic optic neuropathy, central serous chorioretinopathy, retinopathy (e.g., diabetic or hypertensive retinopathy), retinal degeneration, macular degeneration, geographic atrophy, macular edema, Stargardt disease, vitelliform macular 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.

[1446] In certain embodiments, the one or more diseases are selected from the group consisting of hypophosphatasia, achondroplasia, Muenke syndrome, hypertension, osteogenesis imperfecta, and achondroplasia.

[1447] In certain embodiments, the one or more diseases that can be treated with a CNP agonist (particularly CNP) are hypophosphatasia. In certain embodiments, the one or more diseases that can be treated with CNP are achondroplasia. In certain embodiments, the one or more diseases that can be treated with CNP are Muenke syndrome. In certain embodiments, the one or more diseases that can be treated with CNP are hypertension. In certain embodiments, the one or more diseases that can be treated with CNP are osteogenesis imperfecta. In certain embodiments, the one or more diseases that can be treated with CNP are achondroplasia.

[1448] The present application is further described by the following non-limiting items.

[1449] 1. A liquid pharmaceutical formulation comprising a CNP compound, a buffer, an isotonicity agent, a preservative and optionally an antioxidant.

[1450] 2. The liquid pharmaceutical formulation according to item 1, which is intended to be stored at 2 to 8°C for at least 6 months.

[1451] 3. The liquid pharmaceutical formulation according to item 1 or 2, which is for storage at 2 to 8°C for 6 to 48 months.

[1452] 4. The liquid pharmaceutical formulation according to any one of items 1 to 3, which is for storage at 2 to 8°C for 6 to 36 months.

[1453] 5. The liquid pharmaceutical formulation according to any one of items 1 to 4, which is intended for storage at 5°C for 6 to 36 months.

[1454] 6. The liquid pharmaceutical formulation according to any one of items 1 to 5, wherein the CNP compound is a CNP conjugate.

[1455] 7. The liquid pharmaceutical formulation of any one of items 1 to 6, wherein the buffer is selected from the group consisting of acetic acid, succinic acid, citric acid, lactic acid, glutamic acid, fumaric acid, aspartic acid, glutaric acid, phosphoric acid, histidine, gluconic acid, tartaric acid, malic acid, and Tris (tris(hydroxymethyl)aminomethane).

[1456] 8. The liquid pharmaceutical formulation according to any one of items 1 to 7, wherein the buffer is acetic acid.

[1457] 9. The liquid pharmaceutical preparation according to any one of items 1 to 8, wherein the isotonic agent is selected from the group consisting of mannitol, trehalose, sucrose, raffinose, gelatin, lactose, calcium hydrogen phosphate, sorbitol, xylitol, glycine, histidine, ethanol, hydroxyethyl starch, potassium chloride, sodium chloride, glucose, dextran, propylene glycol and glycerol.

[1458] 10. The liquid pharmaceutical formulation according to any one of items 1 to 9, wherein the isotonicity agent is mannitol or trehalose.

[1459] 11. The liquid pharmaceutical preparation according to any one of items 1 to 10, wherein the preservative is selected from the group consisting of m-cresol, benzyl alcohol, benzoic acid, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, potassium sorbate, chlorobutanol, benzyl alcohol, phenylmercuric nitrate, thimerosal, sorbic acid, potassium sorbate, chlorocresol, benzalkonium chloride, ethylene glycol monoethyl ether, chlorhexidine, chlorobutanol, phenylethyl alcohol, and phenylmercuric acetate.

[1460] 12. The liquid pharmaceutical formulation according to any one of items 1 to 11, wherein the preservative is m-cresol or phenol.

[1461] 13. A liquid pharmaceutical formulation according to any one of items 1 to 12, wherein the liquid pharmaceutical formulation comprises an antioxidant selected from the group consisting of methionine, butylated hydroxytoluene, butylated hydroxyanisole, tocopherol, propyl gallate, ascorbic acid, sodium bisulfite, ethylenediaminetetraacetic acid (EDTA), cysteine, glutathione, monothioglycerol, poly(ethyleneimine), vitamin E, ectoine and mulberry yellow.

[1462] 14. The liquid pharmaceutical formulation according to any one of items 1 to 13, wherein the pH of the liquid pharmaceutical formulation is from about pH 3.5 to about pH 5.5.

[1463] 15. The liquid pharmaceutical formulation according to any one of items 1 to 14, wherein the pH of the liquid pharmaceutical formulation is 4.5.

[1464] 16. The liquid pharmaceutical formulation according to any one of items 1 to 15, wherein the liquid pharmaceutical formulation further comprises a pH adjuster.

[1465] 17. The liquid pharmaceutical formulation according to item 16, wherein the pH adjuster is selected from the group consisting of sodium hydroxide, sodium acetate, Tris (tris(hydroxymethyl)aminomethane), potassium hydroxide, lysine, and mixtures thereof.

[1466] 18. The liquid pharmaceutical formulation according to any one of items 1 to 17, wherein the liquid pharmaceutical formulation comprises a CNP compound, acetic acid, an isotonicity agent, a preservative, and optionally an antioxidant.

[1467] 19. The liquid pharmaceutical formulation according to any one of items 1 to 18, wherein the liquid pharmaceutical formulation comprises a CNP compound, acetic acid, trehalose dihydrate, a preservative, and optionally an antioxidant.

[1468] 20. The liquid pharmaceutical formulation according to any one of items 1 to 19, wherein the liquid pharmaceutical formulation comprises a CNP compound, acetic acid, trehalose dihydrate, m-cresol, and optionally an antioxidant.

[1469] 21. The liquid pharmaceutical formulation according to any one of items 1 to 20, wherein the liquid pharmaceutical formulation comprises a CNP compound, acetic acid, trehalose dihydrate, m-cresol and methionine.

[1470] 22. The liquid pharmaceutical formulation according to any one of items 1 to 17, wherein the liquid pharmaceutical formulation comprises a CNP compound, acetic acid, mannitol, a preservative and optionally an antioxidant.

[1471] 23. The liquid pharmaceutical formulation according to any one of items 1 to 17 or 22, wherein the liquid pharmaceutical formulation comprises a CNP compound, acetic acid, mannitol, m-cresol and optionally an antioxidant.

[1472] 24. The liquid pharmaceutical formulation according to any one of items 1-17, 22 or 23, wherein the liquid pharmaceutical formulation comprises a CNP compound, acetic acid, mannitol, m-cresol and methionine.

[1473] 25. The liquid pharmaceutical formulation according to any one of items 1 to 24, wherein the CNP compound is a reversible CNP conjugate comprising a CNP moiety covalently and reversibly conjugated to a carrier moiety.

[1474] 26. A liquid pharmaceutical formulation according to claim 25, wherein the CNP portion has a 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.

[1475] 27. The liquid pharmaceutical formulation according to item 25 or 26, wherein 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, SEQ ID NO: 25 or SEQ ID NO: 30.

[1476] 28. The liquid pharmaceutical formulation according to any one of items 25-27, wherein the CNP portion has the sequence of SEQ ID NO: 24.

[1477] 29. The liquid pharmaceutical formulation according to any one of items 25-28, wherein the carrier moiety is a polymeric moiety.

[1478] 30. The liquid pharmaceutical formulation according to any one of items 1 to 29, wherein the CNP compound is a reversible CNP conjugate of formula (Ia):

[1479]

[1480] in,

[1481] -D is the CNP part;

[1482] -L 1 - is a reversible joint portion;

[1483] -L 2 - is a single chemical bond or spacer moiety;

[1484] -Z is a carrier moiety; and

[1485] 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.

[1486] 31. The liquid pharmaceutical formulation according to item 30, wherein x of formula (Ia) is 1.

[1487] 32. The liquid pharmaceutical formulation of item 30 or 31, wherein said -Z is a branched polymeric moiety.

[1488] 33. The liquid pharmaceutical formulation of any one of items 30-32, wherein said -Z is a branched PEG-based polymeric moiety.

[1489] 34. The liquid pharmaceutical formulation according to any one of items 6-18 or 22-33, wherein the liquid pharmaceutical formulation comprises:

[1490]

[1491] 35. The liquid pharmaceutical formulation according to any one of items 6-21 or 25-33, wherein the liquid pharmaceutical formulation comprises:

[1492]

[1493] 36. The liquid pharmaceutical formulation according to any one of items 30-35, wherein the part-L 1 -is formula (II):

[1494]

[1495] The dashed line indicates the connection to the nitrogen of -D via formation of an amide bond, where -D is a CNP moiety;

[1496] -X is -C(R 4 R 4a )-、-N(R 4 )-、-O-、-C(R 4 R 4a )-C(R 5 R 5a )-、-C(R 5 R 5a )-C(R 4 R 4a )-、-C(R 4 R 4a )-N(R 6 )-、-N(R 6 )-C(R 4 R 4a )-、-C(R 4 R 4a )-O-、-OC(R 4 R 4a )-or-C(R 7 R 7a )-;

[1497] X 1 is C or S(O);

[1498] -X 2 Yes-C(R 8 R 8a )-or-C(R 8 R 8a )-C(R 9 R 9a )-;

[1499] =X 3 is =O, =S or =N-CN;

[1500] -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 Independently selected from the group consisting of: -H and C 1-6 alkyl;

[1501] -R 3 、-R 3a Independently selected from the group consisting of: -H and C 1-6 Alkyl, provided that -R 3 、-R 3a If one or both are not -H, they are passed to sp 3 - hybridized carbon atoms connected to the N atoms to which they are attached;

[1502] -R 7 Yes-N(R 10 R 10a ) or -NR 10 -(C=O)-R 11 ;

[1503] -R 7a 、-R 10 、-R 10a 、-R 11 Independently of each other, -H or C 1-6 alkyl;

[1504] Optionally, -R 1a / -R 4a 、-R 1a / -R 5a 、-R1a / -R 7a 、-R 4a / -R 5a 、-R 8a / -R 9a One or more of the pairs form a chemical bond;

[1505] Optionally, -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 pairs are joined to the atoms to which they are attached to form a C 3-10 cycloalkyl or 3- to 10-membered heterocyclic group;

[1506] Optionally, -R 1 / -R 4 、-R 1 / -R 5 、-R 1 / -R 6 、-R 1 / -R 7a 、-R 4 / -R 5 、-R 4 / -R 6 、-R 8 / -R 9 、-R 2 / -R 3 One or more of the pairs are joined to the atoms to which they are attached to form ring A;

[1507] Optionally, -R 3 / -R 3a is joined to the nitrogen atom to which it is attached to form a 3- to 10-membered heterocycle;

[1509] A is selected from the group consisting of: phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclic group and 8- to 11-membered heterobicyclic group; and

[1510] where -L 1 - is replaced by -L 2 -Z, and -L 1 - is optionally further substituted, provided that the hydrogen atom marked with an asterisk in formula (II) is not replaced by -L 2-Z or a substituent.

[1511] 37. The liquid pharmaceutical formulation according to any one of items 30-36, wherein the part-L 1 -L 2 -is the formula (IId-ii):

[1512]

[1513] in,

[1514] The unlabeled dashed line indicates the connection to the nitrogen of the amino acid side chain of the cyclic portion of -D through the formation of an amide bond; and the dashed line marked with an asterisk indicates the connection to -Z; wherein -D has the sequence of SEQ ID NO: 24 and -L 1 - conjugated to lysine at position 26;

[1515] Wherein, the -Z is formula (h):

[1516]

[1517] in,

[1518] The dotted line indicates the connection to -L 2 -;and

[1519] Each-Z c For part

[1520]

[1521] in,

[1522] Each c1 is independently an integer from about 200 to 250.

[1523] 38. The liquid pharmaceutical formulation according to any one of items 30-37, wherein the part-L 1 -L 2 -is the formula (IId-ii'):

[1524]

[1525] in,

[1526] The unlabeled dashed line indicates the connection to -D defined in item 37; and the labeled dashed line indicates the connection to -Z defined in item 37.

[1527] 39. The liquid pharmaceutical formulation according to any one of items 30-38, wherein the part-L 1 -L 2 - is the formula (hi):

[1528]

[1529] in,

[1530] The dotted line indicates the connection to -L 2 -;and

[1531] Each-Z c For part

[1532]

[1533] in,

[1534] Each c1 is independently an integer from 200 to 250.

[1535] 40. The liquid pharmaceutical formulation of any one of items 37-39, wherein each c1 is about 225.

[1536] 41. The liquid pharmaceutical formulation according to any one of items 1-24, 34 or 35, wherein the CNP compound is a stable CNP conjugate comprising a CNP moiety covalently and stably conjugated to a carrier moiety.

[1537] 42. The liquid pharmaceutical formulation according to any one of items 1-24, 34, 35 or 41, wherein the CNP compound is a stable CNP conjugate comprising a CNP portion covalently and stably conjugated to a fatty acid derivative.

[1538] 43. The liquid pharmaceutical formulation according to any one of items 1-17 or 23-29, wherein the CNP compound is a CNP agonist.

[1539] 44. The liquid pharmaceutical formulation of item 43, wherein the CNP agonist is CNP.

[1540] 45. The liquid pharmaceutical formulation of claim 44, wherein the CNP has a 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.

[1541] 46. ​​The liquid pharmaceutical formulation of item 44 or 45, wherein the CNP has the sequence of 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.

[1542] 47. A method for preparing the liquid pharmaceutical formulation of any one of items 1 to 46, wherein the method comprises the following steps:

[1543] (vi) mixing the CNP compound of any one of items 1 to 46 with at least a buffer, an isotonicity agent, a preservative, and optionally an antioxidant;

[1544] (vii) adjusting the pH of the mixture of step (i);

[1545] (viii) optionally, filtering the mixture of step (ii);

[1546] (ix) transferring an amount of the mixture of step (ii) or (iii) equivalent to the desired dose into a container;

[1547] (x) sealing the container; and

[1548] The order of steps (ii) and (iii) can optionally be reversed.

[1549] 48. A method according to claim 47, wherein the buffer is acetic acid.

[1550] 49. The method according to item 47 or 48, wherein the isotonic agent is selected from the group consisting of mannitol and trehalose.

[1551] 50. A container comprising the liquid pharmaceutical formulation of any one of items 1-46.

[1552] 51. The container of item 50, wherein the container is selected from the group consisting of a vial, a syringe, an ampoule, and a cartridge.

[1553] 52. The liquid pharmaceutical formulation according to any one of items 1 to 46, for use in the treatment of one or more diseases treatable with CNP.

[1554] 53. A liquid pharmaceutical formulation for use in claim 52, wherein the disease that can be treated with CNP is selected from the group consisting of: achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasia, thanatophoric dysplasia, osteogenesis imperfecta, hypochondrodysplasia punctata, homozygous achondroplasia, flexor dysostosis, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly, proximal achondrodysplasia punctata, Jansen metaphyseal dysplasia, congenital spondyloepiphyseal dysplasia, osteodysplasia, malformations, congenital short femur, Langer mesomelic dysplasia, Nievergelt mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acrodysplasia, peripheral skeletal dysplasia, Kniest development dysplasia, fibrocartilage development, Roberts syndrome, acromesomelic dysplasia, phocomelia, Morquio syndrome, Kniest syndrome, osteodysplasia variant, spondyloepiphyseal dysplasia, neurofibromatosis, Lacks syndrome, leopard skin syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, cardiofacial and cutaneous syndrome, Kestrel's elastin deficiency, SHOX gene deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysplasia, craniosynostosis, dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, segmental dysplasia, enchondroma, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jakob-Levi syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis, and systemic osteopetrosis.

[1555] 54. Use of the liquid pharmaceutical formulation according to item 52, wherein the disease that can be treated with CNP is selected from the group consisting of achondroplasia, hypochondroplasia, short stature, Noonan syndrome and SHOX gene deficiency.

[1556] 55. Use of a liquid pharmaceutical formulation according to claim 52, wherein the disease is achondroplasia.

[1557] 56. A method of treating, reducing the risk of, or delaying the progression of a disease treatable by CNP in a human patient, said method comprising the step of administering to said patient a therapeutically effective amount of the liquid pharmaceutical formulation according to any one of items 1-46.

[1558] 57. The method according to item 56, wherein the disease is selected from the group consisting of achondroplasia, hypochondroplasia, short stature, Noonan syndrome and SHOX gene deficiency.

[1559] 58. A method according to item 56 or 57, wherein the disease is achondroplasia.

[1560] 59. Use of the liquid pharmaceutical formulation according to any one of items 1 to 46 in the preparation of a medicament for treating achondroplasia. Example

[1561] Materials and methods

[1562] Peptides: CNP-38 TFA salt, CNP-27 TFA salt, CNP-53 acetate salt, and PG-CNP-37 TFA salt were purchased from Caslo (Lynby, Denmark). Reference peptides for peptide mapping analysis were purchased from Wuxi, China.

[1563] Reagents: TFA (sequencing grade, Thermo), water (ULC / MS grade, Biosolve; and hypotonic, sterile, pyrogen-free, Carl Roth), acetonitrile (ULC / MS grade, Biosolve), poloxamer 188 (cell culture grade, Applichem), Tween-20 (complying with European Pharmacopoeia, Carl Roth), hot lysozyme (Promega), NaH2PO4*H2O (ACS>98%, Sigma), 4M NaOH (Carl Roth), 1M NaOH (European Pharmacopoeia reagent grade, Carl Roth), 1M HCl (European Pharmacopoeia reagent grade, Carl Roth), NaCl (European Pharmacopoeia grade>99%, Carl Roth), phosphoric acid (ACS grade, >85%, Merck), CaCl2 dihydrate (ACS grade>99%), trehalose dihydrate (USP grade, Pfanstiehl), succinic acid (for analysis, ACS, Applichem), acetic acid (ROTIPURAN 100%, analytical grade (pa), Carl Roth; and conforming to the standards of the European Pharmacopoeia, British Pharmacopoeia, Japanese Pharmacopoeia, USP, and American Chemical Society (ACS), Merck), m-cresol (European Pharmacopoeia / USP injection grade, Hedinger), phenol (ultra-pure, ACS reagent, European Pharmacopoeia reagent, VWR), mannitol (conforms to the standards of the European Pharmacopoeia, British Pharmacopoeia, USP, and Japanese Pharmacopoeia, Merck; and conforms to the standards of the European Pharmacopoeia, USP, and British Pharmacopoeia, Carl Roth), citric acid (ACS reagent, >99.5%, Sigma), citric acid monohydrate (conforms to the standards of the European Pharmacopoeia, British Pharmacopoeia, Chinese Pharmacopoeia, Japanese Pharmacopoeia, and USP, E330, FCC), sodium citrate dihydrate (≥99%, European Pharmacopoeia grade, Carl Roth), ethylenediaminetetraacetic acid disodium salt dihydrate (EDTA Disodium Salt Dihydrate) (suitable for electrophoresis and molecular biology, 99.0-101% [titration], Sigma), methionine (compliant with the European Pharmacopoeia and United States Pharmacopoeia standards, AppliChem), tris (hydroxymethylaminomethane) (Tris) (BioPerformance certified, compliant with the European Pharmacopoeia and United States Pharmacopoeia test standards, suitable for cell culture, >99.9% [titration], Sigma), polysorbate 80 (tested according to the European Pharmacopoeia, Sigma Aldrich).

[1564] Preparation of CNP-38 formulations (F1-F23)

[1565] Prepare excipient stock solutions by combining the desired excipients in a volumetric flask and filling to 90% of the desired volume with water. Adjust the pH of the stock solution using 1 M Tris solution or 4 M NaOH solution as needed, and adjust the volume to the desired volume with water. Filter the stock solution through a 0.2 μm PVDF filter. Add phenol and m-cresol to the formulation as needed. Next, dissolve the desired amount of CNP-38 in the desired amount of the desired excipient stock solution, check the pH, and adjust as necessary using 1.0 M Tris or 0.1 M NaOH. Filter the formulation through a 0.22 μm PVDF filter and aliquot for incubation.

[1566] Preparation of CNP preparation (F33-45)

[1567] With the exception of phenol and m-cresol, all excipient stock solutions were prepared by mixing the desired excipients in a volumetric flask and filling to 90% of the desired volume with water. The pH of the stock solutions was adjusted as needed using 1 M NaOH solution, and the volume was adjusted to the desired volume with water. The stock solutions were filtered through a 0.2 μm PVDF filter. After filtration, stock solutions of phenol (1.75%) and m-cresol (1.05%) were prepared by adding the preservatives to the corresponding excipient stock solutions. Next, the desired amount of CNP was dissolved in the desired amount of the desired excipient stock solution, the pH was checked and adjusted as necessary using 1 M NaOH. The formulation was filtered through a 0.22 μm PVDF filter, and the corresponding preservative stock solution was added. The formulation was aliquoted for incubation.

[1568] Preparation of compound (1) preparations (F24-F32)

[1569] The solution of compound (1) was buffer exchanged in the desired buffer using membrane diafiltration and then concentrated by ultrafiltration. Membrane diafiltration and ultrafiltration were performed by TFF using an Akta Flux S TFF system and a Pellicon XL50 Cassette (Biomax 10 kDa). After ultrafiltration, the concentration was checked and the solution was diluted back to a concentration of 3.6 or 6.0 mg CNP-38 equivalents / ml. Next, the stock solution was aliquoted, methionine was added where required, and the pH was checked and adjusted if necessary. Finally, m-cresol was added to the sample, sterile filtered using a 0.22 μm PVDF filter, and aliquoted in preparation for incubation.

[1570] Preparation of compound (1) preparations F46-F50

[1571] The solution of compound (1) was buffer exchanged into the specified buffer using membrane diafiltration and then concentrated by ultrafiltration. Ultra-15, -10K) membrane diafiltration and ultrafiltration. After ultrafiltration, check the concentration and dilute the solution back to a concentration of 2.8mg CNP-38 equivalent / ml. Next, check the pH and adjust with 1M HCl if necessary. The preparation is filtered using a 0.22μm PVDF filter, preservative stock solution is added and the sample is diluted to 2.2mg CNP-38 equivalent / ml. The preparation is aliquoted and prepared for incubation.

[1572] pH: The pH of the formulations was measured with a calibrated pH meter using a standard ionic strength electrode at room temperature.

[1573] Analytical methods

[1574] The purity of CNP-38 was determined by RP-HPLC-MS: mobile phase A consisted of 0.05% TFA in water (v / v), and mobile phase B consisted of 0.04% TFA (v / v) in acetonitrile. A Waters Acquity CSH C18 column ( The column was run at a flow rate of 0.3 ml / min, detection at a wavelength of 215 nm, and a column temperature of 60°C (±1°C). For LCMS, the HPLC system was coupled to an LTQ Orbitrap Discovery XL instrument, and ESI-MS was performed in positive ion mode. A step elution gradient was used, as shown in Table 21.

[1575] Table 21

[1576]

[1577]

[1578] Preparative RP-HPLC was used to separate the succinic acid adduct CNP-38. Mobile phase A consisted of water / acetonitrile / TFA (95 / 5 / 0.05, v / v / v), and mobile phase B consisted of water / acetonitrile / TFA (75 / 25 / 0.05, v / v / v). A Waters Acquity CSH C18 column ( 10 μm, 10 x 150 mm (+10 x 10 mm pre-column). The flow rate was set to 10 ml / min, detection was at a wavelength of 215 nm, and the column was operated at room temperature. The elution gradient is shown in Table 22.

[1579] Table 22

[1580] Time [minutes] ％B 0.0 50 2.5 60 17.5 60 27.5 67.5 30 75 35 100

[1581] 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 (v / v), and mobile phase B consisted of 0.04% TFA (v / v) in acetonitrile. A Waters Acquity CSHC18 column ( 1.7 μm, 2.1x100 mm). The flow rate was set to 0.3 ml / min, detection was at a wavelength of 215 nm, the column operating temperature was 60°C (± 1°C), and the sample injection volume was approximately 3 μg CNP equivalent (20 μL of 0.135 mg / ml sample). A step elution gradient was used, first from 5% B to 54% B in 25.5 minutes, then from 54% B to 59% B in 10.0 minutes. The sample was diluted to 0.15 mg CNP-38 equivalent / ml with dilution buffer (10 mM succinic acid, 84 g / L trehalose dihydrate, adjusted to pH 5 with 1 M Tris), and further diluted to 9:1 (w / w) with dilution buffer containing 0.5% Tween-20 (w / v). The content was determined by comparing the peak area with a reference solution of known content. Free CNP-38 was identified by reference peptides and quantified relative to the total peak area.

[1582] Purity of (1) was determined by SE-HPLC: The mobile phase consisted of an aqueous solution of 15 mM sodium phosphate (pH 7.40), 135 mM sodium chloride, and 0.2% poloxamer 188 (e.g., 2.07 g NaH2PO4*H2O, 7.89 g NaCl, and 2.00 g poloxamer 188 dissolved in 1 L HPLC-grade water; the pH was adjusted to 7.40 using 4 M aqueous NaOH). A GE Superdex 200 Increase 10 / 300 GL column was used. The isocratic flow rate was set at 0.75 ml / min, detection was at a wavelength of 215 nm, the column was run at room temperature, and the sample injection volume was 17 μg CNP-38 equivalent (50 μL of 0.34 mg / ml sample). The sample was diluted to 0.34 mg / mL with dilution buffer (10 mM succinic acid, 84 g / L trehalose dihydrate, adjusted to pH 5 with 1 M Tris) and dilution buffer containing 0.1% poloxamer 188 (w / v), resulting in a final poloxamer 188 concentration of 0.05% (e.g., 20 μL of a 3.6 mg CNP-38 equivalent / mL solution was diluted to 0.68 mg / mL with 86 μL of dilution buffer, and 50 μL of this solution was further diluted with 50 μL of dilution buffer containing 0.1% poloxamer 188).

[1583] Peptide mapping of (1) was performed to assess the conversion of aspartic acid (28) to isoaspartic acid (28) (Isoasp), and the oxidation of methionine (33) to methionine sulfoxide (33) (Met(O)): Thermolysin digestion of (1) was performed at a ratio of thermolysin to CNP-38 = 1:20 (w / w). The pH was adjusted to 7.5 using proteolysis buffer concentrate. After incubation at 37°C for 7 hours, the digestion was terminated by the addition of 5% (v / v) phosphoric acid. The resulting peptide mixture was purified by RP-HPLC on a Waters Acquity UPLC HSS T3 column ( The separation was performed on a 1.8 μm column (2.1 x 150 mm) with 0.1% volume ratio (vol%) TFA (v / v) in water as mobile phase A and 0.09% TFA (v / v) in acetonitrile as mobile phase B, with detection at 210 nm. The flow rate was set at 0.28 ml / min, the column operating temperature was 45°C (±1°C), and the sample injection volume was 19 μg CNP-38 equivalent (e.g., 20 μL of 0.94 mg / ml sample). The eluent gradient was from 0% B to 3% B over 31.0 minutes. Thermolysin fragments containing aspartic acid (28) (LisoDR) and methionine (33) (IGSM(O)SG) were identified by reference peptides and quantified based on their respective peak areas relative to the peak area of ​​the corresponding unmodified fragment (LDR; IGS+MSG).

[1584] Peptide mapping of CNP-38 and its variants was performed using LCMS. Peptide mapping and RP-HPLC were performed as described (1). For LCMS, the HPLC system was coupled to an LTQ Orbitrap Discovery XL instrument, and ESI-MS was performed in positive ion mode.

[1585] IEX-HPLC mobile phase A consisted of 20 mM sodium phosphate buffer (pH 6.5) containing 10% (v / v) ethanol, and mobile phase B consisted of 20 mM sodium phosphate buffer (pH 6.5) and 1 M sodium chloride (containing 10% (v / v) ethanol). A ProPacElite WCX 5 μm; 4 mm x 250 mm column (Thermo Scientific) was used. The flow rate was set at 0.4 ml / min, detection was at a wavelength of 220 nm, the column was run at a temperature of 40°C (± 8°C), and the sample injection volume was approximately 5.3 μg CNP-38 equivalents (7 μL of 0.75 mg / ml sample). The gradient is described in Table 23. The sample was diluted to 0.75 mg / ml with water.

[1586] Table 23

[1587] Time [minutes] ％B 0.0 2 2.0 2 17.0 16 17.1 98 25.0 98 25.1 2 43.0 2

[1588] The content and purity of CNP (F33-45) were determined by RP-HPLC / MS: mobile phase A consisted of 0.05% TFA in water (v / v), and mobile phase B consisted of 0.04% TFA (v / v) in acetonitrile. A Waters Acquity CSH C18 column ( 1.7 μm, 2.1x100 mm). The flow rate was set to 0.3 ml / min, detection was at a wavelength of 215 nm, the column operating temperature was 65 ° C, and the sample injection volume was approximately 3 μg CNP-38 equivalent (20 μL of 0.135 mg / ml sample). The sample was diluted to 0.135 mg / ml with dilution buffer (10 mM succinic acid, 84 g / L trehalose dihydrate, adjusted to pH 5 with 1 M Tris). The gradient is as described in Table 24. For LCMS, the HPLC system was coupled to an LTQ Orbitrap Discovery XL instrument and ESI-MS was performed in positive ion mode. Changes in T1M, T3M, and T6M content were determined by comparing the peak area with that of the T0 sample. To determine purity, all peaks above the limit of quantification (LOQ, 0.10%) and not present in the blank sample were integrated.

[1589] Table 24

[1590]

[1591]

[1592] Example 1 Synthesis of Compound (1)

[1593]

[1594] Compound (1) was synthesized as described for conjugate 11i in WO 2017 / 118693 A1.

[1595] Example 2: Stability of CNP-38 at different pH values ​​in succinate buffer

[1596] The effect of pH of liquid pharmaceutical formulations on the stability of CNP-38 was evaluated. To this end, six different formulations (F1, F2, F3, F4, F5 and F6) containing 2.8 mg / ml CNP-38 were prepared (Table 1).

[1597] Table 1

[1598]

[1599] The formulations were incubated at 37°C for 7 days, after which their overall purity and the presence of succinate adducts were assessed using RP-HPLC and RP-HPLC-MS (molecular weight (MW) of CNP-38 + 100 Da). The results showed that after incubation, the F3 formulation at pH 4.5 had the highest purity (showed the least degradation), and succinate adducts were detected in F1 to F5, but not in F6 (see Table 2).

[1600] Table 2

[1601]

[1602]

[1603] Example 3: Identification of the main CNP-38 succinate adducts

[1604] The major succinate isomers were separated by RP-HPLC using CNP-38 incubated as in Example 2. Separately, succinylated CNP-38 was generated de novo by incubating CNP-38 with succinic anhydride: 2.5 equivalents of succinic anhydride in 0.6 ml of PBS buffer were added to 10 mg of CNP-38 in 5 ml of PBS. The pH of the mixture was adjusted to pH ~6.5 with 4 N NaOH, and the mixture was incubated for 30 minutes. This addition and pH adjustment were repeated three times, and the major isomers were separated by R...

Claims

1. A liquid pharmaceutical formulation comprising a CNP compound, a buffer, an isotonicity agent, a preservative and optionally an antioxidant.

2. The liquid pharmaceutical formulation according to claim 1, which is intended to be stored at 2 to 8°C for at least 6 months.

3. The liquid pharmaceutical formulation according to claim 1 or 2, which is used for storage at 2-8°C for 6 to 48 months.

4. The liquid pharmaceutical formulation according to any one of claims 1 to 3, which is intended to be stored at 2-8°C for 6 to 36 months.

5. The liquid pharmaceutical formulation according to any one of claims 1 to 4, which is intended for storage at 5°C for 6 to 36 months.

6. The liquid pharmaceutical formulation according to any one of claims 1 to 5, wherein the CNP compound is a CNP conjugate.

7. The liquid pharmaceutical formulation according to any one of claims 1 to 6, wherein the buffer is selected from the group consisting of acetic acid, succinic acid, citric acid, lactic acid, glutamic acid, fumaric acid, aspartic acid, glutaric acid, phosphoric acid, histidine, gluconic acid, tartaric acid, malic acid and Tris (tris(hydroxymethyl)aminomethane).

8. The liquid pharmaceutical formulation according to any one of claims 1 to 7, wherein the buffer is acetic acid.

9. The liquid pharmaceutical preparation according to any one of claims 1 to 8, wherein the isotonic agent is selected from the group consisting of mannitol, trehalose, sucrose, raffinose, gelatin, lactose, calcium hydrogen phosphate, sorbitol, xylitol, glycine, histidine, ethanol, hydroxyethyl starch, potassium chloride, sodium chloride, glucose, dextran, propylene glycol and glycerol.

10. The liquid pharmaceutical formulation according to any one of claims 1 to 9, wherein the isotonicity agent is mannitol or trehalose.

11. The liquid pharmaceutical formulation according to any one of claims 1 to 10, wherein the isotonicity agent is mannitol.

12. The liquid pharmaceutical formulation according to any one of claims 1 to 10, wherein the isotonicity agent is trehalose.

13. The liquid pharmaceutical preparation according to any one of claims 1 to 12, wherein the preservative is selected from the group consisting of m-cresol, benzyl alcohol, benzoic acid, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, potassium sorbate, chlorobutanol, benzyl alcohol, phenylmercuric nitrate, thimerosal, sorbic acid, potassium sorbate, chlorocresol, benzalkonium chloride, ethylene glycol monoethyl ether, chlorhexidine, chlorobutanol, phenylethyl alcohol, and phenylmercuric acetate.

14. The liquid pharmaceutical formulation according to any one of claims 1 to 13, wherein the preservative is m-cresol or phenol.

15. The liquid pharmaceutical formulation according to any one of claims 1 to 14, wherein the preservative is m-cresol.

16. The liquid pharmaceutical formulation according to any one of claims 1-14, wherein the preservative is phenol.

17. A liquid pharmaceutical formulation according to any one of claims 1-16, wherein the liquid pharmaceutical formulation comprises an antioxidant selected from the group consisting of methionine, butylated hydroxytoluene, butylated hydroxyanisole, tocopherol, propyl gallate, ascorbic acid, sodium bisulfite, ethylenediaminetetraacetic acid (EDTA), cysteine, glutathione, monothioglycerol, poly(ethyleneimine), vitamin E, ectoine and mulberry yellow.

18. The liquid drug formulation of any one of claims 1-17, wherein the pH of the liquid drug formulation is about pH 3.5 to about pH 5.

5.

19. The liquid drug formulation of any one of claims 1-18, wherein the pH of the liquid drug formulation is about 4.

5.

20. The liquid drug formulation according to any one of claims 1-19, wherein the liquid drug formulation further comprises a pH adjuster.

21. The liquid pharmaceutical formulation according to claim 20, wherein the pH adjuster is selected from the group consisting of sodium hydroxide, sodium acetate, Tris (tris(hydroxymethyl)aminomethane), potassium hydroxide, lysine, and mixtures thereof.

22. The liquid pharmaceutical formulation according to any one of claims 1-21, wherein the CNP compound is a reversible CNP conjugate comprising a CNP moiety covalently and reversibly conjugated to a carrier moiety.

23. The liquid drug formulation of claim 22, wherein the carrier moiety is a polymeric moiety.

24. The liquid pharmaceutical formulation according to any one of claims 1-21, wherein the CNP compound is a CNP agonist.

25. The liquid pharmaceutical formulation according to any one of claims 1 to 23, wherein the CNP compound is a reversible CNP conjugate of formula (Ia): in, -D is the CNP part; -L 1 - is a reversible joint part; -L 2 - is a single chemical bond or a spacer moiety; -Z is a carrier moiety; and 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.

26. The liquid pharmaceutical formulation according to claim 25, wherein x of formula (Ia) is 1.

27. The liquid pharmaceutical formulation of claim 25 or 26, wherein the -Z is a branched polymeric moiety.

28. The liquid drug formulation of any one of claims 25-27, wherein the -Z is a branched PEG-based polymeric moiety.

29. The liquid drug formulation according to any one of claims 6-11, 13-15, 17-23 or 25-28, wherein the liquid drug formulation comprises:

30. The liquid drug formulation according to any one of claims 6-10, 12-15, 17-23 or 25-28, wherein the liquid drug formulation comprises:

31. The liquid pharmaceutical formulation according to any one of claims 25 to 30, wherein the part -L 1 -is formula (II): Wherein the dashed line indicates the connection to the nitrogen of -D via formation of an amide bond, where -D is a CNP moiety; -X is -C(R 4 R 4a )-, -N(R 4 )-, -O-, -C(R 4 R 4a )-C(R 5 R 5a )-, -C(R 5 R 5a )-C(R 4 R 4a )-, -C(R 4 R 4a )-N(R 6 )-, -N(R 6 )-C(R 4 R 4a )-, -C(R 4 R 4a )-O-, -O-C(R 4 R 4a )- or -C(R 7 R 7a )-; X 1 is C or S(O); -X 2 Yes-C(R 8 R 8a )-or-C(R 8 R 8a )-C(R 9 R 9a )-; =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 Independently selected from the group consisting of: -H and C 1-6 alkyl; -R 3 , -R 3a Independently selected from the group consisting of: -H and C 1-6 Alkyl, provided that -R 3 , -R 3a If one or both are not -H, they are passed through sp 3 - hybridized carbon atoms are connected to the N atoms to which they are attached; -R 7 Yes-N(R 10 R 10a ) or -NR 10 -(C=O)-R 11 ; -R 7a , -R 10 , -R 10a , -R 11 Independently of each other, -H or C 1-6 alkyl; Optionally, -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 pairs form a chemical bond; Optionally, -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 pairs join together with the atoms to which they are attached to form C 3-10 Cycloalkyl or 3- to 10-membered heterocyclic group; Optionally, -R 1 / -R 4 , -R 1 / -R 5 , -R 1 / -R 6 , -R 1 / -R 7a , -R 4 / -R 5 , -R 4 / -R 6 , -R 8 / -R 9 , -R 2 / -R 3 One or more of the pairs are linked together with the atoms to which they are attached to form ring A; Optionally, -R 3 / -R 3a are joined together with the nitrogen atom to which they are attached to form a 3- to 10-membered heterocyclic ring; A is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetrahydronaphthyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; and wherein -L 1 - is replaced by -L 2 - is replaced by -Z, and -L 1 - is optionally further substituted, provided that the hydrogen marked with an asterisk in formula (II) is not substituted by -L 2 -Z or a substituent.

32. The liquid pharmaceutical formulation according to any one of claims 25 to 31, wherein the part -L 1 -L 2 -is the formula (IId-ii): in, The unlabeled dashed line indicates the connection to the nitrogen of the amino acid side chain of the cyclic portion of -D through the formation of an amide bond; and the dashed line marked with an asterisk indicates the connection to -Z; wherein -D has the sequence of SEQ ID NO: 24 and -L 1 - conjugated to lysine at position 26; Wherein, the -Z is formula (h): in, The dotted line indicates the connection to -L 2 -;and Each-Z c For the following parts: in, Each c1 is independently an integer of about 200-250.

33. A method for preparing the liquid pharmaceutical formulation of any one of claims 1-32, wherein the method comprises the following steps: (i) mixing the CNP compound according to any one of claims 1 to 32 with at least a buffer, an isotonicity agent, a preservative and optionally an antioxidant; (ii) adjusting the pH of the mixture of step (i); (iii) optionally, filtering the mixture of step (ii); (iv) transferring an amount of the mixture of step (ii) or (iii) equivalent to the desired dose into a container; (v) sealing the container; and The order of steps (ii) and (iii) may optionally be reversed.

34. The method of claim 33, wherein the buffer is acetic acid.

35. The method of claim 33 or 34, wherein the isotonic agent is selected from the group consisting of mannitol and trehalose.

36. The method of any one of claims 33-35, wherein the isotonic agent is mannitol.

37. The method of any one of claims 33-35, wherein the isotonic agent is trehalose.

38. A container comprising the liquid pharmaceutical formulation of any one of claims 1-37.

39. The container of claim 38, wherein the container is selected from the group consisting of a vial, a syringe, an ampoule, and a cartridge.

40. The liquid pharmaceutical formulation according to any one of claims 1-32, for use in treating one or more diseases treatable with CNP.

41. A liquid pharmaceutical formulation for use according to claim 40, wherein the disease that can be treated with CNP is selected from the group consisting of achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasia, thanatophoric dysplasia, osteogenesis imperfecta, hypochondrodysplasia punctata, homozygous achondroplasia, flexor limb dysplasia, congenital lethal hypophosphatasia, perinatal lethal osteogenesis imperfecta, short rib polydactyly, proximal limb punctate chondrodysplasia, Jansen type metaphyseal dysplasia, congenital spondyloepiphyseal dysplasia, osteodysplasia, malformations, congenital short femur, Langer type meloid dysplasia, Nievergelt type meloid dysplasia, Robinow syndrome, Reinhardt syndrome, acrodysplasia, peripheral skeletal dysplasia, Kniest syndrome Dysplasia, fibrochondrogenesis, Roberts syndrome, acrometoplastic dysplasia, phocomelia, Morquio syndrome, Kniest syndrome, variant bone dysplasia, spondyloepiphyseal dysplasia, neurofibromatosis, Lexemetaphyseal dysplasia, Leopard skin syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, cardiofacial and cutaneous syndrome, Kerstiello elastin deficiency, SHOX gene deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysplasia, craniosynostosis, malformations of fingers, brachydactyly, flexor digits, polydactyly, syndactyly, segmental dysplasia, enchondroma, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jakob-Leyden syndrome, Marfan syndrome, McCune-Albright syndrome, osteosclerosis and systemic brittle osteosclerosis.

42. The use of the liquid pharmaceutical formulation according to claim 40, wherein the disease that can be treated with CNP is selected from the group consisting of achondroplasia, hypochondroplasia, short stature, Noonan syndrome and SHOX gene deficiency.

43. Use of the liquid pharmaceutical formulation according to claim 40, wherein the disease is achondroplasia.

44. A method of treating, reducing the risk of, or delaying the progression of a disease treatable by CNP in a human patient, the method comprising the step of administering to the patient a therapeutically effective amount of a liquid pharmaceutical formulation according to any one of claims 1-32.

45. The method of claim 44, wherein the disease is selected from the group consisting of achondroplasia, hypochondroplasia, short stature, Noonan syndrome, and SHOX gene deficiency.

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

47. The liquid pharmaceutical formulation according to any one of claims 1 to 32, for use in the preparation of a medicament for treating achondroplasia.

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