Novel polymeric hGH prodrugs

Abstract

The present invention provides a sustained-release formulation of human growth hormone that can be administered at high concentrations and in injection volumes of less than 1.0 mL for a variety of indications requiring human growth hormone therapy. The present invention relates to human growth hormone polymer prodrugs, as well as dry, liquid and reconstituted pharmaceutical formulations comprising said prodrugs. The invention further relates to their use as medicaments for treating diseases treatable by growth hormone, and to methods of treatment. The invention also relates to methods of administering such human growth hormone polymer prodrugs or pharmaceutical formulations.

Description

[Technical Field]

[0001] The present invention relates to human growth hormone polymer prodrugs, as well as dry, liquid and reconstituted pharmaceutical formulations comprising said prodrugs. The invention further relates to their use as medicaments for treating diseases treatable with growth hormone, and to methods of treatment. The invention also relates to methods of administering such human growth hormone polymer prodrugs or pharmaceutical formulations. [Background technology]

[0002] Human growth hormone (hGH) is a hormone that stimulates growth and cell proliferation in humans and other animals. It is a 191-amino acid, single-chain polypeptide hormone synthesized, stored, and secreted by somatotrophs located in the lateral wing of the anterior pituitary gland.

[0003] Growth hormone has a variety of functions in the body, the most notable of which is the increase in height throughout childhood, and there are several diseases that can be treated by the therapeutic use of hGH, such as childhood and adult growth hormone deficiency (GHD), idiopathic short stature (ISS), short stature homeobox (SHOX) gene mutations, Turner syndrome (TS), Noonan syndrome (NS), Prader-Willi syndrome (PWS), children born small for gestational age (SGA), chronic renal failure (CRI), wasting due to HIV or AIDS or other malignancies, short bowel syndrome (SBS), sarcopenia and frailty.

[0004] The standard treatment for hGH-related disorders is by frequent, usually daily, subcutaneous injections, which is particularly inconvenient for the primarily pediatric patient population. Therefore, various approaches are under development to provide sustained-release depots that require less frequent administration of hGH, such as those described in WO2009 / 133137A2. Summary of the Invention [Problem to be solved by the invention]

[0005] It is also desirable to maintain a small injection volume to ensure drug administration in a manner convenient for patients. Increasing the injection volume to 0.5-1.0 mL significantly increases pain at the injection site, and injection volumes exceeding 1.0 mL should be avoided. Because the majority of patients requiring hGH treatment are children, injection volumes should be kept to a minimum to ensure adequate compliance and promote the desired treatment outcomes. However, there are limitations to the amount of hGH that can be delivered per given volume, and the use of certain excipients, such as covalently and non-covalently bound carriers such as polymers, reduces the amount of hGH. In such cases, the volume administered per injection must be increased, or more than one injection is required. If this is not an option, certain diseases requiring higher doses of hGH, such as ISS, Turner syndrome, Noonan syndrome, chronic kidney disease, Prader-Willi syndrome, and adolescent GHD patients, cannot be treated with a given pharmaceutical formulation. Furthermore, pediatric patients requiring growth hormone treatment grow and gain weight, resulting in the need for increased hGH doses to ensure exposure to relatively consistent hGH concentrations.

[0006] It would therefore be desirable to provide sustained release formulations of hGH that can be administered in high concentrations and in injection volumes of less than 1.0 mL for a variety of indications requiring hGH therapy.

[0007] The viscosity of a pharmaceutical formulation further determines whether it can be injected through a fine gauge needle: as viscosity increases, a larger diameter needle is required to ensure that the pharmaceutical formulation can be injected within an acceptable time frame.

[0008] Because the size of the needle required to inject the hGH formulation affects patient acceptance, it is desirable to provide a sustained release formulation of hGH with a small needle diameter and a viscosity that facilitates administration with an acceptable injection time.

[0009] When a pharmaceutical formulation containing hGH is stored in its dry form, it is desirable that its reconstitution proceed quickly and with as little foaming / bubbling as possible to minimize pre-administration procedures and ensure proper administration of the drug. [Means for solving the problem]

[0010] SUMMARY OF THE INVENTION It is therefore an object of the present invention to at least partially overcome the above-mentioned drawbacks.

[0011] The present invention provides a polymeric human growth hormone (hGH) prodrug of formula (Ia) or (Ib) or a pharmaceutically acceptable salt thereof.

[0012] [ka] [In the formula, -D is the hGH moiety attached to the rest of the molecule via the amine functionality; n is 0, 1, 2, 3 or 4; -X- is a chemical bond or a spacer; =Y1, =Y5 are independently selected from the group consisting of =O and =S; -Y2- is selected from the group consisting of -O- and -S-; -Y3- is selected from the group consisting of -O- and -S-; -Y4- is -O-, -NR 5 - and -C(R 6 R 6a )-, -R 1 is a water-soluble PEG-based moiety comprising at least 40% PEG having a molecular weight in the range of 30-50 kDa, more preferably 35-45 kDa; -R 2 , -R 3 , -R 5 , -R 6 , -R 6aare independently selected from the group consisting of -H, 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; -R 4 is selected from the group consisting of 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; -W- is C 3~10 Cycloalkyl, 8-30 membered carbopolycyclyl, 3-10 membered heterocyclyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 )-, C optionally interrupted by one or more groups selected from the group consisting of 1~20 is selected from the group consisting of alkyl, -Nu is -N(R 7 R 7a ), -N(R 7 OH), -N(R 7 )-N(R 7a R 7b ), -S(R 7 ), -COOH,

[0013] [ka] is a nucleophile selected from the group consisting of -Ar- is

[0014] [ka] (In the formula, The dashed line indicates the bond to the remainder of the prodrug; -Z 1- is -O-, -S- and -N(R 7 )-, -Z 2 - is -N(R 7 )-is) is selected from the group consisting of -R 7 , -R 7a , -R 7b -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl] And, The prodrugs of formula (Ia) and (Ib) are optionally further substituted: This is achieved by a human growth hormone (hGH) polymer prodrug or a pharmaceutically acceptable salt thereof. DETAILED DESCRIPTION OF THE INVENTION

[0015] It has now surprisingly been found that the hGH polymer prodrugs of the present invention exhibit a variety of unexpected properties.

[0016] By reducing the amount of PEG per hGH moiety, it is expected that the amount of hGH equivalent that can be dissolved in pharmaceutical preparations with given viscosity will increase.However, compared with, for example, compound 36 of WO2009 / 133137A2, the prodrug of the present invention can improve the relative hGH concentration higher than that proportional to the reduction in PEG size.In other words, when hGH polymer prodrug is the hGH polymer prodrug of the present invention, the pharmaceutical preparation that contains hGH polymer prodrug with given viscosity can contain a relatively large amount of hGH compared with, for example, compound 36 of WO2009 / 133137A2.

[0017] This is advantageous because only limited volumes can be administered to patients to limit the pain associated with injectable drugs. Thus, the ability to administer more hGH per given injection volume opens up new patient populations, namely, patients with diseases that require larger hGH doses per injection, and patients with diseases that may only require moderate doses per unit weight, but are severe and therefore require more hGH equivalents.

[0018] It was also surprising to find that the hGH polymer prodrugs of the present invention also have surprising advantages with regard to their preparation: the purification of the hGH polymer prodrugs of the present invention can be carried out at loadings at least three times higher than, for example, compound 36 of WO2009 / 133137A2, without compromising the separation efficiency and product quality, thereby significantly reducing the number of purification steps required.

[0019] Furthermore, when the prodrugs of the present invention are contained in a dry pharmaceutical formulation, the dry pharmaceutical formulation can be reconstituted more quickly with less foam formation than, for example, compound 36 of WO2009 / 133137A2. Thus, reconstitution of the dry pharmaceutical formulation of the present invention saves time and ensures administration of the appropriate dosage.

[0020] Within the scope of the present invention, the terms are used in the following sense:

[0021] As used herein, the term "human growth hormone (hGH)" refers to an hGH polypeptide, preferably derived from a mammalian species, more preferably derived from humans and mammalian species, more preferably derived from humans and murine species, characterized by promoting growth and maintaining normal body composition, anabolism, and lipid metabolism during anagen, as well as all of their variants, analogs, orthologs, homologs, and derivatives, and fragments thereof. Preferably, the term "hGH" refers to the hGH polypeptide of SEQ ID NO: 1, as well as its variants, homologs, and derivatives that exhibit essentially the same biological activity, i.e., promoting growth and maintaining normal body composition, anabolism, and lipid metabolism during anagen. More preferably, the term "hGH" refers to the polypeptide of SEQ ID NO: 1.

[0022] SEQ ID NO: 1 has the following sequence: FPTIPLSRLFDNAMLRAHRLHQLAFDTYQEFEEAYIPKEQKYSFLQNPQTSLCFSESIPTPSNREETQQKSNLELLRISLLLIQSWLEPVQFLRSVFANSLVYGASDSNVYDLLKDLEEGIQTLMGRLEDGSPRTGQIFKQTYSKFDTNSHNDDALLKNYGLLYCFRKDMDKVETFLRIVQCRSVEGSCGF

[0023] As used herein, the term "hGH polypeptide variant" refers to a polypeptide derived from the same species but differing from a reference hGH polypeptide. Preferably, such a reference hGH polypeptide sequence is the sequence of SEQ ID NO: 1. Generally, differences are limited, such that the amino acid sequences of the reference and variants are closely similar overall and, in many regions, identical. Preferably, the hGH polypeptide variant is at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a reference hGH polypeptide, preferably the hGH polypeptide of SEQ ID NO: 1. A polypeptide having an amino acid sequence that is at least, e.g., 95%, "identical" to a reference amino acid sequence intends that the amino acid sequence of the reference polypeptide is identical to that of the reference sequence, except that the sequence of the reference polypeptide may contain up to five amino acid modifications per every 100 amino acids of the reference amino acid sequence. Such modifications of the reference sequence may occur at the amino (N-terminal) or carboxy-terminal (C-terminal) positions of the reference amino acid sequence, or anywhere between such terminal positions, interspersed individually among residues either in the reference sequence, or in one or more consecutive groups within the reference sequence. The interrogated sequence may be the entire amino acid sequence of the reference sequence, or may be any fragment designated as described herein. Preferably, the interrogated sequence is the sequence of SEQ ID NO: 1.

[0024] Such hGH polypeptide variants can be naturally occurring variants, such as naturally occurring allelic variants encoded by one of several alternative forms of hGH occupying a given genetic locus in a chromosome or organism, or isoforms encoded by naturally occurring splice variants derived from a single primary transcript. Alternatively, the hGH polypeptide variants can be variants that are not known to occur naturally and can be made using mutagenesis techniques known in the art.

[0025] It is known in the art that one or more amino acids can be deleted from the N-terminus or C-terminus of a biologically active peptide or protein without substantial loss of biological function.

[0026] It is also recognized by those skilled in the art that some amino acid sequences of hGH polypeptides can be changed without significantly affecting the structure or function of the protein.Such mutations include deletions, insertions, inversions, repeats and substitutions, which are selected according to general rules known in the art so as to have little effect on activity.For example, guidance on how to make silent amino acid substitutions phenotypically is provided in Bowie et al. (1990), Science 247:1306-1310, the entire contents of which are incorporated herein by reference, in which the authors indicate that there are two main approaches to studying tolerance to amino acid sequence changes.

[0027] The term hGH polypeptide also encompasses all hGH polypeptides encoded by hGH analogs, orthologs and / or species homologs.As used herein, the term "hGH analog" refers to the hGH of different, unrelated organisms that perform the same function in each organism, but do not originate from the ancestral structure that the ancestors of the organisms shared.Instead, similar hGHs have developed independently, and then subsequently evolved to perform the same or similar functions.In other words, similar hGH polypeptides are polypeptides that have completely different amino acid sequences but exhibit the same biological activity, i.e., promote growth during adolescence, and maintain normal body composition, anabolism, and lipid metabolism.

[0028] As used herein, the term "hGH ortholog" refers to hGH in two different species whose sequences are related to each other through a common homologue of hGH in an ancestral species, but have evolved to become divergent from each other.

[0029] As used herein, the term "hGH homolog" refers to hGH from different organisms that perform the same function in each organism and originate from an ancestral structure shared by the ancestors of the organisms. In other words, a homologous hGH polypeptide is a polypeptide with a completely similar amino acid sequence that exhibits the same biological activity, i.e., promotes growth during adolescence and maintains normal body composition, anabolism, and lipid metabolism. Preferably, an hGH polypeptide homolog can be defined as a polypeptide that exhibits at least 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to a reference hGH polypeptide, preferably the hGH polypeptide of SEQ ID NO: 1.

[0030] Thus, hGH polypeptides according to the present invention can, for example, be: (i) those in which at least one of the amino acid residues has been substituted with a conserved or non-conserved amino acid residue, preferably a conserved amino acid residue, and such substituted amino acid residue may or may not be one encoded by the genetic code; and / or (ii) those in which at least one of the amino acid residues includes a substituent group; and / or (iii) those in which the hGH 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) those in which additional amino acids have been fused to the hGH polypeptide, such as an IgG Fc fusion region peptide, or a leader or secretory sequence, or a sequence used for purifying the polypeptide in the above forms, or a preprotein sequence.

[0031] The hGH polypeptide may be a monomer or a multimer. The multimer may be a dimer, trimer, tetramer, or a multimer comprising at least five monomeric polypeptide units. The multimer may also be a homodimer or heterodimer. The multimers of the present invention may be the result of hydrophobic, hydrophilic, ionic, and / or covalent association and / or may be indirectly linked, for example, by the formation of a liposome. Preferably, the hGH polypeptide is monomeric.

[0032] As used herein, the term "hGH polypeptide fragment" refers to any peptide or polypeptide that comprises a contiguous span of a portion of the amino acid sequence of an hGH polypeptide, preferably the polypeptide of SEQ ID NO:1.

[0033] More specifically, an hGH polypeptide fragment comprises at least 6, preferably at least 8 or 10, more preferably at least 12, 15, 20, 25, 30, 35, 40, 50, 60, 75, 100, 125, 150, 175, or 191 consecutive amino acids of an hGH polypeptide, more preferably the polypeptide of SEQ ID NO: 1. hGH polypeptide fragments are further described as a subgenus of hGH 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 an hGH polypeptide, preferably the polypeptide of SEQ ID NO: 1. Further included are species of hGH polypeptide fragments that are at least 6 amino acids in length as described above, further specified with respect to N- and C-terminal positions. Similarly, all of the hGH polypeptide fragments that are at least 6 amino acids in length as described above, which may be specified by N- and C-terminal positions, are encompassed as individual species by the term "hGH polypeptide fragment." That is, for any given amino acid sequence of an hGH polypeptide, preferably an hGH polypeptide of SEQ ID NO: 1, each combination of N-terminal and C-terminal positions that can be occupied by a fragment at least 6 consecutive amino acid residues in length is encompassed by the present invention.

[0034] It is noted that the above species of polypeptide fragments can alternatively be described by the formula "a to b," where "a" equals the position of the most amino acid in the polynucleotide that will be the N-terminus, "b" equals the position of the most amino acid that will be the C-terminus, and further, "a" equals an integer between 1 and the number of amino acids in the hGH polypeptide sequence minus 6, "b" equals an integer between 7 and the number of amino acids in the hGH polypeptide sequence, and "a" is preferably an integer less than "b" by at least 6 for the hGH polypeptide sequence of SEQ ID NO: 1.

[0035] The term "drug," as used herein, refers to a substance used in the treatment, cure, prevention, or diagnosis of disease, or otherwise used to enhance physical or mental well-being. When a drug is conjugated to another moiety, the portion of the resulting product that is derived from the drug is called a "biologically active moiety."

[0036] As used herein, the term "prodrug" refers to a biologically active moiety that is reversibly and covalently bound to a specific protecting group via a reversible prodrug linker moiety, which contains a reversible linking group to the biologically active moiety, in order to modify or eliminate undesirable properties in the parent molecule. This includes enhancing the desired properties of the drug and suppressing undesirable properties. Certain non-toxic protecting groups are called "carriers." A prodrug reversibly releases the biologically active moiety to which it is covalently bound in the form of its corresponding drug.

[0037] As used herein, the term "free form" of a drug means the drug in its unmodified, pharmacologically active form.

[0038] As used herein, the term "liquid formulation" refers to a formulation comprising an hGH polymer prodrug of the present invention and at least one solvent, the preferred solvent being water.

[0039] As used herein, the term "dried formulation" means that a formulation containing an hGH polymer prodrug of the present invention is provided in a dried form. Suitable methods for drying are spray drying and lyophilization, also known as freeze-drying. Such dried formulations containing an hGH polymer prodrug have a residual water content of up to 10%, preferably less than 5%, and more preferably less than 2%, as determined by Karl Fischer. A preferred method of drying is lyophilization. A "lyophilized formulation" means that a formulation containing an hGH polymer prodrug of the present invention is first frozen and then subjected to a reduction in water content by vacuum means. This term does not exclude additional drying steps that may be performed during manufacturing before filling the formulation into final containers.

[0040] As used herein, the term "reconstituted formulation" refers to the result of adding a solvent, also called a "reconstitution solution," to a dry formulation. Preferably, the amount of solvent is such that the dry formulation is completely dissolved in the resulting reconstituted formulation.

[0041] As used herein, the term "excipient" refers to a diluent, adjuvant, or vehicle with which a therapeutic agent is administered.

[0042] The term "water-soluble" as in "water-soluble portion" refers to a portion that dissolves in water at room temperature. Typically, a solution of the water-soluble portion will transmit at least about 75%, more preferably at least about 95%, of the light transmitted by the same solution after filtration. On a weight basis, the water-soluble portion or portion thereof will preferably be at least about 35% (by weight) water-soluble, more preferably at least about 50% (by weight) water-soluble, even more preferably about 70% (by weight) water-soluble, and even more preferably about 85% (by weight) water-soluble. However, it is most preferred that the water-soluble portion or portion thereof be about 95% (by weight) water-soluble or completely soluble in water.

[0043] As used herein, the term "hydrogel" refers to a hydrophilic or amphiphilic polymer network composed of homopolymers or copolymers that is insoluble due to the presence of covalent chemical crosslinks. The crosslinks provide the network structure and physical integrity. Hydrogels exhibit thermodynamic compatibility with water, allowing them to swell in aqueous media.

[0044] As used herein, the term "functional group" refers to a group of atoms capable of reacting with another functional group, including, but not limited to, the following groups: carboxylic acid (-(C=O)OH), primary or secondary amine (-NH, -NH-), maleimide, thiol (-SH), sulfonic acid (-(O=S=O)OH), carbonate, carbamate (-O(C=O)N<), hydroxy (-OH), aldehyde (-(C=O)H), ketone (-(C=O)-), hydrazine (>NN<), isocyanate, isothiocyanate, phosphate (-O(P=O)OHOH), phosphonate (-O(P=O)OHH), haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine, disulfide, vinyl sulfone, vinyl ketone, diazoalkane, oxirane, and aziridine.

[0045] As used herein, the term "moiety" refers to a portion of a molecule that lacks at least one atom compared to the corresponding reagent. For example, when a reagent of the formula "HXH" reacts with another reagent and becomes part of a reaction product, the corresponding moiety in the reaction product has the structure "HX-" or "-X-," where each "-" indicates a bond to another moiety. Thus, the biologically active moiety is released from the prodrug as a drug.

[0046] When an arrangement or chemical structure of atoms is presented that connects two moieties or interrupts a moiety, it is understood that the arrangement or chemical structure can connect the two moieties in either direction, unless expressly stated otherwise. For example, the moiety "-C(O)N(R)-" can connect two moieties or interrupt a moiety as either "-C(O)N(R)-" or "-N(R)C(O)-". Similarly, a moiety

[0047] [ka] can be joined into two parts, or

[0048] [ka] You can interrupt a section either as:

[0049] When the compounds according to formulas (Ia) and (Ib) contain one or more acidic or basic groups, the present invention also includes their corresponding pharmaceutically or toxicologically acceptable salts, in particular their pharmaceutically usable salts. Thus, compounds of formulas (Ia) and (Ib) containing acidic groups can be used according to the present invention, for example, as alkali metal salts, alkaline earth metal salts, or ammonium salts. More precise examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts, or salts with ammonia or organic amines (e.g., ethylamine, ethanolamine, triethanolamine, or amino acids). Compounds of formulas (Ia) and (Ib) containing one or more basic groups, i.e., groups that can be protonated, can exist and can be used according to the present invention in the form of addition salts with inorganic or organic acids. Examples of suitable acids include hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, and other acids known to those skilled in the art. Additional methods for converting basic groups into cations, such as alkylation of amine groups, to form suitable counterions for positively charged ammonium groups and their salts are known to those skilled in the art. When the compounds of formulas (Ia) and (Ib) contain both acidic and basic groups in the molecule, the present invention encompasses not only the salt forms described, but also internal salts or betaines (zwitterions). The respective salts according to formula (Ia) and (Ib) can be obtained by conventional methods known to those skilled in the art, such as, for example, by contacting them in a solvent or dispersion with an organic or inorganic acid or base, or by anion or cation exchange with other salts. The present invention also includes all salts of the compounds of formula (Ia) and (Ib) which are not directly suitable for use in medicines due to their low physiological compatibility, but which can be used, for example, as intermediates in chemical reactions or for preparing pharmaceutically acceptable salts.

[0050] The term "pharmaceutically acceptable" means approved by a regulatory body such as the EMA (Europe) and / or the FDA (USA), and / or any other national regulatory body, for use in animals, preferably in humans.

[0051] As used herein, the term "polymer" refers to a molecule comprising repeating structural units, i.e., monomers, linked by chemical bonds in a linear, cyclic, branched, cross-linked, or dendrimeric configuration, or a combination thereof, which may be of synthetic or biological origin, or a combination thereof. It is understood that a polymer can also include other moieties / moieties, such as, for example, one or more other chemical groups and / or one or more functional groups. Preferably, a soluble polymer has a molecular weight of at least 0.5 kDa, e.g., at least 1 kDa, at least 2 kDa, at least 3 kDa, or at least 5 kDa. If the polymer is soluble, it preferably has a molecular weight of at most 1000 kDa, such as at most 750 kDa, at most 500 kDa, at most 300 kDa, at most 200 kDa, at most 100 kDa, etc. It is understood that in the case of insoluble polymers, such as cross-linked hydrogels, a significant molecular weight range cannot be presented.

[0052] As used herein, the term "polymeric" means an agent or moiety that includes one or more polymers.

[0053] Those skilled in the art will understand that the polymerization products resulting from a polymerization reaction do not all have the same molecular weight, but rather exhibit a molecular weight distribution. Thus, as used herein, the molecular weight range, molecular weight, range of the number of monomers in a polymer, and number of monomers in a polymer refer to number average molecular weight and number average of monomers. As used herein, the term "number average molecular weight" refers to the ordinary arithmetic average of the molecular weights of the individual polymers.

[0054] As used herein, the term "PEG-based comprising at least X% PEG" in reference to a moiety or reagent means that the moiety or reagent comprises at least X% (w / w) ethylene glycol units (-CHCHO-), which may be in blocks, alternating arrangement, or randomly distributed within the moiety or reagent, preferably all of the ethylene glycol units of the moiety or reagent are present in one block, with the remaining weight percentage of the PEG-based moiety or reagent being other moieties preferably selected from the following moieties and linking groups: C 1~50 Alkyl, C 2~50 Alkenyl, C 2~50 Alkynyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl, and A linking group selected from the group including:

[0055] [ka] (In the formula, The dashed line indicates the bond to the remainder of the moiety or reagent; R and R a are independently selected from the group consisting of H, methyl, ethyl, propyl, butyl, pentyl and hexyl).

[0056] The term "substituted," as used herein, means that one or more -H atoms of a molecule or moiety are replaced with a different atom or group of atoms, referred to as "substituents."

[0057] Preferably, the one or more further optional substituents are halogen, —CN, —COOR x1 , -OR x1 , -C(O)R x1 , -C(O)N(R x1 R x1a ), -S(O)2N(R x1R x1a ), -S(O)N(R x1 R x1a ), -S(O)R x1 , -S(O)R x1 , -N(R x1 )S(O)2N(R x1a R x1b ), -SR x1 , -N(R x1 R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R x1a , -N(R x1 )S(O)R x1a , -N(R x1 )C(O)OR x1a , -N(R x1 )C(O)N(R x1a R x1b ), -OC(O)N(R x1 R x1a ), -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be substituted with one or more of the same or different R x2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(R x3 )C(O)N(Rx3a )- and -OC(O)N(R x3 )-, R x1 , R x1a , R x1b -H, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be substituted with one or more of the same or different R x2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-; -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(R x3 )C(O)N(R x3a )- and -OC(O)N(R x3 )-, T 0 are phenyl, naphthyl, indenyl, indanyl, tetralinyl, and C 3~10 independently selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; T 0 each independently represents one or more of the same or different R x2 and optionally replaced by R x2 are halogen, -CN, oxo (=O), and -COOR, respectively. x4, -OR x4 , -C(O)R x4 , -C(O)N(R x4 R x4a ), -S(O)2N(R x4 R x4a ), -S(O)N(R x4 R x4a ), -S(O)R x4 , -S(O)R x4 , -N(R x4 )S(O)2N(R x4a R x4b ), -SR x4 , -N(R x4 R x4a ), -NO2, -OC(O)R x4 , -N(R x4 )C(O)R x4a , -N(R x4 )S(O)2R x4a , -N(R x4 )S(O)R x4a , -N(R x4 )C(O)OR x4a , -N(R x4 )C(O)N(R x4a R x4b ), -OC(O)N(R x4 R x4a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more of the same or different halogens; R x3 , R x3a , R x4 , R x4a , R x4b are -H and C, respectively. 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more of the same or different halogens.

[0058] More preferably, the one or more further optional substituents are halogen, —CN, —COOR x1 , -OR x1 , -C(O)R x1 , -C(O)N(R x1 R x1a), -S(O)2N(R x1 R x1a ), -S(O)N(R x1 R x1a ), -S(O)R x1 , -S(O)R x1 , -N(R x1 )S(O)2N(R x1a R x1b ), -SR x1 , -N(R x1 R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R x1a , -N(R x1 )S(O)R x1a , -N(R x1 )C(O)OR x1a , -N(R x1 )C(O)N(R x1a R x1b ), -OC(O)N(R x1 R x1a ), -T 0 , C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl; 0 , C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Alkynyl may be substituted with one or more of the same or different R x2 is optionally replaced by C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(Rx3 )C(O)N(R x3a )- and -OC(O)N(R x3 )-, R x1 , R x1a , R x1b , R x3 , R x3a are -H, halogen, and C, respectively. 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl; T 0 are phenyl, naphthyl, indenyl, indanyl, tetralinyl, and C 3~10 independently selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; T 0 each independently represents one or more of the same or different R x2 and optionally replaced by R x2 are halogen, -CN, oxo (=O), and -COOR, respectively. x4 , -OR x4 , -C(O)R x4 , -C(O)N(R x4 R x4a ), -S(O)2N(R x4 R x4a ), -S(O)N(R x4 R x4a ), -S(O)R x4 , -S(O)R x4 , -N(R x4 )S(O)2N(R x4a R x4b ), -SR x4 , -N(R x4 R x4a ), -NO2, -OC(O)R x4 , -N(R x4 )C(O)R x4a , -N(R x4 )S(O)2R x4a , -N(R x4 )S(O)R x4a , -N(R x4 )C(O)ORx4a , -N(R x4 )C(O)N(R x4a R x4b ), -OC(O)N(R x4 R x4a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more of the same or different halogens; R x4 , R x4a , R x4b are -H, halogen, and C, respectively. 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl.

[0059] Even more preferably, the one or more further optional substituents are halogen, —CN, —COOR x1 , -OR x1 , -C(O)R x1 , -C(O)N(R x1 R x1a ), -S(O)2N(R x1 R x1a ), -S(O)N(R x1 R x1a ), -S(O)R x1 , -S(O)R x1 , -N(R x1 )S(O)2N(R x1a R x1b ), -SR x1 , -N(R x1 R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R x1a , -N(R x1 )S(O)R x1a , -N(R x1 )C(O)OR x1a , -N(R x1 )C(O)N(R x1a R x1b ), -OC(O)N(R x1 R x1a ), -T0 , C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl; 0 , C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl may be substituted with one or more of the same or different R x2 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(R x3 )C(O)N(R x3a )- and -OC(O)N(R x3 )-, R x1 , R x1a , R x1b , R x2 , R x3 , R x3a are -H, halogen, and C, respectively. 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl; T 0 are phenyl, naphthyl, indenyl, indanyl, tetralinyl, and C 3~10 independently selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; T 0 each independently represents one or more of the same or different R x2 is optionally replaced by

[0060] Preferably, up to six -H atoms of an optionally substituted molecule or moiety are independently replaced by substituents, e.g., five -H atoms are independently replaced by substituents, four -H atoms are independently replaced by substituents, three -H atoms are independently replaced by substituents, two -H atoms are independently replaced by substituents, or one -H atom is replaced by a substituent.

[0061] The term "spacer" as used herein includes -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z1 )-, -S(O)2N(R z1 )-, -S(O)N(R z1 )-, -S(O)2-, -S(O)-, -N(R z1 )S(O)2N(R z1a )-, -S-, -N(R z1 )-, -OC(OR z1 )(R z1a )-, -N(R z1 )C(O)N(R z1a )-, -OC(O)N(R z1 )-, C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl, -T-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be substituted with one or more of the same or different R z2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z3 )-, -S(O)2N(R z3 )-, -S(O)N(R z3 )-, -S(O)2-, -S(O)-, -N(R z3 )S(O)2N(R z3a )-, -S-, -N(R z3)-, -OC(OR z3 )(R z3a )-, -N(R z3 )C(O)N(R z3a )- and -OC(O)N(R z3 )-, R z1 and R z1a -H, -T, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl, -T, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be substituted with one or more of the same or different R z2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z4 )-, -S(O)2N(R z4 )-, -S(O)N(R z4 )-, -S(O)2-, -S(O)-, -N(R z4 )S(O)2N(R z4a )-, -S-, -N(R z4 )-, -OC(OR z4 )(R z4a )-, -N(R z4 )C(O)N(R z4a )- and -OC(O)N(R z4 )-, T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, 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; each T is independently selected from the group consisting of one or more of the same or different R z2 and optionally replaced by R z2are halogen, -CN, oxo (=O), and -COOR, respectively. z5 , -OR z5 , -C(O)R z5 , -C(O)N(R z5 R z5a ), -S(O)2N(R z5 R z5a ), -S(O)N(R z5 R z5a ), -S(O)R z5 , -S(O)R z5 , -N(R z5 )S(O)2N(R z5a R z5b ), -SR z5 , -N(R z5 R z5a ), -NO2, -OC(O)R z5 , -N(R z5 )C(O)R z5a , -N(R z5 )S(O)2R z5a , -N(R z5 )S(O)R z5a , -N(R z5 )C(O)OR z5a , -N(R z5 )C(O)N(R z5a R z5b ), -OC(O)N(R z5 R z5a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more of the same or different halogens; R z3 , R z3a , R z4 , R z4a , R z5 , R z5a and R z5b are -H and C, respectively. 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more of the same or different halogens.

[0062] More preferably, the term "spacer" is selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(Rz1 )-, -S(O)2N(R z1 )-, -S(O)N(R z1 )-, -S(O)2-, -S(O)-, -N(R z1 )S(O)2N(R z1a )-, -S-, -N(R z1 )-, -OC(OR z1 )(R z1a )-, -N(R z1 )C(O)N(R z1a )-, -OC(O)N(R z1 )-, C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl refers to a moiety selected from the group consisting of -T-, -C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be substituted with one or more of the same or different R z2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z3 )-, -S(O)2N(R z3 )-, -S(O)N(R z3 )-, -S(O)2-, -S(O)-, -N(R z3 )S(O)2N(R z3a )-, -S-, -N(R z3 )-, -OC(OR z3 )(R z3a )-, -N(R z3 )C(O)N(R z3a )- and -OC(O)N(R z3 )-, R z1 and R z1a -H, -T, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl, -T, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50Alkynyl may be substituted with one or more of the same or different R z2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z4 )-, -S(O)2N(R z4 )-, -S(O)N(R z4 )-, -S(O)2-, -S(O)-, -N(R z4 )S(O)2N(R z4a )-, -S-, -N(R z4 )-, -OC(OR z4 )(R z4a )-, -N(R z4 )C(O)N(R z4a )- and -OC(O)N(R z4 )-, T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, 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; each T is independently selected from the group consisting of one or more of the same or different R z2 and optionally replaced by R z2 are halogen, -CN, oxo (=O), and -COOR, respectively. z5 , -OR z5 , -C(O)R z5 , -C(O)N(R z5 R z5a ), -S(O)2N(R z5 R z5a ), -S(O)N(R z5 R z5a ), -S(O)R z5 , -S(O)R z5 , -N(R z5 )S(O)2N(R z5a R z5b ), -SR z5 , -N(R z5 R z5a), -NO2, -OC(O)R z5 , -N(R z5 )C(O)R z5a , -N(R z5 )S(O)2R z5a , -N(R z5 )S(O)R z5a , -N(R z5 )C(O)OR z5a , -N(R z5 )C(O)N(R z5a R z5b ), -OC(O)N(R z5 R z5a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more of the same or different halogens; R z3 , R z3a , R z4 , R z4a , R z5 , R z5a and R z5b are -H and C, respectively. 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more of the same or different halogens.

[0063] Even more preferably, the term "spacer" is selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z1 )-, -S(O)2N(R z1 )-, -S(O)N(R z1 )-, -S(O)2-, -S(O)-, -N(R z1 )S(O)2N(R z1a )-, -S-, -N(R z1 )-, -OC(OR z1 )(R z1a )-, -N(R z1 )C(O)N(R z1a )-, -OC(O)N(R z1 )-, C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl refers to a moiety selected from the group consisting of -T-, -C1~20 Alkyl, C 2~20 Alkenyl and C 2~20 Alkynyl may be substituted with one or more of the same or different R z2 is optionally replaced by C 1~20 Alkyl, C 2~20 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z3 )-, -S(O)2N(R z3 )-, -S(O)N(R z3 )-, -S(O)2-, -S(O)-, -N(R z3 )S(O)2N(R z3a )-, -S-, -N(R z3 )-, -OC(OR z3 )(R z3a )-, -N(R z3 )C(O)N(R z3a )- and -OC(O)N(R z3 )-, R z1 and R z1a -H, -T, C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 alkynyl; T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 independently selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; R z2 are halogen and C, respectively. 1~6 independently selected from the group consisting of alkyl, R z3 , R z3a , R z4 , R z4a , R z5 , R z5a and R z5b are -H and C, respectively. 1~6 alkyl, C 1~6The alkyl is optionally substituted with one or more of the same or different halogens.

[0064] The term "interrupted" means that a group of atoms is inserted in a moiety between two carbon atoms, or, if the insertion is at one of the moieties' termini, between a carbon atom and a hydrogen atom. When a moiety is interrupted at one of its termini by a group of atoms, and the interrupted moiety is attached to a second moiety, it is understood that the interrupting group of atoms may also be positioned so as to be located between the last atom of the moiety and the first atom of the second moiety.

[0065] As used herein, the term "C 1~4 "Alkyl," alone or in combination, means a straight-chain or branched alkyl moiety having 1 to 4 carbon atoms. When present at the terminal end of a molecule, it is a straight-chain or branched C 1~4 Examples of alkyl are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. 1~4 When linked by alkyl, such C 1~4 Examples of alkyl groups are -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -CH(C2H5)-, and -C(CH3)2-. 1~4 Each hydrogen of an alkyl carbon may be optionally replaced by a substituent as defined above. 1~4 The alkyl may be interrupted by one or more moieties as defined below.

[0066] As used herein, the term "C 1~6 "Alkyl," alone or in combination, means a straight-chain or branched alkyl moiety having 1 to 6 carbon atoms. When present at the terminal end of a molecule, it is 1~6Examples 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 linked by alkyl groups, such as C 1~6 Examples of alkyl groups are -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -CH(C2H5)- and -C(CH3)2-. 1~6 Each hydrogen atom of the carbon may be optionally replaced by a substituent as defined above. 1~6 The alkyl may be interrupted by one or more moieties as defined below.

[0067] Therefore, "C 1~10 Alkyl," "C 1~20 Alkyl" or "C 1~50 "Alkyl" means an alkyl chain having 1 to 10, 1 to 20, or 1 to 50 carbon atoms, respectively; C 1~10 , C 1~20 or C 1~50 Each hydrogen atom of the carbon may be optionally replaced by a substituent as defined above, optionally C 1~10 Alkyl or C 1~50 The alkyl may be interrupted by one or more moieties as defined below.

[0068] As used herein, the term "C 2~6 "Alkenyl," alone or in combination, means a straight-chain or branched hydrocarbon moiety containing at least one carbon-carbon double bond having 2 to 6 carbon atoms. When present at the end of the molecule, examples are -CH=CH2, -CH=CH-CH3, -CH2-CH=CH2, -CH=CHCH2-CH3, and -CH=CH-CH=CH2. When two portions of the molecule are C 2~6 When linked by an alkenyl group, such C2~6 An example of an alkenyl is -CH=CH-. 2~6 Each hydrogen atom of the alkenyl moiety may be optionally replaced by a substituent as defined above. 2~6 The alkenyl may be interrupted by one or more moieties as defined below.

[0069] Hence the term "C 2~10 alkenyl," "C 2~20 alkenyl" or "C 2~50 The term "alkenyl," alone or in combination, refers to a straight-chain or branched hydrocarbon moiety containing at least one carbon-carbon double bond, having 2 to 10 carbon atoms, 2 to 20 carbon atoms, or 2 to 50 carbon atoms. 2~10 Alkenyl, C 2~20 Alkenyl or C 2~50 Each hydrogen atom of the alkenyl group may be optionally replaced by a substituent as defined above. 2~10 Alkenyl, C 2~20 Alkenyl or C 2~50 The alkenyl may be interrupted by one or more moieties as defined below.

[0070] As used herein, the term "C 2~6 "Alkynyl," alone or in combination, means a straight-chain or branched hydrocarbon moiety containing at least one carbon-carbon triple bond having 2 to 6 carbon atoms. When present at the end of the molecule, examples are -C≡CH, -CH2-C≡CH, CH2-CH2-C≡CH, and CH2-C≡C-CH3. When two parts of the molecule are connected by an alkynyl group, an example is -C≡C-. C 2~6 Each hydrogen atom of the alkynyl group may be optionally replaced by a substituent as defined above. Optionally, one or more double bonds may occur. Optionally, C 2~6 The alkynyl may be interrupted by one or more moieties as defined below.

[0071] Therefore, as used herein, the term "C 2~10 alkynyl", "C 2~20 alkynyl" and "C 2~50 The term "alkynyl," alone or in combination, refers to a straight-chain or branched hydrocarbon moiety containing at least one carbon-carbon triple bond, having from 2 to 10 carbon atoms, from 2 to 20 carbon atoms, or from 2 to 50 carbon atoms, respectively. 2~10 Alkynyl, C 2~20 Alkynyl or C 2~50 Each hydrogen atom of the alkynyl group may be optionally replaced by a substituent as defined above. Optionally, one or more double bonds may occur. Optionally, C 2~10 Alkynyl, C 2~20 Alkynyl or C 2~50 The alkynyl may be interrupted by one or more moieties as defined below.

[0072] As mentioned above, C 1~4 Alkyl, C 1~6 Alkyl, C 1~10 Alkyl, C 1~20 Alkyl, C 1~50 Alkyl, C 2~6 Alkenyl, C 2~10 Alkenyl, C 2~20 Alkenyl, C 2~50 Alkenyl, C 2~6 Alkynyl, C 2~10 Alkynyl, C 2~20 Alkenyl or C 2~50 Alkynyl is one or more of the following moieties:

[0073] [ka] (In the formula, The dashed line indicates the bond to the remainder of the moiety or reagent; R and R a may be optionally interrupted by H, methyl, ethyl, propyl, butyl, pentyl and hexyl).

[0074] As used herein, the term "C 3~10 "Cycloalkyl" means a cyclic alkyl chain having 3 to 10 carbon atoms, which can be saturated or unsaturated, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl. 3~10 Each hydrogen atom of a cycloalkyl carbon may be replaced by a substituent as defined above. 3~10 "Cycloalkyl" also includes bridged bicycles such as norbornane or norbornene.

[0075] The term "8- to 30-membered carbopolycyclyl" or "8- to 30-membered carbopolycycle" refers to a cyclic moiety consisting of two or more rings having 8 to 30 ring atoms, where two adjacent rings share at least one ring atom and can contain up to a maximum number of double bonds (aromatic or fully, partially, or unsaturated non-aromatic rings). Preferably, 8- to 30-membered carbopolycyclyl refers to a cyclic moiety consisting of two, three, four, or five rings, more preferably two, three, or four rings.

[0076] As used herein, the term "3- to 10-membered heterocyclyl" or "3- to 10-membered heterocycle" means a ring (aromatic or fully, partially, or unsaturated non-aromatic ring) having 3, 4, 5, 6, 7, 8, 9, or 10 ring atoms, which may include up to a maximum number of double bonds, in which at least one and up to four ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and the ring is connected to the remainder of the molecule via a carbon or nitrogen atom. Examples of 3-10 membered heterocycles include, but are not limited to, aziridine, oxirane, thiirane, azirine, oxirene, thiylene, azetidine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazole, isothiazol, thiazoline, thiadiazole, thiadiazoline, tetrahydrofuran, Examples include tetrahydrothiophene, pyrrolidine, imidazolidine, pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, thiadiazolidine, sulfolane, pyran, dihydropyran, tetrahydropyran, imidazolidine, pyridine, pyridazine, pyrazine, pyrimidine, piperazine, piperidine, morpholine, tetrazole, triazole, triazolidine, tetrazolidine, diazepane, azepine, and homopiperazine. Each hydrogen atom of the 3- to 10-membered heterocyclyl or 3- to 10-membered heterocyclic group may be replaced by a substituent defined below.

[0077] As used herein, the term "8- to 11-membered heterobicyclyl" or "8- to 11-membered heterobicycle" means a two-ring heterocyclic moiety having 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 (aromatic or fully, partially, or unsaturated non-aromatic rings), and wherein at least one ring atom and up to six ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and the ring is linked to the remainder of the molecule through a carbon or nitrogen atom. Examples of 8- to 11-membered heterobicycles are indole, indoline, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzisothiazole, benzimidazole, benzimidazoline, quinoline, quinazoline, dihydroquinazoline, quinoline, dihydroquinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, decahydroisoquinoline, tetrahydroisoquinoline, dihydroisoquinoline, benzazepine, purine, and pteridine. The term 8- to 11-membered heterobicycle also includes two-ring spiro structures such as 1,4-dioxa-8-azaspiro[4.5]decane or bridged heterocycles such as 8-aza-bicyclo[3.2.1]octane. Each hydrogen atom of the carbon of an 8- to 11-membered heterobicyclyl or 8- to 11-membered heterobicycle may be replaced by a substituent as defined below.

[0078] Similarly, the term "8-30 membered heteropolycyclyl" or "8-30 membered heteropoly ring" means a heterocyclic moiety of more than two rings, preferably three, four, or five rings, having 8 to 30 ring atoms, where two adjacent rings share at least one ring atom and can contain up to a maximum number of double bonds (aromatic or fully, partially, or unsaturated non-aromatic rings), where at least one ring atom and up to ten ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and the ring is linked to the remainder of the molecule via a carbon or nitrogen atom.

[0079] As used herein, "halogen" means fluoro, chloro, bromo, or iodo. It is generally preferred that the halogen is fluoro or chloro.

[0080] In general, the terms "comprise" or "comprising" also encompass "consist of" or "consisting of."

[0081] In a preferred embodiment, =Y in formula (Ia) and (Ib) 1 is =O.

[0082] In a preferred embodiment, -Y in formula (Ia) and (Ib) 2 - is -O-.

[0083] In a preferred embodiment, -Y in formula (Ia) and (Ib) 3 - is -O-.

[0084] In a preferred embodiment, -Y in formula (Ia) and (Ib) 4 -, -NR 5 -It is.

[0085] In a preferred embodiment, =Y in formula (Ia) and (Ib) 5 is =O.

[0086] In a preferred embodiment, n in formulas (Ia) and (Ib) is 0 or 1. Most preferably, n in formulas (Ia) and (Ib) is 0.

[0087] Preferably, R in formula (Ia) and (Ib) 1 is branched and comprises at least three polymeric moieties.

[0088] More preferably, R in formula (Ia) and (Ib) 1contains at least one branch point, preferably at least two branch points, and preferably at least three polymer chains based on PEG, each branch point being -N<, -CR 8 and R is preferably selected from the group consisting of < and >C<. 8 -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl may be substituted with one or more of the same or different R 9 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl is -C(O)O-, -O-, -C(O)-, -C(O)N(R 10 )-, -S(O)2N(R 10 )-, -S(O)N(R 10 )-, -S(O)2-, -S(O)-, -N(R 10 )S(O)2N(R 10a )-, -S-, -N(R 10 )-, -OC(OR 10 )(R 10a )-, -N(R 10 )C(O)N(R 10a )- and -OC(O)N(R 10 )- and optionally interrupted by R 9 , R 10 and R 10a -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl.

[0089] In one preferred embodiment, R of formula (Ia) and (Ib) 1 is the first branch point BP 1 and from this branch point there are at least two parts C 1 and C 2 is extended, and C 1 and C 2 At least one of the two branches is at least the second branch point BP 2and from this branch point there are at least two parts P 1 and P 2 More preferably, R 1 is the first branch point BP 1 From this branch point, two parts C 1 and C 2 is extended, and this part C 1 is the branch point BP 2 From this branch point, there are at least two parts P 1 and P 2 extends, and part C 2 is the third branch point BP 3 and from this branch point there are at least two parts P 3 and P 4 is extended.

[0090] In another preferred embodiment, R 1 is the first branch point BP 1 , the second branch point BP 2 and the third branch point BP 3 Part C containing 1 and at least one portion P 1 is BP 1 and at least one portion P 2 is BP 2 and at least one portion P 3 is BP 3 More preferably, R 1 is the first branch point BP 1 , the second branch point BP 2 , the third branch point BP 3 and the fourth branch point BP 4 Part C containing 1 and at least one portion P 1 is BP 1 and at least one portion P 2 is BP 2 and at least one portion P 3 is BP 3 and at least one portion P 4 is BP 4 It extends from

[0091] Preferably, BP 1 , B.P. 2 , B.P. 3 and BP 4 -CR 8 R is independently selected from -C<, -C< and -N<; 8 -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl may be substituted with one or more of the same or different R 9 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl is -C(O)O-, -O-, -C(O)-, -C(O)N(R 10 )-, -S(O)2N(R 10 )-, -S(O)N(R 10 )-, -S(O)2-, -S(O)-, -N(R 10 )S(O)2N(R 10a )-, -S-, -N(R 10 )-, -OC(OR 10 )(R 10a )-, -N(R 10 )C(O)N(R 10a )- and -OC(O)N(R 10 )- and optionally interrupted by R 9 , R 10 and R 10a -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl.

[0092] Preferably, C 1 and C 2 is C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be substituted with one or more of the same or different R11 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 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 )-, -T- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, 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; each -T- is independently selected from the group consisting of one or more of the same or different R 11 and optionally replaced by R 11 are halogen, -CN, oxo, (=O), and -COOR, respectively. 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 R 12a ), -S(O)2N(R 12 R 12a ), -S(O)N(R 12 R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a R 12b ), -SR 12 , -N(R 12 R 12a ), -NO2, -OC(O)R 12 , -N(R12 )C(O)R 12a , -N(R 12 )S(O)2R 12a , -N(R 12 )S(O)R 12a , -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a R 12b ), -OC(O)N(R 12 R 12a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more of the same or different halogens; R 12 , R 12a and R 12b are -H and C, respectively. 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 The alkynyl is optionally substituted with one or more of the same or different halogens.

[0093] Preferably, P 1 , P 2 , P 3 , P 4 are, independently of each other, polymer moieties, more preferably PEG-based chains comprising at least 40% PEG, even more preferably at least 50% PEG, even more preferably at least 60% PEG, even more preferably at least 70% PEG, even more preferably at least 80% PEG, even more preferably at least 90% PEG and most preferably at least 95% PEG.

[0094] Preferably, P 1 , P 2 , P 3 , P 4have, independently of each other, a molecular weight of at least 5 kDa, such as 7.5 kDa, 10 kDa, 12 kDa or 15 kDa.

[0095] In a preferred embodiment, -R 1 is the moiety of formula (II)

[0096] [ka] (In the formula, -BP 1 <, -BP 2 <, -BP 3 < is -N< and -C(R 8 )<, wherein: R 8 is H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, -P 1 , -P 2 , -P 3 , -P 4 are, independently of each other, PEG-based chains containing at least 40% PEG and having a molecular weight in the range of 8-12 kDa; -C 1 -, -C 2 -C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be substituted with one or more of the same or different R 9 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 10 )-, -S(O)2N(R 10 )-, -S(O)N(R 10 )-, -S(O)2-, -S(O)-, -N(R10 )S(O)2N(R 10a )-, -S-, -N(R 10 )-, -OC(OR 10 )(R 10a )-, -N(R 10 )C(O)N(R 10a )- and -OC(O)N(R 10 )-, T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, 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; each T is independently selected from the group consisting of one or more of the same or different R 9 and optionally independently substituted by R 9 are halogen, -CN, oxo (=O), and -COOR, respectively. 11 , -OR 11 , -C(O)R 11 , -C(O)N(R 11 R 11a ), -S(O)2N(R 11 R 11a ), -S(O)N(R 11 R 11a ), -S(O)R 11 , -S(O)R 11 , -N(R 11 )S(O)2N(R 11a R 11b ), -SR 11 , -N(R 11 R 11a ), -NO2, -OC(O)R 11 , -N(R 11 )C(O)R 11a , -N(R 11 )S(O)2R 11a , -N(R 11 )S(O)R 11a , -N(R 11 )C(O)OR 11a , -N(R 11 )C(O)N(R 11a R 11b), -OC(O)N(R 11 R 11a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more of the same or different halogens; R 10 , R 10a , R 11 , R 11a and R 11b are -H and C, respectively. 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more of the same or different halogens. Includes.

[0097] In a preferred embodiment, the BP of formula (II) 1 is -N<.

[0098] In a preferred embodiment, the BP of formula (II) 2 and BP 2 are both -CH<.

[0099] First branch point BP 1 It is advantageous if the binding sites of X and X are separated by no more than a certain number of atoms.

[0100] Preferably, critical distances in the prodrugs of the present invention are less than 60 atoms, more preferably less than 50 atoms, even more preferably less than 40 atoms, even more preferably less than 30 atoms, even more preferably less than 20 atoms, and most preferably less than 10 atoms.

[0101] The term "significant distance" refers to the distance between R 1 The first branch point BP included in 1 and the atom marked by an asterisk in formula (a), or, when the prodrug of the invention is of formula (Ib), R 1 The first branch point BP included in1 and the atom indicated by the asterisk in formula (b):

[0102] [ka] (where the dashed line indicates the bond to the remainder of the prodrug of formula (Ia) in case (a) and the bond to the remainder of the prodrug of formula (Ib) in case (b)).

[0103] In a preferred embodiment, C of formula (II) 1 and C 2 -O-, -C(O)N(R 10 )- and 3- to 10-membered heterocyclyl 1~50 alkyl, and the 3- to 10-membered heterocyclyl is substituted with at least one oxo (=O).

[0104] Most preferably, C of formula (II) 1 and C 2 is represented by formula (IIa)

[0105] [ka] (In the formula, The dashed lines indicated by asterisks represent the BP 1 indicates the bond to The unmarked dashed lines represent BPs, respectively. 2 or BP 3 indicates the bond to q1 is 1, 2, 3, 4, 5, 6, 7 or 8, preferably q1 is 4, 5, 6, 7 or 8, more preferably q1 is 5, 6 or 7, and most preferably q1 is 6; q2 is 1, 2, 3, 4 or 5, preferably q2 is 1, 2 or 3, most preferably q2 is 2; q3 is 1, 2, 3, 4, 5, 6, 7 or 8, preferably q3 is 2, 3, 4 or 5, more preferably q3 is 2, 3 or 4, and most preferably q3 is 3; q4 is 1, 2 or 3, and most preferably q4 is 1. is.

[0106] In a preferred embodiment, P of formula (II) 1 , P 2 , P 3 and P 4 are, independently of one another, of the formula (IIb)

[0107] [ka] (In the formula, The dashed line indicates R 1 The remainder, i.e., BP 2 or BP 3 indicates the bond to m is 0 or 1; p is an integer ranging from 180 to 270, more preferably from 200 to 250, even more preferably from 210 to 240, and most preferably from 220 to 240; q is selected from the group consisting of 1, 2, 3, 4, 5 and 6).

[0108] More preferably, -R 1 is the moiety of formula (IIc):

[0109] [ka] (In the formula, p1, p2, p3, p4 are independently integers in the range of 180 to 270, preferably 200 to 250, even more preferably 210 to 240 and most preferably 220 to 240).

[0110] In a preferred embodiment, -R 2 is selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. More preferably, -R in formula (Ib) 2is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. Even more preferably, -R in formula (Ib) 2 is selected from -H, methyl and ethyl. Most preferably, -R in formula (Ib) 2 is -H.

[0111] In a preferred embodiment, -R 3 is selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. More preferably, -R in formula (Ia) and (Ib) 3 is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. Even more preferably, -R in formula (Ia) and (Ib) 3 is selected from -H, methyl and ethyl. Most preferably, -R in formula (Ia) and (Ib) 3 is -H.

[0112] In a preferred embodiment, -R 4 are each independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. More preferably, -R in formula (Ia) and (Ib) 4 is selected from the group consisting of methyl, ethyl, n-propyl and isopropyl. Even more preferably, -R in formula (Ia) and (Ib) 4 is selected from methyl and ethyl.

[0113] In a preferred embodiment, -R 5 is selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. More preferably, -R in formula (Ia) and (Ib) 5 is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. Even more preferably, -R in formula (Ia) and (Ib) 5is selected from methyl and ethyl. Most preferably, -R in formula (Ia) and (Ib) 5 is methyl.

[0114] In a preferred embodiment, -R 6 and -R 6a is independently selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. More preferably, -R in formula (Ia) and (Ib) 6 and -R 6a is independently selected from the group consisting of -H, methyl, ethyl, n-propyl, and isopropyl. Even more preferably, -R in formula (Ia) and (Ib) 6 and -R 6a is independently selected from -H, methyl and ethyl. Most preferably, -R in formula (Ia) and (Ib) 6 and -R 6a are both -H.

[0115] In a preferred embodiment, X in formula (Ia) and (Ib) is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z1 )-, -S(O)2N(R z1 )-, -S(O)N(R z1 )-, -S(O)2-, -S(O)-, -N(R z1 )S(O)2N(R z1a )-, -S-, -N(R z1 )-, -OC(OR z1 )(R z1a )-, -N(R z1 )C(O)N(R z1a )-, -OC(O)N(R z1 )-, C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl, -T-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be substituted with one or more of the same or different R z2 is optionally replaced by C1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z3 )-, -S(O)2N(R z3 )-, -S(O)N(R z3 )-, -S(O)2-, -S(O)-, -N(R z3 )S(O)2N(R z3a )-, -S-, -N(R z3 )-, -OC(OR z3 )(R z3a )-, -N(R z3 )C(O)N(R z3a )- and -OC(O)N(R z3 )-, R z1 and R z1a -H, -T, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl, -T, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 The alkynyl is optionally substituted by one or more identical or different Rz2, and C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z4 )-, -S(O)2N(R z4 )-, -S(O)N(R z4 )-, -S(O)2-, -S(O)-, -N(R z4 )S(O)2N(R z4a )-, -S-, -N(R z4 )-, -OC(OR z4 )(R z4a )-, -N(R z4 )C(O)N(R z4a )- and -OC(O)N(R z4 )-, T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, 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; each T is independently selected from the group consisting of one or more of the same or different R z2 and optionally replaced by R z2 are halogen, -CN, oxo (=O), and -COOR, respectively. z5 , -OR z5 , -C(O)R z5 , -C(O)N(R z5 R z5a ), -S(O)2N(R z5 R z5a ), -S(O)N(R z5 R z5a ), -S(O)R z5 , -S(O)R z5 , -N(R z5 )S(O)2N(R z5a R z5b ), -SR z5 , -N(R z5 R z5a ), -NO2, -OC(O)R z5 , -N(R z5 )C(O)R z5a , -N(R z5 )S(O)2R z5a , -N(R z5 )S(O)R z5a , -N(R z5 )C(O)OR z5a , -N(R z5 )C(O)N(R z5a R z5b ), -OC(O)N(R z5 R z5a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more of the same or different halogens; R z3 , R z3a , R z4 , R z4a , R z5 , Rz5a and R z5b are -H and C, respectively. 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more of the same or different halogens.

[0116] More preferably, X in formula (Ia) and (Ib) is C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 alkynyl, C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Alkynyl may be substituted with one or more of the same or different R z2 is optionally replaced by C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R z3 )-, -S(O)2N(R z3 )-, -S(O)N(R z3 )-, -S(O)2-, -S(O)-, -N(R z3 )S(O)2N(R z3a )-, -S-, -N(R z3 )-, -OC(OR z3 )(R z3a )-, -N(R z3 )C(O)N(R z3a )- and -OC(O)N(R z3 )-, T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, 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; each T is independently selected from the group consisting of one or more of the same or different R z2 and optionally replaced by R z2 are respectively, C 1~6 alkyl, and C1~6 the alkyl is optionally substituted with one or more of the same or different halogens; R z3 , R z3a are -H and C, respectively. 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more of the same or different halogens.

[0117] Even more preferably, X in formula (Ia) and (Ib) is —C(O)O—, —O—, —C(O)—, —C(O)N(R z3 )-, -S-, -N(R z3 )-, -OC(OR z3 )(R z3a )- and -OC(O)N(R z3 C optionally interrupted by one or more groups selected from the group consisting of 1~10 is alkyl, R z3 , R z3 are -H and C, respectively. 1~6 alkyl.

[0118] Most preferably, X in formula (Ia) and (Ib) is a group of formula (III)

[0119] [ka] (In the formula, The dashed lines indicated by asterisks are 1 indicates the bond to The unmarked dashed line indicates the bond to the remainder of the prodrug; q5 is 1, 2, 3, 4, 5, 6, 7 or 8, preferably q5 is 1, 2, 3, 4 or 5, more preferably q5 is 2, 3 or 4, and most preferably q5 is 3).

[0120] Preferably, Ar in formula (Ia) and (Ib) is phenyl. Most preferably, Ar in formula (Ia) and (Ib) is

[0121] [ka] where the dashed line indicates the bond to the remainder of the prodrug of formula (Ia) or (Ib).

[0122] Preferably, W in formula (Ia) and (Ib) is C 3~10 Cycloalkyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 C optionally interrupted by )- 1~20 Even more preferably, W in formula (Ia) and (Ib) is C 3~10 Cycloalkyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 C optionally interrupted by )- 1~10 Even more preferably, W in formula (Ia) and (Ib) is C 3~10 Cycloalkyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 C optionally interrupted by )- 1-6 Most preferably, W in formula (Ia) and (Ib) is

[0123] [ka] (In the formula, The dashed line indicates the bond to the rest of the molecule).

[0124] Preferably, -Nu in formula (Ia) and (Ib) is -N(R 7 R 7a )

[0125] Preferably, -R in formula (Ia) and (Ib) 7 and -R 7aare independently selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. More preferably, -R in formula (Ia) and (Ib) 7 and -R 7a are independently selected from -H, methyl, ethyl, n-propyl and isopropyl. Even more preferably, -R in formula (Ia) and (Ib) 7 and -R 7a are each independently selected from methyl or ethyl. Most preferably, -R in formula (Ia) and (Ib) 7 and -R 7a Both are methyl.

[0126] Most preferably, the hGH polymer prodrugs of the present invention have the formula (IV):

[0127] [ka] (In the formula, D is the hGH moiety that is attached to the rest of the molecule via the amine functionality; p1, p2, p3, and p4 are independently integers ranging from 180 to 270, preferably from 200 to 250, even more preferably from 210 to 240, and most preferably from 220 to 240.

[0128] Most preferably, D in formula (IV) is attached to the remainder of the molecule via the amine provided by the lysine side chain.

[0129] Another aspect of the present invention is a pharmaceutical formulation comprising at least one hGH polymer prodrug of the present invention and at least one excipient. Preferably, at least one hGH polymer prodrug of the present invention is of formula (IV).

[0130] In one embodiment, the pharmaceutical formulation is a liquid formulation comprising at least one hGH polymer prodrug of the present invention and at least one excipient. Preferably, the at least one hGH polymer prodrug of the present invention is of formula (IV).

[0131] Preferably, such liquid formulations contain 3-300 mg / mL of the hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV) (corresponding to 1-100 mg of hGH equivalents / mL). More preferably, the liquid formulations contain 9-150 mg / mL of the hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV) (corresponding to 3-50 mg of hGH equivalents / mL). Even more preferably, the liquid formulations contain 15-120 mg / mL of the hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV) (corresponding to 5-40 mg of hGH equivalents / mL). Even more preferably, the liquid formulation contains 30-45 mg / mL of an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of Formula (IV), corresponding to 10-15 mg of hGH equivalents / mL, or equivalently, preferably, the liquid formulation contains 75-105 mg / mL of an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of Formula (IV), corresponding to 25-30 mg of hGH equivalents / mL. In particularly preferred embodiments thereof, the liquid formulation contains 42 or 84 mg / mL of an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of Formula (IV), corresponding to 14 or 28 mg of hGH equivalents / mL.

[0132] The liquid formulation of the hGH polymer prodrug according to the present invention may contain one or more excipients. Excipients used in parenteral formulations can be classified as, for example, buffers, tonicity adjusters, preservatives, stabilizers, anti-adsorption agents, oxidation protectants, viscosifiers / viscosity enhancers, or other adjuvants. However, in some cases, one excipient may have two or three functions. The liquid formulation may contain one or more of the following excipients: (i) Buffering agents: physiologically acceptable buffers that maintain the pH in the desired range (sodium phosphate, sodium bicarbonate, sodium succinate, sodium histidine, sodium citrate, sodium acetate, sodium sulfate, sodium nitrate, sodium chloride, sodium pyruvate, etc.). Antacids such as Mg(OH)2 or ZnCO3 can also be used. (ii) Tonicity adjusters: To minimize pain that may result from cell damage due to differences in osmotic pressure in the injectable depot. Examples are glycerin and sodium chloride. Effective concentrations can be determined by osmometry using an estimated osmolality of 285-315 mOsmol / kg for serum. (iii) Preservatives and / or Antimicrobials: Multi-dose parenteral formulations must contain preservatives in concentrations sufficient to minimize the patient's risk of infection upon injection, and corresponding regulatory requirements have been established. Typical preservatives include m-cresol, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, chlorobutanol, benzyl alcohol, phenylmercuric nitrate, thimerosal, sorbic acid, potassium sorbate, benzoic acid, chlorocresol, and benzalkonium chloride. (iv) Stabilizers: Stabilization can be achieved by enhancing the protein stabilizing power, destabilizing the denatured state, or directly binding an excipient to the protein. Stabilizers can be amino acids such as alanine, arginine, aspartic acid, glycine, histidine, lysine, and proline; sugars such as glucose, sucrose, and trehalose; polyols such as glycerol, mannitol, and sorbitol; salts such as potassium phosphate and sodium sulfate; chelating agents such as EDTA and hexaphosphate; ligands such as divalent metal ions (zinc, calcium, and the like); other salts; or organic molecules such as phenol derivatives. Additionally, oligomers or polymers such as cyclodextrin, dextran, dendrimers, PEG, PVP, protamine, or HSA can be used.

[0133] (v) Anti-adsorption agents: Ionic or non-ionic surfactants or other proteins or soluble polymers are primarily used to competitively coat or adsorb to the inner surface of the formulation container. Examples include poloxamer (Pluronic F-68), PEG dodecyl ether (Brij35), polysorbate 20 and 80, dextran, polyethylene glycol, PEG-polyhistidine, BSA and HSA, and gelatin. The concentration and type of excipient selected depend on the effect to be avoided, but typically a surfactant monolayer forms at the interface just above the CMC value. (vi) Oxidative protectants: Antioxidants such as ascorbic acid, ectoine, methionine, glutathione, monothioglycerol, morin, polyethyleneimine (PEI), propyl gallate, and vitamin E, and chelating agents such as citric acid, EDTA, hexaphosphate, and thioglycolic acid can also be used. (vii) Spreading or Diffusing Agents: Modifies the permeability of connective tissue by hydrolysis of components of the extracellular matrix in the interstitial space, such as, but not limited to, hyaluronic acid, a polysaccharide found in the intercellular spaces of connective tissue. Spreading agents, such as, but not limited to, hyaluronidase, temporarily reduce the viscosity of the extracellular matrix, facilitating the diffusion of injected drugs. (viii) Other adjuvants: wetting agents, viscosity modifiers, antibiotics, hyaluronidase, etc. Acids and bases such as hydrochloric acid and sodium hydroxide are necessary adjuvants for pH adjustment during production.

[0134] Liquid formulations of hGH polymer prodrugs according to the present invention contain one or more buffering agents. Buffers with sufficient pharmaceutically acceptable buffering capacity in the desired pH range are preferred. In a preferred embodiment, the buffering agent is selected from the group consisting of sodium phosphate, sodium bicarbonate, sodium succinate, sodium histidine, sodium citrate, and sodium acetate. Most preferably, the buffering agent is a succinate salt. Typically, the pH is adjusted using succinic acid at a concentration of 5-50 mM, more preferably 10 mM, and titrating the solution to the desired pH with Tris base, more preferably a 1 molar Tris base solution.

[0135] In a preferred embodiment, the pH of the liquid formulation of the present invention ranges from pH 1 to pH 10, more preferably from pH 3 to pH 7, even more preferably from pH 4 to pH 6, even more preferably from pH 4.5 to pH 5.5, and most preferably has a pH of 5.0. Preferably, the buffer concentration and pH are selected to minimize release of hGH during storage and to minimize deamidation, aggregation, and precipitation of hGH.

[0136] Preferably, the liquid formulations of the hGH polymer prodrugs of the present invention contain one or more oxidation protection agents, such as antioxidants or chelating agents. A preferred antioxidant is methionine.

[0137] In one embodiment, a liquid formulation of the present invention comprises trehalose.

[0138] In one embodiment, the liquid formulations of the present invention include one or more preservatives and / or antimicrobial agents, such as, for example, benzyl alcohol and / or cresol.

[0139] In one embodiment, the liquid formulation of the present invention comprises an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV), an oxidative protectant and a buffering agent, and even more preferably an hGH polymer prodrug of formula (IV), an oxidative protectant, a stabilizer and a buffering agent.

[0140] Preferably, the liquid formulations of the present invention comprise an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV), methionine and succinate, and even more preferably an hGH polymer prodrug of formula (IV), methionine, succinate and trehalose (optionally as a dihydrate).

[0141] Optionally, the liquid formulations of the present invention also include benzyl alcohol and / or cresol.

[0142] Preferably, the liquid formulation of the present invention comprises: hGH polymer prodrug 3~300mg / ml Succinic acid 5-50mM Optionally, trehalose dihydrate 25-150mg / ml Optionally, methionine 1-50mM Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0, titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 1 to 100 mg of hGH equivalents / ml.

[0143] More preferably, the liquid formulation of the present invention comprises: hGH polymer prodrug 3~300mg / ml Succinic acid 5-50mM Optionally, trehalose dihydrate 50-90mg / ml Optionally, methionine 1-50mM Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0, titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 1 to 100 mg of hGH equivalents / m.

[0144] More preferably, the liquid formulation of the present invention comprises: hGH polymer prodrug 9~150mg / ml Succinic acid 5-50mM Optionally, trehalose dihydrate 50-90mg / ml Optionally, methionine 1-50mM Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0, titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 1 to 100 mg of hGH equivalents / ml.

[0145] Even more preferably, the liquid formulation of the present invention comprises: hGH polymer prodrug 15~120mg / ml Succinic acid 5~40mM Optionally, trehalose dihydrate 60-86mg / ml Optionally, methionine 5-40mM Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0, titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 5 to 40 mg of hGH equivalents / ml.

[0146] Even more preferably, the liquid formulation of the present invention comprises: hGH polymer prodrug 30~45mg / ml Succinic acid 5-20mM Optionally, trehalose dihydrate 75-86mg / ml Optionally, methionine 5-20mM Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 10 to 15 mg of hGH equivalents / ml.

[0147] In an equally preferred embodiment, the liquid formulation of the present invention comprises: hGH polymer prodrug 75~105mg / ml Succinic acid 5-20mM Optionally, trehalose dihydrate 60-81mg / ml Optionally, methionine 5-20mM Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 25 to 35 mg of hGH equivalents / ml.

[0148] In a preferred embodiment, a liquid formulation comprising at least one hGH polymer prodrug of the present invention, preferably of formula (IV), is hGH polymer prodrug 42mg / ml Succinic acid 10mM Optionally, trehalose dihydrate 79-86mg / ml Optionally, methionine 10mM Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably a polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of hGH polymer prodrug corresponds to 14 mg of hGH equivalents / ml.

[0149] In another preferred embodiment, the liquid formulation of the present invention comprising at least one polymeric prodrug, preferably of formula (IV), comprises: hGH polymer prodrug 84mg / ml Succinic acid 10mM Optionally, trehalose dihydrate 70-80mg / ml Optionally, methionine 10mM Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of hGH polymer prodrug corresponds to 28 mg of hGH equivalents / ml.

[0150] In one embodiment, the liquid formulations of the present invention comprise at least one additional bioactive agent in its free form or as a prodrug, preferably selected from the group consisting of IGF-1, ghrelin and ghrelin-like compounds, gonadotropin-releasing hormone agonists, gonadotropin-releasing hormone analogs, growth hormone-releasing factor, growth hormone-releasing factor analogs, gonadal steroids, antiandrogens, nonsteroidal aromatase inhibitors, HIV combination therapies, free fatty acid regulators, anabolic steroids, estrogen agonists and antagonists, propranolol, appetite suppressants, osteoporosis medications (including bisphosphonates, bone-building agents, estrogens, parathyroid hormone, selective receptor modulators, and / or antidiabetic agents, such as insulin, thiazolidinediones, sulfonylureas, incretin mimetics, meglitinides, biguanides, alpha-glucosidase inhibitors, and amylin analogs). Preferably, the at least one additional bioactive agent is in its free form.

[0151] In another embodiment, the pharmaceutical formulation of the present invention is a dry formulation.

[0152] Preferably, such dry pharmaceutical formulations comprise 1-99.9% (w / w), more preferably 1.9-89% (w / ), even more preferably 3-83% (w / w), even more preferably 9.0-71% (w / w), even more preferably 15-63% (w / w), even more preferably 26-36% (w / w) or 48-62% (w / w), and most preferably 32-34% (w / w) or 50-54% (w / w) of the hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of Formula (IV).

[0153] Preferably, the dry pharmaceutical formulation of the present invention comprises at least one lyoprotectant, preferably selected from the group consisting of amino acids, methylamines, lyotropic salts, polyols, propylene glycol, polyethylene glycol, pluronics, hydroxyalkyl starches, and combinations thereof.

[0154] When the lyoprotectant is an amino acid, it is preferably selected from the group consisting of monosodium glutamate and histidine.

[0155] When the lyoprotectant is a polyol, it is preferably selected from the group consisting of sucrose, trehalose, glycerin, erythritol, glycerol, arabitol, xylitol, sorbitol and mannitol.

[0156] When the lyoprotectant is a methylamine, it is preferably a betaine.

[0157] When the lyoprotectant is a lyotropic salt, it is preferably magnesium sulfate.

[0158] When the lyoprotectant is a hydroxyalkyl starch, it is preferably a hydroxyethyl starch.

[0159] In a preferred embodiment, the lyoprotectant is a non-reducing sugar. Even more preferably, the lyoprotectant is trehalose or sucrose. Most preferably, the lyoprotectant is trehalose.

[0160] Preferably, the dry pharmaceutical formulation of the present invention comprises 8-97% (w / w), more preferably 14-96% (w / w), even more preferably 24-90% (w / w), even more preferably 32-84% (w / w), even more preferably 60-73% (w / w) or 35-52% (w / w), and most preferably 64-66% (w / w) or 45-48% (w / w) of at least one lyoprotectant, preferably trehalose dihydrate.

[0161] Preferably, the dry formulation of the present invention comprises at least one buffering agent. Preferably, the buffering agent is selected from the group consisting of sodium phosphate, sodium bicarbonate, sodium succinate, sodium histidine, sodium citrate, and sodium acetate. Most preferably, the buffering agent is succinate. Preferably, the pH is adjusted using succinic acid at a concentration of 5-50 mM, more preferably 10 mM, and titrating the solution to the desired pH with Tris base, more preferably with a 1 molar Tris base solution.

[0162] Preferably, the dry formulation comprises: (a) hGH polymer prodrug 3-300 mg / ml Succinic acid 5-50mM Optionally, trehalose dihydrate 25-150mg / ml A liquid formulation comprising: providing a liquid formulation having a pH in the range of pH 4.0 to pH 6.0 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, wherein the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV); (b) drying the liquid formulation of step (a). The compound is obtained by a method comprising:

[0163] More preferably, the liquid formulation of step (a) comprises: hGH polymer prodrug 3~300mg / ml Succinic acid 5-50mM Optionally, trehalose dihydrate 50-90mg / ml Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the invention, preferably an hGH polymer prodrug of formula (IV).

[0164] More preferably, the liquid formulation of step (a) comprises: hGH polymer prodrug 9~150mg / ml Succinic acid 5-50mM Optionally, trehalose dihydrate 50-90mg / ml Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the invention, preferably an hGH polymer prodrug of formula (IV).

[0165] Even more preferably, the liquid formulation of step (a) comprises hGH polymer prodrug 15~120mg / ml Succinic acid 5~40mM Optionally, trehalose dihydrate 60-86mg / ml Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the invention, preferably an hGH polymer prodrug of formula (IV).

[0166] Even more preferably, the liquid formulation of step (a) comprises hGH polymer prodrug 30~45mg / ml Succinic acid 5-20mM Optionally, trehalose dihydrate 75-86mg / ml Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the invention, preferably an hGH polymer prodrug of formula (IV).

[0167] Even more preferably, the liquid formulation of step (a) comprises hGH polymer prodrug 75~105mg / ml Succinic acid 5-20mM Optionally, trehalose dihydrate 60-81mg / ml Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the invention, preferably an hGH polymer prodrug of formula (IV).

[0168] Most preferably, the liquid formulation of step (a) comprises: hGH polymer prodrug 42mg / ml Succinic acid 10mM Optionally, trehalose dihydrate 79-86mg / ml Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the invention, preferably an hGH polymer prodrug of formula (IV).

[0169] In an equally preferred embodiment, the liquid formulation of step (a) comprises: hGH polymer prodrug 84mg / ml Succinic acid 10mM Optionally, trehalose dihydrate 70-80mg / ml Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and the hGH polymer prodrug is an hGH polymer prodrug of the invention, preferably an hGH polymer prodrug of formula (IV).

[0170] Preferably, in step (b), the liquid formulation is dried by lyophilization.

[0171] In one embodiment, the formulation of step (a) comprises at least one additional bioactive agent in its free form or as a prodrug, the at least one additional bioactive agent being selected from the group consisting of IGF-1, ghrelin and ghrelin-like compounds, gonadotropin-releasing hormone agonists, gonadotropin-releasing hormone analogs, growth hormone-releasing factor, growth hormone-releasing factor analogs, gonadal steroids, antiandrogens, nonsteroidal aromatase inhibitors, HIV combination therapies, free fatty acid regulators, anabolic steroids, estrogen agonists and antagonists, propranolol, appetite suppressants, osteoporosis medications (including bisphosphonates, bone-forming agents, estrogens, parathyroid hormone, selective receptor modulators, and / or antidiabetic agents (such as insulin, thiazolidinediones, sulfonylureas, incretin mimetics, meglitinides, biguanides, alpha-glucosidase inhibitors, and amylin analogs)). Preferably, the at least one additional bioactive agent is in its free form.

[0172] Another aspect of the present invention is a pharmaceutical composition comprising, based on the total weight of the formulation: hGH polymer prodrug 2~89%(w / w) Succinic acid 0.4~1.8%(w / w) Trehalose dihydrate 7~97%(w / w) Tris 0.4-2% (w / w) A dry formulation comprising: The hGH polymer prodrug is a dry formulation of an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV).

[0173] In a preferred embodiment, the dry formulation of the present invention comprises, based on the total weight of the formulation: hGH polymer prodrug 3~83%(w / w) Succinic acid 0.6~1.6%(w / w) Trehalose dihydrate 14~96%(w / w) Tris 0.6-1.7% (w / w) Including, The hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV).

[0174] In an even more preferred embodiment, the dry formulation of the present invention comprises 9.0-71% (w / w) hGH polymer prodrug, based on the total weight of the formulation. Succinic acid 0.6~2.8%(w / w) Trehalose dihydrate 24~90%(w / w) Tris 0.6-2.9% (w / w) Including, The hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV).

[0175] In an even more preferred embodiment, the dry formulation of the present invention comprises 15-63% (w / w) hGH polymer prodrug, based on the total weight of the formulation. Succinic acid 0.6~2.5%(w / w) Trehalose dihydrate 32~84%(w / w) Tris 0.6-2.6% (w / w) Including, The hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV).

[0176] In an even more preferred embodiment, the dry formulation of the present invention comprises 26-36% (w / w) hGH polymer prodrug, based on the total weight of the formulation. Succinic acid 0.5-1.9% (w / w) Trehalose dihydrate 60-73% (w / w) Tris 0.5-1.9% (w / w) Including, The hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV).

[0177] In an equally preferred embodiment, the dry formulation of the present invention comprises, based on the total weight of the formulation: hGH polymer prodrug 48~62%(w / w) Succinic acid 0.4~1.4%(w / w) Trehalose dihydrate 35~52%(w / w) Tris 0.4-1.4% (w / w) Including, The hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV).

[0178] Most preferably, the dry formulation of the present invention comprises, based on the total weight of the formulation: hGH polymer prodrug 32-34% (w / w) Succinic acid 0.9-1.0% (w / w) Trehalose dihydrate 64~66%(w / w) Tris 0.5-1.4% (w / w) Including, The hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV).

[0179] In an equally preferred embodiment, the dry formulation of the present invention comprises, based on the total weight of the formulation: hGH polymer prodrug 50-54 (w / w) Succinic acid 0.7-0.8% (w / w) Trehalose dihydrate 45-48% (w / w) Tris 0.4-1.1% (w / w) Including, The hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV).

[0180] In one embodiment, the dry formulation of the present invention comprises at least one additional bioactive agent in its free form or as a prodrug, the at least one additional bioactive agent being selected from the group consisting of IGF-1, ghrelin and ghrelin-like compounds, gonadotropin-releasing hormone agonists, gonadotropin-releasing hormone analogs, growth hormone-releasing factor, growth hormone-releasing factor analogs, gonadal steroids, antiandrogens, nonsteroidal aromatase inhibitors, HIV combination therapies, free fatty acid regulators, anabolic steroids, estrogen agonists and antagonists, propranolol, appetite suppressants, osteoporosis medications (including bisphosphonates, bone-building agents, estrogens, parathyroid hormone, selective receptor modulators, and / or antidiabetic agents, such as insulin, thiazolidinediones, sulfonylureas, incretin mimetics, meglitinides, biguanides, alpha-glucosidase inhibitors, and amylin analogs). Preferably, the at least one additional bioactive agent is in its free form.

[0181] Preferably, the dry formulation of the present invention is obtained by lyophilization.

[0182] Preferably, the dry formulation of the present invention is lyophilized in a vial, syringe, dual-chamber syringe, ampoule, cartridge or dual-chamber cartridge.

[0183] A preferred vial is a glass vial.

[0184] In one embodiment, the dry formulation of the present invention is lyophilized in a cartridge for use in a pen injector.

[0185] In another embodiment, the dry formulation is lyophilized in the first chamber of a dual chamber cartridge, and the second chamber of the cartridge is filled with the reconstitution solution.

[0186] Before administering the dry formulation of the present invention to a patient in need thereof, the dry formulation is reconstituted. Reconstitution can be carried out in a container (such as a vial, syringe, dual-chamber syringe, ampoule, cartridge, and dual-chamber cartridge) in which the dry formulation of the hGH polymer prodrug of the present invention is provided, or the dry formulation of the present invention can be transferred to a different container and then reconstituted.

[0187] Reconstitution is achieved by adding a predetermined amount of a reconstitution solution to the dry formulation, which is a sterile liquid such as water or a buffer, which may contain additional additives such as preservatives and / or antimicrobial agents.

[0188] In one embodiment, the reconstitution solution is sterile water containing 0.7-1.1% benzyl alcohol, more preferably 0.9% benzyl alcohol. In another embodiment, the reconstitution solution is sterile water containing 0.2-0.4% cresol, more preferably 0.3% cresol. Preferably, the reconstitution solution is sterile water.

[0189] Preferably, the pH of the reconstituted formulation of the present invention is in the range of pH 1 to pH 10, more preferably in the range of pH 3 to pH 7, even more preferably in the range of pH 4 to pH 6, even more preferably in the range of pH 4.5 to 5.5, and most preferably has a pH of 5.0.

[0190] Another aspect of the present invention is a method for preparing a reconstituted formulation comprising an hGH polymer prodrug of the present invention, comprising: contacting the dry pharmaceutical formulation of the present invention with a reconstitution solution The method includes:

[0191] Another aspect of the present invention is a reconstituted formulation obtainable from the process for preparing a reconstituted formulation of the present invention.

[0192] Preferably, the reconstituted formulation of the present invention comprises: hGH polymer prodrug 3~300mg / ml Succinic acid 5-50mM Trehalose dihydrate 25~150mg / ml Tris 1~50mM Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 1 to 100 mg of hGH equivalents / ml.

[0193] Even more preferably, the reconstituted formulation of the present invention comprises: hGH polymer prodrug 3~300mg / ml Succinic acid 5-50mM Trehalose dihydrate 50-90mg / ml Tris 5-50mM Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 1 to 100 mg of hGH equivalents / ml.

[0194] In an even more preferred embodiment, the reconstituted formulation of the invention comprises: hGH polymer prodrug 9~150mg / ml Succinic acid 5-50mM Trehalose dihydrate 50-90mg / ml Tris 5-50mM Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 3 to 50 mg of hGH equivalents / ml.

[0195] In an even more preferred embodiment, the reconstituted formulation of the invention comprises: hGH polymer prodrug 15~120mg / ml Succinic acid 5~40mM Trehalose dihydrate 60~86mg / ml Tris 5-40mM Including, The hGH polymer prodrug has a pH in the range of pH 4.0 to pH 6.0, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 5 to 40 mg of hGH equivalents / ml.

[0196] Even more preferably, the reconstituted formulation of the present invention comprises: hGH polymer prodrug 30~45mg / ml Succinic acid 5-20mM Trehalose dihydrate 75~86mg / ml Tris 5-20mM Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 10 to 15 mg of hGH equivalent / ml.

[0197] In an equally preferred embodiment, the reconstituted formulation of the invention comprises: hGH polymer prodrug 75~105mg / ml Succinic acid 5-20mM Trehalose dihydrate 60~81mg / ml Tris 5-20mM Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 25 to 35 mg of hGH equivalents / ml.

[0198] Most preferably, the reconstituted formulation of the present invention comprises: hGH polymer prodrug 42mg / ml Succinic acid 10mM Trehalose dihydrate 79~86mg / ml Tris 5-15mM Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 14 mg of hGH equivalents / ml.

[0199] In an equally preferred embodiment, the reconstituted formulation of the invention comprises: hGH polymer prodrug 84mg / ml Succinic acid 10mM Trehalose dihydrate 70-80mg / ml Tris 5-15mM Including, The hGH polymer prodrug has a pH in the range of pH 4.5 to pH 5.5, and the hGH polymer prodrug is an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV). When the hGH polymer prodrug is formula (IV), the amount of the hGH polymer prodrug corresponds to 28 mg of hGH equivalents / ml.

[0200] The reconstituted formulation optionally includes one or more preservatives and / or antimicrobial agents. Preferably, the one or more preservatives and / or antimicrobial agents is benzyl alcohol at a concentration of 0.7-1.1% (w / v), more preferably 0.9% (w / v). In another embodiment, the one or more preservatives and / or antimicrobial agents is cresol at a concentration of 0.2-0.4% (w / v), more preferably 0.3% (w / v).

[0201] Those skilled in the art are well aware that whenever a dry, liquid, or reconstituted formulation of the present invention contains trehalose dihydrate, this dihydrate form can also be replaced by other hydrated forms of trehalose, including anhydrous trehalose. Those skilled in the art would have no difficulty calculating the corresponding amounts of these other hydrated forms of trehalose, including anhydrous trehalose, contained in the corresponding dry, liquid, or reconstituted formulation. Therefore, it is understood that dry, liquid, or reconstituted formulations containing trehalose in hydrated forms other than the dihydrate are also within the scope of the present invention.

[0202] Another aspect of the present invention is a liquid pharmaceutical formulation, a dry pharmaceutical formulation or a reconstituted pharmaceutical formulation which is an hGH polymer prodrug of the present invention, preferably of formula (IV), or which comprises at least one hGH polymer prodrug of the present invention, preferably of formula (IV), for use as a pharmaceutical.

[0203] Another aspect of the present invention is the use of an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV), or a liquid pharmaceutical formulation, a dry pharmaceutical formulation, or a reconstituted pharmaceutical formulation comprising at least one hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV), in a method for treating a disease that can be treated with hGH.

[0204] Preferably, the disease treatable with hGH is selected from the group consisting of growth hormone deficiency (GHD) in children, idiopathic short stature (ISS), short stature homeobox (SHOX) gene mutations, Turner syndrome (TS), Noonan syndrome (NS), Prader-Willi syndrome (PWS), small for gestational age (SGA) in children, chronic renal failure (CRI), growth hormone deficiency (GHD) in adults, wasting due to HIV or AIDS or other malignant diseases, short bowel syndrome (SBS), sarcopenia and frailty.

[0205] In one embodiment, the disease that can be treated with hGH is GHD in children.

[0206] In another embodiment, the disease that can be treated with hGH is GHD in adults.

[0207] In another embodiment, the disease that can be treated with hGH is ISS.

[0208] In another embodiment, the disease that can be treated with hGH is a SHOX gene mutation.

[0209] In another embodiment, the disease that can be treated with hGH is TS.

[0210] In another embodiment, the disease that can be treated with hGH is NS.

[0211] In another embodiment, the disease that can be treated with hGH is PWS.

[0212] In another embodiment, the disease that can be treated with hGH is SGA.

[0213] In another embodiment, the disease that can be treated with hGH is CRI.

[0214] In another embodiment, the disease that can be treated with hGH is wasting due to HIV or AIDS or other malignant diseases.

[0215] In another embodiment, the disease that can be treated with hGH is SBS.

[0216] In another embodiment, the disease that can be treated with hGH is sarcopenia.

[0217] In another embodiment, the disease that can be treated with hGH is frailty.

[0218] Another aspect of the present invention is the use of an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid pharmaceutical formulation, a dry pharmaceutical formulation, or a reconstituted pharmaceutical formulation comprising at least one hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV), for the manufacture of a medicament for treating a disease treatable with hGH.

[0219] Preferably, the diseases that can be treated with hGH are selected from the group consisting of GHD, ISS, SHOX gene mutations, TS, NS, PWS, SGA, CRI in children, GHD in adults, wasting due to HIV or AIDS or other malignant diseases, SBS, sarcopenia and frailty.

[0220] In one embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the invention is used in the manufacture of a medicament for treating GHD in children.

[0221] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the invention is used in the manufacture of a medicament for treating GHD in adults.

[0222] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the invention is used in the manufacture of a medicament for treating ISS.

[0223] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dried or reconstituted pharmaceutical formulation of the present invention is used in the manufacture of a medicament for treating a SHOX gene mutation.

[0224] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the invention is used in the manufacture of a medicament for treating TS.

[0225] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the invention is used in the manufacture of a medicament for treating NS.

[0226] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the invention is used in the manufacture of a medicament for treating PWS.

[0227] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the invention is used in the manufacture of a medicament for treating SGA.

[0228] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the present invention is used in the manufacture of a medicament for treating CRI.

[0229] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid pharmaceutical formulation, a dried pharmaceutical formulation, or a reconstituted pharmaceutical formulation of the present invention is used in the manufacture of a medicament for treating wasting caused by HIV or AIDS or other malignant diseases.

[0230] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the invention is used in the manufacture of a medicament for treating SBS.

[0231] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the present invention is used in the manufacture of a medicament for treating sarcopenia.

[0232] In another embodiment, an hGH polymer prodrug, preferably an hGH polymer prodrug of formula (IV), or a liquid, dry or reconstituted pharmaceutical formulation of the invention is used in the manufacture of a medicament for treating frailty.

[0233] Another aspect of the present invention is a method for treating, controlling, delaying or preventing, preferably treating, at least one disease that can be treated, controlled, delayed or prevented by hGH in a mammalian patient, preferably a human patient, in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of an hGH polymer prodrug of the present invention, preferably of formula (IV), or a liquid formulation, a dry formulation or a reconstituted formulation comprising at least one hGH polymer prodrug of the present invention, preferably of formula (IV).

[0234] Preferably, the at least one disease treatable with hGH is selected from the group consisting of GHD, ISS, SHOX gene mutations, TS, NS, PWS, SGA, CRI in children, GHD in adults, wasting due to HIV or AIDS or other malignant diseases, SBS, sarcopenia and frailty.

[0235] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is GHD in children.

[0236] In another embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is GHD in adults.

[0237] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is ISS.

[0238] In one embodiment, the disease that can be treated with hGH using the methods of treating, regulating, delaying or preventing of the present invention is a SHOX gene mutation.

[0239] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is TS.

[0240] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is NS.

[0241] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is PWS.

[0242] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is SGA.

[0243] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is CRI.

[0244] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, slowing or preventing of the present invention is wasting due to HIV or AIDS or other malignant diseases.

[0245] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is SBS.

[0246] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is sarcopenia.

[0247] In one embodiment, the disease that can be treated with hGH using the methods of treating, controlling, delaying or preventing of the present invention is infirmity.

[0248] Another aspect of the present invention is a method of administering an hGH polymer prodrug, or liquid formulation or reconstituted formulation of the present invention, comprising the step of administering the hGH polymer prodrug, or liquid formulation or reconstituted formulation of the present invention via topical, enteral, or parenteral administration, or by external application, injection, or infusion method, including intra-articular, peri-articular, intradermal, subcutaneous, intramuscular, intravenous, intraosseous, intraperitoneal, intrathecal, intracapsular, intraorbital, intravitreal, intratympanic, intravesical, intracardiac, transtracheal, subcuticular, subcapsular, subarachnoid, intrathecal, intraventricular, intrasternal injection or infusion, direct delivery to the brain via an implanted device (e.g., an Ommaya reservoir) that allows delivery of the present invention or the like to brain tissue or brain fluid, direct intraventricular injection or infusion, injection or infusion into the brain or brain-related regions, injection into the subchoroidal space, retroorbital injection, and eye drop.

[0249] Preferably, the method comprises administering an hGH polymer prodrug of the present invention, or a liquid or reconstituted formulation, by injection, more preferably by subcutaneous injection.

[0250] In a preferred embodiment, the present invention relates to an hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV), or a liquid formulation or reconstituted formulation comprising at least one hGH polymer prodrug of the present invention, preferably an hGH polymer prodrug of formula (IV), for use in the treatment of GHD in children by subcutaneous injection.

[0251] Another aspect of the invention is a container containing an hGH polymer prodrug, or liquid or reconstituted formulation, of the invention for administration by injection, preferably by subcutaneous injection.

[0252] Preferred containers are syringes, dual chamber syringes, vials, stoppered and sealed vials, ampoules, cartridges and dual chamber cartridges.

[0253] [Example] method Cation exchange chromatography Purification of the conjugate by cation exchange chromatography was performed using an AKTA Pure system (GE Healthcare) equipped with a Macrocap SP column with a column volume of 279 mL. Each reaction mixture was applied to a column pre-equilibrated in 20 mM sodium acetate, 10 mM L-methionine buffer, pH 4.0 (Buffer A). After loading, the column was washed with Buffer A for three column volumes to remove any unreacted PEG reagent. The mono-conjugate was eluted using a gradient of 0 to 30% Buffer B (20 mM sodium acetate, 1 M sodium chloride, pH 4.5) over a total of 15 column volumes. Unreacted growth hormone was eluted using a gradient of 30 to 80% B over a total of three column volumes. The column was cleaned with 100% Buffer B for three column volumes. The flow rate was 20 mL / min for loading and 25 mL / min during elution. Elution was monitored by detection at 280 nm.

[0254] SDS-PAGE analysis The mPEG-hGH conjugates were analyzed by SDS-PAGE using NuPAGE® Novex 4-12% Bis-Tris gels (1.0 mm thick, 12 lanes), NuPAGE MOPS SDS-Running Buffer, HiMark™ Pre-stained High Molecular Weight Protein Standard, and Coomassie Colloidal Blue™ Staining Kit (Invitrogen). An amount of conjugate equivalent to 1 μg of hGH was applied per lane, and electrophoresis and subsequent staining were performed according to the supplier's protocol. Images of the gels were captured using a Digi Image System (Kisker Biotech) and a Power Shot G10 camera (Canon).

[0255] Dialysis / Ultrafiltration The dialysis and ultrafiltration steps were performed over a 50 cm 2 The experiments were performed using a laboratory-scale TFF system (Millipore) equipped with Pellicon XL Biomax membranes with a membrane area of ​​100 μg / ml and a molecular weight cutoff of 5 or 10 kDa for hGH alone, 10 kDa for 4 × 10 kDa mPEG-linker-hGH monoconjugate 2, and 50 kDa for 4 × 20 kDa mPEG-linker-hGH monoconjugate 1.

[0256] RP-HPLC The following RP-HPLC parameters were used: Mobile phase A consisted of 0.05% aqueous TFA, and mobile phase B consisted of 0.04% TFA in acetonitrile. A Waters UPLC C18 BEH 300Å 1.7 μm 2.1 × 50 mm column was used. The flow rate was set to 0.2–0.4 mL / min, detection was set at a wavelength of 215 nm, and the column operating temperature was set to 30°C (±5°C). The autosampler temperature was set to 4°C, and the sample injection load volume was 20 μL. For peak separation, the gradient shown in Table 1 was used.

[0257] [Table 1]

[0258] buffer exchange Buffer exchange was performed using an AKTA explorer system (GE Healthcare) equipped with a HiPrep 26 / 10 desalting column or a HiTrap desalting column.

[0259] Example 1: Transient synthesis of 4 x 20 kDa mPEG-linker-hGH monoconjugate 1 (standard product, not according to the present invention)

[0260] [ka]

[0261] Following a procedure similar to that described in WO2009 / 133137A2, 4 x 20 kDa mPEG-linker-hGH monoconjugate 1 were synthesized. The formulations of 4 x 20 kDa mPEG-linker-hGH monoconjugate 1 shown in Table 2 were prepared.

[0262] [Table 2]

[0263] Example 2: Synthesis of high-potency transient 4 x 10 kDa mPEG-linker-hGH monoconjugate 2

[0264] [ka] Following a procedure similar to that described in WO2009 / 133137A2, 4 x 10 kDa mPEG-linker-hGH monoconjugate 2 was synthesized. In detail, this preparation method was carried out as follows.

[0265] hGH was buffer exchanged into 100 mM sodium borate, pH 9, and the hGH concentration was adjusted to 10 mg / mL. A molar excess of a four-arm, branched 40 kDa mPEG-pentafluorophenyl carbonate derivative relative to the amount of hGH was dissolved in water to form a 6% (w / w) reagent solution. This reagent solution was added to the hGH solution in a 1:1 ratio (by weight) and mixed. The reaction mixture was incubated with stirring at 12–16°C for 105 minutes. Subsequently, the reaction mixture was quenched by adding 4 volumes of a solution containing 27 mM acetic acid and 12.5 mM L-methionine to 1 volume of the reaction mixture, lowering the pH of the solution to 4–4.5. After sterile filtration, the reaction mixture was incubated at room temperature for 16 ± 4 hours. The 4 × 10 kDa mPEG-linker-hGH monoconjugate 2 was purified by cation exchange chromatography.

[0266] Buffer exchange and adjustment of the 4 × 10 kDa mPEG-linker-hGH monoconjugate 2 to the desired concentration were achieved using a tangential flow filtration system. Thus, the eluate from the cation exchange chromatography was ultrafiltered and diafiltered into formulation buffer (10 mM succinic acid, 85 g / L trehalose dihydrate, pH 5.0 with 1 M Tris-solution). Using the same system, the trehalose concentration was reduced to 65 g / L, and the concentration of this stock solution was adjusted to 105 ± 3 mg / mL of 4 × 10 kDa mPEG-linker-hGH monoconjugate 2 (corresponding to 35 ± 1 mg hGH equivalents / mL). The formulations shown in Table 3 were prepared from this stock solution of Compound 2 by diluting the stock solution with a high-strength formulation buffer (10 mM succinic acid, 89 g / L trehalose dihydrate, pH 5.0 with 1 M Tris-base).

[0267] [Table 3]

[0268] Individual batches were analyzed by RP-HPLC, SE-HPLC, peptide mapping, and SDS-PAGE. SDS-PAGE showed that all formulations had comparable product quality, similar to the reference. During method development, it was discovered that the loading capacity of the cation exchange chromatography column used to purify 4 × 10 kDa mPEG-linker-hGH monoconjugate 2 could be significantly improved compared to the purification procedure for 4 × 20 kDa mPEG-linker-hGH monoconjugate 1.

[0269] Conclusion: The 4 × 10 kDa mPEG-linker-hGH monoconjugate 2 could be synthesized using a manufacturing method with only minor modifications compared to that described in EP-A 2 113 256, demonstrating improved handling and product properties. The load on the CIEX column for purification could be increased by at least three times without compromising separation efficiency and product quality. Furthermore, the content of the final product could be increased to over 100 mg / mL for 4 × 10 kDa mPEG-linker-hGH conjugate 2, corresponding to approximately 35 mg hGH equivalents / mL.

[0270] Example 3: Injectability of high-strength formulations of 4 x 10 kDa mPEG-linker-hGH monoconjugate 2 compared to 4 x 20 kDa mPEG-linker-hGH monoconjugate 1 The individual formulations from Examples 1 and 2 were investigated for their injectability through needles with various internal diameters. The tests were carried out on a Mecmesin Multitest 1-d stand equipped with a measuring device BFG 200N, using Emperor Lite software (version 1.16-015). The tested needles included a 27G needle 0.4 x 13 mm 27G x 1 / 2" from BD (reference 300635, lot 101009), a 29G needle 0.33 x 13 mm from Transcoject, and a 30G needle 0.30 x 12 mm 30G x 1 / 2" from Sterican (lot 2G13258811). The measuring device was set up to measure the plunger depressing force for a given constant plunger speed. The applied plunger speeds, which correspond to the applied injection speeds, were as follows: Injection speed 688mm / min 5 seconds / mL 12mL / min 344mm / min 10sec / mL 6mL / min 229mm / min 15sec / mL 4mL / min 172mm / min 20sec / mL 3mL / min 138mm / min 25sec / mL 2.4mL / min 115mm / min 30sec / mL 2mL / min

[0271] The test was carried out using the following steps: Fill a 1.1 ml Luer lock syringe (BD, reference 309628) with the sample (using a 20 G needle, 0.90 x 40 mm, 20 G x 1 1 / 2 inches, manufactured by Sterican). 2. Remove air bubbles 3. Attach the test needle to the syringe (starting with the largest inner diameter) 4. Clamp this syringe into the holder 5. Choose the right measurement settings 6. Start the measurement and collect the sample in a glass vial (place it under the syringe) 7. Remove the syringe from the holder 8. Refill the syringe with the test sample and measure the set point. ->These steps were repeated for all needles (with decreasing needle diameter) and for each test sample.

[0272] The formulation buffer without mPEG-linker-hGH monoconjugate 1 or 2 was used as a reference solution.

[0273] The injection force was determined for 4 x 10 kDa mPEG-linker-hGH monoconjugate 2 for all different injection needles and all injection rates and compared to the results for 4 x 20 kDa mPEG-linker-hGH monoconjugate 1. Table 4 shows a comparison of the injection force between 4 x 10 kDa mPEG-linker-hGH monoconjugate 2 and 4 x 20 kDa mPEG-linker-hGH monoconjugate 1 using a 27G needle, 0.4 x 13 mm, 27G x 1 / 2" from BD (reference 300635, lot 101009).

[0274] [Table 4]

[0275] Table 5 shows a comparison of the injection force between 4 x 10 kDa mPEG-linker-hGH monoconjugate 2 and 4 x 20 kDa mPEG-linker-hGH monoconjugate 1 using a 29G needle 0.33 x 13 mm from Transcoject.

[0276] [Table 5]

[0277] Table 6 shows a comparison of injection force between 4 x 10 kDa mPEG-linker-hGH monoconjugate 2 and 4 x 20 kDa mPEG-linker-hGH monoconjugate 1 using a 30G needle 0.30 x 12 mm, 30G x 1 / 2" manufactured by Sterican (Lot 2G13258811).

[0278] [Table 6]

[0279] Conclusion: The injectability of the 4 × 10 kDa mPEG-linker-hGH monoconjugate 2 was significantly improved, and its injectability could be reduced by 3.5 to 4–5 times compared with that of the 4 × 20 kDa mPEG-linker-hGH monoconjugate 1.

[0280] Example 4: Viscosity Measurement of 4 x 10 kDa mPEG-linker-hGH Monoconjugate 2 Compared to 4 x 20 kDa mPEG-linker-hGH Monoconjugate 1 The dynamic viscosity of the test samples was determined at Infraserv Knapsack (now Synlab Pharma Institute) using a method conforming to European Pharmacopoeia Method 2.2.10. All measurements were performed using a cone / plate measuring system (CP50 / 1) at 23.0 ± 0.1 °C with approximately 1–5 mL of test sample. The shear rate was 100 s -1 ~10s -1 The range was set as follows.

[0281] All test formulations of 4 x 10 kDa mPEG-linker-hGH monoconjugate 2 and 4 x 20 kDa mPEG-linker-hGH monoconjugate 1 were adjusted to an equiosmolarity of approximately 290 mOsmol / kg by increasing or decreasing the amount of trehalose in the formulation. The dynamic viscosity values ​​measured for all test samples are summarized in Table 7.

[0282] [Table 7]

[0283] Conclusion: The dynamic viscosity of the 4×10 kDa mPEG-linker-hGH monoconjugate 2 could be significantly reduced by about 4-5 times compared to that of the 4×20 kDa mPEG-linker-hGH monoconjugate 1.

[0284] Example 5 Reconstitution Time of Lyophilized 4 × 10 kDa mPEG-Linker-hGH Monoconjugate 2 1 mL of 4 × 10 kDa mPEG-Linker-hGH Monoconjugate 2 was lyophilized in a Din2R vial. After lyophilization, the lyo cake was dissolved in 1 mL of water for injection. This reconstitution time was compared to the dissolution time of the lyophilized 4 × 20 kDa mPEG-Linker-hGH Monoconjugate 1. During reconstitution, many air bubbles were detected with 4 × 20 kDa mPEG-Linker-hGH Monoconjugate 1. The lyophilized cake itself dissolved fairly quickly, and the time to obtain a clear solution with minimal residual air bubbles was significantly shorter for 4 × 10 kDa mPEG-Linker-hGH Monoconjugate 2. The results of this reconstitution procedure are summarized in Table 8.

[0285] [Table 8]

[0286] Conclusion: The reconstitution time to achieve a clear, substantially bubble-free solution was significantly shorter for the 4 x 10 kDa mPEG-linker-hGH monoconjugate 2 compared to the 4 x 20 kDa mPEG linker-hGH monoconjugate 1.

[0287] Example 6: In vitro hydrolysis of 4 x 10 kDa mPEG-linker-hGH monoconjugate 2 To determine the in vitro linker cleavage rates of 4 × 10 kDa mPEG-linker-hGH monoconjugate 2 or 4 × 20 kDa mPEG-linker-hGH monoconjugate 1, the compounds were buffer-exchanged into PBST buffer at pH 7.4, and the eluted solution was filtered through a 0.22 μm filter and incubated at 37°C for 1 week. Samples were taken at time intervals and analyzed by RP-HPLC. All peaks were integrated and assigned, and the area of ​​the relevant peak was plotted against incubation time. Curve-fitting software was applied to determine first-order cleavage rates. Table 9 shows the in vitro hydrolysis rates of 4 × 10 kDa mPEG-linker-hGH monoconjugate 2 and 4 × 20 kDa mPEG-linker-hGH monoconjugate 1 at pH 7.4 and 37°C.

[0288] [Table 9]

[0289] Conclusion: The in vitro hydrolysis rates of conjugates 1 and 2 at pH 7.4 and 37° C. were in the range of 105±5 h. Both half-lives were fairly comparable and fell within the 95% confidence interval.

[0290] Example 7: Quantification of conjugates 1 and 2 in serum samples from animal studies An ELISA-based method was used to quantify conjugates 1 and 2 in serum samples from animal studies. The same sandwich ELISA format was used for both conjugates 1 and 2, utilizing a sheep anti-hGH polyclonal antibody (Abcam, catalog no. ab64499) as the capture antibody and a biotinylated rabbit anti-PEG antibody (Epitomics, catalog no. 2137-1) as the detection antibody. Readout was performed using streptavidin-HRP (Jackson ImmunoResearch, catalog no. 016-030-084) and a commercially available TMB liquid substrate system (Sigma, catalog no. T0440). Prior to measurement, serum standards and samples were diluted 1:50 with pH 7.0 buffer (50 mM HEPES, 1 mM CaCl2, 0.05% Tween-20, and 1% BSA). Samples were incubated in the ELISA plate at 37°C for 2 hours with shaking.

[0291] Example 8: Quantification of total mPEG 40 and 80 in serum samples from animal studies An ELISA-based method was used to quantify mPEG40 and mPEG80 in serum samples from animal studies. The same sandwich ELISA format was used for both test substances, mPEG40 and mPEG80, utilizing an anti-PEG (methoxy group) rabbit monoclonal antibody (Epitomics, catalog no. 2061-1) as the capture antibody and a biotinylated anti-PEG mouse monoclonal IgM antibody (ANP Tech, catalog no. 90-1052) as the detection antibody. Readout was performed using streptavidin-HRP (Jackson ImmunoResearch, catalog no. 016-030-084) and a commercially available TMB liquid substrate system (Sigma, catalog no. T0440). Prior to measurement, serum standards and samples were diluted 1:50 with pH 7.0 buffer (50 mM HEPES, 1 mM CaCl2, 0.05% Tween-20, and 1% BSA). The samples were incubated in the ELISA plate at 37°C for 2 hours under shaking.

[0292] Example 9: Comparative pharmacokinetic study in cynomolgus monkeys treated with conjugates 1 and 2 Two groups of five healthy, non-naive male cynomolgus monkeys each received a single subcutaneous dose of Conjugate 1 or Conjugate 2 at a target dose level of 1 mg hGH equivalent per kg (equivalent to 3 mg Conjugate 2 / kg and 5 mg Conjugate 1 / kg, respectively). Blood samples were collected and serum generated up to 336 hours after administration for PK determination (for mPEG quantification, serum samples were collected up to 56 days). Pharmacokinetic analysis according to Example 7 showed that both compounds produced comparable maximum conjugate levels (9,200 ng hGH equivalents / mL for Conjugate 1 and 7,400 ng hGH equivalents / mL for Conjugate 2), which were reached approximately 36 hours after administration. mPEG concentration levels were determined according to Example 8. The PK profiles of both mPEGs had their maximum concentration levels at 48 hours after administration. The clearance of mPEG40 was faster than that of mPEG80, as indicated by the terminal elimination half-lives (300 hours for mPEG80 and 260 hours for mPEG40). This resulted in significantly lower overall mPEG exposure for conjugate 2 than for conjugate 1 in this comparative PK study.

[0293] The following are examples of aspects of the present invention. [Section 1] A human growth hormone (hGH) polymer prodrug of formula (Ia) or (Ib) or a pharmaceutically acceptable salt thereof [ka] [In the formula, -D is the hGH moiety attached to the rest of the molecule via the amine functionality; n is 0, 1, 2, 3 or 4; -X- is a chemical bond or a spacer; =Y1, =Y5 are independently selected from the group consisting of =O and =S; -Y2- is selected from the group consisting of -O- and -S-; - Y3- is selected from the group consisting of -O- and -S-; -Y4- is -O-, -NR 5 - and -C(R 6 R 6a )-, -R 1 is a water-soluble PEG-based moiety containing at least 40% PEG having a molecular weight in the range of 30-50 kDa; -R 2 , -R 3 , -R 5 , -R 6 , -R 6a are independently selected from the group consisting of -H, 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; -R 4 is selected from the group consisting of 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; -W- is C 3~10 Cycloalkyl, 8-30 membered carbopolycyclyl, 3-10 membered heterocyclyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 )-, C 1~20 is selected from the group consisting of alkyl, -Nu is -N(R 7 R 7a ), -N(R 7OH), -N(R 7 )-N(R 7a R 7b ), -S(R 7 ), -COOH, [ka] is a nucleophile selected from the group consisting of -Ar- is [ka] (In the formula, The dashed line indicates the bond to the remainder of the prodrug; -Z 1 - is -O-, -S- and -N(R 7 )-, -Z 2 - is -N(R 7 )-is) is selected from the group consisting of -R 7 , -R 7a , -R 7b -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl] And, The prodrugs of formula (Ia) and (Ib) are optionally further substituted human growth hormone (hGH) polymer prodrugs or pharmaceutically acceptable salts thereof. [Section 2] -R 1 However, the moiety of formula (II): [ka] (In the formula, -BP 1 <, -BP 2 <, -BP 3 < is -N< and -C(R 8 )<, wherein: R 8 is H, C 1~6 Alkyl, C 2~6 Alkenyl and C2~6 alkynyl, -P 1 , -P 2 , -P 3 , -P 4 are, independently of each other, PEG-based chains containing at least 40% PEG and having a molecular weight in the range of 8-12 kDa; -C 1 -, -C 2 -C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be substituted with one or more of the same or different R 9 and C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 10 )-, -S(O)2N(R 10 )-, -S(O)N(R 10 )-, -S(O)2-, -S(O)-, -N(R 10 )S(O)2N(R 10a )-, -S-, -N(R 10 )-, -OC(OR 10 )(R 10a )-, -N(R 10 )C(O)N(R 10a )- and -OC(O)N(R 10 )-, T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, 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; each T is independently selected from the group consisting of one or more of the same or different R 9 and optionally substituted by R 9are halogen, -CN, oxo (=O), and -COOR, respectively. 11 , -OR 11 , -C(O)R 11 , -C(O)N(R 11 R 11a ), -S(O)2N(R 11 R 11a ), -S(O)N(R 11 R 11a ), -S(O)R 11 , -S(O)R 11 , -N(R 11 )S(O)2N(R 11a R 11b ), -SR 11 , -N(R 11 R 11a ), -NO2, -OC(O)R 11 , -N(R 11 )C(O)R 11a , -N(R 11 )S(O)2R 11a , -N(R 11 )S(O)R 11a , -N(R 11 )C(O)OR 11a , -N(R 11 )C(O)N(R 11a R 11b ), -OC(O)N(R 11 R 11a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl may be optionally substituted with one or more of the same or different halogens; R 10 , R 10a , R 11 , R 11a and R 11b are -H and C, respectively. 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl may be substituted with one or more of the same or different halogens. Item 1. The prodrug according to item 1, comprising: [Section 3] -R 1 But the part of formula (IIc): [ka] (In the formula, p1, p2, p3, and p4 are independently integers ranging from 180 to 270, preferably from 200 to 250, even more preferably from 210 to 240, and most preferably from 220 to 240. Item 3. The prodrug according to Item 1 or 2, comprising: [Section 4] The hGH polymer prodrugs of the present invention have the formula (IV): [ka] (In the formula, D is the hGH moiety that is attached to the rest of the molecule via the amine functionality; p1, p2, p3, and p4 are independently integers ranging from 180 to 270, preferably from 200 to 250, even more preferably from 210 to 240, and most preferably from 220 to 240. The prodrug according to any one of items 1 to 3, wherein [Section 5] A pharmaceutical formulation comprising the prodrug of any one of items 1 to 4 and at least one excipient. [Section 6] Item 6. The pharmaceutical formulation according to Item 5, which is a liquid formulation and contains 3 to 300 mg / mL of the prodrug according to any one of Items 1 to 4. [Section 7] The liquid formulation hGH polymer prodrug 3~300mg / ml Succinic acid 5-50mM Any trehalose dihydrate 25-150mg / ml Optional methionine 1-50mM Including, Item 7. The liquid pharmaceutical formulation of item 6, having a pH in the range of pH 4.0 to pH 6.0 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, and wherein the hGH polymer prodrug is the prodrug of any one of items 1 to 4. [Section 8] Item 6. The pharmaceutical formulation according to Item 5, which is a dry formulation and contains 1 to 99.9% (w / w) of the prodrug according to any one of Items 1 to 4. [Section 9] (a) hGH polymer prodrug 3-300 mg / ml Succinic acid 5-50mM Any trehalose dihydrate 25-150mg / ml A liquid formulation comprising: providing a liquid formulation having a pH in the range of pH 4.0 to pH 6.0 titrated with a suitable buffer, preferably with Tris base, more preferably with a 1 molar Tris base solution, wherein the hGH polymer prodrug is a prodrug according to any one of paragraphs 1 to 4; and (b) drying the liquid formulation of step (a). Item 9. A dried pharmaceutical formulation according to Item 8, obtained by a method comprising: [Section 10] The dry formulation comprises, based on the total weight of the formulation: hGH polymer prodrug 14-65% (w / w) Succinic acid 0.5-2.5% (w / w) Trehalose dihydrate 31~84%(w / w) Tris 0.4-4% (w / w) Including, Item 10. The dry pharmaceutical formulation according to item 8 or 9, wherein the hGH polymer prodrug is a prodrug according to any one of items 1 to 4. [Section 11] A method for preparing a reconstituted formulation containing the prodrug according to any one of items 1 to 4, comprising: A method comprising contacting the dried pharmaceutical formulation according to any one of paragraphs 8 to 10 with a reconstitution solution. [Section 12] Item 12. A reconstituted formulation obtained by the method according to Item 11. [Section 13] The reconstituted formulation hGH polymer prodrug 3~300mg / ml Succinic acid 5-50mM Trehalose dihydrate 25~150mg / ml Tris 1~50mM Including, The pH ranges from pH 4.0 to pH 6.0, and the hGH polymer prodrug is the prodrug described in any one of items 1 to 4. Item 13. The reconstituted formulation according to Item 12. [Section 14] Use of the prodrug according to any one of clauses 1 to 4 or the pharmaceutical formulation according to any one of clauses 5 to 13 as a medicine. [Section 15] Use of the prodrug of any one of clauses 1 to 4 or the pharmaceutical formulation of any one of clauses 5 to 13 in a method for treating a disease that can be treated with hGH. [Section 16] [Item 17] Use of the prodrug of any one of Items 1 to 4 or the pharmaceutical formulation of any one of Items 5 to 13 for the manufacture of a medicament for treating a disease that can be treated with hGH. Item 17. Use according to item 16, wherein the disease that can be treated with hGH is selected from the group consisting of growth hormone deficiency in children, idiopathic short stature, short stature homeobox gene mutations, Turner syndrome, Noonan syndrome, Prader-Willi syndrome, small for gestational age in children, chronic renal failure, growth hormone deficiency in adults, wasting due to HIV or AIDS or other malignant diseases, short bowel syndrome, sarcopenia and frailty. [Section 18] A method for treating, controlling, delaying or preventing at least one disease that can be treated, controlled, delayed or prevented by hGH in a mammalian patient in need thereof, comprising administering to said patient in need thereof a therapeutically effective amount of a prodrug according to any one of paragraphs 1 to 4 or a pharmaceutical formulation according to any one of paragraphs 5 to 13. [Section 19] A method for administering the prodrug of any one of paragraphs 1 to 4 or the pharmaceutical formulation of any one of paragraphs 5 to 13, comprising administering the hGH polymer prodrug of the present invention, or the liquid formulation or reconstituted formulation, via topical, enteral, or parenteral administration, or by external application, injection, or infusion, including intra-articular, peri-articular, intradermal, subcutaneous, intramuscular, intravenous, intraosseous, intraperitoneal, intrathecal, intracapsular, intraorbital, intravitreal, intratympanic, intravesical, intracardiac, transtracheal, subcuticular, subcapsular, subarachnoid, intrathecal, intraventricular, or intrasternal injection or infusion, direct delivery to the brain via an implantable device enabling delivery to brain tissue or brain fluid, such as the product of the present invention, direct intraventricular injection or infusion, injection or infusion into the brain or brain-related regions, injection into the subchoroidal space, retroorbital injection, and eye drop. Abbreviation: AIDS Acquired Immune Deficiency Syndrome CRI chronic renal failure DF Diafiltration ELISA enzyme-linked immunosorbent assay EP European Pharmacopoeia eq stoichiometric equivalent G gauge GHD Growth Hormone Deficiency HIV human immunodeficiency virus ISS Idiopathic Short Stature MW molecular weight NS Noonan syndrome PEG polyethylene glycol PWS Prader-Willi syndrome PK Pharmacokinetics RP-HPLC Reversed-phase high-performance liquid chromatography rt room temperature SBS short bowel syndrome SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis SEC size exclusion chromatography SHOX Short Height Homeobox SGA small for gestational age TFF Tangential Flow Filtration Tris Tris(hydroxymethyl)aminomethane TS Turner syndrome UF Ultrafiltration

[0294] SEQUENCE LISTING <110> Ascendis Pharma Endocrinology Division A / S <120> Novel Polymeric hGH Prodrugs <130> PA25-542 <150> EP14193603.9 <151> 2014-11-18 <160> 1 <170> PatentIn version 3.5 <210> 1 <211> 191 <212> PRT <213> Homo sapiens <00> <400> 1 Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg 1 5 10 15 Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu 20 25 30<00~ Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro 35 40 45 Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg 50 55 60 Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu 65 70 75 80 Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val 85 90 95 Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp 100 105 110 Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu 115 120 125 Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser 130 135 140 Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr 145 150 155 160 Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe 165 170 175 Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 180 185 190

Claims

1. A human growth hormone (hGH) polymer prodrug of formula (Ia) or (Ib) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating growth hormone deficiency in children. 【Chemistry 1】 [In the formula, -D is the hGH moiety attached to the rest of the molecule via the amine functionality; n is 0, 1, 2, 3 or 4; -X- is a chemical bond or a spacer; =Y 1 , =Y 5 are independently selected from the group consisting of =O and =S; -Y 2 - is selected from the group consisting of -O- and -S-; -Y 3 - is selected from the group consisting of -O- and -S-; -Y 4 -, -O-, -NR 5 - and -C(R 6 R 6a )-, -R 1 is a water-soluble PEG-based moiety comprising at least 40% PEG having a molecular weight in the range of 30-50 kDa; -R 2 , -R 3 , -R 5 , -R 6 , -R 6a are independently selected from the group consisting of -H, 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; -R 4 is selected from the group consisting of 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; -W- is C 3～10 Cycloalkyl, 8- to 30-membered carbopolycyclyl, 3- to 10-membered heterocyclyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 )-, C 1～20 is selected from the group consisting of alkyl, -Nu is -N(R 7 R 7a ), -N(R 7 OH), -N(R 7 )-N(R 7a R 7b ), -S(R 7 ), -COOH, 【Chemistry 2】 is a nucleophile selected from the group consisting of -Ar- is 【Transformation 3】 (In the formula, The dashed line indicates the bond to the remainder of the prodrug; -Z 1 - is -O-, -S- and -N(R 7 )-, -Z 2 - is -N(R 7 )-is) is selected from the group consisting of -R 7 , -R 7a , -R 7b -H, C 1～6 Alkyl, C 2～6 Alkenyl and C 2～6 alkynyl] The use of The prodrugs of formula (Ia) and (Ib) may be further substituted.

2. -R 1 However, the moiety of formula (II): 【Chemistry 4】 (In the formula, -BP 1 <, -BP 2 <, -BP 3 < is selected independently of each other from the group consisting of -N< and -C(R 8 )< R 8 is H, C 1～6 Alkyl, C 2～6 Alkenyl and C 2～6 alkynyl, -P 1 , -P 2 , -P 3 , -P 4 are, independently of each other, PEG-based chains containing at least 40% PEG and having a molecular weight in the range of 8-12 kDa; -C 1 -, -C 2 -C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 alkynyl, C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 Alkynyl may be substituted with one or more of the same or different R 9 and C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 10 )-, -S(O) 2 N(R 10 )-, -S(O)N(R 10 )-, -S(O) 2 -, -S(O)-, -N(R 10 )S(O) 2 N(R 10a )-, -S-, -N(R 10 )-, -OC(OR 10 )(R 10a )-, -N(R 10 )C(O)N(R 10a )- and -OC(O)N(R 10 )-, T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, 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; each T is independently selected from the group consisting of one or more of the same or different R 9 and optionally substituted by R 9 are halogen, -CN, oxo (=O), and -COOR, respectively. 11 , -OR 11 , -C(O)R 11 , -C(O)N(R 11 R 11a ), -S(O) 2 N(R 11 R 11a ), -S(O)N(R 11 R 11a ), -S(O) 2 R 11 , -S(O)R 11 , -N(R 11 )S(O) 2 N(R 11a R 11b ), -SR 11 , -N(R 11 R 11a ), -NO 2 , -OC(O)R 11 , -N(R 11 )C(O)R 11a , -N(R 11 )S(O) 2 R 11a , -N(R 11 )S(O)R 11a , -N(R 11 )C(O)OR 11a , -N(R 11 )C(O)N(R 11a R 11b ), -OC(O)N(R 11 R 11a ), and C 1～6 alkyl, independently selected from the group consisting of C 1～6 The alkyl may be optionally substituted with one or more of the same or different halogens; R 10 , R 10a , R 11 , R 11a and R 11b are -H and C, respectively. 1～6 alkyl, independently selected from the group consisting of C 1～6 The alkyl may be substituted with one or more of the same or different halogens.

2. The use according to claim 1, comprising:

3. -P 1 , -P 2 , -P 3 , -P 4 are independently of each other of the formula (IIb) 【Transformation 5】 (In the formula, The dashed line indicates R 1 The remainder, i.e., BP 2 or BP 3 indicates the bond to m is 0 or 1; p is an integer ranging from 180 to 270, more preferably from 200 to 250, even more preferably from 210 to 240, and most preferably from 220 to 240; q is selected from the group consisting of 1, 2, 3, 4, 5 and 6 3. The use according to claim 2, wherein

4. BP of formula (II) 1 The use according to claim 2 or 3, wherein -N<.

5. BP of formula (II) 2 and BP 3 and are both -CH<.

6. First branch point BP 1 6. The use according to any one of claims 2 to 5, wherein the binding sites of X and X are separated by less than 60 atoms.

7. First branch point BP 1 7. The use according to any one of claims 2 to 6, wherein the binding sites of X and X are separated by less than 30 atoms.

8. First branch point BP 1 8. The use according to any one of claims 2 to 7, wherein the binding sites of X and X are separated by less than 20 atoms.

9. First branch point BP 1 9. The use according to any one of claims 2 to 8, wherein the binding sites of X and X are separated by less than 10 atoms.

10. C in formula (II) 1 and C 2 However, -O-, -C(O)N(R 10 )- and 3- to 10-membered heterocyclyl 1～50 10. The use according to any one of claims 2 to 9, wherein the 3- to 10-membered heterocyclyl is alkyl and is substituted by at least one oxo (=O).

11. C 1 and C 2 is represented by the formula (IIa) 【Transformation 6】 (In the formula, The dashed lines indicated by asterisks represent the BP 1 indicates the bond to The unmarked dashed lines represent BPs, respectively. 2 or BP 3 indicates the bond to q1 is 1, 2, 3, 4, 5, 6, 7 or 8, preferably q1 is 4, 5, 6, 7 or 8, more preferably q1 is 5, 6 or 7, and most preferably q1 is 6; q2 is 1, 2, 3, 4 or 5, preferably q2 is 1, 2 or 3, most preferably q2 is 2; q3 is 1, 2, 3, 4, 5, 6, 7 or 8, preferably q3 is 2, 3, 4 or 5, more preferably q3 is 2, 3 or 4, and most preferably q3 is 3; q4 is 1, 2 or 3, and most preferably q4 is 1.

11. The use according to any one of claims 2 to 10, wherein

12. -R 1 But the part of formula (IIc): 【Transformation 7】 (In the formula, p1, p2, p3, p4 are independently integers ranging from 180 to 270, preferably from 200 to 250, even more preferably from 210 to 240, and most preferably from 220 to 240.

12. The use according to any one of claims 1 to 11, comprising:

13. The hGH polymer prodrugs of the present invention have the formula (IV): 【Transformation 8】 (In the formula, D is the hGH moiety that is attached to the rest of the molecule via the amine functionality; p1, p2, p3, p4 are independently integers ranging from 180 to 270, preferably from 200 to 250, even more preferably from 210 to 240, and most preferably from 220 to 240.

13. The use according to any one of claims 1 to 12, wherein

14. 14. The prodrug of any one of claims 1 to 13, wherein -D is the hGH portion of SEQ ID NO:1.

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