Conjugates undergoing intramolecular rearrangement

Abstract

The present invention relates to conjugates and pharmaceutically acceptable salts thereof, reagents, intermediates, methods of synthesizing said conjugates, pharmaceutical compositions comprising said conjugates, and uses of said conjugates.

Description

[Technical Field]

[0001] The present invention relates to conjugates and pharmaceutically acceptable salts thereof, reagents, intermediates, methods for synthesizing the conjugates, pharmaceutical compositions comprising the conjugates, and uses of the conjugates. [Background technology]

[0002] To improve the physicochemical or pharmacokinetic properties of a drug, such as its in vivo circulating half-life, such drugs may be conjugated to a carrier, such as a polymer. Typically, polymers in drug delivery are used by non-covalent complexation of the drug with the polymer, embedding of the drug in the polymer, or covalent conjugation of the drug to a polymer portion.

[0003] However, non-covalent approaches require highly efficient drug encapsulation to prevent uncontrolled burst release of drugs due to the degradation of drug-polymer complexes after administration. Suppressing the diffusion of unbound water-soluble drug molecules requires strong van der Waals contact, frequently mediated through hydrophobic and charged moieties for electrostatic bonding. Many conformationally sensitive drugs, such as proteins or peptides, become dysfunctional during the complexation process and / or during the subsequent storage of non-covalently bound drugs.

[0004] Alternatively, the drug may be covalently conjugated to the polymer portion via the linker portion, so the linkage between the drug and the linker is stable, or the linkage between the drug and the linker portion is reversible via the linker portion. When the drug is conjugated to the linker portion via a stable linkage, such a conjugate must exhibit sufficient residual activity to have a medicinal effect, and therefore the conjugate is always in an active form.

[0005] The synthesis of conjugates containing a drug covalently conjugated to a linker moiety via reversible linkage typically involves the reaction of a reagent containing the linker moiety with a functional group on the drug moiety, such as an amine functional group. For example, WO2009 / 095479A2 discloses a conjugate containing a linker moiety conjugated to a drug via an amide bond, thereby making the amide bond reversible, for example, through the neighboring group involvement of a functional group or other group contained within the linker moiety, such as an amine and the amide group. More specifically, a nucleophilic amine within the linker moiety enhances the nucleophilicity of the nitrogen atom contained in the amide or thioamide, which then attacks the carbonyl portion of the amide group connecting the drug to the linker moiety, resulting in the cleavage of the amide bond and the release of the drug in its unmodified form. The synthesis of such conjugates can be difficult, for example, because neighboring group involvement must be deactivated to avoid premature cyclization of the reagent containing the linker moiety and the formation of by-products during the conjugation of the reagent containing the linker moiety to the drug.

[0006] One way to avoid the aforementioned premature cyclization is to inactivate the nucleophilicity of one of the neighboring groups, for example, an amide group, by using an amide protecting group. Since such an amide protecting group needs to be removed after the conjugation of the drug into the reagent containing the linker moiety, such process synthesis may require the use of a protecting group moiety that should be easily removed, for example, under mild conditions. The selection of such a protecting group is limited to those skilled in the art, especially when the drug moiety is a protein moiety, as the deprotection of the protecting group moiety must preferably occur under aqueous conditions with limited use of organic solvents and reagents to avoid protein inactivation or damage.

[0007] Therefore, it is necessary to identify solutions to the challenge of chemical synthesis of conjugates, particularly those containing a protein-drug moiety, in which the linkage between the drug and the linker is reversible because the drug is linked to the support via the linker moiety. [Overview of the project]

[0008] Therefore, it is an object of the present invention to at least partially overcome the drawbacks described above.

[0009] This object is achieved by a conjugate comprising at least one -D part conjugated to at least one Z part via at least one -L 1 -L 2 - part, or a pharmaceutically acceptable salt thereof, wherein the -L 1 - part is conjugated to the nitrogen of a primary or secondary amine of the -D part, and the linkage between -D and -L 1 - is reversible, and the -L 2 - part is conjugated to Z, each -D is independently a primary or secondary amine-containing part of a drug D-H, each -L 2 - is independently a single bond or a spacer part, each Z is independently a polymer part or C 8~24 alkyl, each -L 1 - is independently of the formula (I):

[0010]

Chemical formula

[0011] Another aspect of the present invention is a reagent comprising a -L*- moiety, wherein the -L*- is conjugated to -Q, -Q is -OH or -LG, and -LG is the leaving group portion. -L*- is equation (II):

[0012] [ka] [In the ceremony The dashed line indicates a connection to -Q. v is selected from the group consisting of 0 or 1. -X 1 - is -C(R 8 )(R 8a )-,-N(R 9 Selected from the group consisting of )- and -O-, =X 2 is =O and =N(R 10 Selected from the group consisting of, -X 3 - is selected from the group consisting of -O-, -S-, and -Se-, Each p is independently selected from the group consisting of 0 or 1, provided that at most one p is 0. -R 6 is -PG and -R 6a is -H, -C(R 11 )(R 11a )(R 11b), selected from the group consisting of -T and -PG, or -R 6 and -R 6a -C(R 11 )(R 11a )(R 11b Independently selected from the group consisting of ) and -T, -R A and -R B It is independently selected from the group consisting of -H and -PG, except for -R A or -R B The condition is that one or less of them can be -H, -PG is the amine protecting group moiety, -R 9 -C(R 11 )(R 11a )(R 11b Selected from the group consisting of ) and -T, -R 10 is -H, -C(R 11 )(R 11a )(R 11b Selected from the group consisting of ) and -T, -R 1 ,-R 1a ,-R 2 ,-R 2a ,-R 3 ,-R 3a ,-R 4 ,-R 4a ,-R 5 ,-R 5a ,-R 7 ,-R 8 ,-R 8a ,-R 11 ,-R 11a and -R 11b -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12)S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkynnyls, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is one or more identical or different -R 13 In some cases, it is replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R) 14 )-,-S(O)2N(R 14 )-,-S(O)N(R 14 )-, -S(O)2-, -S(O)-, -N(R 14 )S(O)2N(R 14a )-, -S-, -N(R 14 )-, -OC(OR 14 )(R 14a )-,-N(R 14 )C(O)N(R 14a )- and -OC(O)N(R 14 )- is optionally interrupted by one or more groups selected from the group consisting of, -R 12 ,-R 12a ,-R 12b -H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkynyls, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is one or more identical or different -R 13In some cases, it is replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R) 14 )-,-S(O)2N(R 14 )-,-S(O)N(R 14 )-, -S(O)2-, -S(O)-, -N(R 14 )S(O)2N(R 14a )-, -S-, -N(R 14 )-, -OC(OR 14 )(R 14a )-,-N(R 14 )C(O)N(R 14a )- and -OC(O)N(R 14 )- is optionally interrupted by one or more groups selected from the group consisting of, Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Independently selected from the group consisting of cycloalkyls, 3-10 membered heterocyclyls, and 8-11 membered heterobicyclyls, each T is one or more identical or different -R groups. 13 It is independently substituted depending on the case, -R 13 These are halogen, -CN, oxo, and -C(O)OR 15 , -OR 15 , -C(O)R 15 ,-C(O)N(R 15 )(R 15a ), -S(O)2N(R 15 )(R 15a ), -S(O)N(R 15 )(R 15a ), -S(O)2R 15 ,-S(O)R 15 , -N(R 15 )S(O)2N(R 15a )(R 15b ), -SR 15 , -N(R 15 )(R 15a ), -NO2, -OC(O)R 15 , -N(R 15 )C(O)R 15a , -N(R 15)S(O)2R 15a , -N(R 15 )S(O)R 15a , -N(R 15 )C(O)OR 15a , -N(R 15 )C(O)N(R 15a )(R 15b ), -OC(O)N(R 15 )(R 15a ) and C 1~6 Selected from the group consisting of alkyl groups, C 1~6 Alkyl is optionally substituted with one or more halogens, which are the same or different. -R 14 ,-R 14a ,-R 15 ,-R 15a and -R 15b -H and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is optionally substituted with one or more halogens, which are the same or different. Depending on the case, -R 6 / -R 6a ,-R A / -R B or -R 6 / -R A One or more of the pairs form the -PG portion, Depending on the case, -R 1 / -R 1a ,-R 2 / -R 2a ,-R 3 / -R 3a ,-R 4 / -R 4a ,-R 5 / -R 5a or -R 8 / -R 8a One or more of the pairs, together with the atom they are bonded to, become C 3~10 Forming cycloalkyl, 3-10 membered heterocyclyl, or 8-11 membered heterobicyclyl, Depending on the case, -R 1 / -R 2 ,-R 1 / -R 8 ,-R 1 / -R 9 ,-R 2 / -R 9 or -R 2 / -R 10 One or more of the pairs, together with the atom to which they are bonded, form a ring-A-. -A- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl, Depending on the case, -R 3 / -R 6 ,-R 4 / -R 6 ,-R 5 / -R 6 ,-R 6 / -R 6a or -R 6 / -R 7 One or more of the pairs form a ring -A'- together with the atom to which they are bonded. -A'- is selected from the group consisting of 3- to 10-membered heterocyclyls and 8- to 11-membered heterobicyclyls. This is the linker part, -L*- is at least one -L 2 -Z portion or at least one -L 2 -The Y portion is sometimes replaced, and sometimes further replaced. -L 2 - indicates a single bond or spacer portion. Z independently forms a polymer portion or C 8~24 It is alkyl, -Y is a functional group that may exist in its protected form. This is the aforementioned reagent.

[0013] In a particular embodiment, -L*- in formula (II) is at least one -L 2 -Y portion or at least one -L 2 -Z is substituted, and may be further substituted. In a particular embodiment, -L*- in formula (II) is replaced by at least one -L 2 -Y portion or at least one -L2 -Z is replaced, but -X 3 - is not -S-, and in some cases, -L*- in equation (II) is further substituted.

[0014] In a particular embodiment, -L*- in formula (II) is at least one -L 2 -It is replaced by the Y part. In a particular embodiment, -L*- in formula (II) is replaced by one -L 2 -Y is substituted. In a particular embodiment, -L*- in formula (II) is replaced by two -L 2 -It is replaced by the Y part. In a particular embodiment, -L*- in formula (II) is replaced by three -L 2 -The Y portion is being replaced.

[0015] In a particular embodiment, -L*- in formula (II) is at least one -L 2 -Z is substituted. In a particular embodiment, -L*- in formula (II) is replaced with one -L 2 -Z is substituted. In a particular embodiment, -L*- in formula (II) is replaced by two -L 2 -Z is substituted. In a particular embodiment, -L*- in formula (II) is replaced by three -L 2 -Z is being replaced.

[0016] The part of the structure below

[0017] [ka] Regarding "-R 6 / -R 6a The phrase "The pair forms the -PG part" is -R 6 and -R 6a However, these, along with the nitrogen atom to which they are bonded, form an imine functional group, for example:

[0018] [ka] [In the formula, -R xand -R y is -H, C 1~4 [Independently selected from the group consisting of alkyl, phenyl, and methoxyphenyl] to form or -R 6 and -R 6a However, along with the nitrogen to which these are bonded, azide functional groups, for example [ka] It is understood that this means forming something.

[0019] The following structural parts

[0020] [ka] Regarding this, "Depending on the case, -R A and -R B The phrase "The pair can form the -PG portion" is -R A and -R B However, along with the nitrogen atom to which these are bonded, the imine functional group, for example

[0021] [ka] [In the formula, -R x and -R y is -H, C 1~4 [Independently selected from the group consisting of alkyl, phenyl, and methoxyphenyl] to form or -R A and -R B However, along with the nitrogen to which these are bonded, azide functional groups, for example

[0022] [ka] It is also understood to mean forming something.

[0023] The part of the structure below

[0024] [ka] Regarding this, "Depending on the case, -R 6 / -R A The phrase "The pair can form the -PG portion" is -R 6 and -R A However, together with the nitrogen atom to which these are bonded,

[0025] [ka] [In the formula, -R t and -R z is -H, C 1~4 A variable group -R is independently selected from the group consisting of alkyl, phenyl, and methoxyphenyl. 3 ,-R 3a ,-R 5 ,-R 5a ,-R 6a ,-R 7 ,-R B And p are as defined in equation (II). It is also understood to mean forming something.

[0026] Another aspect of the present invention is an intermediate (A) for a reagent of the present invention comprising the -L*- portion of formula (II) defined above, wherein the -L*- portion is conjugated to at least one -D portion, Each -D is independently a primary or secondary amine-containing portion of drug DH. The dashed line in equation (II) indicates the bonding of the primary or secondary amine of -D to the nitrogen. The -L*- in equation (II) is at least one -L 2 -Z portion or at least one -L 2 -The Y portion is sometimes replaced, and sometimes further replaced. -L 2 - is an independent single bond or spacer portion, Z independently forms a polymer portion or C 8-24 It is alkyl, -Y is a functional group that may exist in its protected form depending on the circumstances. It is an intermediate (A).

[0027] The expression "-Y is a functional group that may exist in its protected form depending on the circumstances" is understood to mean that -Y can be reversibly attached to the protecting group portion.

[0028] Intermediate (A) containing the linker moiety of formula -L*- can be obtained by the reaction of a reagent containing the linker moiety of formula -L*- with the drug DH, for example, by the displacement of -Q. When -Q is -OH, the reaction of the reagent with the drug moiety DH in the presence of a coupling reagent can yield, for example, (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium, hexafluorophosphate, (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, N,N,N',N'-tetramethyl-O-(1H-benzotriazole-1- Those skilled in the art will understand that this can be carried out in the presence of a coupling reagent selected from the group consisting of (yl)uronium hexafluorophosphate, O-(benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, (benzotriazole-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate, and N,N,N',N'-tetramethyl-O-(N-succinimidyl)uronium tetrafluoroborate.

[0029] Another aspect of the present invention is a method for synthesizing the conjugates or pharmaceutically acceptable salts thereof as defined above. The conjugates or conjugate intermediates of the present invention may be prepared by known methods or according to the synthetic methods described below.

[0030] A method for synthesizing a conjugate according to the present invention, (a) A step of preparing a reagent containing the linker-L*- of formula (II), (b) A step of conjugating the reagent from step (a) with a primary or secondary amine-containing drug to obtain intermediate (A), (c) The intermediate (A) of step (b) is exposed to deprotection conditions to form the intermediate (C') or the linker-L of formula (I). 1 - A step to obtain a conjugate or intermediate (B) containing, (d) Depending on the case, the intermediate (B) or (C') obtained in step (c) is subjected to shift conditions. (e) optionally a step of deprotecting intermediate (B) or (C') of step (d), and (f) A step of isolating the conjugate resulting from step (c), (d), or (e). Includes, Depending on the case, at least one Z portion may bond to at least one intermediate (A), (B), or (C') between steps (b) and (c), (c) and (d), (d) and (e), or (e) and (f). It is a method.

[0031] The intermediate (C') of step (c) is conjugated to at least one Z portion to form the linker L of equation (I). 1 -It is understood that this results in a conjugate containing -. In a particular embodiment, the intermediate (C') of step (c) conjugates to one Z portion to form the linker -L of formula (I). 1 - produces a conjugate containing this.

[0032] Another embodiment is a method for synthesizing a conjugate according to the present invention, (a) A step of preparing a reagent containing the linker-L*- of formula (II), (b) A step of conjugating the reagent from step (a) with a primary or secondary amine-containing drug to obtain intermediate (A), (c) The intermediate (A) of step (b) is exposed to deprotection conditions to form the linker L of formula (I) 1 - A step to obtain a conjugate or intermediate (B) containing, (d) Depending on the case, the intermediate (B) obtained in step (c) is subjected to shift conditions. (e) optionally a step of deprotecting the intermediate (B) of step (d), and (f) A step of isolating the conjugate resulting from step (c), (d), or (e). Includes, Depending on the case, at least one Z portion may bond to at least one intermediate (A) or (B) between steps (b) and (c), (c) and (d), (d) and (e), or (e) and (f). It is a method.

[0033] A reagent containing the linker -L*- of formula (II) contains at least one -L 2 -Z or -L 2 -It is understood that if the Y portion is not substituted, at least one Z portion may bond to intermediate (B) after step (d). In a particular embodiment, one Z portion bonds to intermediate (B) after step (d).

[0034] In the method described above, the binding or conjugation of at least one Z moiety to at least one intermediate (A) or (B) is such that the reagent of step (a) already contains the linker -L*- of formula (II), and -L*- is at least one -L 2 It is also understood that if the -Z portion has already been replaced, it may be necessary.

[0035] For example, if the -Y portion of the -L*-Y section exists in its protective form, that is, if it is reversibly connected to the protective base portion, it will also be apparent to those skilled in the art that the portion needs to be exposed to deprotection conditions before conjugation to Z. [Brief explanation of the drawing]

[0036] [Figure 1]Figure 1 shows an example of a rearrangement that takes place during the steps of a method for synthesizing a conjugate according to the present invention, the conjugate corresponding to structure (C) in Figure 1. Step (a) comprises preparing a reagent containing the linker -L*- of formula (II), which corresponds to structure (R) in Figure 1. Step (b) comprises a conjugation of a primary or secondary amine-containing drug, i.e., DH, with the reagent containing the linker -L*- of formula (II), i.e., structure (R), which is carried out by nucleophilic attack of the primary or secondary amine functional group of the drug on the carbonyl group directly bonded to the -Q moiety. This results in the release of QH and the formation of intermediate (A), corresponding to structure (A) in Figure 1. In step (c), intermediate (A) is subjected to deprotection conditions, thereby converting the variable groups -RA and -RB, which are independently selected from the group consisting of -H and -PG, provided that one or less of the -RA or -RB atoms can be -H, to -H atoms. Simultaneously, the nitrogen atom marked with "#" undergoes an intramolecular shift with the variable group -X3- to produce the conjugate according to the present invention corresponding to structure (C) in Figure 1. Figure 1 also shows the optimal conditions under which, after intermediate (A) is subjected to deprotection conditions, the variable groups -RA and -RB are converted to -H atoms to provide intermediate (B) corresponding to structure (B) in Figure 1. In step (d), intermediate (B) obtained in step (c) is subjected to shift conditions, under which the nitrogen atom marked with "#" undergoes an intramolecular shift with the variable group -X3- to produce the conjugate according to the present invention corresponding to structure (C). For simplicity, the -L2-Z or -L2-Y portions are not shown. For simplicity, step (e), if necessary, is also not shown in Figure 1. In structures (R), (A), and (B) of Figure 1, the variable groups -R6 and -R6a are defined by formula (II) described above, and it is understood that if at least one of -R6 and -R6a is -PG, the variable group can be converted to an -H atom under the conditions of step (c) or (d), preferably under the conditions of step (e). [Modes for carrying out the invention]

[0037] Within the scope of the meaning of this invention, the terms are used as follows:

[0038] As used herein, the term “drug” refers to a substance used to treat, cure, prevent or diagnose a disease, or otherwise to enhance the physical or mental health of a patient. When a drug is conjugated into another part, the part of the resulting product derived from the drug is called the “drug part.”

[0039] As used herein, the term “primary or secondary amine-containing portion of drug DH” means a portion of a drug comprising at least one primary or secondary amine functional group, wherein the drug may optionally have one or more further functional groups comprising one or more additional primary and / or secondary amine functional groups.

[0040] As used herein, the term “part” means a portion of a molecule that lacks one or more atoms compared to the corresponding reagent. For example, when a reagent of the formula “HXH” reacts with another reagent and becomes part of the reaction product, the corresponding part of the reaction product has the structure “HX-” or “-X-”, where each “-” indicates bonding to another part. Thus, the drug part is released as a drug from reversible bonding.

[0041] If an arrangement or chemical structure of a group of atoms is provided, and this group of atoms is bonded to or interrupts two parts, it is understood that the arrangement or chemical structure may be bonded to the two parts in either direction unless otherwise explicitly stated. For example, "-C(O)N(R x The part )-" is "-C(O)N(R x )-" or "-N(R x It can be bonded to two parts or interrupted to one part as either )C(O)-.

[0042] As used herein, the term “protecting group moiety” refers to a moiety that reversibly attaches to a functional group, preventing it from reacting with, for example, another functional group. Suitable alcohol (-OH) protecting groups include, for example, acetyl, benzoyl, benzyl, β-methoxyethoxymethyl ether, dimethoxytrityl, methoxymethyl ether, methoxytrityl, p-methoxybenzyl ether, methylthiomethyl ether, pivaloyl, tetrahydropyranyl, trityl, trimethylsilyl, tert-butyldimethylsilyl, tri-isopropylsilyloxymethyl, triisopropylsilyl ether, methyl ether, and ethoxyethyl ether. Suitable carbonyl protecting groups include, for example, acetals and ketals, acylal, and dithiane. Suitable carboxylic acid protecting groups include, for example, methyl esters, benzyl esters, tert-butyl esters, 2,6-dimethylphenol, 2,6-diisopropylphenol, 2,6-di-tert-butylphenol, silyl esters, ortho esters, and oxazolines. Suitable phosphate protecting groups include, for example, 2-cyanoethyl and methyl.

[0043] As used herein, the term “amine protecting group portion” refers to a portion used for the reversible protection of an amine protecting group during a chemical reaction process, thereby preventing the amine from reacting with, for example, another functional group.

[0044] As used herein, the term “leaving group moiety” refers to an atom or group of atoms that is separated from the rest of a molecule, for example, from a reagent, during a chemical reaction with another functional group.

[0045] As used herein, the term “deprotection conditions” refers to conditions under which at least one protecting group moiety, for example, an amine protecting group moiety of an intermediate, is separated or cleaved from a functional group, for example, from an amine group, under conditions that may include the use of an acid, a base, a reducing agent, an oxidizing agent, hydrogenation, or light and optionally a scavenger reagent.

[0046] As used herein, the term “reducing agent” refers to a chemical compound or element that loses or donates electrons to an electron acceptor, such as an oxidizing agent in a redox chemical reaction.

[0047] As used herein, the term “oxidizing agent” refers to a chemical compound that can oxidize other chemical compounds.

[0048] As used herein, the term “capture reagent” refers to a chemical compound that captures another reaction intermediate, such as a reactive reaction intermediate.

[0049] As used herein, the term “shift conditions” refers to conditions under which a primary amine intermediate, such as intermediate (B), can undergo intramolecular rearrangement, which may include the use of a buffer or an organic solvent.

[0050] As used herein, the terms “buffer” or “buffering agent” refer to chemical compounds that maintain pH within a desired range. Physiologically tolerable buffers include, for example, acetate, adipate, alanine, ammonium, arginine, ascorbate, aspartate, benzoate, bicarbonate, carbonate, citrate, diethanolamine, edetate, ethylenediamine, fumarate, and gluconate. These include (gluconate), glutamate, glycine, guanidine, histidine, lactate, lysine, malate, metaphosphate, pentetate, phosphate, pyruvate, sorbate, succinate, tartrate, tromethamine, and α-ketoglutarate.

[0051] As used herein, the term “polar protic solvent” refers to a solvent that contains bonds between atoms having different electronegativity, has a large dipole moment, and has at least one hydrogen atom directly bonded to an electronegative atom, such as oxygen, nitrogen, or sulfur.

[0052] As used herein, the term “polar aprotic solvent” refers to a solvent that contains bonds between atoms with different electronegativity, has a large dipole moment, and does not contain hydrogen atoms directly bonded to electronegative atoms, such as oxygen, nitrogen, or sulfur atoms.

[0053] As used herein, the term “reagent” means a chemical compound containing at least one functional group that reacts with the functional group of another chemical compound or drug. A drug containing a functional group is also understood to be a reagent.

[0054] Those skilled in the art will recognize that the conjugate of the present invention is a prodrug. As used herein, the term “prodrug” means at least one -L 1 -L 2 - refers to a drug moiety that is reversibly and covalently conjugated to a fatty acid-derived moiety or polymer moiety, e.g., Z, via a moiety. The prodrug releases the reversibly and covalently conjugated drug moiety-D in the form of its corresponding drug DH. In other words, the prodrug contains at least one -L 1 -L 2 - A conjugate containing a drug moiety covalently and reversibly conjugated to a polymer moiety via a portion. Such a prodrug or conjugate releases the previously conjugated drug moiety in the form of a free drug.

[0055] As used herein, the terms “reversible linkage” or “biodegradable linkage” refer to a linkage that is cleaved under physiological conditions in the absence of enzymes, where physiological conditions are aqueous buffer at pH 7.4 and 37°C, and the half-life ranges from 1 hour to 6 months, e.g., 10 hours to 4 months, e.g., 1 day to 3 months, 2 days to 2 months, or 3 days to 1 month. However, reversible linkages can also be cleaved under other conditions, e.g., different pH or different temperature, with a half-life ranging from 1 hour to 6 months, but it is understood that tests to determine reversibility are performed under the physiological conditions described above (aqueous buffer, pH 7.4, 37°C). Thus, a “stable linkage” is a linkage with a half-life of more than 6 months under physiological conditions.

[0056] When used herein, the term “about” in combination with a number is used to indicate a range from the number plus or minus 10% or less of the number (including the number), in a particular embodiment, 8% or less of the number (including the number), in a particular embodiment, 5% or less of the number (including the number), and in a particular embodiment, 2% or less of the number (including the number). For example, the phrase “about 200” is used to mean a range from 200+ / -10% (including this value), i.e., a range from 180 to 220 (including these values), in a particular embodiment, a range from 200+ / -8%, i.e., a range from 184 to 216 (including these values), in a particular embodiment, a range from 200+ / -5% (including this value), i.e., a range from 190 to 210 (including these values), and in a particular embodiment, a range from 200+ / -2%, i.e., a range from 196 to 204 (including these values). When a percentage is presented as "approximately 20%", it is understood that it does not mean "20% + / - 10%", i.e., it does not mean a range from 10 to 30% (including these values), but rather that "approximately 20%" means a range from 18 to 22% (including these values), i.e., plus or minus 10% of the number 20.

[0057] When used herein, the term "C 1~4"Alkyl" refers to a linear or branched alkyl moiety having 1 to 4 carbon atoms, either alone or in combination. When present at the end of a molecule, it refers to a linear or branched C 1~4 Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Two parts of the molecule are C 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 atom in the alkyl carbon may optionally be replaced by a substituent as defined below. 1~4 Alkyl can be interrupted by one or more parts as defined below.

[0058] When used herein, the term "C 1~6 "Alkyl" refers to a linear or branched alkyl moiety having 1 to 6 carbon atoms, either alone or in combination. When present at the end of a molecule, it refers to linear and branched C atoms. 1~6 Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethylpropyl. Two parts of the molecule are C 1~6 When linked by an alkyl group, such C 1~6 Examples of alkyl groups are -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -CH(C2H5)-, and -C(CH3)2-. 1~6 Each hydrogen atom in carbon may be optionally replaced by a substituent as defined below. 1~6 Alkyl can be interrupted by one or more parts as defined below.

[0059] Therefore, "C 1~10"Alkyl", "C 1~20 "Alkyl", "C 8~24 "Alkyl" or "C 1~50 "Alkyl" refers to an alkyl chain having 1 to 10, 1 to 20, 8 to 24, or 1 to 50 carbon atoms, respectively. 1~10 , C 1~20 , C 8~24 or C 1~50 Each hydrogen atom in carbon may be optionally replaced by a substituent as defined below. 1~10 Alkyl, C 1~20 Alkyl, C 8~24 Alkyl or C 1~50 Alkyl can be interrupted by one or more parts as defined below.

[0060] When used herein, the term "C 2~6 An "alkenyl" refers to a straight-chain or branched-chain hydrocarbon moiety, either alone or in combination, that contains at least one carbon-carbon double bond having 2 to 6 carbon atoms. When present at the ends of a molecule, examples include -CH=CH2, -CH=CH-CH3, -CH2-CH=CH2, -CH=CHCH2-CH3, and -CH=CH-CH=CH2. Two parts of a molecule are C 2~6 When linked by an alkenyl group, such C 2~6 An example of an alkenil is -CH=CH-. 2~6 Each hydrogen atom in the alkenyl moiety may optionally be replaced by a substituent as defined below. 2~6 An alkenil can be interrupted in one or more parts as defined below.

[0061] Therefore, the term "C 2~10 Alkenil, "C 2~20 "Alkenil" or "C 2~50 "Alkenyl" means a straight-chain or branched-chain hydrocarbon moiety, either alone or in combination, containing at least one carbon-carbon double bond having 2 to 10, 2 to 20, or 2 to 50 carbon atoms, respectively. 2~10 Alkenil, C 2~20 Alkenyl or C 2~50Each hydrogen atom of the alkenyl group may optionally be replaced by a substituent as defined below. 2~10 Alkenil, C 2~20 Alkenyl or C 2~50 An alkenil can be interrupted in one or more parts as defined below.

[0062] When used herein, the term "C 2~6 "Alkynyl" refers to a straight-chain or branched-chain hydrocarbon moiety, either alone or in combination, that contains at least one carbon-carbon triple bond having 2 to 6 carbon atoms. When present at the end of a molecule, examples include -C≡CH, -CH2-C≡CH, CH2-CH2-C≡CH, and CH2-C≡C-CH3. When two parts of a molecule are linked by an alkynyl group, an example is -C≡C-. 2~6 Each hydrogen atom of the alkynyl group may optionally be replaced by a substituent as defined below. One or more double bonds may be formed. C may be present. 2~6 Alkinyl can be interrupted in one or more parts as defined below.

[0063] Therefore, when used herein, the term "C 2~10 Alkinyl, C 2~20 "Alkinyl" and "C 2~50 "Alkynyl" means a linear or branched hydrocarbon moiety, either alone or in combination, that contains at least one carbon-carbon triple bond having 2 to 10, 2 to 20, or 2 to 50 carbon atoms, respectively. 2~10 Alkinyl, C 2~20 Alkinyl or C 2~50 Each hydrogen atom of the alkynyl group may optionally be replaced by a substituent as defined below. One or more double bonds may be formed. C may be present. 2~10 Alkinyl, C 2~20 Alkinyl or C 2~50 Alkinyl can be interrupted in one or more parts as defined below.

[0064] As described above, C 1~4 Alkyl, C1~6 Alkyl, C 1~10 Alkyl, C 1~20 Alkyl, C 1~50 Alkyl, C 8~24 Alkyl, C 2~6 Alkenil, C 2~10 Alkenil, C 2~20 Alkenil, C 2~50 Alkenil, C 2~6 Alkinyl, C 2~10 Alkinyl, C 2~20 Alkenyl or C 2~50 Alkinyl, in certain embodiments, is as follows:

[0065] [ka] [In the formula, The dashed line indicates binding to a portion or the rest of the reagent. -R and -R a [These 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] It may be interrupted by one or more parts selected from the group consisting of; These parts and connections are sometimes further replaced.

[0066] When used herein, the term "C 3~10 "Cycloalkyl" refers to a cyclic alkyl chain having 3 to 10 carbon atoms, which may be saturated or unsaturated. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl. 3~10 Each hydrogen atom in the cycloalkyl carbon may be replaced by a substituent as defined below. Term "C 3~10 "Cycloalkyl" also includes cross-linked birings such as norbornane or norbornene.

[0067] As used herein, the terms “8-30 membered carbopolycyclil” or “8-30 membered carbopolycycle” mean two or more ring-shaped portions having 8 to 30 ring atoms, where two adjacent rings share at least one ring atom and may contain up to a maximum number of double bonds (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic rings). In certain embodiments, an 8-30 membered carbopolycyclil means a ring-shaped portion with 2, 3, 4, or 5 rings. In certain embodiments, an 8-30 membered carbopolycyclil means a ring-shaped portion with 2, 3, or 4 rings.

[0068] As used herein, the terms “3- to 10-membered heterocyclyl” or “3- to 10-membered heterocycle” mean a ring having 3, 4, 5, 6, 7, 8, 9, or 10 ring atoms, which may contain up to a maximum number of double bonds (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic rings), wherein at least one to four ring atoms are replaced by heteroatoms selected from the group consisting of sulfur (including -S(O)-, -S(O)2-), oxygen, and nitrogen (including =N(O)-), and the ring is linked to the rest of the molecule via carbon or nitrogen atoms. Examples of 3- to 10-membered heterocycles include, but are not limited to, aziridine, oxirane, thiirane, azirine, oxilen, thiirane, azetidine, oxetane, thietan, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazole, isothiazoline, thiadiazole, thiadiazoleline, tetrahydrofuran, tetrahydrothiophene, pyrrolidine, imidazolidine, pyrazolidine, oxazolidine, isooxazolidine, thiazoline, isothiazoline, thiadiazole, thiadiazoleline, tetrahydrothiophene, pyrrolidine, imidazolidine, pyrazolidine, oxazolidine, isooxazolidine, thiadiazolidine, sulforane, pyran, dihydropyran, tetrahydropyran, imidazolidine, pyridine, pyridazine, pyridine, piperazine, piperidine, morpholine, tetrazole, triazole, triazolidine, tetrazolidin, diazepane, azepine, and homopiperazine. Each hydrogen atom of a 3- to 10-membered heterocyclyl or 3- to 10-membered heterocyclic group may be replaced by a substituent as defined below.

[0069] As used herein, the terms “8-11 member heterobicyryl” or “8-11 member heterobicycle” mean a bicyclic heterocyclic portion having 8-11 ring atoms, where at least one ring atom is shared by both rings and may contain up to a maximum number of double bonds (fully saturated, partially saturated, or unsaturated, aromatic or aromatic rings), where at least one to six ring atoms are replaced by heteroatoms selected from the group consisting of sulfur (including -S(O)-, -S(O)2-), oxygen, and nitrogen (including =N(O)-), and the rings are linked to the rest of the molecule via carbon or nitrogen atoms. Examples of 8- to 11-membered heterobicyclic rings include indole, indoline, benzofuran, benzothiophene, benzoxazole, benzoisoxazole, benzothiazole, benzoisothiazole, benzimidazole, benzimidazolin, quinoline, quinazoline, dihydroquinazoline, quinoline, dihydroquinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, decahydroisoquinoline, tetrahydroisoquinoline, dihydroisoquinoline, benzoazepine, purine, and pteridine. The term 8- to 11-membered heterobicyclic ring also includes bicyclic spiro structures such as 1,4-dioxa-8-azaspiro[4.5]decane or bridging heterocyclic rings such as 8-aza-bicyclo[3.2.1]octane. Each hydrogen atom in an 8- to 11-membered heterobicyclyl or 8- to 11-membered heterobicyclic carbon may be replaced by a substituent as defined below.

[0070] Similarly, the term “8-30 membered heteropolycyclil” or “8-30 membered heteropolycycle” means two or more rings having 8-30 ring atoms, in certain embodiments a heterocyclic portion of 3, 4, or 5 rings, where at least 1 to 10 ring atoms are replaced by heteroatoms selected from the group consisting of sulfur (including -S(O)-, -S(O)2-), oxygen, and nitrogen (including =N(O)-), and the rings are linked to the rest of the molecule via carbon or nitrogen atoms.

[0071] The part of the structure below

[0072] [ka] Regarding "-R x / -R y The pair, together with the atom to which they are bonded, is C 3~10 The phrase "forms cycloalkyl, 3-10 membered heterocyclyl, or 8-11 membered heterobicyclyl" is -R x and -R y The structure is as follows:

[0073] [ka] (In the formula, R is C 3~10 (They are cycloalkyl, 3-10 membered heterocyclyl, or 8-11 membered heterobicyclyl.) It is understood that this means forming something.

[0074] The part of the structure below

[0075] [ka] Regarding "-R x / -R yThe phrase "These pairs, together with the atoms they are bonded to, form a ring -A-" is expressed as -R x and -R y The structure is as follows:

[0076] [ka] It is also understood to mean forming something.

[0077] As used herein, the term “excipient” refers to a diluent, adjuvant, or vehicle with which a therapeutic agent, such as a drug or conjugate, is administered. Such pharmaceutical excipients may be sterile liquids, such as water, or oils, including peanut oil, soybean oil, mineral oil, and sesame oil, or oils of petroleum, animal, plant, or synthetic origin. When the pharmaceutical composition is administered orally, water is a preferred excipient. When the pharmaceutical composition is administered intravenously, saline and aqueous dextrose are preferred excipients. Salt solutions, as well as aqueous dextrose and glycerol solutions, are preferably used as liquid excipients for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, mannitol, trehalose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene glycol, hyaluronic acid, propylene glycol, water, and ethanol. The pharmaceutical composition may, if desired, contain trace amounts of wetting agents or emulsifiers, pH buffers, etc., such as acetate, succinate, tris, carbonate, phosphate, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), MES (2-(N-morpholino)ethanesulfonic acid), or detergents, such as Tween®, poloxamer, poloxamine, CHAPS, Igepal®, or amino acids, such as glycine, lysine, or histidine. These pharmaceutical compositions may take the form of liquids, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations, etc. The pharmaceutical composition may be formulated as a suppository using traditional binders and excipients, such as triglycerides. Oral formulations may contain standard excipients, such as pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Such compositions contain a therapeutically effective amount of the drug or drug portion together with appropriate amounts of excipients to provide a form for appropriate administration to the patient. The formulation should be suitable for the mode of administration.

[0078] As used herein, the term “free form” of a drug refers to the unmodified, pharmacologically active form of the drug after it has been released, for example, from a conjugate.

[0079] As used herein, the term “functional group” means a group of atoms that can react with other groups of atoms. Exemplary functional groups include carboxylic acids, primary amines, secondary amines, tertiary amines, maleimides, thiols, sulfonic acids, carbonates, carbamates, hydroxyls, aldehydes, ketones, hydrazines, isocyanates, isothiocyanates, phosphoric acids, phosphonic acids, haloacetyls, alkyl halides, acryloyls, aryl fluorides, hydroxylamines, disulfides, sulfonamides, sulfuric acids, vinyl sulfones, vinyl ketones, diazoalkanes, oxiranes, and aziridines.

[0080] As used herein, the term "halogen" means fluoro, chloro, bromo, or iodine. In certain embodiments, the halogen is fluoro or chloro.

[0081] As used herein, the term “interrupted” means that a portion is inserted between two carbon atoms, or, if the insertion is at one end of the portion, between a carbon or heteroatom and a hydrogen atom, or in a particular embodiment, between a carbon atom and a hydrogen atom.

[0082] If the conjugate of the present invention contains one or more acidic or basic groups, the present invention also includes corresponding pharmaceutically or toxicologically acceptable salts thereof, in particular pharmaceutically useful salts thereof. Thus, the conjugate of the present invention containing acidic groups can be used, for example, as alkali metal salts, alkaline earth metal salts, or ammonium salts according to the present invention. More precise examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts, or ammonia or organic amines, such as ethylamine, ethanolamine, triethanolamine, or amino acids, or quaternary ammonium salts, such as tetrabutylammonium and cetyltrimethylammonium. The conjugate of the present invention containing one or more basic groups, i.e., protonable groups, may exist in the form of addition salts with inorganic or organic acids, and may be used according to the present invention in the form of such addition salts. 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, trifluoroacetic acid, and other acids known to those skilled in the art. Further methods are known to those skilled in the art, such as alkylation of amine groups, to convert basic groups into cations to produce positively charged ammonium groups and suitable counterions for salts. When the conjugate of the present invention contains both an acidic and a basic group, the present invention also includes internal salts or betaines (zwitterionic) in addition to the described salt forms. The corresponding salts can be obtained by conventional methods known to those skilled in the art, for example, by contacting these prodrugs with organic or inorganic acids or bases in a solvent or dispersion, or by anion exchange or cation exchange with other salts. The present invention also includes all salts of the conjugates of the present invention, which are not directly suitable for pharmaceutical use due to their low physiological compatibility, but can be used, for example, as intermediates in chemical reactions or in the preparation of pharmaceutically acceptable salts.

[0083] As used herein, the term “pharmaceutically acceptable” means a substance that does not cause harm when administered to a patient and is preferably approved by a regulatory authority, e.g., the EMA (Europe) and / or the FDA (US), and / or any other national regulatory authority, for use in animals, and preferably for use in humans.

[0084] As used herein, the term "peptide" refers to a chain of at least two to 50 amino acid monomer moieties, sometimes called "amino acid residues" when linked by peptide (amide) linkages. Amino acid monomers may be selected from the group consisting of proteogenic and non-proteogenic amino acids, and may be D- or L-amino acids. The term "peptide" also includes peptide mimetics, such as peptoids, beta-peptides, cyclic peptides, and depsipeptides, and encompasses peptide mimetic chains having up to 50 monomer moieties. Cyclic peptides may be monocyclic, dicyclic, tricyclic, or tetracyclic peptides. The term "peptide" also includes lassopeptides.

[0085] As used herein, the term "protein" refers to a chain of more than 50 amino acid monomers, which may also be called "amino acid residues" linked by peptide linkages, preferably consisting of 12,000 or fewer amino acid monomers linked by peptide linkages, for example, a portion with 10,000 or fewer amino acid monomers, a portion with 8,000 or fewer amino acid monomers, a portion with 5,000 or fewer amino acid monomers, or a portion with 2,000 or fewer amino acid monomers.

[0086] As used herein, the term “small molecule drug” refers to a drug that is an organic compound having a molecular weight of less than 1000 Da, for example, less than 900 Da or less than 800 Da. It is understood that nucleic acid base-based drug moieties, such as adenine or guanine analogs, may also be a type of small molecule drug.

[0087] As used herein, the term “medium-molecule drug” refers to a drug that is not a peptide or a protein, but an organic compound having a molecular weight in the range of 1 kDa to 7.5 kDa (including these values).

[0088] As used herein, the term “oligonucleotide” preferably refers to double- or single-stranded RNA and DNA having 2 to 1000 nucleotides and any modifications thereof. Modifications include, for example, those that provide other chemical groups that incorporate additional charge, polarity, hydrogen bonding, electrostatic interactions, and mobility into the nucleic acid ligand base or the nucleic acid ligand as a whole. Such modifications include, for example, 2'-position sugar modifications, 5-position pyrimidine modifications, 8-position purine modifications, modifications in extracyclic amines, 4-thiouridine substitutions, 5-bromo or 5-iodouracil substitutions; skeletal modifications, methylation, unusual base-pairing combinations, e.g., isobases, isocytidine and isoguanidine. Modifications may also include 3' and 5' modifications, e.g., capping and stereochemical changes. Modifications also include aptamers.

[0089] As used herein, the term “peptide nucleic acid” refers to an organic polymer having a peptide backbone, i.e., a backbone to which monomers are linked to one another via peptide linkages, to which nucleic acid bases, such as adenine, cytosine, guanine, thymine, and uracil are bound. In certain embodiments, the peptide backbone comprises N-(2-aminoethyl)-glycine.

[0090] As used herein, the term “polymer” means a molecule containing repeating structural units, i.e., monomers, that are chemically linked in linear, cyclic, branched, crosslinked, or dendrimeric, or combinations thereof, and can be synthetic, biologically derived, or a combination of both. The monomers may be identical, in which case the polymer is a homopolymer; or they may be different, in which case the polymer is a heteropolymer. Heteropolymers are sometimes called “copolymers,” and include, for example, alternating copolymers in which different types of monomers are arranged alternately, periodic copolymers in which different types of monomers are arranged in a repeating sequence, statistical copolymers in which different types of monomers are arranged randomly, block copolymers in which blocks of different homopolymers consisting of only one type of monomer are covalently linked, and gradient copolymers in which the composition of different monomers gradually changes along the polymer chain. It is understood that polymers may also contain one or more other parts, for example, one or more functional groups. Similarly, peptides or proteins are understood to be polymers even if the side chains of individual amino acid residues are different. It is understood that covalently crosslinked polymers, such as hydrogels, cannot provide a meaningful molecular weight range.

[0091] As used herein, the terms “polymeric” or “polymeric part” refer to a reagent or part comprising one or more polymers or polymeric parts. A polymeric reagent or part may also comprise one or more other parts, which in certain embodiments are as follows: - C 1~50 Alkyl, C 2~50 Alkenil, C 2~50 Alkinyl, C 3~10 Cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl, - Branch points, e.g., -CR<, >C< or -N<, and - the below described:

[0092] [ka] [In the formula, The dashed line indicates binding to a portion or the rest of the reagent. -R and -R a [These 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] A concatenation selected from the group including, Selected from the group consisting of, These parts and connections are sometimes further replaced.

[0093] It will be understood by those skilled in the art that polymerization products obtained by polymerization reactions do not all have the same molecular weight, but rather exhibit a molecular weight distribution. Therefore, when used herein, the molecular weight range, molecular weight, range of monomer numbers, and number of monomers in a polymer refer to the number-average molecular weight and the average number of monomers, i.e., the arithmetic mean of the molecular weight of the polymer or polymer part and the arithmetic mean of the number of monomers in the polymer or polymer part.

[0094] Therefore, in a polymer portion containing "x" monomer units, any integer presented as "x" corresponds to the arithmetic mean (arithmetic mean number) of the monomers. Any range of integers presented as "x" provides a range of integers that is the arithmetic mean number of the monomers. An integer "x" presented as "about x" means that the arithmetic mean number of the monomers is in the range of integers x+ / -10%, in a particular embodiment it is in the range of integers x+ / -8%, in a particular embodiment it is in the range of integers x+ / -5%, and in a particular embodiment it is in the range of integers x+ / -2%.

[0095] As used herein, the term “number-mean molecular weight” means the usual arithmetic mean of the molecular weights of individual polymers.

[0096] When used herein, the term “PEG-based” in relation to a portion or reagent means that the portion or reagent contains PEG. In certain embodiments, such a PEG-based portion or reagent contains at least 10% (w / w) PEG, e.g., at least 20% (w / w) PEG, e.g., at least 30% (w / w) PEG, e.g., at least 40% (w / w) PEG, e.g., at least 50% (w / w) PEG, e.g., at least 60% (w / w) PEG, e.g., at least 70% (w / w) PEG, e.g., at least 80% (w / w) PEG, e.g., at least 90% (w / w) PEG, e.g., at least 95% (w / w) PEG. The remaining weight percentage of the PEG-based portion or reagent is that of the other portions, e.g., as follows: - C 1~50 Alkyl, C 2~50 Alkenil, C 2~50 Alkinyl, C 3~10 Cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl, - Branch points, e.g., -CR<, >C< or -N<, and - the below described:

[0097] [ka] [In the formula, The dashed line indicates binding to a portion or the rest of the reagent. -R and -R a [These 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] A concatenation selected from the group including, It may also be selected from the group consisting of, These parts and connections are sometimes further replaced.

[0098] When used herein, the term “PEG-based containing at least X% PEG” in relation to a portion or reagent means that the portion or reagent contains at least X% (w / w) ethylene glycol units (-CH2CH2O-), which may be arranged in blocks and alternatingly, or randomly distributed within the portion or reagent. In a particular embodiment, all ethylene glycol units of the portion or reagent are present in a single block, and the remaining weight percentage of the PEG-based portion or reagent is as follows in a particular embodiment: - C 1~50 Alkyl, C 2~50 Alkenil, C 2~50 Alkinyl, C 3~10 Cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl, - Branch points, e.g., -CR<, >C< or -N<, and - the below described:

[0099] [ka] [In the formula, The dashed lines indicate binding to a portion or the rest of the reagent, and -R and -R a [These 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] A concatenation selected from the group including, It is a part of the group consisting of other parts selected from the group, These parts and connections are sometimes further replaced.

[0100] When used herein, the term “hyaluronic acid base” in relation to a portion or reagent means that the portion or reagent contains hyaluronic acid. Such a hyaluronic acid base portion or reagent contains at least 10% (w / w) hyaluronic acid, e.g., at least 20% (w / w) hyaluronic acid, e.g., at least 30% (w / w) hyaluronic acid, e.g., at least 40% (w / w) hyaluronic acid, e.g., at least 50% (w / w) hyaluronic acid, e.g., at least 60% (w / w) hyaluronic acid, e.g., at least 70% (w / w) hyaluronic acid, e.g., at least 80% (w / w) hyaluronic acid, e.g., at least 90% (w / w) hyaluronic acid, or e.g., at least 95% (w / w) hyaluronic acid. The remaining weight percentage of the hyaluronic acid base portion or reagent is that of the other portions, e.g., as follows: - C 1~50 Alkyl, C 2~50 Alkenil, C 2~50 Alkinyl, C 3~10 Cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl, - Branch points, e.g., -CR<, >C< or -N<, and - the below described:

[0101] [ka] [In the formula, The dashed line indicates binding to a portion or the rest of the reagent. -R and -R a [These 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] A connection selected from the group consisting of, It may also be selected from the group consisting of, These parts and connections are sometimes further replaced.

[0102] As used herein, the term "hydrogel" means a hydrophilic or amphiphilic polymer network composed of homopolymers or copolymers that is insoluble due to the presence of hydrophobic interactions, hydrogen bonds, ionic interactions and / or covalent chemical crosslinks. Crosslinks provide the network structure and physical integrity.

[0103] As used herein, the term “random coil” refers to a peptide or protein that, in certain embodiments, adopts / has / forms a conformation substantially lacking defined secondary and tertiary structures, as determined by circular dichroism spectroscopy performed in aqueous buffer at ambient temperature and pH 7.4. In certain embodiments, the ambient temperature is approximately 20°C, i.e., between 18°C ​​and 22°C, while in certain embodiments, the ambient temperature is 20°C.

[0104] As used herein, the term “spacer” or “spacer portion” refers to a portion suitable for connecting two parts. A suitable spacer is C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Selected from the group consisting of alkynyl, these C 1~50 Alkyl, C2~50 Alkenyl or C 2~50 Alkinyl is -NH-, -N(C 1~4 Alkyl)-, -O-, -S-, -C(O)-, -C(O)NH-, -C(O)N(C 1~4 It may be optionally interrupted and optionally substituted with one or more groups selected from alkyl)-, -OC(O)-, -S(O)-, -S(O)2-, 4- to 7-membered heterocyclyl, phenyl, and naphthyl.

[0105] As used herein, the term “substituted” means that one or more -H atoms in a molecule or part are replaced by different atoms or groups of atoms called “substituents.”

[0106] As used herein, the term “substituent” means, in a particular embodiment, a halogen, -CN, -C(O)OR x1 , -OR x1 , -C(O)R x1 ,-C(O)N(R x1 )(R x1a ), -S(O)2N(R x1 )(R x1a ), -S(O)N(R x1 )(R x1a ), -S(O)2R x1 ,-S(O)R x1 , -N(R x1 )S(O)2N(R x1a )(R x1b ), -SR x1 , -N(R x1 )(R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R x1a , -N(R x1 )S(O)R x1a , -N(R x1 )C(O)OR x1a , -N(R x1 )C(O)N(R x1a )(R x1b ), -OC(O)N(R x1 )(Rx1a ), -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 This refers to the part selected from the group consisting of alkynyls, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is one or more identical or different -R x2 In some cases, it is replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl 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 )- is optionally interrupted by one or more groups selected from the group consisting of, -R x1 ,-R x1a ,-R x1b -H, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Independently selected from the group consisting of alkynnyls, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is one or more identical or different -R x2 In some cases, it is replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl 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 )- is optionally interrupted by one or more groups selected from the group consisting of, Each T 0 Phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl, each T 0 This refers to one or more -Rs that are identical or different. x2 It is independently substituted depending on the case, Each-R x2 These are halogen, -CN, oxo (=O), -C(O)OR x4 , -OR x4 , -C(O)R x4 ,-C(O)N(R x4 )(R x4a ), -S(O)2N(R x4 )(R x4a ), -S(O)N(R x4 )(R x4a ), -S(O)2R x4 ,-S(O)R x4 , -N(R x4 )S(O)2N(R x4a )(R x4b ), -SR x4 , -N(R x4 )(R x4a ), -NO2, -OC(O)R x4 , -N(R x4 )C(O)R x4a , -N(R x4 )S(O)2R x4a , -N(R x4 )S(O)Rx4a , -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 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is optionally substituted with one or more halogens, which are the same or different. Each-R x3 ,-R x3a ,-R x4 ,-R x4a ,-R x4b -H and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is sometimes substituted with one or more halogens, which are either the same or different.

[0107] In certain embodiments, the term "substituent" can refer to halogens, -CN, -C(O)OR x1 , -OR x1 , -C(O)R x1 ,-C(O)N(R x1 )(R x1a ), -S(O)2N(R x1 )(R x1a ), -S(O)N(R x1 )(R x1a ), -S(O)2R x1 ,-S(O)R x1 , -N(R x1 )S(O)2N(R x1 )(R x1a ), -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 x1 )(R x1a ), -OC(O)N(R x1 )(R x1a ), -T 0 , C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 This refers to the part selected from the group consisting of alkynyls, -T 0 , C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Alkinyl is one or more identical or different -R x2 In some cases, it is replaced by C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Alkinyl 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 )- is optionally interrupted by one or more groups selected from the group consisting of, Each-R x1 ,-R x1a ,-R x1b ,-R x3 ,-R x3a -H, halogen, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkinyls, Each T 0 Phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl, each T 0 This refers to one or more -Rs that are identical or different. x2 It is independently substituted depending on the case, Each-R x2 These are halogen, -CN, oxo (=O), -C(O)OR x4 , -OR x4 , -C(O)R x4 ,-C(O)N(R x4 )(R x4a ), -S(O)2N(R x4 )(R x4a ), -S(O)N(R x4 )(R x4a ), -S(O)2R x4 ,-S(O)R x4 , -N(R x4 )S(O)2N(R x4a )(R x4b ), -SR x4 , -N(R x4 )(R x4a ), -NO2, -OC(O)R x4 , -N(R x4 )C(O)R x4a , -N(R x4 )S(O)2R x4a , -N(R x4 )S(O)R x4a , -N(R x4 )C(O)OR x4a , -N(R x4 )C(O)N(R x4a )(R x4b ), -OC(O)N(R x4 )(R x4a ) and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is optionally substituted with one or more halogens, which are the same or different. Each-R x4 ,-R x4a ,-R x4b -H, halogen, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6It is independently selected from the group consisting of alkynnyls.

[0108] In certain embodiments, the term "substituent" can refer to halogens, -CN, -C(O)OR x1 , -OR x1 , -C(O)R x1 ,-C(O)N(R x1 )(R x1a ), -S(O)2N(R x1 )(R x1a ), -S(O)N(R x1 )(R x1a ), -S(O)2R x1 ,-S(O)R x1 , -N(R x1 )S(O)2N(R x1a )(R x1b ), -SR x1 , -N(R x1 )(R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R x1a , -N(R x1 )S(O)R x1a , -N(R x1 )C(O)OR x1a , -N(R x1 )C(O)N(R x1a )(R x1b ), -OC(O)N(R x1 )(R x1a ), -T 0 , C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 This refers to the part selected from the group consisting of alkynyls, -T 0 , C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is one or more identical or different -R x2 In some cases, it is replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl 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 )- is optionally interrupted by one or more groups selected from the group consisting of, Each-R x1 ,-R x1a ,-R x1b ,-R x2 ,-R x3 ,-R x3a -H, halogen, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkinyls, Each T 0 Phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl, each T 0 This refers to one or more -Rs that are identical or different. x2 They are independently substituted depending on the situation.

[0109] In certain embodiments, in optionally substituted molecules, up to six -H atoms may be independently replaced by substituents, for example, five -H atoms may be independently replaced by substituents, four -H atoms may be independently replaced by substituents, three -H atoms may be independently replaced by substituents, two -H atoms may be independently replaced by substituents, or one -H atom may be independently replaced by substituents.

[0110] As used herein, the term “therapeutic dose” means an amount sufficient to cure, alleviate, or partially prevent the clinical symptoms of a given disease and its complications. The effective dose for each purpose depends on the severity of the disease or injury, as well as the subject’s weight and overall health.

[0111] As used herein, the term "water-insoluble" refers to a compound in which the amount that dissolves in 1 liter of water at 20°C to form a homogeneous solution is less than 1 g. Conversely, the term "water-soluble" refers to a compound in which the amount that dissolves in 1 liter of water at 20°C to form a homogeneous solution is 1 g or more.

[0112] Generally, the terms "comprise(s)" or "comprising" also include "consist of" or "consisting of".

[0113] In certain embodiments, all -D portions of the conjugate are identical, i.e., have the same chemical structure. In such cases, all -D portions of the conjugate originate from the same type of drug molecule.

[0114] In certain embodiments, the conjugate of the present invention comprises different -D moieties, i.e., -D moieties having different chemical structures. These different structures originate from different types of drug molecules. In certain embodiments, the conjugate of the present invention comprises two different types of -D moieties. In certain embodiments, the conjugate of the present invention comprises three different types of -D moieties. In certain embodiments, the conjugate of the present invention comprises four different types of -D moieties. In certain embodiments, the conjugate of the present invention comprises five different types of -D moieties.

[0115] If the conjugate of the present invention includes more than one type of -D, all -D portions are of the same type of -L. 1 -It may be conjugated to -L of a different type. 1- may be conjugated to -D of the first type, i.e., -L of the first type 1 - may be conjugated to -D, and the second type -L 1 -It can also be conjugated to other things. Different types of -L 1 -The use of - in certain embodiments may allow for different release kinetics with different types of -D, for example, fast release with the first type of -D, moderate release with the second type of -D, and slow release with the third type of -D. Thus, in certain embodiments, the conjugate of the present invention may be one type of -L 1 -Includes. In a particular embodiment, the conjugate of the present invention is of two types -L 1 -Includes. In a particular embodiment, the conjugate of the present invention is of three types -L 1 -Includes. In a particular embodiment, the conjugate of the present invention is of four types -L 1 -Includes

[0116] In a particular embodiment, the conjugate of the present invention comprises one type -D and one type -L 1 -Includes. In a particular embodiment, the conjugate of the present invention includes two types of -D and two types of -L 1 -Includes. In a particular embodiment, the conjugate of the present invention includes three types of -D and three types of -L 1 -Includes. In a particular embodiment, the conjugate of the present invention includes four types -D and four types -L 1 -Includes. In a particular embodiment, the conjugate of the present invention includes two types of -D and one type of -L 1 -Includes. In a particular embodiment, the conjugate of the present invention includes three types of -D and one type of -L 1 -Includes. In a particular embodiment, the conjugate of the present invention includes four types of -D and one type of -L 1 -Includes

[0117] In a particular embodiment, all -L of the conjugate 1 -The parts have the same structure. In a particular embodiment, the conjugate is two or more different types of -L 1 - Parts, e.g., 2, 3, 4 or 5 different types -L 1 -Includes a portion. Two or more different types of -L 1 -The - part may be conjugated to the same or different types of -D. Different types of -L 1 - is used, for example, in the first group of -L with a short emission half-life. 1 -The second group -L, which releases a portion with a longer half-life 1 -When combined with a portion, it enables the release of the same or different types of drug DH from the conjugate of the present invention with different release half-lives.

[0118] In a particular embodiment, -D is selected from the group consisting of small molecules, medium-sized molecules, oligonucleotides, peptide nucleic acids, peptides, and protein drug moieties.

[0119] In a particular embodiment, -D is selected from the group consisting of small molecules, medium molecules, peptides, and protein drug moieties.

[0120] In certain embodiments, -D is a small molecule drug moiety. In certain embodiments, such a small molecule drug moiety is a nucleic acid base-based drug moiety.

[0121] In certain embodiments, -D is a medium-molecule drug moiety. In certain embodiments, -D is an oligonucleotide drug moiety. In certain embodiments, -D is a peptide nucleic acid protein drug moiety.

[0122] The -D portion is understood to include at least one primary or secondary amine group, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 primary or secondary amine groups, and may also include one or more primary amine groups and one or more secondary amine groups.

[0123] In a particular embodiment, -D is the peptide drug portion.

[0124] In a particular embodiment, -D is a peptide drug moiety selected from the group consisting of type C natriuretic peptide, parathyroid hormone, W peptide, memnopeptide A, and G1 peptide.

[0125] In a particular embodiment, -D is a bicyclic peptide drug moiety.

[0126] In certain embodiments, -D is a protein drug moiety. In certain embodiments, such a protein moiety is a monoclonal or polyclonal antibody or a fragment or fusion thereof.

[0127] Surprisingly, it has been found that the conjugates of the present invention can be obtained through synthetic methods that avoid the use of amide or amidine protecting groups, for example, while providing the benefit of obtaining stable reagents. This is particularly beneficial for protein drug moieties, as some proteins are more susceptible to degradation under deprotection conditions of amide or amidine protecting groups than, for example, small, medium, and peptide drug moieties.

[0128] In a particular embodiment, -X of formula (I) 3 - is -O-. In a particular embodiment, -X of formula (I) 3 - is -S-. In a particular embodiment, -X of formula (I) 3 - is -Se-

[0129] In a particular embodiment, the -R of formula (I) 6 is -H-. In a particular embodiment, -R of formula (I)6 is -C(R 11 )(R 11a )(R 11b ). In certain embodiments, -R 6 of formula (I) is -T.

[0130] In certain embodiments, -R 6a of formula (I) is -H. In certain embodiments, -R 6a of formula (I) is -C(R 11 )(R 11a )(R 11b ). In certain embodiments, -R 6a of formula (I) is -T.

[0131] In certain embodiments, both -R 6 and -R 6a of formula (I) are -H. <000418​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​In a particular embodiment, -R of formula (II) 6 is -C(R 11 )(R 11a )(R 11b ) and -R of equation (II) 6a is -T. In a particular embodiment, -R of equation (II) 6 and -R 6a Both are -C(R 11 )(R 11a )(R 11b ) is. In a particular embodiment, -R of formula (II) 6 and -R 6a Both are -T.

[0135] In a particular embodiment, -R of formula (II) A is -H, and -R in equation (II) B It is -PG.

[0136] In a particular embodiment, -R of formula (II) 6 -R in equation (II) 6a Together, they form the -PG portion.

[0137] In a particular embodiment, -R of formula (II) 6 -R in equation (II) A Together, they form the -PG portion.

[0138] In a particular embodiment, -R of formula (II) A -R in equation (II) B Together, they form the -PG portion.

[0139] In a particular embodiment, v in formula (I) or (II) is 0. In a particular embodiment, v in formula (I) or (II) is 1.

[0140] In a particular embodiment, -X of formula (I) or (II) 1 - is -C(R 8 )(R 8a )-. In a particular embodiment, -X of formula (I) or (II) 1 - is -N(R9 )-. In a particular embodiment, -X of formula (I) or (II) 1 - is -O-

[0141] In a particular embodiment, equation (I) or (II) = X 2 is = O. In a particular embodiment, equation (I) or (II) = X 2 is = N(R 10 )

[0142] In a particular embodiment, -R of formula (I) or (II) 9 is -C(R 11 )(R 11a )(R 11b ) is. In certain embodiments, -R of formula (I) or (II) 9 is -T.

[0143] In a particular embodiment, -R of formula (I) or (II) 10 is -H. In certain embodiments, -R of formula (I) or (II) 10 is -C(R 11 )(R 11a )(R 11b ) is. In certain embodiments, -R of formula (I) or (II) 10 is -T.

[0144] In a particular embodiment, -R of formula (I) or (II) 1 -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR12 , -NO2, -N(R 12 )(C(O)OR 12a , -N(R 12 )(C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 alkyl, C 2~6 alkenyl and C 2~6 alkynyl selected from the group consisting of. In certain embodiments, -R 1 in formula (I) or (II) is -H. In certain embodiments, -R 1 in formula (I) or (II) is halogen. In certain embodiments, -R 1 in formula (I) or (II) is -T. In certain embodiments, -R 1 in formula (I) or (II) is C 1~6 alkyl. In certain embodiments, -R 1 in formula (I) or (II) is C 2~6 alkenyl. In certain embodiments, -R 1 in formula (I) or (II) is C 2~6 alkynyl. In certain embodiments, -R 1 in formula (I) or (II) is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl and 1-ethylpropyl.

[0145] In certain embodiments, -R 1a in formula (I) or (II) is -H, halogen, -CN, -C(O)OR<00,00989>, -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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 1a is -H. In certain embodiments, -R of formula (I) or (II) 1a is a halogen. In certain embodiments, -R of formula (I) or (II) 1a is -T. In certain embodiments, -R of formula (I) or (II) 1a is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 1a is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 1a is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 1a The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0146] In a particular embodiment, -R of formula (I) or (II) 2 -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 ,-C(O)N(R12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 2 is -H. In certain embodiments, -R of formula (I) or (II) 2 is a halogen. In certain embodiments, -R of formula (I) or (II) 2 is -T. In certain embodiments, -R of formula (I) or (II) 2 is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 2 is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 2 is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 2 The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0147] In a particular embodiment, -R of formula (I) or (II) 2a -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 2a is -H. In certain embodiments, -R of formula (I) or (II) 2a is a halogen. In certain embodiments, -R of formula (I) or (II) 2a is -T. In certain embodiments, -R of formula (I) or (II) 2a is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 2a is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 2a is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 2aThe group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0148] In a particular embodiment, -R of formula (I) or (II) 3 -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 3 is -H. In certain embodiments, -R of formula (I) or (II) 3 is a halogen. In certain embodiments, -R of formula (I) or (II) 3 is -T. In certain embodiments, -R of formula (I) or (II) 3 is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 3 is C 2~6It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 3 is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 3 The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0149] In a particular embodiment, -R of formula (I) or (II) 3a -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 3a is -H. In certain embodiments, -R of formula (I) or (II) 3a is a halogen. In certain embodiments, -R of formula (I) or (II) 3ais -T. In certain embodiments, -R of formula (I) or (II) 3a is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 3a is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 3a is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 3a The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0150] In a particular embodiment, -R of formula (I) or (II) 4 -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II).4 is -H. In a particular embodiment, -R of formula (I) 4 is a halogen. In certain embodiments, -R of formula (I) or (II) 4 is -T. In certain embodiments, -R of formula (I) or (II) 4 is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 4 is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 4 is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 4 The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0151] In a particular embodiment, -R of formula (I) or (II) 4a -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 4a is -H. In certain embodiments, -R of formula (I) or (II) 4a is a halogen. In certain embodiments, -R of formula (I) or (II) 4a is -T. In certain embodiments, -R of formula (I) or (II) 4a is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 4a is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 4a is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 4a The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0152] In a particular embodiment, -R of formula (I) or (II) 5 -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a, -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 5 is -H. In certain embodiments, -R of formula (I) or (II) 5 is a halogen. In certain embodiments, -R of formula (I) or (II) 5 is -T. In certain embodiments, -R of formula (I) or (II) 5 is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 5 is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 5 is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 5 The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0153] In a particular embodiment, -R of formula (I) or (II) 5a -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 5a is -H. In certain embodiments, -R of formula (I) or (II) 5a is a halogen. In certain embodiments, -R of formula (I) or (II) 5a is -T. In certain embodiments, -R of formula (I) or (II) 5a is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 5a is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 5a is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 5a The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0154] In a particular embodiment, -R of formula (I) or (II) 7 -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 7 is -H. In certain embodiments, -R of formula (I) or (II) 7 is a halogen. In certain embodiments, -R of formula (I) or (II) 7 is -T. In certain embodiments, -R of formula (I) or (II) 7 is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 7 is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 7 is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 7 The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0155] In a particular embodiment, -R of formula (I) or (II) 8 -H, halogen, -CN, -C(O)OR12 , -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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 8 is -H. In certain embodiments, -R of formula (I) or (II) 8 is a halogen. In certain embodiments, -R of formula (I) or (II) 8 is -T. In certain embodiments, -R of formula (I) or (II) 8 is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 8 is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 8 is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 8 The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0156] In a particular embodiment, -R of formula (I) or (II) 8a -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 8a is -H. In certain embodiments, -R of formula (I) or (II) 8a is a halogen. In certain embodiments, -R of formula (I) or (II) 8a is -T. In certain embodiments, -R of formula (I) or (II) 8 is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 8a is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 8a is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 8aThe group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0157] In a particular embodiment, -R of formula (I) or (II) 11 -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 11 is -H. In certain embodiments, -R of formula (I) or (II) 11 is a halogen. In certain embodiments, -R of formula (I) or (II) 11 is -T. In certain embodiments, -R of formula (I) or (II) 11 is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 11 is C 2~6It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 11 is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 11 The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0158] In a particular embodiment, -R of formula (I) or (II) 11a -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 11a is -H. In certain embodiments, -R of formula (I) or (II) 11a is a halogen. In certain embodiments, -R of formula (I) or (II) 11ais -T. In certain embodiments, -R of formula (I) or (II) 11a is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 11a is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 11a is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 11a The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0159] In a particular embodiment, -R of formula (I) or (II) 11b -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II).11b is -H. In certain embodiments, -R of formula (I) or (II) 11b is a halogen. In certain embodiments, -R of formula (I) or (II) 11b is -T. In certain embodiments, -R of formula (I) or (II) 11b is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 11b is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 11b is C 2~6 It is an alkynyl. In certain embodiments, it is -R of formula (I) or (II). 11b The group is selected from -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0160] In a particular embodiment, -R of formula (I) or (II) 12 -H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 12 is -H. In certain embodiments, -R of formula (I) or (II) 12 is -T. In certain embodiments, -R of formula (I) or (II) 12 is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 12 is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 12 is C 2~6 It is alkinyl.

[0161] In a particular embodiment, -R of formula (I) or (II) 12a-H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 12a is -H. In certain embodiments, -R of formula (I) or (II) 12a is -T. In certain embodiments, -R of formula (I) or (II) 12a is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 12a is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 12a is C 2~6 It is alkinyl.

[0162] In a particular embodiment, -R of formula (I) or (II) 12b -H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Selected from the group consisting of alkynyls. In certain embodiments, -R of formula (I) or (II). 12b is -H. In certain embodiments, -R of formula (I) or (II) 12b is -T. In certain embodiments, -R of formula (I) or (II) 12b is C 1~6 It is alkyl. In certain embodiments, it is -R of formula (I) or (II). 12b is C 2~6 It is an alkenyl. In certain embodiments, it is -R of formula (I) or (II). 12b is C 2~6 It is alkinyl.

[0163] In certain embodiments, T in formula (I) or (II) is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10Selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl. In certain embodiments, T in formula (I) or (II) is phenyl. In certain embodiments, T in formula (I) or (II) is naphthyl. In certain embodiments, T in formula (I) or (II) is indenyl. In certain embodiments, T in formula (I) or (II) is indanyl. In certain embodiments, T in formula (I) or (II) is tetralinyl. In certain embodiments, T in formula (I) or (II) is tetralinyl. In certain embodiments, T in formula (I) or (II) is C 3~10 It is a cycloalkyl compound. In certain embodiments, T in formula (I) or (II) is a 3- to 10-membered heterocyclyl compound. In certain embodiments, T in formula (I) or (II) is an 8- to 11-membered heterobicyclyl compound.

[0164] In a particular embodiment, T in formula (I) or (II) is one or more -R in formula (I) or (II) that are identical or different. 13 It has been replaced with.

[0165] In a particular embodiment, T in formula (I) or (II) is one of the -R in formula (I) or (II). 13 It has been replaced with.

[0166] In a particular embodiment, T in formula (I) or (II) is -R 13 It has not been replaced.

[0167] In a particular embodiment, -R of formula (I) or (II) 13 These are halogen, -CN, oxo, and -C(O)OR 15 , -OR 15 , -C(O)R 15 ,-C(O)N(R 15 )(R 15a ), -S(O)2N(R 15 )(R 15a ), -S(O)N(R 15 )(R 15a ), -S(O)2R15 ,-S(O)R 15 , -N(R 15 )S(O)2N(R 15a )(R 15b ), -SR 15 , -N(R 15 )(R 15a ), -NO2, -OC(O)R 15 , -N(R 15 )C(O)R 15a , -N(R 15 )S(O)2R 15a , -N(R 15 )S(O)R 15a , -N(R 15 )C(O)OR 15a , -N(R 15 )C(O)N(R 15a )(R 15b ), -OC(O)N(R 15 )(R 15a ) and C 1~6 Selected from the group consisting of alkyl groups. In certain embodiments, -R of formula (I) or (II) 13 is a halogen. In certain embodiments, -R of formula (I) or (II) 13 is -CN. In certain embodiments, -R of formula (I) or (II) 13 is -oxo. In certain embodiments, -R of formula (I) or (II) 13 is -C(O)OR 15 In a particular embodiment, -R of formula (I) or (II) 13 は-OR 15 In a particular embodiment, -R of formula (I) or (II) 13 -C(O)R 15 In a particular embodiment, -R of formula (I) or (II) 13 is -C(O)N(R 15 )(R 15a ) is. In certain embodiments, -R of formula (I) or (II) 13 is -S(O)2N(R 15 )(R 15a ) is. In certain embodiments, -R of formula (I) or (II) 13 is -S(O)N(R15 )(R 15a ) is. In certain embodiments, -R of formula (I) or (II) 13 -S(O)2R 15 In a particular embodiment, -R of formula (I) or (II) 13 -S(O)R 15 In a particular embodiment, -R of formula (I) or (II) 13 is -N(R 15 )S(O)2N(R 15a )(R 15b ) is. In certain embodiments, -R of formula (I) or (II) 13 Ha-SR 15 In a particular embodiment, -R of formula (I) or (II) 13 is -N(R 15 )(R 15a ) is. In certain embodiments, -R of formula (I) or (II) 13 is -NO2. In certain embodiments, -R of formula (I) or (II) 13 -OC(O)R 15 In a particular embodiment, -R of formula (I) or (II) 13 is -N(R 15 )C(O)R 15a In a particular embodiment, -R of formula (I) or (II) 13 is -N(R 15 )S(O)2R 15a In a particular embodiment, -R of formula (I) or (II) 13 is -N(R 15 )C(O)OR 15a In a particular embodiment, -R of formula (I) or (II) 13 is -N(R 15 )C(O)N(R 15a )(R 15b ) is. In certain embodiments, -R of formula (I) or (II) 13 -OC(O)N(R 15 )(R 15a ) is. In certain embodiments, -R of formula (I) or (II) 13 is C 1~6It is alkyl.

[0168] In a particular embodiment, -R of formula (I) or (II) 14 -H and C 1~6 Selected from the group consisting of alkyl groups. In certain embodiments, -R of formula (I) or (II) 14 is -H. In certain embodiments, -R of formula (I) or (II) 14 is C 1~6 It is alkyl.

[0169] In a particular embodiment, -R of formula (I) or (II) 14a -H and C 1~6 Selected from the group consisting of alkyl groups. In certain embodiments, -R of formula (I) or (II) 14a is -H. In certain embodiments, -R of formula (I) or (II) 14a is C 1~6 It is alkyl.

[0170] In a particular embodiment, -R of formula (I) or (II) 15 -H and C 1~6 Selected from the group consisting of alkyl groups. In certain embodiments, -R of formula (I) or (II) 15 is -H. In certain embodiments, -R of formula (I) or (II) 15 is C 1~6 It is alkyl.

[0171] In a particular embodiment, -R of formula (I) or (II) 15a -H and C 1~6 Selected from the group consisting of alkyl groups. In certain embodiments, -R of formula (I) or (II) 15a is -H. In certain embodiments, -R of formula (I) or (II) 15a is C 1~6 It is alkyl.

[0172] In a particular embodiment, -R of formula (I) or (II) 15b -H and C1~6 Selected from the group consisting of alkyl groups. In certain embodiments, -R of formula (I) or (II) 15b is -H. In certain embodiments, -R of formula (I) or (II) 15b is C 1~6 It is alkyl.

[0173] In a particular embodiment, -R of formula (I) or (II) 1 and -R 1a These, together with the atoms to which they are bonded, form C 3~10 Forms a cycloalkyl group. In certain embodiments, -R of formula (I) or (II) 1 and -R 1a These, together with the atoms to which they are bonded, form 3- to 10-membered heterocyclines. In certain embodiments, -R of formula (I) or (II) 1 and -R 1a These atoms, together with the atoms to which they are bonded, form 8-11 member heterobicyryls.

[0174] In a particular embodiment, -R of formula (I) or (II) 2 and -R 2a These, together with the atoms to which they are bonded, form C 3~10 Forms a cycloalkyl group. In certain embodiments, -R of formula (I) or (II) 2 and -R 2a These, together with the atoms to which they are bonded, form 3- to 10-membered heterocyclines. In certain embodiments, -R of formula (I) or (II) 2 and -R 2a These atoms, together with the atoms to which they are bonded, form 8-11 member heterobicyryls.

[0175] In a particular embodiment, -R of formula (I) or (II) 3 and -R 3a These, together with the atoms to which they are bonded, form C 3~10 Forms a cycloalkyl group. In certain embodiments, -R of formula (I) or (II) 3 and -R3a These, together with the atoms to which they are bonded, form 3- to 10-membered heterocyclines. In certain embodiments, -R of formula (I) or (II) 3 and -R 3a These atoms, together with the atoms to which they are bonded, form 8-11 member heterobicyryls.

[0176] In a particular embodiment, -R of formula (I) or (II) 4 and -R 4a These, together with the atoms to which they are bonded, form C 3~10 Forms a cycloalkyl group. In certain embodiments, -R of formula (I) or (II) 4 and -R 4a These, together with the atoms to which they are bonded, form 3- to 10-membered heterocyclines. In certain embodiments, -R of formula (I) or (II) 4 and -R 4a These atoms, together with the atoms to which they are bonded, form 8-11 member heterobicyryls.

[0177] In a particular embodiment, -R of formula (I) or (II) 5 and -R 5a These, together with the atoms to which they are bonded, form C 3~10 Forms a cycloalkyl group. In certain embodiments, -R of formula (I) or (II) 5 and -R 5a These, together with the atoms to which they are bonded, form 3- to 10-membered heterocyclines. In certain embodiments, -R of formula (I) or (II) 5 and -R 5a These atoms, together with the atoms to which they are bonded, form 8-11 member heterobicyryls.

[0178] In a particular embodiment, -R of formula (I) or (II) 8 and -R 8a These, together with the atoms to which they are bonded, form C 3~10 Forms a cycloalkyl group. In certain embodiments, -R of formula (I) or (II)8 and -R 8a These, together with the atoms to which they are bonded, form 3- to 10-membered heterocyclines. In certain embodiments, -R of formula (I) or (II) 8 and -R 8a These atoms, together with the atoms to which they are bonded, form 8-11 member heterobicyryls.

[0179] In a particular embodiment, -R of formula (I) or (II) 1 and -R 2 These, together with the atoms to which they are bonded, form the ring-A- of formula (I) or (II).

[0180] In a particular embodiment, -R of formula (I) or (II) 1 and -R 8 These, together with the atoms to which they are bonded, form the ring-A- of formula (I) or (II).

[0181] In a particular embodiment, -R of formula (I) or (II) 1 and -R 9 These, together with the atoms to which they are bonded, form the ring-A- of formula (I) or (II).

[0182] In a particular embodiment, -R of formula (I) or (II) 2 and -R 9 These, together with the atoms to which they are bonded, form the ring-A- of formula (I) or (II).

[0183] In a particular embodiment, -R of formula (I) or (II) 2 and -R 10 These, together with the atoms to which they are bonded, form the ring-A- of formula (I) or (II).

[0184] In certain embodiments, -A- in formula (I) or (II) is phenyl. In certain embodiments, -A- in formula (I) or (II) is naphthyl. In certain embodiments, -A- in formula (I) or (II) is indenyl. In certain embodiments, -A- in formula (I) or (II) is indanyl. In certain embodiments, -A- in formula (I) or (II) is tetralinyl. In certain embodiments, -A- in formula (I) or (II) is C 3~10 It is a cycloalkyl compound. In certain embodiments, the -A- in formula (I) or (II) is a 3- to 10-membered heterocyclyl compound. In certain embodiments, the -A- in formula (I) or (II) is an 8- to 11-membered heterobicyclyl compound.

[0185] In a particular embodiment, -R of formula (I) or (II) 1 and -R 9 These, together with the atoms to which they are bonded, form the ring -A'- of formula (I) or (II).

[0186] In a particular embodiment, -R of formula (I) or (II) 2 and -R 9 These, together with the atoms to which they are bonded, form the ring -A'- of formula (I) or (II).

[0187] In a particular embodiment, -R of formula (I) or (II) 3 and -R 6 These, together with the atoms to which they are bonded, form the ring -A'- of formula (I) or (II).

[0188] In a particular embodiment, -R of formula (I) or (II) 4 and -R 6 These, together with the atoms to which they are bonded, form the ring -A'- of formula (I) or (II).

[0189] In a particular embodiment, -R of formula (I) or (II) 5 and -R 6These, together with the atoms to which they are bonded, form the ring -A'- of formula (I) or (II).

[0190] In a particular embodiment, -R of formula (I) or (II) 6 and -R 6a These, together with the atoms to which they are bonded, form the ring -A'- of formula (I) or (II).

[0191] In a particular embodiment, -R of formula (I) or (II) 6 and -R 7 These, together with the atoms to which they are bonded, form the ring -A'- of formula (I) or (II).

[0192] In certain embodiments, -A'- in formula (I) or (II) is a 3- to 10-membered heterocyclyl. In certain embodiments, -A'- in formula (I) or (II) is an 8- to 11-membered heterobicyclyl.

[0193] In a particular embodiment, -PG is as follows:

[0194] [ka] TIFF0007851853000021.tif251160[In the formula, The dashed line indicates the bond to the nitrogen atom in formula (II) that can be substituted with -PG, where -R is C 1~6 It is alkyl. It is selected from the group consisting of the following.

[0195] In a particular embodiment, -PG is the linker portion of a reversible prodrug disclosed in WO2005 / 099768A2. Thus, -PG is given by formula (bi):

[0196] [ka] [In the formula, The dashed line indicates the bond to the nitrogen atom in formula (II) that can be substituted with -PG. n is 0, 1, 2, 3, or 4. -X- is a chemical bond or spacer, =Y1 and =Y5 are independently selected from the group consisting of =O and =S. -Y2- and -Y3- were independently selected from the group consisting of -O- and -S-. -Y4- is -O-, -NR 5 - and -C(R 6 R 6a Selected from the group consisting of )-, -R 2 ,-R 3 ,-R 5 ,-R 6 ,-R 6a These 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 This 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, -CO-, -C(O)N(R) 7 )-, -O-, -S- and -N(R 7 C is interrupted in one or more of the groups selected from the group consisting of )- 1~20 Selected from the group consisting of alkyl groups, -Nu is a nucleophile, -Ar- is a polysubstituted aromatic hydrocarbon or polysubstituted aromatic heterocycle. It belongs to them.

[0197] The dashed line in equation (bi) indicates the bond of the -PG portion to one of the nitrogen atoms in equation (II) to which it can bond.

[0198] In some cases, -PG in equation (bi) is further substituted.

[0199] In a particular embodiment, =Y5 in equation (bi) equals =O.

[0200] In a particular embodiment, -Y3- in equation (bi) is -O-.

[0201] In a particular embodiment, -R2, -R3 and -R4 in formula (bi) are -H, C 1~6 Alkyl, C 2~6 Alkenyl or C 2~6 Selected independently from alkinyl, C 1~6 Alkyl, C 2~6 Alkenyl or C 2~6 Alkinyl is sometimes further substituted.

[0202] In a particular embodiment, =Y1 in equation (bi) equals =O.

[0203] In a particular embodiment, -Y2- in equation (bi) is -O-.

[0204] In a particular embodiment, -Y4- in formula (bi) is -NR 5 That is the case.

[0205] In a particular embodiment, the -R of formula (bi) 5 is -H or C 1~6 It is alkyl.

[0206] In a particular embodiment, Ar of formula (bi) is as follows:

[0207] [ka] TIFF0007851853000024.tif54160[In the formula, -Z 1 - are -O-, -S- and -N(R 7 Selected from the group consisting of )-, -Z 2 - is -N(R 7 )- and, -R 7 is -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 [Selected from the group consisting of alkynnyls] It is selected from the group consisting of the following.

[0208] In a particular embodiment, part of formula (bi)

[0209] [ka] The following:

[0210] [ka] [In the formula, -W-, -R 5 ,-R 6 ,-R 6a It is used as defined above. m is 2, 3, 4, 5, 6, 7, 8, 9, or 10. -R 9 ,-R 10 ,-R 11 and -R 12 is -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkynnyls, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is sometimes further substituted. It is selected from the group consisting of the following.

[0211] In a particular embodiment, -PG is formula (b-i')

[0212] [ka] [In the formula, The dashed line indicates the bond to the nitrogen atom in formula (II) that can be substituted by -PG. It belongs to them.

[0213] In a particular embodiment, -Q is -OH.

[0214] In a particular embodiment, -Q is -LG.

[0215] In certain embodiments, -LG is selected from the group consisting of chloride, bromide, fluoride, nitrophenoxy, imidazolyl, N-hydroxysuccinimidyl, N-hydroxybenzotriazolyl, N-hydroxyazobenzotriazolyl, pentafluorophenoxy, N-hydroxysulfosuccinimidyl, diphenylphosphinomethaneethyl, 2-diphenylphosphinophenoxy, norbornene-N-hydroxysuccinimidyl, N-hydroxyphthalimide, pyridinoxy, nonafluorotert-butyloxy, and hexafluoroisopropyloxy.

[0216] In certain embodiments, -LG is a chloride. In certain embodiments, -LG is a bromide. In certain embodiments, -LG is a fluoride. In certain embodiments, -LG is nitrophenoxy. In certain embodiments, -LG is imidazolyl. In certain embodiments, -LG is N-hydroxysuccinimidyl. In certain embodiments, -LG is N-hydroxybenzotriazolyl. In certain embodiments, -LG is pentafluorophenoxy. In certain embodiments, -LG is N-hydroxysulfosuccinimidyl. In certain embodiments, -LG is diphenylphosphinomethaneethyl. In certain embodiments, -LG is 2-diphenylphosphinophenoxy. In certain embodiments, -LG is norbornene-N-hydroxysuccinimidyl. In certain embodiments, -LG is N-hydroxyphthalimide. In certain embodiments, -LG is pyridinoxy. In certain embodiments, -LG is nonafluorotert-butyloxy. In certain embodiments, -LG is hexafluoroisopropyloxy.

[0217] In certain embodiments, -Y of formula (II) is as disclosed in WO2016 / 020373A1. Thus, -Y of formula (II) is thiols, maleimides, amines, hydroxyls, carboxylic acids and derivatives, carbonates and derivatives, carbamates and derivatives, isothiocyanates, disulfides, pyridyl disulfides, methylthiosulfonyl, vinyl sulfones, aldehydes, ketones, haloacetyls, selenides, azides, -NH-NH2, -O-NH2, terminal alkynes, formula (z'i)

[0218] [ka] [In the formula, Y 1 , Y 2 These are independently C or N, R a , Ra' , R a1 , R a1' These are independently -H or C 1~6 It is alkyl, Y 2 If N, then ax1 is 0, and Y 2 If C, then ax1 is 1, Depending on the circumstances, Y 2 If C, R a / R a1 The pair forms a chemical bond, Depending on the circumstances, Y 2 If C, R a' / R a1' The pairs, together with the atoms to which they are bonded, form ring A'. A' is cyclopropyl or phenyl. The compound, Equation (z'ii)

[0219] [ka] [In the formula, Y 3 [is C or N] The compound, Formula (z'iii)

[0220] [ka] The compound, Expression (z'iv)

[0221] [ka] [In the formula, R a2 , R a2' , R a3 , R a3' is -H,

[0222] [ka] This indicates a single bond or a double bond. Depending on the case, Ra2' / R a3' The pair, together with the atom to which they are bonded, forms ring A 1 ' form, A 1 ' is a 5-membered heterocyclyl.' The compound, Equation (z'v)

[0223] [ka] [In the formula, R a4 , R a4' , R a5 , R a5' is -H,

[0224] [ka] This indicates a single bond or a double bond. Depending on the case, R a4 / R a5 The pair forms a chemical bond, Depending on the case, R a4' / R a5' The pair, together with the atom to which they are bonded, forms ring A 2 ' form, A 2 ' is a 5-membered heterocycline.' The compound, Equation (z'vi)

[0225] [ka] [In the formula, R a6 , R a6' Both are C 1~6 Alkyl, or R a6 , R a6' One of them is -H, and the other is C 1~6 Alkyl, -COOR a7 , -CONHR a7' and CH2OR a7'' Selected from, R a7 , Ra7' , R a7'' -H or C 1~4 It is alkyl. The compound, expression (z'vii)

[0226] [ka] The compound, Expression (z'viii)

[0227] [ka] [In the formula, R a8 , R a8' , R a8'' -H and C 1~4 [Independently selected from the group consisting of alkyl groups] The compound, Expression(z'ix)

[0228] [ka] [In the formula, R a9 is -H or C 1~4 It is alkyl. The compound, Expression(z'x)

[0229] [ka] [In the formula, R a9 -COOR a11 , -CONHR a11 And below:

[0230] [ka] (In the ceremony Y 4 is C or N, R a12 -H, -COOR a13 ,-CONRa13 R a13' -CH2NR a13 R a13' and -NR a13 COR a13' Selected from the group consisting of, R a13 , R a13' -H and C 1~4 (Independently selected from the group consisting of alkyl groups) Selected from, A a3 -H, methyl, tert-butyl, -CF3, -COOR, below:

[0231] [ka] (In the formula, Each Y 5 , Y 6 , Y 7 , Y 8 is independently C or N, except Y 5 , Y 6 , Y 7 , Y 8 The condition is that three or fewer of them are N, Each Y 9 , Y 10 , Y 11 , Y 12 , Y 13 is one of C, N, S, or O, except Y 9 , Y 10 , Y 11 , Y 12 , Y 13 (The condition is that four or fewer of them are N, S, or O) Selected from the group consisting of: The compound, Equation (z'xi)

[0232] [ka] The compound, Equation (z'xii)

[0233] [ka] [In the formula, R a19 , R a19' is -H, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~8 [Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl, and tetralinyl] The compound, Equation (z'xiii)

[0234] [ka] [In the formula, R a20 is -H, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~8 [Selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl, and tetralinyl] The compound, Expression(z'xiv)

[0235] [ka] [In the formula, Ar is selected from phenyl, naphthyl, indenyl, indanyl, and tetralinyl. Y 14 It is a halogen, R a22 , R a23 , R a23' is -H, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~8 [Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl, and tetralinyl] The compound, Formula(z'xv)

[0236] [ka] [Ar is selected from phenyl, naphthyl, indenyl, indanyl and tetralinyl, R a24 , R a24' , R a24'' , R a24''' is -H, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~8 [Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl, and tetralinyl] The compound, Equation (z'xvi)

[0237] [ka] [In the formula, R a25 is -H, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~8 [Selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl, and tetralinyl] The compound, Expression(z'xvii)

[0238] [ka] [In the formula, R a27 , R a27' These are independently -H or C 1~6 It is alkyl. The compound, Formula(z'xviii)

[0239] [ka] The compound, Expression(z'xix)

[0240] [ka] [In the formula, -PPh2 is expressed by the following formula

[0241] [ka] (In the equation, the dashed line indicates the connection of part (z'xix) to the rest of the equation.) Represents a group having, R a12 The following:

[0242] [ka] (In the formula, The unmarked dashed line indicates the joining of part (z'xix) to the rest of the equation. The dashed line indicated by the asterisk is -L 2 - indicates binding to q is either 1 or 2. Y 16 (is O or S) Selected from the group consisting of: The compound, Formula (z'xx)

[0243] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] Compounds of, and Equation (z'xxi)

[0244] [ka] Selected from the group consisting of the following compounds, Formulas (z'i), (z'ii), (z'iii), (z'iv), (z'v), (z'vi), (z'vii), (z'viii), (z'ix), (z'x), (z'x i), (z'xii), (z'xiii), (z'xiv), (z'xv), (z'xvi), (z'xvii), (z'xviii) and (z'xxi) are 2 -It is partially substituted, and in some cases further substituted. -If Y is a compound of formula (z'i), (z'ii), (z'iii), (z'iv), (z'v), (z'vi), (z'vii), (z'viii), (z'ix), (z'x), (z'xi), (z'xii), (z'xiii), (z'xiv), (z'xv), (z'xvi), (z'xvii), (z'xviii), or (z'xxi), any hydrogen atom of the above formula is -L 2 -It can be replaced in parts.

[0245] In a particular embodiment, Y of formula (z'i) 1 C is C.

[0246] In a particular embodiment, R of formula (z'i) a , R a' , R a1 , R a1' is -H.

[0247] In a particular embodiment, equation (z'i) is as follows:

[0248] [ka] [In the formula, the dashed line represents -L] 2 - indicates binding to R a , R a1 , R a1' [This is used as defined in equation (z'i)] It is selected from the group consisting of the following.

[0249] In a particular embodiment, formula (z'ii) is as follows:

[0250] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] It is selected from the group consisting of the following.

[0251] In a particular embodiment, formula (z'iii) is as follows:

[0252] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] It is selected from the group consisting of the following.

[0253] In a particular embodiment, equation (z'iv) is as follows:

[0254] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] It is selected from the group consisting of the following.

[0255] In a particular embodiment, equation (z'v) is as follows:

[0256] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] That is the case.

[0257] In a particular embodiment, equation (z'vi) is as follows:

[0258] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] It is selected from the group consisting of the following.

[0259] In a particular embodiment, equation (z'vii) is as follows:

[0260] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] That is the case.

[0261] In a particular embodiment, equation (z'viii) is as follows:

[0262] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] It is selected from the group consisting of the following.

[0263] In a particular embodiment, equation (z'ix) is as follows:

[0264] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] That is the case.

[0265] In a particular embodiment, the A of equation (z'x) a3 The following

[0266] [ka] [In the equation, the dashed line indicates the connection to the rest of (z'x)] It is selected from the group consisting of the following.

[0267] In a particular embodiment, part of equation (z'x)

[0268] [ka] The following

[0269] [ka] [In the formula, The unmarked dashed line indicates the connection to the rest of (z'x), The dashed line indicated by the asterisk is -L 2 [Indicates a connection to -] It is selected from the group consisting of the following.

[0270] In a particular embodiment, equation (z'xii) is as follows:

[0271] [ka] [In the formula, the dashed line represents -L] 2 - indicates binding to R a19' [Selected from the group consisting of H, methyl, ethyl, propyl, and butyl] It is selected from the group consisting of the following.

[0272] In a particular embodiment, equation (z'xiii) is as follows:

[0273] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] That is the case.

[0274] In a particular embodiment, equation (z'xiv) is as follows:

[0275] [ka] [In the formula, the dashed line represents -L] 2 - indicates binding to Ar is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, and tetralinyl. Y 14 It is a halogen. That is the case.

[0276] In a particular embodiment, equation (z'xv) is as follows:

[0277] [ka] [In the formula, the dashed line represents -L] 2 - indicates binding to Ar is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, and tetralinyl. R a24' , R a24'' , R a24''' [The element is independently selected from the group consisting of H, methyl, ethyl, propyl, and butyl.] That is the case.

[0278] In a particular embodiment, equation (z'xvi) is as follows:

[0279] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] That is the case.

[0280] In a particular embodiment, equation (z'xvii) is as follows:

[0281] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] That is the case.

[0282] In a particular embodiment, -Y is a substituted acyl borate disclosed in WO2018 / 011266A1. Therefore, in a particular embodiment, -Y is as follows:

[0283] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] That is the case.

[0284] In a particular embodiment, -Y is a hydroxylamine disclosed in WO2018 / 011266A1. Therefore, in a particular embodiment, -Y is as follows:

[0285] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] That is the case.

[0286] In a particular embodiment, equation (z'xxi) is as follows:

[0287] [ka] [In the formula, the dashed line represents -L] 2 [Indicates a connection to -] That is the case.

[0288] In a particular embodiment, -Y is as follows:

[0289] [ka] TIFF0007851853000077.tif199162TIFF0007851853000078.tif58162[In the formula, the dashed line is -L 2 - indicates binding to The dashed line indicated by the asterisk shows the connection of part (z'xix) to the rest of the equation. R a , R a1 , R a1' It is used as defined in equation (z'i), R a19' It is used as defined in equation (z'xii), Y 4 It is used as defined in equation (z'xiv), and R a24' , Ra24'' , R a24''' [This is used as defined in the formula (z'xv)] It is selected from the group consisting of the following.

[0290] In a particular embodiment, -Y exists in its protective form.

[0291] In certain embodiments, -Y is a thiol that connects to a moiety used for reversible protection of the thiol functional group. In certain embodiments, -Y is as follows:

[0292] [ka] [In the formula, The dashed line indicates a connection to -Y. Ar is an aromatic moiety that may be further substituted in some cases. R 01 , R 03 , R 04 These are chemical bonds that are independent of each other, or C 1~50 Alkyl, C 2~50 Alkenil or C 2~50 It is alkinyl, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is one or more R that are the same or different. 3 In some cases, it is replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is -Q-, -C(O)O-, -O-, -C(O)-, -C(O)N(R) 4 )-,-S(O)2N(R 4 )-,-S(O)N(R 4 )-, -S(O)2-, -S(O)-, -N(R 4 )S(O)2N(R 4a )-, -S-, -N(R 4 )-,-OC(O)R 4 , -N(R 4 )C(O)-, -N(R 4 )S(O)2-, -N(R 4)S(O)-, -N(R 4 )C(O)O-, -N(R 4 )C(O)N(R 4a )- and -OC(O)N(R 4 R 4a ) may be interrupted by one or more groups selected from the group consisting of, R 02 is -H, C 1~50 Alkyl, C 2~50 Alkenyl or C 2~50 It is alkinyl, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is one or more R that are the same or different. 3 In some cases, it is replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is -Q-, -C(O)O-, -O-, -C(O)-, -C(O)N(R) 4 )-,-S(O)2N(R 4 )-,-S(O)N(R 4 )-, -S(O)2-, -S(O)-, -N(R 4 )S(O)2N(R 4a )-, -S-, -N(R 4 )-,-OC(O)R 4 , -N(R 4 )C(O)-, -N(R 4 )S(O)2-, -N(R 4 )S(O)-, -N(R 4 )C(O)O-, -N(R 4 )C(O)N(R 4a )- and -OC(O)N(R 4 R 4a ) may be interrupted by one or more groups selected from the group consisting of, Q is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Selected from the group consisting of cycloalkyls, 4- to 7-membered heterocyclines, and 8- to 11-membered heterobicyclines, T is the same or different one or more -R 3 It is sometimes replaced by, R 3These are halogen, -CN, oxo (=O), and -COOR. 5 , -OR 5 , -C(O)R 5 ,-C(O)N(R 5 R 5a ), -S(O)2N(R 5 R 5a ), -S(O)N(R 5 R 5a ), -S(O)2R 5 ,-S(O)R 5 , -N(R 5 )S(O)2N(R 5a R 5b ), -SR 5 , -N(R 5 R 5a ), -NO2, -OC(O)R 5 , -N(R 5 )C(O)R 5a , -N(R 5 )S(O)2R 5a , -N(R 5 )S(O)R 5a , -N(R 5 )C(O)OR 5a , -N(R 5 )C(O)N(R 5a R 5b ), -OC(O)N(R 5 R 5a ) or C 1~6 It is alkyl, C 1~6 Alkyl is optionally substituted with one or more halogens, which are the same or different. R 4 , R 4a , R 5 , R 5a , R 5b is -H or C 1~6 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is sometimes substituted with one or more halogens, which are the same or different. It is a thiol that connects to a portion selected from the group consisting of the following.

[0293] -L 1- connects to -D via an amide linkage. This linkage may not be reversible in itself, but in this invention, -L 1 It is understood that adjacent groups present in the linkage, such as amides, primary amines, secondary amines, and tertiary amines, make these linkages reversible.

[0294] In a particular embodiment, the reagent of the present invention is of formula (II')

[0295] [ka] [In the formula, the dashed line indicates a connection to -Q, -R 1 ,-R 1a ,-R 2 ,-R 2a ,-R 3 ,-R 3a ,-R 5 ,-R 5a And -PG is used as defined in formula (II). Includes linker-L*-, -L*- is at least one -L 2 -Z portion or at least one -L 2 -The Y portion is sometimes replaced, and sometimes further replaced. -L 2 -, -Y, and Z are used as defined in equation (II).

[0296] In a particular embodiment, -L*- in formula (II') is at least one -L 2 -Y portion or at least one -L 2 -Z is substituted, and may be further substituted. In a particular embodiment, -L*- in formula (II') is replaced by at least one -L 2 -Y portion or at least one -L 2 -Z is replaced, and in some cases further replaced, except -X 3 The condition is that - is not -S-.

[0297] In a particular embodiment, -L*- in formula (II') is at least one -L 2 -It is replaced by the Y part. In a particular embodiment, -L*- in formula (II') is replaced by one -L 2 -Y is substituted. In a particular embodiment, -L*- in formula (II') is replaced by two -L 2 -Y is substituted. In a particular embodiment, -L*- in formula (II') is replaced by three -L 2 -The Y portion is being replaced.

[0298] In a particular embodiment, -L*- in formula (II') is at least one -L 2 -Z is substituted. In a particular embodiment, -L*- in formula (II') is replaced with one -L 2 -Z is substituted. In a particular embodiment, -L*- in formula (II') is replaced by two -L 2 -Z is substituted. In a particular embodiment, -L*- in formula (II') is replaced by three -L 2 -Z is being replaced.

[0299] In a particular embodiment, -L 1 - is further substituted with one or more substituents.

[0300] In a particular embodiment, -L 1 - has not been further replaced.

[0301] In certain embodiments, -L*- is further substituted with one or more substituents.

[0302] In certain embodiments, -L*- is not further substituted.

[0303] In a particular embodiment, -L 1 - is equation (Ia)

[0304] [ka] [In the formula, The dashed line indicates the bonding of a primary or secondary amine to the nitrogen of -D in formula (I). -R 1 ,-R 1a ,-R 2 ,-R 2a ,-R 5 ,-R 5a ,-R 6 and -R 6a [It is used as defined in formula (I)] It belongs to them.

[0305] In a particular embodiment, the -R of formula (Ia) 1 ,-R 1a ,-R 2 ,-R 2a ,-R 5 ,-R 5a ,-R 6 and -R 6a The compounds are independently selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0306] In a particular embodiment, -R 1 ,-R 1a ,-R 2 is -H and -R 2a is -N(R 12 )C(O)H.

[0307] In a particular embodiment, the -R of formula (Ia) 1 The is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. In a particular embodiment, the -R of formula (Ia) 1a The is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. In a particular embodiment, the -R of formula (Ia) 2The is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. In a particular embodiment, the -R of formula (Ia) 2a The is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. In a particular embodiment, the -R of formula (Ia) 5 The is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. In a particular embodiment, the -R of formula (Ia) 5a The is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. In a particular embodiment, the -R of formula (Ia) 6 The is selected from the group consisting of -H, methyl, ethyl, n-propyl and isopropyl. In a particular embodiment, the -R of formula (Ia) 6a The compound is selected from the group consisting of -H, methyl, ethyl, n-propyl, and isopropyl.

[0308] In a particular embodiment, the -R of formula (Ia) 1 is -H. In a particular embodiment, -R of equation (Ia) 1a is -H. In a particular embodiment, -R of equation (Ia) 2 is -H. In a particular embodiment, -R of equation (Ia) 2a is -H. In a particular embodiment, -R of equation (Ia) 5 is -H. In a particular embodiment, -R of equation (Ia) 5a is -H. In a particular embodiment, -R of equation (Ia) 6 is -H. In a particular embodiment, -R of equation (Ia) 6a is -H.

[0309] In a particular embodiment, the -R of formula (Ia) 1 is -H, and -H is -L 2 It is replaced by -. In a particular embodiment, the -R in formula (Ia) 1a is -H, and -H is -L 2It is replaced by -. In a particular embodiment, the -R in formula (Ia) 2 is -H, and -H is -L 2 It is replaced by -. In a particular embodiment, the -R in formula (Ia) 2a is -H, and -H is -L 2 It is replaced by -. In a particular embodiment, the -R in formula (Ia) 5 is -H, and -H is -L 2 It is replaced by -. In a particular embodiment, the -R in formula (Ia) 5a is -H, and -H is -L 2 It is replaced by -. In a particular embodiment, the -R in formula (Ia) 6 is -H, and -H is -L 2 It is replaced by -. In a particular embodiment, the -R in formula (Ia) 6a is -H, and -H is -L 2 It is replaced with -.

[0310] In a particular embodiment, -L 1 - is equation (Ib)

[0311] [ka] [In the formula, The dashed line indicates the bonding of a primary or secondary amine to nitrogen in -D of formula (I). It belongs to them.

[0312] In a particular embodiment, -L 1 - is equation (Ic)

[0313] [ka] [In the formula, A dashed line marked with an asterisk indicates the bonding of a primary or secondary amine to the nitrogen of -D in formula (I), while an unmarked dashed line indicates -L 2 - indicates binding to -L 2 - is used as defined in formula (I). It belongs to them.

[0314] In a particular embodiment, -L 1 - is expression (Id)

[0315] [ka] [In the formula, dashed lines marked with an asterisk indicate the bonding of a primary or secondary amine to the nitrogen of -D in formula (I), and unmarked dashed lines indicate -L] 2 - indicates binding to -L 2 - and Z are used as defined in formula (I). It belongs to them.

[0316] In a particular embodiment, all -L of the conjugate of formula (I) 2 -The parts are identical. In a particular embodiment, the conjugate of formula (I) is one or more types of -L 2 - For example, 2, 3, 4 or 5 different -L 2 -Includes a portion. -L of more than one such type. 2 - is one type of -L 1 -May be connected to only one or more types of -L 1 - You may connect it to

[0317] In a particular embodiment, -L 2 - represents a chemical bond.

[0318] In a particular embodiment, -L 2 - indicates the spacer portion.

[0319] In a particular embodiment, -L 2 - stands for -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-,-S(O)2N(R y1 )-,-S(O)N(R y1 )-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(R y1a)-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-,-N(R y1 )C(O)N(R y1a )-,-OC(O)N(R y1 )-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Selected from the group consisting of alkynnyls, -T'-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is one or more identical or different -R y2 In some cases, it is replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-,-S(O)2N(R y3 )-,-S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-,-N(R y3 )C(O)N(R y3a )- and -OC(O)N(R y3 )- is optionally interrupted by one or more groups selected from the group consisting of, -R y1 and -R y1a -H, -T', C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Independently selected from the group consisting of alkynnyls, -T', C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is one or more identical or different -R y2 In some cases, it is replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50Alkinyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 )-,-S(O)2N(R y4 )-,-S(O)N(R y4 )-, -S(O)2-, -S(O)-, -N(R y4 )S(O)2N(R y4a )-, -S-, -N(R y4 )-, -OC(OR y4 )(R y4a )-,-N(R y4 )C(O)N(R y4a )- and -OC(O)N(R y4 )- is optionally interrupted by one or more groups selected from the group consisting of, Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl, each T' is identical or distinct from one or more -R y2 It is independently substituted depending on the case, Each-R y2 These are halogen, -CN, oxo (=O), -C(O)OR y5 , -OR y5 , -C(O)R y5 ,-C(O)N(R y5 )(R y5a ), -S(O)2N(R y5 )(R y5a ), -S(O)N(R y5 )(R y5a ), -S(O)2R y5 ,-S(O)R y5 , -N(R y5 )S(O)2N(R y5 )(R y5a ), -SR y5 , -N(R y5 )(R y5a ), -NO2, -OC(O)R y5 , -N(R y5 )C(O)R y5a , -N(R y5 )S(O)2R y5a , -N(Ry5 )S(O)R y5a , -N(R y5 )C(O)OR y5a , -N(R y5 )C(O)N(R y5 )(R y5a ), -OC(O)N(R y5 )(R y5a ) and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is optionally substituted with one or more halogens, which are the same or different. Each-R y3 ,-R y3a ,-R y4 ,-R y4a ,-R y5 ,-R y5a and -R y5b -H and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is sometimes substituted with one or more halogens, which are either the same or different.

[0320] In a particular embodiment, -L 2 - stands for -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-,-S(O)2N(R y1 )-,-S(O)N(R y1 )-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-,-N(R y1 )C(O)N(R y1a )-,-OC(O)N(R y1 )-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Selected from the group consisting of alkynnyls, -T'-, C 1~20 Alkyl, C 2~20 Alkenyl and C 2~20 Alkinyl is one or more identical or different -R y2In some cases, it is replaced by C 1~20 Alkyl, C 2~20 Alkenyl and C 2~20 Alkinyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-,-S(O)2N(R y3 )-,-S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-,-N(R y3 )C(O)N(R y3a )- and -OC(O)N(R y3 )- is optionally interrupted by one or more groups selected from the group consisting of, -R y1 and -R y1a -H, -T', C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Independently selected from the group consisting of alkynnyls, -T', C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Alkinyl is one or more identical or different -R y2 In some cases, it is replaced by C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Alkinyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 )-,-S(O)2N(R y4 )-,-S(O)N(R y4 )-, -S(O)2-, -S(O)-, -N(R y4 )S(O)2N(R y4a )-, -S-, -N(R y4 )-, -OC(OR y4 )(R y4a )-,-N(R y4 )C(O)N(R y4a )- and -OC(O)N(R y4)- is optionally interrupted by one or more groups selected from the group consisting of, Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl, each T' is identical or distinct from one or more -R y2 It is independently substituted depending on the case, -R y2 These are halogen, -CN, oxo (=O), -C(O)OR y5 , -OR y5 , -C(O)R y5 ,-C(O)N(R y5 )(R y5a ), -S(O)2N(R y5 )(R y5a ), -S(O)N(R y5 )(R y5a ), -S(O)2R y5 ,-S(O)R y5 , -N(R y5 )S(O)2N(R y5a )(R y5b ), -SR y5 , -N(R y5 )(R y5a ), -NO2, -OC(O)R y5 , -N(R y5 )C(O)R y5a , -N(R y5 )S(O)2R y5a , -N(R y5 )S(O)R y5a , -N(R y5 )C(O)OR y5a , -N(R y5 )C(O)N(R y5a )(R y5b ), -OC(O)N(R y5 )(R y5a ) and C 1~6 Selected from the group consisting of alkyl groups, C 1~6 Alkyl is optionally substituted with one or more halogens, which are the same or different. Each-R y3 ,-Ry3a ,-R y4 ,-R y4a ,-R y5 ,-R y5a and -R y5b -H and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is sometimes substituted with one or more halogens, which are either the same or different.

[0321] In a particular embodiment, -L 2 - stands for -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-,-S(O)2N(R y1 )-,-S(O)N(R y1 )-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-,-N(R y1 )C(O)N(R y1a )-,-OC(O)N(R y1 )-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Selected from the group consisting of alkynnyls, -T'-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is one or more identical or different -R y2 In some cases, it is replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-,-S(O)2N(R y3 )-,-S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a)-,-N(R y3 )C(O)N(R y3a )- and -OC(O)N(R y3 )- is optionally interrupted by one or more groups selected from the group consisting of, -R y1 and -R y1a -H, -T', C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Independently selected from the group consisting of alkinyls, Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl, Each-R y2 is halogen and C 1~6 Independently selected from the group consisting of alkyl groups, each -R y3 ,-R y3a ,-R y4 ,-R y4a ,-R y5 ,-R y5a and -R y5b -H and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is sometimes substituted with one or more halogens, which are either the same or different.

[0322] In a particular embodiment, -L 2 - is -O-, -T'- and -C(O)N(R y1 )- and one or more groups independently selected from the group consisting of C which may be interrupted 1~20 It is an alkyl chain, C 1~20 The alkyl chain consists of -OH, -T', and -C(O)N(R) y6 R y6a ) is optionally substituted with one or more groups independently selected from the group consisting of -R y1 ,-R y6 ,-R y6a H and C 1~4Independently selected from the group consisting of alkyls, T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 The group is selected from cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl.

[0323] In a particular embodiment, -L 2 - has a molecular weight in the range of 14 g / mol to 750 g / mol.

[0324] In a particular embodiment, -L 2 - is below:

[0325] [ka] [In the formula, the dashed lines represent -L] 1 -, -L 2 - indicates the remainder or bonding to Z, -R and -R a [These 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] Includes a portion selected from the group consisting of

[0326] In a particular embodiment, -L 2 - is below:

[0327] [ka] Includes the portion selected from.

[0328] In a particular embodiment, -L 2 - has a chain length of 1 to 20 atoms.

[0329] When used herein, -L 2 -Regarding the part, the term "chain length" is -L 1 -L exists in the shortest connection between - and -Z 2 - Refers to the number of atoms.

[0330] Generally, -L 2 - is -R in equation (I) or (II). 1 ,-R 1a ,-R 2 ,-R 2a ,-R 3 ,-R 3a ,-R 4 ,-R 4a ,-R 5 ,-R 5a ,-R 6 ,-R 6a ,-R 7 ,-R 8 ,-R 8a ,-R 9 ,-R 10 ,-R 11 ,-R 11a ,-R 11b ,-R 12 ,-R 12a ,-R 13 ,-R 14 ,-R 14a ,-R 15 ,-R 15a and -R 15b One hydrogen presented by is -L 2 -L at any position where it is replaced by - 1 It can bind to - or -L*-.

[0331] In a particular embodiment, -R of formula (I) or formula (II) 1 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 1a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 2 One hydrogen presented by is -L 2- is replaced by -R in formula (I) or formula (II) in certain embodiments. 2a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 3 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 3a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 4 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 4a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 5 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 5a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 6 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 6a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 7 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 8 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 8a One hydrogen presented by is -L 2- is replaced by -R in formula (I) or formula (II) in certain embodiments. 9 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 10 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 11 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 11a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 11b One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 12 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 12a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 12b One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 13 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 14 One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 14a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 15 One hydrogen presented by is -L 2- is replaced by -R in formula (I) or formula (II) in certain embodiments. 15a One hydrogen presented by is -L 2 - is replaced by -R in formula (I) or formula (II) in certain embodiments. 15b One hydrogen presented by is -L 2 It can be replaced with -.

[0332] In a particular embodiment, Z and -L 2 The connection of - is a stable connection.

[0333] In a particular embodiment, Z is C 8~24 This is the alkyl portion.

[0334] In a particular embodiment, Z is water-soluble.

[0335] In a particular embodiment, Z is a water-soluble polymer portion.

[0336] If Z is a water-soluble polymer portion, such a polymer portion has a molecular weight in the range of 1 kDa to 1000 kDa (including these values). In certain embodiments, Z has a molecular weight in the range of 5 kDa to 1000 kDa (including these values). In certain embodiments, Z has a molecular weight in the range of 5 kDa to 500 kDa (including these values). In certain embodiments, Z has a molecular weight in the range of 10 kDa to 250 kDa (including these values). In certain embodiments, Z has a molecular weight in the range of 10 kDa to 150 kDa (including these values). In certain embodiments, Z has a molecular weight in the range of 12 kDa to 100 kDa (including these values). In certain embodiments, Z has a molecular weight in the range of 15 kDa to 80 kDa (including these values). In certain embodiments, Z has a molecular weight in the range of 10 kDa to 80 kDa (including these values).

[0337] In a particular embodiment, Z has a molecular weight of approximately 80 kDa. In a particular embodiment, Z has a molecular weight of approximately 70 kDa. In a particular embodiment, Z has a molecular weight of approximately 60 kDa. In a particular embodiment, Z has a molecular weight of approximately 50 kDa. In a particular embodiment, Z has a molecular weight of approximately 40 kDa. In a particular embodiment, Z has a molecular weight of approximately 30 kDa. In a particular embodiment, Z has a molecular weight of approximately 20 kDa. In a particular embodiment, Z has a molecular weight of approximately 10 kDa. In a particular embodiment, Z has a molecular weight of approximately 5 kDa.

[0338] In certain embodiments, Z is 2-methacryloyloxyethyl phosphorylcholine, poly(acrylic acid), poly(acrylate), poly(acrylamide), poly(alkyloxy)polymer, poly(amide), poly(amideamine), poly(amino acid), poly(anhydride), poly(aspartamide), poly(butyric acid), poly(glycolic acid), polybutylene terephthalate, poly(caprolactone), poly(carbonate), poly(cyanoacrylate), poly(dimethylacrylamide), poly(ester), poly(ethylene), poly(ethylene glycol), poly(ethylene oxide), poly(ethyl phosphate), poly(ethyl oxazoline), poly(glycolic acid), poly(hydroxyethyl acrylate), poly(hydroxyethyl oxazoline), poly(hydroxymethyl methacrylate), poly(hydroxypropyl methacrylamide), poly(hydroxypropyl methacrylate), poly(hydroxypropyl The water-soluble polymer portion comprises polymers selected from the group consisting of oxazoline, poly(iminocarbonate), poly(lactic acid), poly(lactic acid-coglycolic acid), poly(methacrylamide), poly(methacrylate), poly(methyloxazoline), poly(organophosphazene), poly(orthoester), poly(oxazoline), poly(propylene glycol), poly(siloxane), poly(urethane), poly(vinyl alcohol), poly(vinylamine), poly(vinyl methyl ether), poly(vinylpyrrolidone), silicone, cellulose, carbomethylcellulose, hydroxypropylmethylcellulose, chitin, chitosan, dextran, dextrin, gelatin, hyaluronic acid and derivatives, functionalized hyaluronic acid, mannan, pectin, rhamnogalacturonan, starch, hydroxyalkyl starch, hydroxyethyl starch and other carbohydrate-based polymers, xylan, and copolymers thereof.

[0339] In a particular embodiment, Z is a water-soluble polymer moiety comprising a protein selected from the group consisting of proteins, for example, carboxyl-terminal peptides of chorionic gonadotropins described in US2012 / 0035101A1 incorporated herein by reference; albumin; XTEN sequences described in WO2011123813A2 incorporated herein by reference; proline / alanine random coil sequences described in WO2011 / 144756A1 incorporated herein by reference; proline / alanine / serine random coil sequences described in WO2008 / 155134A1 and WO2013 / 024049A1 incorporated herein by reference; and Fc fusion proteins.

[0340] In a particular embodiment, Z is polysarcosine.

[0341] In a particular embodiment, Z comprises poly(N-methylglycine).

[0342] In a particular embodiment, Z includes a random coil protein moiety.

[0343] In certain embodiments, such a random coil protein portion contains at least 25 amino acid residues and up to 2000 amino acids. In certain embodiments, such a random coil protein portion contains at least 30 amino acid residues and up to 1500 amino acid residues. In certain embodiments, such a random coil protein portion contains at least 50 amino acid residues and up to 500 amino acid residues.

[0344] In certain embodiments, Z comprises a random coil protein moiety, where at least 80%, in certain embodiments at least 85%, in certain embodiments at least 90%, in certain embodiments at least 95%, in certain embodiments at least 98%, and in certain embodiments at least 99% of the total number of amino acids forming the random coil protein moiety are selected from alanine and proline. In certain embodiments, at least 10%, but less than 75%, and in certain embodiments less than 65%, of the total number of amino acid residues in such a random coil protein moiety are proline residues. In certain embodiments, such a random coil protein moiety is as described in WO2011 / 144756A1, the whole thereof is incorporated herein by reference.

[0345] In certain embodiments, Z comprises a random coil protein moiety, where at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, and at least 99% of the total number of amino acids forming the random coil protein moiety are selected from alanine, serine, and proline. In certain embodiments, at least 4%, but less than 40%, of the total number of amino acid residues in such a random coil protein moiety are proline residues. In certain embodiments, such a random coil protein moiety is as described in WO2008 / 155134A1, the whole of which is incorporated herein by reference.

[0346] In certain embodiments, Z comprises a random coil protein moiety, wherein at least 80%, in certain embodiments at least 85%, in certain embodiments at least 90%, in certain embodiments at least 95%, in certain embodiments at least 98%, and in certain embodiments 99% of the total number of amino acids forming the random coil protein moiety are selected from alanine, glycine, serine, threonine, glutamate, and proline. In certain embodiments, such a random coil protein moiety is such as that described in WO2010 / 091122A1, which is incorporated herein by reference.

[0347] In a particular embodiment, Z is a hyaluronic acid-based polymer.

[0348] In a particular embodiment, Z is a polymer portion disclosed in WO2013 / 024047A1, which is incorporated herein by reference.

[0349] In a particular embodiment, Z is a polymer portion disclosed in WO2013 / 024048A1, which is incorporated herein by reference.

[0350] In certain embodiments, Z is a PEG-based polymer, such as a linear, branched, or multi-armed PEG-based polymer.

[0351] In a particular embodiment, Z is a linear PEG-based polymer.

[0352] In a particular embodiment, Z is a branched chain C having 1, 2, 3, 4, 5, or 6 branching points. 8~24 It is alkyl. In a particular embodiment, Z is a branched chain C having 1, 2, or 3 branching points. 8~24 It is alkyl. In a particular embodiment, Z is a branched chain C having one branch point. 8~24 It is alkyl. In a particular embodiment, Z is a branched chain C having two branching points.8~24 It is alkyl. In a particular embodiment, Z is a branched chain C having three branching points. 8~24 It is alkyl.

[0353] In certain embodiments, Z is a branched-chain polymer. In certain embodiments, Z is a branched-chain polymer having 1, 2, 3, 4, 5, or 6 branch points. In certain embodiments, Z is a branched-chain polymer having 1, 2, or 3 branch points. In certain embodiments, Z is a branched-chain polymer having 1 branch point. In certain embodiments, Z is a branched-chain polymer having 2 branch points. In certain embodiments, Z is a branched-chain polymer having 3 branch points.

[0354] In a particular embodiment, the branch point is selected from the group consisting of -N<, -CH<, and >C<.

[0355] In a particular embodiment, such a branched chain Z portion is PEG-based.

[0356] In a particular embodiment, Z is a multi-arm PEG-based polymer.

[0357] In a particular embodiment, Z is a multi-arm PEG-based polymer having at least two PEG-based arms, for example, 2, 3, 4, 5, 6, 7, or 8 PEG-based arms.

[0358] In a particular embodiment, Z is a branched-chain PEG-based polymer containing at least 10% PEG, having one branch point and two PEG-based polymer arms, and having a molecular weight of about 40 kDa. Thus, each of the two PEG-based polymer arms has a molecular weight of about 20 kDa. In a particular embodiment, the branch point is -CH<.

[0359] In a particular embodiment, Z is a branched-chain PEG-based polymer containing at least 10% PEG, having three branching points and four PEG-based polymer arms, and having a molecular weight of about 40 kDa. Thus, each of the four PEG-based polymer arms has a molecular weight of about 10 kDa. In a particular embodiment, each of the three branching points is -CH<.

[0360] In a particular embodiment, Z is water-insoluble.

[0361] In a particular embodiment, Z is a water-insoluble polymer portion.

[0362] In certain embodiments, Z is 2-methacryloyloxyethyl phosphorylcholine, poly(acrylic acid), poly(acrylate), poly(acrylamide), poly(alkyloxy)polymer, poly(amide), poly(amideamine), poly(amino acid), poly(anhydride), poly(aspartamide), poly(butyric acid), poly(glycolic acid), polybutylene terephthalate, poly(caprolactone), poly(carbonate), poly(cyanoacrylate), poly(dimethylacrylamide), poly(ester), poly(ethylene), poly(ethylene glycol), poly(ethylene oxide), poly(ethyl phosphate), poly(ethyl oxazoline), poly(glycolic acid), poly(hydroxyethyl acrylate), poly(hydroxyethyl oxazoline), poly(hydroxymethyl methacrylate), poly(hydroxypropyl methacrylamide), poly(hydroxypropyl methacrylate), poly(hydroxypropyl The water-insoluble polymer portion comprises polymers selected from the group consisting of oxazoline, poly(iminocarbonate), poly(lactic acid), poly(lactic acid-coglycolic acid), poly(methacrylamide), poly(methacrylate), poly(methyloxazoline), poly(organophosphazene), poly(orthoester), poly(oxazoline), poly(propylene glycol), poly(siloxane), poly(urethane), poly(vinyl alcohol), poly(vinylamine), poly(vinyl methyl ether), poly(vinylpyrrolidone), silicone, cellulose, carbomethylcellulose, hydroxypropylmethylcellulose, chitin, chitosan, dextran, dextrin, gelatin, hyaluronic acid and its derivatives, functionalized hyaluronic acid, mannan, pectin, rhamnogalacturonan, starch, hydroxyalkyl starch, hydroxyethyl starch and other carbohydrate-based polymers, xylan, and copolymers thereof.

[0363] In a particular embodiment, Z is a hydrogel.

[0364] In certain embodiments, Z is a PEG-based or hyaluronic acid-based hydrogel. In certain embodiments, Z is a PEG-based hydrogel. In certain embodiments, Z is a hyaluronic acid-based hydrogel.

[0365] In certain embodiments, Z is a hydrogel described in WO2006 / 003014A2, WO2011 / 012715A1, or WO2014 / 056926A1, all of which are incorporated herein by reference.

[0366] In certain embodiments, Z is a hydrogel disclosed in WO2013 / 036847A1. In particular, in certain embodiments, Z is a hydrogel produced by a method comprising the step of reacting at least a first reactive polymer with a crosslinked compound to be cleaved, wherein the crosslinked compound to be cleaved has a first functional group-Y that reacts with the first reactive polymer. 1 The polymer comprises a portion which is cleaved by elimination under physiological conditions, the portion which contains a second functional group -Y that reacts with a second reactive polymer. 2 It includes. In a particular embodiment, the crosslinking compound to be cleaved is of formula (PL-1):

[0367] [ka] [In the formula, m is either 0 or 1; -X is a functional group that can be attached to a reactive polymer and is readily detached under physiological conditions, and the second functional group -Y 2 Includes; -R 1 ,-R 2 and -R 5 At least one of the first functional group-Y can be attached to the polymer. 1 Includes; -R 1 and -R 2One or only one of these is selected from the group consisting of -H, alkyl, arylalkyl, and heteroarylalkyl; Depending on the case, -R 1 and -R 2 They can combine to form a 3-8 member ring; -R 1 and -R 2 At least one or both of the following are -CN, -NO2, aryl, heteroaryl, alkenyl, alkynyl, -COR 3 -SOR 3 , -SO2R 3 and -SR 4 Independently selected from the group consisting of; -R 3 -H, alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -OR 9 and -NR 9 Selected from a group consisting of 2; -R 4 This is selected from the group consisting of alkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; Each-R 5 (OCH2CH2) is an integer where -H is alkyl, alkenylalkyl, or alkynylalkyl, and p is an integer in the range of 1 to 1000. p Independently selected from the group consisting of O-alkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; Each-R 9 is independently selected from the group consisting of -H and alkyl, or -R 9 Both of them, together with the nitrogen they are bonded to, form a heterocyclic ring; The part of equation (PL-1) may be further substituted depending on the circumstances. It belongs to them.

[0368] The following sections describe such hydrogels in more detail.

[0369] In certain embodiments, -X in formula (PL-1) is succinimidyl carbonate, sulfosuccinimidyl carbonate halogenate, thioether, ester, nitrophenyl carbonate, chloroformic acid, fluoroformic acid, optionally substituted phenol, and formula (PL-2):

[0370] [ka] [In the formula, The dashed line indicates the connection to the rest of equation (PL-1); -T * - are -O-, -S-, and -NR 6 -Selected from the group consisting of; z is an integer selected from the group consisting of 1, 2, 3, 4, 5, and 6; -X'- is nonexistent or -OR 7 - and -SR 7 -Selected from the group consisting of; -Y 2 It is a functional group that can be bonded to reactive polymers; -R 6 is selected from the group consisting of -H, alkyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl; -R 7 (OCH2CH2) is an integer where alkylene, phenylene, and p are in the range of 1 to 1000. p Selected from the group consisting of: It is selected from the group consisting of the following.

[0371] In certain embodiments, -X in formula (PL-1) comprises an activated carbonate, such as succinimidyl carbonate, sulfosuccinimidyl carbonate, or nitrophenyl carbonate. In certain embodiments, -X in formula (PL-1) comprises a carbonyl halogen, such as O(C=O)Cl or O(C=O)F. In certain embodiments, -X in formula (PL-1) has formula (PL-2). In certain embodiments, -X in formula (PL-1) is OR 7 or SR 7 And here R 7Depending on the case, substituted alkylene, substituted phenylene or (OCH2CH2) p Here, p is between 1 and 1000.

[0372] In a particular embodiment, p in formula (PL-2) is an integer in the range of 1 to 100. In a particular embodiment, p in formula (PL-2) is an integer in the range of 1 to 10.

[0373] In a particular embodiment, the -Y of formula (PL-1) 1 and -Y in equation (PL-2) 2 These are independently N3, NH2, and NH-CO2 t Bu, SH, S t Bu, maleimide, CO2H, CO2 t Bu, 1,3-diene, cyclopentadiene, furan, alkyne, cyclooctin, acrylate or acrylamide, wherein t Bu is tert-butyl, and -Y 1 or -Y 2 If one of them contains N3, the other does not contain alkyne or cyclooctin;-Y 1 or -Y 2 If one of them contains SH, the other does not contain maleimide, acrylate, or acrylamide;-Y 1 or -Y 2 If one of them contains NH2, the other does not contain CO2H; -Y 1 or -Y 2 If one of them contains 1,3-diene or cyclopentadiene, the other does not contain furan.

[0374] In a particular embodiment, the crosslinking compound to be cleaved is of formula (PL-3):

[0375] [ka] [In the formula, m is either 0 or 1; n is an integer selected from 1 to 1000; s is 0, 1, or 2; t is selected from the group consisting of 2, 4, 8, 16, and 32; -W- stands for -O(C=O)O-, -O(C=O)NH-, -O(C=O)S-, -O(C=O)NR 6 CH2O- and -O(C=O)NR 6 Selected from the group consisting of S-; -Q is a core group with valence = t; it connects to multiple arms of the crosslinked compound that is cleaved; t is an integer selected from 2, 4, 8, 16, and 32; -R 1 ,-R 2 and -R 5 This is as defined in equation (PL-1). It belongs to them.

[0376] In a particular embodiment, t in formula (PL-3) is 2. In a particular embodiment, t in formula (PL-3) is 4. In a particular embodiment, t in formula (PL-3) is 8. In a particular embodiment, t in formula (PL-3) is 16. In a particular embodiment, t in formula (PL-3) is 32.

[0377] In a particular embodiment, -Q in formula (PL-3) is:

[0378] [ka] The structure is selected from the group consisting of the following, where the dashed line indicates the bond to the remaining part of the crosslinked compound that is cleaved.

[0379] In a particular embodiment, -Q in formula (PL-3) has the structure of (PL-3-i). In a particular embodiment, -Q in formula (PL-3) has the structure of (PL-3-ii). In a particular embodiment, -Q in formula (PL-3) has the structure of (PL-3-iii).

[0380] In a particular embodiment, the crosslinked compound to be cleaved is of formula (PL-3), where m is 0, n is approximately 100, s is 0, t is 4, -W- is -O(C=O)NH-, -Q has the structure of (PL-3i), and -R 2 H is H, while -R is H 5 is -H, and the other is -R 5 It is (CH2)5N3, and -R 1 The elements are (4-chlorophenyl)SO2, phenyl substituted with -SO2, morpholino-SO2, or -CN.

[0381] In a particular embodiment, the -Y of formula (PL-3) 1 These are N3, NH2, and NH-CO2. t Bu, SH, S t Bu, maleimide, CO2H, CO2 t Bu, 1,3-diene, cyclopentadiene, furan, alkyne, cyclooctin, acrylate or acrylamide, wherein t Bu is tert-butyl.

[0382] In a particular embodiment, each -Y of formula (PL-1) or (PL-3) 1 and -Y in equation (PL-2) 2 N 2 NH2, NH-CO2 t Bu, SH, S t Bu, maleimide, CO2H, CO2 t This includes Bu, 1,3-diene, cyclopentadiene, furan, alkyne, cyclooctin, acrylate, or acrylamide.

[0383] In a particular embodiment, -Y 1 and -Y 2 One of them is an azide, and the other is a reactive functional group selected from the group consisting of acetylene, cyclooctin, and maleimide. In a particular embodiment, -Y 1 and -Y 2One of the components is a thiol, and the other is a reactive functional group selected from the group consisting of maleimide, acrylate, acrylamide, vinyl sulfone, vinyl sulfonamide, and halocarbonyl. In certain embodiments, -Y 1 and -Y 2 One of the components is an amine, and the other is a selectively reactive functional group selected from carboxylic acids and activated carboxylic acids. In a particular embodiment, -Y 1 and -Y 2 One of the components is maleimide, and the other is a selective reactive functional group selected from the group consisting of 1,3-diene, cyclopentadiene, and furan.

[0384] In certain embodiments, the first and optional second polymers are selected from the group consisting of polyethylene glycol, polypropylene glycol, poly(N-vinylpyrrolidone), polymethacrylate, polyphosphazene, polylactide, polyacrylamide, polyglycolate, polyethyleneimine, agarose, dextran, gelatin, collagen, polylysine, chitosan, alginate, hyaluronan, pectin, and carrageenan homopolymers or copolymers, and contain appropriate reactive functionalities or formula [Y 3 -(CH2) s (CH2CH2O) n ] t It is Q, and here -Y 3 is a reactive functional group, where s is 0, 1, or 2, n is an integer selected from the group in the range of 10 to 1000, and -Q is a core group having a valence t, where t is an integer selected from the group consisting of 2, 4, 8, 16, and 32.

[0385] In a particular embodiment, the first polymer includes a multi-arm polymer. In a particular embodiment, the first polymer includes at least three arms. In a particular embodiment, the first polymer includes at least four arms. In a particular embodiment, the first polymer includes at least five arms. In a particular embodiment, the first polymer includes at least six arms. In a particular embodiment, the first polymer includes at least seven arms. In a particular embodiment, the first polymer includes at least eight arms.

[0386] In certain embodiments, the second polymer includes a multi-arm polymer. In certain embodiments, the second polymer includes at least three arms. In certain embodiments, the second polymer includes at least four arms. In certain embodiments, the second polymer includes at least five arms. In certain embodiments, the second polymer includes at least six arms. In certain embodiments, the second polymer includes at least seven arms. In certain embodiments, the second polymer includes at least eight arms.

[0387] In certain embodiments, the first polymer comprises a polyethylene glycol polymer with two arms. In certain embodiments, the first polymer comprises a polyethylene glycol polymer with four arms. In certain embodiments, the first polymer comprises a polyethylene glycol polymer with eight arms. In certain embodiments, the first polymer comprises a polyethylene glycol polymer with sixteen arms. In certain embodiments, the first polymer comprises a polyethylene glycol polymer with thirty-two arms.

[0388] In certain embodiments, the second polymer comprises a polyethylene glycol polymer with two arms. In certain embodiments, the second polymer comprises a polyethylene glycol polymer with four arms. In certain embodiments, the second polymer comprises a polyethylene glycol polymer with eight arms. In certain embodiments, the second polymer comprises a polyethylene glycol polymer with sixteen arms. In certain embodiments, the second polymer comprises a polyethylene glycol polymer with thirty-two arms.

[0389] In a particular embodiment, the first and second reactive polymers are reacted with the crosslinked compound to be cleaved, either successively or simultaneously.

[0390] In a particular embodiment, the first functional group and the second functional group are the same.

[0391] The terms used only in the context of formulas (PL-1), (PL-2), and (PL-3) have the following meanings:

[0392] The term "the part that can be cleaved by desorption under physiological conditions" is HC-(CH=CH) m -This refers to a structure containing a C-X' group, where m is 0 or 1 and X' is a leaving group. The elimination reaction to remove the HX' elements described above can occur at a rate such that the reaction half-life is between 1 and 10,000 hours under physiological conditions of pH and temperature. Preferably, the reaction half-life is between 1 and 5,000 hours, more preferably between 1 and 1,000 hours, under physiological conditions of pH and temperature. Physiological conditions of pH and temperature mean a pH between 7 and 8 and a temperature between 30 and 40 degrees Celsius.

[0393] The term “reactive polymers and reactive oligomers” refers to polymers or oligomers containing functional groups that are reactive to other functional groups under mild conditions that can meet the stability requirements of peptides, proteins, and other biomolecules. Suitable functional groups found in reactive polymers include maleimides, thiols or protected thiols, alcohols, acrylates, acrylamides, amines or protected amines, carboxylic acids or protected carboxylic acids, azides, alkynes including cycloalkynes, 1,3-dienes including cyclopentadienes and furans, alpha-halocarbonyls, and N-hydroxysuccinimidyl, N-hydroxysulfosuccinimidyl, or nitrophenyl esters or carbonates.

[0394] The term "functional group that can be attached to a reactive polymer" refers to a functional group that reacts with the corresponding functional group of a reactive polymer to form a covalent bond with the polymer. Suitable functional groups that can be attached to a reactive polymer include maleimides, thiols or protected thiols, acrylates, acrylamides, amines or protected amines, carboxylic acids or protected carboxylic acids, azides, alkynes including cycloalkynes, 1,3-dienes including cyclopentadienes and furans, alpha-halocarbonyls, and N-hydroxysuccinimidyl, N-hydroxysulfosuccinimidyl, or nitrophenyl esters or carbonates.

[0395] The term "substituted" refers to an alkyl, alkenyl, alkynyl, aryl, or heteroaryl group containing one or more substituents replacing one or more hydrogen atoms. Substituents generally include halogens containing F, CI, Br, and I; lower alkyls containing linear, branched, and cyclic structures; lower haloalkyls containing fluoroalkyls, chloroalkyls, bromoalkyls, and iodoalkyls; OH; lower alkoxys containing linear, branched, and cyclic structures; SH; lower alkylthios containing linear, branched, and cyclic structures; silyls containing aminos, alkylaminos, dialkylaminos, alkylsilyls, alkoxysilyls, and arylsilyls; nitro; cyano; carbonyl; carboxylic acids, carboxylic acid esters, carboxylic acid amides; aminocarbonyls; aminoacyls; carbamates; ureas; thiocarbamates; thioureas; ke Heteroaryls can be selected from ton; sulfone; sulfonamide; aryls containing phenyl, naphthyl, and anthracenyl; 5-membered heteroaryls containing pyrrole, imidazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, thiadiazole, triazole, oxadiazole, and tetrazole; 6-membered heteroaryls containing pyridine, pyrimidine, and pyrazine; and heteroaryls containing condensed heteroaryls containing benzofuran, benzothiophene, benzoxazole, benzimidazole, indole, benzothiazole, benzoisoxazole, and benzoisothiazole.

[0396] R 1 and R 2 The properties can be modulated by the addition of electron-donating or electron-withdrawing substituents. The term "electron-donating group" is R 1 R 2 This refers to substituents that reduce the acidity of CH; electron-donating groups are typically negative Hammett σ or Tuft σ. * The constants are well known in the field of physical organic chemistry (Hammett constants refer to aryl / heteroaryl substituents, and Tuft constants refer to substituents on the non-aromatic moieties). Examples of suitable electron-donating substituents include lower alkyl, lower alkoxy, lower alkylthio, amino, alkylamino, dialkylamino, and silyl substituents.

[0397] The term "electron-withdrawing group" is R 1 R 2 This refers to substituents that increase the acidity of the CH group; electron-withdrawing groups are typically positive Hammett σ or Tuft σ * The constant is well known in the field of physical organic chemistry. Examples of suitable electron-withdrawing substituents include halogens, difluoromethyl, trifluoromethyl, nitro, cyano, and -R. x C(=O)-R is H x , lower alkyl, lower alkoxy, or amino, or m is 1 or 2 and -R Y S(O) is a lower alkyl group m R Y This includes aryl or heteroaryl groups. As is well known in the art, the electronic effect of substituents can depend on the position of the substituent. For example, alkoxy substituents at the ortho- or para position of an aryl ring are electron-donating and characterized by a negative Hammett σ constant, while alkoxy substituents at the meta position of an aryl ring are electron-withdrawing and characterized by a positive Hammett σ constant.

[0398] The terms "alkyl," "alkenyl," and "alkynyl" refer to linear, branched, or cyclic hydrocarbon groups comprising 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Alkyls are saturated hydrocarbons, alkenyls contain one or more carbon-carbon double bonds, and alkynyls contain one or more carbon-carbon triple bonds. Unless otherwise specified, these groups contain 1 to 6 carbon atoms.

[0399] The term "aryl" includes an aromatic hydrocarbon group comprising 6 to 18 carbon atoms, preferably 6 to 10 carbon atoms, such as phenyl, naphthyl, and anthracenyl groups. The term "heteroaryl" includes an aromatic ring comprising 3 to 15 carbon atoms, preferably 3 to 7 carbon atoms, containing at least one N, O, or S atom, such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl, and similar groups.

[0400] The term "halogen" includes fluoro, chloro, bromo, and iodine.

[0401] The term "maleimide" is derived from the formula

[0402] [ka] It is the basis of.

[0403] In certain embodiments, Z is a hydrogel disclosed in WO2020 / 206358A1. In particular, in certain embodiments, Z is (a) Multiarm polymer-P 2 A step of preparing a first prepolymer comprising, wherein the first prepolymer is of formula (PL-4)

[0404] [ka] [In the formula, n is an integer selected from 0, 1, 2, 3, 4, 5, and 6; r is an integer greater than 2; -Y is a reactive functional group that connects the first prepolymer to the second prepolymer; -R 1 and -R 2 is independently an electron-withdrawing group, an alkyl group, or -H, and at least one -R 1 and -R 2 It is an electron-withdrawing group; Each-R 4 These are independently C1-C3 alkyl groups, or two -R groups. 4 These, together with the carbon atoms to which they are bonded, form a 3- to 6-membered ring; -W- is nonexistent or

[0405] [ka] And, Dashed lines marked with an asterisk indicate binding to -NH-, while unmarked dashed lines indicate binding to -P. 2 This shows a connection with; x, y, and z are each an integer independently selected from 0, 1, 2, 3, 4, 5, and 6; -B' is a group containing -NH2, -ONH2, ketone, aldehyde, -SH, -OH, -CO2H, carboxamide, or cyclooctin or bicyclononine; -C * [These are carboxamide, thioether, thiosuccinimidyl, triazole, or oxime.] It is a step; (b) A multi-arm polymer-P in which each arm is terminated by a reactive functional group -Y'" that reacts with -Y in step (a). 1 The steps include: preparing a second prepolymer containing; (c) -Y and -Y'' react and combine to form -Y * - under conditions that form the two prepolymers of step (a) and (b), and; if applicable (d) A step of isolating the obtained hydrogel and This is a hydrogel produced by a method that includes [a specific component].

[0406] Therefore, -Z is a hydrogel obtained from the method described above. In certain embodiments, the hydrogel produced by the prior method is degradable.

[0407] In a particular embodiment, -Y and -Y'' react under step (c) and formula (PL-4'):

[0408] [ka] [where n, r, -P 1 -Y * -, -R 4 ,-R 1 ,-R 2 , -W- and -P 2 This is as defined above. It forms an insoluble hydrogel matrix containing crosslinks.

[0409] In a particular embodiment, n in formula (PL-4) or (PL-4') is an integer selected from 1, 2, 3, 4, 5, and 6. In a particular embodiment, n in formula (PL-4) or (PL-4') is an integer selected from 1, 2, and 3. In a particular embodiment, n in formula (PL-4) or (PL-4') is an integer selected from 0, 1, 2, and 3. In a particular embodiment, n in formula (PL-4) or (PL-4') is 1. In a particular embodiment, n in formula (PL-4) is 2. In a particular embodiment, n in formula (PL-4) or (PL-4') is 3.

[0410] In a particular embodiment, the multi-arm-P of formula (PL-4) or (PL-4') 2 r is a polymer with r arms, where r is an integer selected from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12. In certain embodiments, r in formula (PL-4) or (PL-4') is an integer selected from 2, 3, 4, 5, 6, 7, and 8. In certain embodiments, r in formula (PL-4) or (PL-4') is an integer selected from 2, 4, 6, and 8. In certain embodiments, r in formula (PL-4) or (PL-4') is 2. In certain embodiments, r in formula (PL-4) or (PL-4') is 4. In certain embodiments, r in formula (PL-4) or (PL-4') is 6. In certain embodiments, r in formula (PL-4) or (PL-4') is 8.

[0411] In a particular embodiment, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of at least 1 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of 1 to 100 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2It has a molecular weight of 1 to 80 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of 1 to 60 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of 1 to 40 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of 1 to 20 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of 1 to 10 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of 1 to 5 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of approximately 20 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of approximately 40 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of approximately 60 kDa. In certain embodiments, the -P of formula (PL-4) or (PL-4') 2 It has a molecular weight of approximately 80 kDa.

[0412] In a particular embodiment, step (b) multi-arm polymer-P 1 r is a polymer with r arms, where r is an integer selected from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12. In a particular embodiment, the multi-arm polymer-P of step (b) 1 r is a polymer with r arms, where r is an integer selected from 2, 3, 4, 5, 6, 7, and 8. In a particular embodiment, the multi-arm polymer-P of step (b) 1 r is a polymer with r arms, where r is an integer selected from 2, 4, 6, and 8. In a particular embodiment, the multi-arm polymer-P of step (b) 1is a polymer with r arms, where r is 2. In a particular embodiment, the multi-arm polymer-P of step (b) 1 is a polymer with r arms, where r is 4. In a particular embodiment, the multi-arm polymer-P of step (b) 1 r is a polymer with r arms, where r is 6. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It is a polymer with r arms, where r is 8.

[0413] In a particular embodiment, step (b) -P 1 It has a molecular weight of at least 1 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of 1 to 100 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of 1 to 80 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of 1 to 60 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of 1 to 40 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of 1 to 20 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of 1 to 10 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of 1 to 5 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of approximately 20 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of approximately 40 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1It has a molecular weight of approximately 60 kDa. In a particular embodiment, the multi-arm polymer-P of step (b) 1 It has a molecular weight of approximately 80 kDa.

[0414] In a particular embodiment, step (b) -P 1 and -P in formula (PL-4) or (PL-4') 2 This includes poly(ethylene glycol) (PEG), poly(ethylene oxide) (PEO), poly(ethyleneimine) (PEI), dextran, hyaluronic acid, or copolymers thereof. In certain embodiments, step (b) -P 1 and P in formula (PL-4) or (PL-4') 2 It is a PEG-based polymer. In a particular embodiment, step (b) -P 1 and -P in formula (PL-4) or (PL-4') 2 It is a hyaluronic acid-based polymer.

[0415] In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 is independently an electron-withdrawing group, an alkyl group, or -H, and at least one -R 1 and -R 2 It is an electron-withdrawing group.

[0416] In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing groups are -CN and -NO 2、 Optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkenyl, optionally substituted alkynyl, -COR 3 -SOR 3 , or -SO2R 3 And here -R 3-H, optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -OR 8 or -NR 8 2, where each -R 8 is independently an alkyl group -H or optionally substituted alkyl group, or both -R groups. 8 The groups, together with the nitrogen atoms to which they are bonded, form a heterocyclic ring; or -SR 9 And here -R 9 This is optionally a substituted alkyl, optionally a substituted aryl, optionally a substituted arylalkyl, optionally a substituted heteroaryl, or optionally a substituted heteroarylalkyl.

[0417] In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing group is -CN. In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing group is -NO2. In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing group is an aryl compound that contains 6 to 10 carbon atoms. In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing group is optionally substituted phenyl, naphthyl, or anthracenyl. In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing group is a heteroaryl compound containing 3 to 7 carbon atoms and substituted with at least one N, O, or S atom. In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2The electron-withdrawing group is optionally substituted pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, or indenyl. In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing group is an alkenyl which contains 2 to 20 carbon atoms. In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing group is an alkynyl which contains 2 to 20 carbon atoms. In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing group is -COR 3 -SOR 3 , or -SO2R 3 And here R 3 -H, optionally substituted alkyl containing 1 to 20 carbon atoms, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -OR 8 or -NR 8 2, where each -R 8 These are alkyl groups that are independently substituted with -H or 1 to 20 carbon atoms, or both -R 8 The groups, together with the nitrogen atoms to which they are bonded, form a heterocyclic ring. In certain embodiments, the -R of formula (PL-4) or (PL-4') 1 and -R 2 The electron-withdrawing group is -SR 9 And here -R 9 is an optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl containing 1 to 20 carbon atoms. In a particular embodiment, at least one -R1 and -R 2 -CN or -SO2R 3 That is the case.

[0418] In a particular embodiment, at least one -R of formula (PL-4) or (PL-4') 1 and -R 2 -CN, -SOR 3 or -SO2R 3 In a particular embodiment, at least one -R of formula (PL-4) or (PL-4') 1 and -R 2 -CN or -SO2R 3 In a particular embodiment, at least one -R of formula (PL-4) or (PL-4') 1 and -R 2 -CN or -SO2R 3 And here -R 3 This may be a substituted alkyl, may be a substituted aryl, or -NR 8 2. In a particular embodiment, at least one -R of formula (PL-4) or (PL-4') 1 and -R 2 These are -CN, -SO2N(CH3)2, -SO2CH3, phenyl substituted with -SO2, phenyl substituted with -SO2 and -Cl, -SO2N(CH2CH2)2O, -SO2CH(CH3)2, -SO2N(CH3)(CH2CH3), or -SO2N(CH2CH2OCH3)2.

[0419] In a particular embodiment, each -R of formula (PL-4) or (PL-4') 4 These are independently C1-C3 alkyl groups, or together they can form a 3-6 membered ring. In certain embodiments, each -R of formula (PL-4) or (PL-4') 4 These are independently C1-C3 alkyl groups. In certain embodiments, the -R of formula (PL-4) or (PL-4') 4 Both are methyl.

[0420] In certain embodiments, -Y and -Y'' are independently selected from the group consisting of amines, aminooxys, ketones, aldehydes, maleidyls, thiols, alcohols, azides, 1,2,4,6-tetradinyls, trans-cyclooctenyls, bicyclononinyls, cyclooctinyls, and protected varieties thereof.

[0421] In certain embodiments, Y and Y'' may react with each other, for example, selectively. For example, if -Y is an amine, then -Y'' is a carboxylic acid, an active ester, or an active carbonate, and the residual connecting functional group -Y is an amide or a carbamate. * -Obtained. As another example, if -Y is an azide, then -Y'' is an alkynyl, bicyclononinyl, or cyclooctinyl, and the residual connecting functional group -Y is a 1,2,3-triazole. * - is obtained. As another example, if -Y is NH2O, then -Y'' is a ketone or aldehyde and the residual connecting functional group -Y is an oxime. * - is obtained. As another example, if -Y is SH, then -Y'' is a maleimide or halocarbonyl, and the residual connecting functional group -Y is a thiosuccinimidyl or thioether. * - Obtain. Similarly, reverse these roles of -Y and -Y'' and -Y in the opposite direction. * - You can obtain this.

[0422] In a particular embodiment, -Y * - comprises amides, oximes, 1,2,3-triazoles, thioethers, thiosuccinimides, or ethers. In certain embodiments, -Y * - is -L 2 - is

[0423] These conjugation reactions can be carried out under conditions known in the art, for example, when -Y is an azide and -Y'' is cyclooctane, and the conjugation occurs in any solvent in which both components exhibit adequate solubility, although it is known that aqueous solutions exhibit a more favorable reaction rate. When mixed in a suitable solvent, typically in an aqueous buffer at pH 2–7 when -Y and -Y'' are azide / cyclooctane, or at pH 6–9 when -Y and -Y'' are activated esters and amines, the -Y and -Y'' groups react to form an insoluble hydrogel matrix containing the crosslink of formula (PL-4'). This process can be carried out in bulk phase or under conditions of emulsification in a mixed organic / aqueous system, such as forming microspheres suitable for injection, in a particulate suspension.

[0424] The terms used only in the context of formulas (PL-4) and (PL-4') have the following meanings:

[0425] The term "alkyl" refers to a linear, branched, or cyclic saturated hydrocarbon group having 1 to 20, 1 to 12, 1 to 8, 1 to 6, or 1 to 4 carbon atoms. In certain embodiments, alkyl groups are linear or branched. Examples of linear or branched alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and n-decyl. In certain embodiments, alkyl groups are cyclic. Examples of cyclic alkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentadienyl, and cyclohexyl.

[0426] The term "alkoxy" refers to an alkyl group bonded to oxygen, and includes methoxy, ethoxy, isopropoxy, cyclopropoxy, and cyclobutoxy.

[0427] The term "alkenyl" refers to a non-aromatic unsaturated hydrocarbon having a carbon-carbon double bond and 2-20, 2-12, 2-8, 2-6, or 2-4 carbon atoms.

[0428] The term "alkynyl" refers to a non-aromatic unsaturated hydrocarbon having a carbon-carbon triple bond and 2-20, 2-12, 2-8, 2-6, or 2-4 carbon atoms.

[0429] The term "aryl" refers to an aromatic hydrocarbon group having 6 to 18 carbon atoms, preferably 6 to 10 carbon atoms, including groups such as phenyl, naphthyl, and anthracenyl. The term "heteroaryl" refers to an aromatic ring having 3 to 15 carbon atoms, preferably 3 to 7 carbon atoms, including at least one N, O, or S atom, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, and indenyl.

[0430] In certain embodiments, an alkenyl, alkynyl, aryl, or heteroaryl moiety may be linked to the rest of the molecule via alkyl linkage. In these circumstances, the substituent is referred to as an alkenylalkyl, alkynylalkyl, arylalkyl, or heteroarylalkyl, indicating that the alkylene moiety lies between the alkenyl, alkynyl, aryl, or heteroaryl moiety and the molecule to which the alkenyl, alkynyl, aryl, or heteroaryl is linked.

[0431] The term "halogen" or "halo" refers to bromo, fluoro, chloro, or iodine.

[0432] The term “heterocyclic ring” or “heterocyclyl” refers to a 3- to 15-membered aromatic or non-aromatic ring containing at least one N, O, or S atom. Examples include piperidinyl, piperazinyl, tetrahydropyranyl, pyrrolidine, and tetrahydrofuranyl, as well as exemplary groups provided above with respect to the term “heteroaryl.” In certain embodiments, the heterocyclic ring or heterocyclyl is non-aromatic. In certain embodiments, the heterocyclic ring or heterocyclyl is aromatic.

[0433] The term "optionally substituted" refers to a group that may be unsubstituted or substituted with one or more identical or different substituents (e.g., 1, 2, 3, 4, or 5). Examples of substituents include alkyl, alkenyl, alkynyl, halogen, -CN, and -OR. aa , -SR aa , -NR aa R bb -NO2, -C=NH(OR aa ), -C(O)R aa -OC(O)R aa , -C(O)OR aa -C(O)NR aa R bb -OC(O)NR aa R bb , -NR aa C(O)R bb , -NR aa C(O)OR bb ,-S(O)R aa -S(O)2R aa , -NR aa S(O)R bb -C(O)NR aa S(O)R bb , -NR aa S(O)2R bb -C(O)NR aa S(O)2R bb -S(O)NR aa R bb -S(O)2NR aa R bb , -P(O)(OR aa )(OR bb ), heterocyclyl, heteroaryl, or aryl, where alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl, and aryl are each independently and may be -R cc It is replaced by -R aa and -R bb Each of these is independently -H, alkyl, alkenyl, alkynyl, heterocyclyl, heteroaryl, or aryl, or -R aa and -R bbThese, together with the nitrogen atom to which they are bonded, form a heterocycline, which may be substituted with alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkoxy, or -CN, where: each -R cc These are independently alkyl, alkenyl, alkynyl, halogen, heterocyclyl, heteroaryl, aryl, -CN, or -NO2.

[0434] In certain embodiments, Z is a polymer network formed by the physical aggregation of polymer chains, which is preferably caused by hydrogen bonding, crystallization, helix formation, or complexation. In certain embodiments, such a polymer network is a thermogelling polymer.

[0435] In a particular embodiment, Z is as follows:

[0436] [ka] Includes a portion selected from the group consisting of

[0437] In certain embodiments, the conjugate of the present invention or a pharmaceutically acceptable salt thereof is of formula (Ia), (Ib), (Ic), or (Id):

[0438] [ka] [In the formula, -D, -L each 2 - and Z are as defined above, and each -L 1 - is independently of equation (I); x is an integer of at least 1; y is an integer selected from the group consisting of 2, 3, 4, and 5. It belongs to them.

[0439] One -D is equivalent to multiple -L 1-Even if it can be conjugated to a part, the corresponding drug part is understood to be represented as "-D" for simplification, and the drug as "DH".

[0440] In certain embodiments, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a hydrogel. In such cases, Z is a plurality of -L 2 -L 1 -It is understood that part D is included and no upper limit can be set for x.

[0441] In a particular embodiment, the conjugate is of equation (Ia). In a particular embodiment, the conjugate is of equation (Ib). In a particular embodiment, the conjugate is of equation (Ic). In a particular embodiment, the conjugate is of equation (Id). In a particular embodiment, the conjugate is of equation (Ia) where x=1.

[0442] In a particular embodiment, the conjugate is of formula (Ia), where Z is a hydrogel.

[0443] In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion, and x is in the range of 2 to 1000, e.g., 2 to 1500, e.g., 2 to 1000, e.g., 2 to 500, e.g., 2 to 250, or e.g., 2 to 100. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion, and x is 20.

[0444] In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 19. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 18. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 17. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 16. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 15. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 14. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 13. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 12. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 11. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 10. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 9. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 8. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer moiety and x is 7. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer moiety and x is 6.In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 5. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 4. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 3. In a particular embodiment, the conjugate is of formula (Ia), (Ic), or (Id), where Z is a water-soluble polymer portion and x is 2.

[0445] In a particular embodiment, the conjugate is of formula (Ib), where Z is a water-soluble polymer portion and y is 1. In a particular embodiment, the conjugate is of formula (Ib), where Z is a water-soluble polymer portion and y is 2. In a particular embodiment, the conjugate is of formula (Ib), where Z is a water-soluble polymer portion and y is 3. In a particular embodiment, the conjugate is of formula (Ib), where Z is a water-soluble polymer portion and y is 4. In a particular embodiment, the conjugate is of formula (Ib), where Z is a water-soluble polymer portion and y is 5.

[0446] The conjugates of the present invention release one or more drugs over a long period of time; that is, they are sustained-release conjugates. In certain embodiments, the release occurs with a release half-life ranging from 1 to 4 months. In certain embodiments, the release occurs with a release half-life ranging from 2 to 2 months. In certain embodiments, the release occurs with a release half-life ranging from 4 to 2 months. In certain embodiments, the release half-life may be in the range of 6 days to 1 month, 7 days to 40 days, 4 days to 15 days, or 3 days to 7 days.

[0447] In a particular embodiment, steps (d) and (e) of the method for synthesizing the conjugate of the present invention are not optional.

[0448] In a particular embodiment, the conjugation of at least one Z portion is not optional.

[0449] In a particular embodiment, at least one Z portion is conjugated into at least one intermediate (A) between steps (b) and (c). In a particular embodiment, at least one Z portion is conjugated into at least one intermediate (A) between steps (c) and (d). In a particular embodiment, at least one Z portion is conjugated into at least one intermediate (A) between steps (c) and (f). In a particular embodiment, steps (d) and (e) are not optional, and at least one Z portion is conjugated into at least one intermediate (B) between steps (d) and (e). In a particular embodiment, step (e) is not optional, and at least one Z portion is conjugated into at least one intermediate (B) between steps (e) and (f).

[0450] In a particular embodiment, at least one Z portion is conjugated during step (b). In a particular embodiment, at least one Z portion is conjugated during step (c). In a particular embodiment, step (d) is not optional, and at least one Z portion is conjugated during step (d). In a particular embodiment, step (e) is not optional, and at least one Z portion is conjugated during step (e).

[0451] In a particular embodiment, one Z portion is conjugated during step (b). In a particular embodiment, one Z portion is conjugated during step (c). In a particular embodiment, step (d) is optional, and one Z portion is conjugated during step (d). In a particular embodiment, step (e) is optional, and one Z portion is conjugated during step (e).

[0452] In a particular embodiment, intermediate (A) is isolated before step (c). In a particular embodiment, step (d) is optional, and intermediate (B) is isolated before step (d). In a particular embodiment, steps (d) and (e) are optional, and intermediate (B) is isolated before step (e).

[0453] In a particular embodiment, the conjugate or intermediate obtained from step (c), (d), or (e) is purified by ion exchange chromatography.

[0454] In a particular embodiment, step (a) of this method, the reagent is the linker-L of formula (II). * -Includes -L * - is one part - L 2 -Y is substituted, and step (b) results in the formation of intermediates (A) and Z, which are isolated and then subjected to deprotection conditions.

[0455] In a particular embodiment, step (a) of this method, the reagent is the linker-L of formula (II). * -Includes -L * - is one part - L 2 -It is substituted with Y, and step (b) results in the formation of an intermediate (A) that is conjugated to Z.

[0456] In certain embodiments, the primary or secondary amine-containing drug in step (b) of the method is a peptide or a protein.

[0457] In certain embodiments, step (d) of the method of the present invention is optional. In certain embodiments, step (e) of the method of the present invention is optional.

[0458] In certain embodiments, the deprotection condition refers to a solution containing a buffer.

[0459] In a particular embodiment, the shift condition refers to a solution containing a buffer.

[0460] Examples of buffering agents include histidine, 1,3-diaminopropane, 2-(N-morpholino)ethanesulfonic acid (MES), 2-bis(2-hydroxyethyl)amino-2-(hydroxymethyl)-1,3-propanediol (BIS-TRIS), acetic acid, adipic acid, ammonia, arginine, boric acid, carbonic acid, citric acid, diethanolamine, ethanolamine, ethylenediamine, formic acid, gluconic acid, glutaric acid, glycine, glycinamide, guanidine, histamine, imidazole, lysine, malic acid, N- (2-hydroxy-1,1-bis(hydroxymethyl)ethyl)glycine (TRICINE), N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES), N-[tris(hydroxymethyl)methyl]-2-aminoethanesulfonic acid (TES), phosphoric acid, piperazine, propionic acid, pyruvate, spermidine, spermine, succinic acid, tartonic acid, triethanolamine (TEA), tromethamine (Tris), tyrosine, and mixtures thereof may be selected from the group.

[0461] In certain embodiments, the deprotection condition refers to a solution containing a buffer with a pH of 10 or less. In certain embodiments, the deprotection condition refers to a solution containing a buffer with a pH of about 3 to about 9. In certain embodiments, the deprotection condition refers to a solution containing a buffer with a pH of about 4 to about 8. In certain embodiments, the deprotection condition refers to a solution containing a buffer with a pH of about 5 to about 7. In certain embodiments, the deprotection condition refers to a solution containing a buffer with a pH of about 6. In certain embodiments, the deprotection condition refers to a solution containing a buffer with a pH of about 7. In certain embodiments, the deprotection condition refers to a solution containing a buffer with a pH of about 7.4. In certain embodiments, the deprotection condition refers to a solution containing a buffer with a pH of 7.4.

[0462] In certain embodiments, the shift condition refers to a solution containing a buffer with a pH of about 5 to about 9. In certain embodiments, the shift condition refers to a solution containing a buffer with a pH of about 6 to about 8. In certain embodiments, the shift condition refers to a solution containing a buffer with a pH of about 7. In certain embodiments, the shift condition refers to a solution containing a buffer with a pH of about 7.4. In certain embodiments, the shift condition refers to a solution containing a buffer with a pH of 7.4.

[0463] In certain embodiments, the deprotection conditions refer to a solution containing a buffer and a scavenger.

[0464] Exemplary scavengers may be selected from the group consisting of ammonium phosphate, acetyllysine, m-cresol, dithiothreitol, 1,2-ethanedithiol, hydrazine, hydroxylamine, imidazole, 2-mercaptopyridine, 4-mercaptopyridine, 2-methoxyphenol, 4-methoxyphenol, morpholine, phenol, piperazine, proline, thioaniline, thioanisole, N,N,N'-trimethylethylenediamine, triethylsilane, triisopropylsilane, and tris(hydroxymethyl)methaneamine.

[0465] In certain embodiments, the deprotection condition refers to a solution containing a buffer and N,N,N'-trimethylethylenediamine.

[0466] In certain embodiments, the deprotection conditions refer to a solution containing an organic solvent, such as a polar protic solvent or a polar aprotic solvent.

[0467] In certain embodiments, the deprotection conditions refer to a solution containing an organic solvent and an acid.

[0468] In certain embodiments, the acid is trifluoroacetic acid. In certain embodiments, the acid is hydrochloric acid.

[0469] In certain embodiments, the deprotection conditions refer to a solution containing an organic solvent and a base.

[0470] In one particular embodiment, the base is 1,8-diazabicyclo[5.4.0]undeca-7-ene (DBU). In other embodiments, the base is piperidine.

[0471] In certain embodiments, the deprotection condition refers to a solution containing a polar aprotic solvent.

[0472] Exemplary nonpolar protic solvents may be selected from the group consisting of dimethyl sulfoxide, 1,2-dimethoxy ether, 1,3-dimethyl-2-imidazolidinone, 1,3-dioxolane, 1,4-dioxane, 2,5-dimethyltetrahydrofuran, 2-methyltetrahydrofuran, 4-acetylmorpholine, 4-propionylmorpholine, acetone, acetonitrile, diethyl carbonate, diethyl ether, dimethyl carbonate, ethyl acetate, ethyl formate, ethyl lactate, ethylene carbonate, gamma-butyrolactone, gamma-valerolactone, hexamethylphosphoramide, methyl acetate, methyl carbonate, monomethyl ether acetate, N,N'-dimethylpropylene urea, N,N-dimethylacetamide, N,N-dimethylformamide, N-formylmorpholine, N-methyl-2-pyrrolidone, propylene carbonate, sulfolane, tetrahydrofuran, tetrahydropyran, tripyrrolidinophosphate triamide, and mixtures thereof.

[0473] In certain embodiments, the deprotection condition refers to a solution containing a polar protic solvent.

[0474] Exemplary polar protic solvents may be selected from the group consisting of ethanol, 1,4-butanediol, acetic acid, cyclohexanol, diethylene glycol, diethylene glycol monoethyl ether, diethylene glycol monomethyl ether, ethylenediamine, ethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monomethyl ether, formamide, formic acid, glycerin, isobutanol, isopropanol, methanesulfonic acid, methanol, n-butanol, n-hexanol, n-pentanol, n-propanol, propionic acid, propylenediamine, propylene glycol, propylene glycol monoethyl ether, propylene glycol monomethyl ether, sec-butanol, t-butanol, triethylene glycol monoethyl ether, triethylene glycol monomethyl ether, triethylene glycol, trifluoroacetic acid, water, and mixtures thereof.

[0475] In certain embodiments, the deprotection condition also provides a shift condition.

[0476] Another aspect of the present invention is a pharmaceutical composition comprising at least one conjugate or a pharmaceutical salt thereof of the present invention.

[0477] In a particular embodiment, the pharmaceutical composition comprises at least one conjugate or a pharmaceutical salt thereof of the present invention, for example, one conjugate. In a particular embodiment, the pharmaceutical composition comprises two conjugates of the present invention. In a particular embodiment, the pharmaceutical composition comprises three conjugates of the present invention.

[0478] Such pharmaceutical compositions may have a pH in the range of pH 3 to pH 8, for example, in the range of pH 4 to pH 6 or pH 4 to pH 5. In a particular embodiment, the pH of the pharmaceutical composition is about 4. In a particular embodiment, the pH of the pharmaceutical composition is about 4.5. In a particular embodiment, the pH of the pharmaceutical composition is about 5. In a particular embodiment, the pH of the pharmaceutical composition is about 5.5.

[0479] In a particular embodiment, the pH of the pharmaceutical composition is 4. In a particular embodiment, the pH of the pharmaceutical composition is 4.5. In a particular embodiment, the pH of the pharmaceutical composition is 5. In a particular embodiment, the pH of the pharmaceutical composition is 5.5.

[0480] In a particular embodiment, such a pharmaceutical composition is a suspension formulation.

[0481] In certain embodiments, such a pharmaceutical is a dry composition. It is understood that such a dry composition can be obtained by drying a suspension composition, for example, by freeze-drying.

[0482] When the pharmaceutical composition is a parenteral composition, suitable excipients may be classified as, for example, buffers, isotonic modifiers, preservatives, stabilizers, anti-adsorption agents, oxidation protection agents, viscosifiers / viscosity enhancers, anti-agglomeration agents, or other auxiliary agents. However, in some cases, a single excipient may have two or three functions. Excipients may be selected from the group consisting of the following: (i) Buffering agents: physiologically tolerable buffers that maintain the pH within a desired range, such as sodium phosphate, bicarbonate, succinate, histidine, citrate and acetate, sulfate, nitrate, chloride, and pyruvate; antacids, such as Mg(OH)2 or ZnCO3, may also be used; (ii) Isotonic modifiers: These are used to minimize pain that may result from cell damage caused by osmotic pressure differences in the injection depot, and examples include glycerin and sodium chloride. The effective concentration can be determined by osmotic measurement using the estimated osmotic pressure of serum between 285 and 315 mOsmol / kg; (iii) Preservatives and / or antimicrobial agents: Multi-dose parenteral formulations require the addition of preservatives at concentrations sufficient to minimize the risk of patient infection at injection, and corresponding regulatory requirements have been established; typical preservatives include m-cresol, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, chlorobutanol, benzyl alcohol, phenylmercury nitrate, thimerosal, sorbic acid, potassium sorbate, benzoic acid, chlorocresol, and benzalkonium chloride; (iv) Stabilizers: Stabilization is achieved by enhancing the protein stabilizing power, by destabilizing the denatured state, or by directly binding excipients to the protein. Stabilizers may include amino acids, such as alanine, arginine, aspartic acid, glycine, histidine, lysine, proline; sugars, such as glucose, sucrose, trehalose; polyols, such as glycerol, mannitol, sorbitol; salts, such as potassium phosphate, sodium sulfate; chelating agents, such as EDTA, hexaphosphate; ligands, such as divalent metal ions (zinc, calcium, etc.); other salts or organic molecules, such as phenol derivatives; in addition, oligomers or polymers, such as cyclodextrin, dextran, dendrimers, PEG, or PVP; or protamine or HSA may be used. (v) Anti-adsorption agents: Primarily ionic or nonionic surfactants, or other proteins or soluble polymers, are used to coat the inner surface of the formulation container or to competitively adsorb thereon, such as poloxamer (Pluronic F-68), PEG dodecyl ether (Brij 35), polysorbate 20 and 80, dextran, polyethylene glycol, PEG-polyhistidine, BSA and HSA, and gelatin. The concentration and type of excipient selected are determined by the effects to be avoided, but typically a single layer of surfactant is formed at a boundary slightly above the CMC value; (vi) Antioxidants: Antioxidants such as ascorbic acid, ectoin, methionine, glutathione, monothioglycerol, morin, polyethyleneimine (PEI), propyl gallate, and vitamin E may be used, as well as chelating agents such as citric acid, EDTA, hexaphosphate, and thioglycolic acid; (vii) Thickeners or viscosity enhancers: Used to slow the settling of particles in vials and syringes, to facilitate mixing and resuspending of particles, and to make it easier to inject the suspension (i.e., to reduce the force on the syringe plunger). Suitable thickeners or viscosity enhancers include, for example, carbomer thickeners such as Carbopol 940 and Carbopol Ultrez 10, cellulose derivatives such as hydroxypropyl methylcellulose (hypromellose, HPMC) or diethylaminoethylcellulose (DEAE or DEAE-C), colloidal magnesium silicate (Veegum) or sodium silicate, hydroxyapatite gel, tricalcium phosphate gel, xanthan gum, and Satia gum UTC. Carrageenan, aliphatic poly(hydroxy acids), e.g., poly(D,L- or L-lactic acid) (PLA) and poly(glycolic acid) (PGA), and copolymers thereof (PLGA), terpolymers of D,L-lactide, glycolide and caprolactone, poloxamers, hydrophilic poly(oxyethylene) blocks and hydrophobic poly(oxypropylene) blocks constituting the triblock of poly(oxyethylene)-poly(oxypropylene)-poly(oxyethylene) (e.g., Pluronic®), polyether ester copolymers, e.g., polyethylene glycol terephthalate / polybutylene terephthalate copolymer, sucrose isobutyrate acetate (SAIB), dextran or its derivatives, dextran and PE The combination of G, polydimethylsiloxane, collagen, chitosan, polyvinyl alcohol (PVA) and its derivatives, polyalkylimide, poly(acrylamide-co-diallyldimethylammonium (DADMA)), polyvinylpyrrolidone (PVP), glycosaminoglycans (GAG), such as dermatan sulfate, chondroitin sulfate, keratan sulfate, heparin, heparan sulfate, hyaluronan, a hydrophobic A block, such as polylactide (PLA) or poly(lactide-co-glycoside) (PLGA), and a hydrophilic B block, such as polyethylene glycol (PEG) or polyvinylpyrrolidone, is an ABA triblock or AB block copolymer, and such block copolymers, as well as the poloxamers mentioned above, are reversed. It may exhibit thermal gelation behavior (it remains in a fluid state at room temperature, facilitating administration, and after injection, the body temperature exceeds the sol-gel transition temperature, causing it to become a gel); (viii) Expanding agents or diffusing agents: These modify the permeability of connective tissue by hydrolysis of components of the extracellular matrix in the interstitial space, such as hyaluronic acid, a polysaccharide found in the intercellular spaces of connective tissue. Expanding agents, such as hyaluronidase, temporarily reduce the viscosity of the extracellular matrix and promote the diffusion of injected drugs; (ix) Anti-flocculating agent: for example, propylene glycol; and (x) Other auxiliary agents: Wetting agents, viscosity modifiers, antibiotics, hyaluronidase, etc. Acids and bases, such as hydrochloric acid and sodium hydroxide, are auxiliary agents necessary for pH adjustment during manufacturing.

[0483] In another embodiment, the present invention relates to a conjugate of the present invention or a pharmaceutical composition comprising a conjugate of the present invention for use as a pharmaceutical.

[0484] In another embodiment, the present invention relates to a pharmaceutical composition comprising the conjugate of the present invention or a pharmaceutically acceptable salt thereof, or the conjugate of the present invention, for use in a method of treating a disease that can be treated with DH or a pharmaceutically acceptable salt thereof.

[0485] In a further embodiment, the present invention relates to a method for preventing a disease that can be prevented or treated by DH, or for treating a patient suffering from such a disease, comprising administering to the patient an effective amount of the conjugate of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the conjugate.

[0486] Since the present invention is applicable to all drug molecules containing primary or secondary amines, it is impossible to further specify the diseases that can be treated. However, it will be obvious to those skilled in the art which diseases can be treated with a particular conjugate.

[0487] [Examples] Materials and methods chemicals Unless otherwise stated, all materials were obtained commercially. The monoclonal antibody CTLA-4 mAB (AMO-M6104, CAS number 477202-00-9) was obtained from AbMole Bioscience Inc., Houston, Texas, US. HHC MET ( TIFF0007851853000097.tif36160 (Sequence ID 1) was procured from an external supplier on a custom-made basis, with protein expression performed from Escherichia coli (E. coli) followed by standard purification procedures known to those skilled in the art.

[0488] reaction The reaction was carried out using anhydrous solvents (CH2Cl2, DMSO, DMF, THF, acetonitrile) purchased from Sigma-Aldrich Chemie GmbH, Munich, Germany. Generally, the reactants were stirred at room temperature and monitored by LC-MS.

[0489] RP-HPLC purification Preparative RP-HPLC purification was performed using a Waters XBridge BEH300 Prep C18 10 μm, 150 × 30 mm column as the stationary phase, with a Waters 600 controller and either a 2487 Dual Absorbance Detector or an Agilent Infinity 1260 preparative system. The product was detected at 215 nm. Linear gradients were used for solvent system A (water containing 0.1% TFA v / v) and solvent system B (acetonitrile containing 0.1% TFA v / v). The HPLC fractions containing the product were pooled and freeze-dried unless otherwise specified.

[0490] Flash chromatography Flash chromatography purification was performed using the Isolera One system at Biotage AB, Sweden, with a Biotage KP-Sil silica cartridge. The product was detected at 254 nm or 280 nm.

[0491] RP-LPLC purification Low-pressure RP chromatography purification was performed using a Biotage AB, Sweden Isolera One system with a Biotage SNAP C18 cartridge. The product was detected at 215 nm or 280 nm. Linear gradients were taken for solvent system A (water containing 0.1% TFA v / v) and solvent system B (acetonitrile containing 0.1% TFA v / v). The fractions containing the product were pooled and lyophilized unless otherwise specified.

[0492] UPLC-MS analysis Analytical ultrafast LC (UPLC)-MS was performed using a Waters BEH300 C18 column (2.1 × 50 mm, 1.7 μm particle size or 2.1 × 100 mm, 1.7 μm particle size) connected to a Waters Micromass ZQ or an Agilent Single Quad MS system; solvent A: water containing 0.05% TFA (v / v); solvent B: acetonitrile containing 0.04% TFA (v / v); or an Agilent 1290 Infinity II.

[0493] OPA assay The amine content of amine-HA was determined by reacting free amino groups with o-phthalaldehyde (OPA) and N-acetylcysteine ​​under alkaline conditions, as systematically described in Molnar-Perl (Ed.) (2015), Journal of Chromatography Library 70:405~444, and then determining the photometric intensity of the resulting chromophore.

[0494] Content of dried hydrogel in hydrogel suspension The hydrogel content was determined by continuously washing representative aliquots of the suspension with water and anhydrous ethanol in a syringe reactor equipped with PE frit, and then drying the solid hydrogel portion under vacuum. The hydrogel content was calculated from the mass of hydrogel residue per syringe and the volume of each aliquot of the hydrogel suspension.

[0495] MTS load quantification The MTS load of appropriate hydrogels was determined by removing MTS groups using TCEP reduction, followed by quantitative determination of free thiols on the hydrogel using the Ellman assay. The determination was performed using aliquots of appropriate MTS-hydrogel suspensions in a syringe reactor equipped with PE frit. The MTS load of the dried hydrogel was calculated using the hydrogel content of the suspension.

[0496] Ellman assay The thiol content of compounds that may be soluble or insoluble in aqueous systems is determined by the reaction of the free compound thiol group with the DTNB reagent at neutral pH and the photometric determination of the released 5-thio-2-nitrobenzoic acid (TNB), as systematically described in GLEllman (1959), Archives of Biochemistry and Biophysics 82:70-77.

[0497] Reverse Ellman assay The maleimide content of a compound that may be soluble or insoluble in aqueous systems is determined by quenching the maleimide group with an excess of 2-mercaptoethanol under neutral conditions. The amount of residual unreacted thiol reagent is determined by the Ellman assay. The maleimide content of the compound is calculated using the difference between the total amount of 2-mercaptoethanol added and the residual thiol after the reaction.

[0498] Protein concentration determination The concentration of the protein solution was determined using a Tecan Infinite M200 with a UV cuvette micrometer (neoLAB) under the following conditions: path length 1 cm; absorbance wavelength 280 nm; absorbance wavelength band 5 nm; reference wavelength 338 nm; reference wavelength band 25 nm; number of flashes 25. HHC MET Extinction coefficient: ε = 2.052 mL / (mg × cm). HHC MET The concentration of the conjugate mixture containing HHC MET The concentration was determined by using the extinction coefficient of CTLA-4 mAB. Extinction coefficient of CTLA-4 mAB: ε = 1.53 mL / (mg × cm). The concentration of the conjugate mixture containing CTLA-4 mAB was determined using the extinction coefficient of CTLA-4 mAB.

[0499] [Example 1] Synthesis of Protected Diamino Alcohol 1c 1c was synthesized according to the following scheme:

[0500] [ka]

[0501] 1b was synthesized according to WO2018 / 175788A1 Example 1A and used as the TFA salt. 1b (352 mg, 0.61 mmol) was dissolved in acetonitrile (2.50 mL), and the solution was cooled in an ice bath. DIPEA (242 μL, 1.39 mmol) was added, and the reaction mixture was mixed. 1,3-diamino-2-propanol (25 mg, 0.28 mmol) was dissolved in acetonitrile (1.00 mL) and added to the reaction mixture. The reaction mixture was mixed and incubated in an ice bath. Reaction control after 5 minutes indicated that the reaction had ended. Approximately 15 minutes later, TFA (106 μL, 1.39 mmol) was added to the ice-cooled reaction mixture. The reaction mixture was diluted with 4 ml of water containing 0.1% TFA. Product 1c was purified by RP-HPLC. Yield: 204 mg (84%, 2x TFA salt) MS: m / z 647.34 = [M+H] + (Calculated value = 647.34).

[0502] Synthesis of linker reagent 1i 1i was synthesized according to the following scheme:

[0503] [ka] TIFF0007851853000100.tif119162

[0504] 1d was synthesized according to WO2018 / 175788A1 Example 1E.

[0505] 1d (800 mg, 3.53 mmol), HOSu (610 mg, 5.30 mmol), and EDC (1.02 g, 5.30 mmol) were dissolved in dichloromethane (9.6 mL). DIPEA (616 μL, 3.53 mmol) was added. After 1 hour, the reaction showed incomplete conversion to the product, and after 75 minutes, HOSu (203 mg, 1.77 mmol) and EDC (339 mg, 1.77 mmol) were added to the reaction mixture. After 135 minutes, the reaction mixture was diluted with 100 ml of ethyl acetate and washed three times with 130 ml of 1 M hydrochloric acid. The organic phase was separated, dried over MgSO4, filtered, and concentrated under reduced pressure. The resulting crude product was dried overnight under high vacuum. The crude product was purified by RP-HPLC to obtain 1e. Yield: 913 mg (80%) MS: m / z 346.02 = [M+Na] + (Calculated value = 346.04).

[0506] N-Me-Asp(OtBu)-OH (100 mg, 0.49 mmol) was suspended in dichloromethane (0.5 mL), and a solution of 1e in dichloromethane (0.5 mL) (239 mg, 0.74 mmol) was added. DIPEA (171 μL, 0.98 mmol) was added. After 95 min, the reaction mixture was quenched with 171 μL of acetic acid, and volatile substances were removed under an argon stream. The crude product was purified by RP-HPLC to obtain 1f. Yield: 235 mg (97%) MS: m / z 412.07 = [M+H] + (Calculated value = 412.15).

[0507] 1c (150 mg, 171 μmol) was dissolved in acetonitrile (4 mL). 1f (85 mg, 206 μmol) and DMAP (42 mg, 343 μmol) were added and dissolved. While stirring, DIC (106 μL, 686 μmol) was added, and the reaction mixture was stirred for 30 minutes. The reaction mixture was quenched with 4 ml of 1% TFA in water and filtered. The filtrate was purified by RP-HPLC to obtain 1 g. Yield: 151 mg (69%, 2x TFA salt) MS: m / z 520.74 = [M+2H] 2+ (Calculated value = 520.74).

[0508] Dissolve 1 g (90 mg, 71 μmol) in 1 mL of dichloromethane, and vigorously stir the solution in an open flask. Add 1 mL of TFA all at once. Stir the reaction mixture for 45 min, and remove volatile substances under a nitrogen stream. Dissolve the residue in 250 μL of acetonitrile and precipitate in 10 ml of diethyl ether. Rinse the reaction flask with 250 μL of anhydrous acetonitrile and add it to the ether suspension. Centrifuge the suspension and remove the supernatant. Immediately dry the precipitate under a nitrogen stream to obtain 1h (2 × TFA salt).

[0509] 1h was dissolved in 1 ml of dichloromethane and 1 ml of acetonitrile. HOSu (25 mg, 213 μmol), EDC (41 mg, 213 μmol) and DMAP (0.87 mg, 7.1 μmol) were added to the reaction. After 30 min, the reaction volume was reduced to 1 ml in a nitrogen stream. After 1 h, HOSu (25 mg, 213 μmol) and EDC (41 mg, 213 μmol) were added. After 2 h, 3 ml of water containing 0.1% TFA was added to the reaction. The pH was adjusted to <pH 2 by adding 60 μL of 10% TFA in acetonitrile / water 1:1. The product was purified from the crude reaction mixture by RP-HPLC to give 1i. Yield: 83 mg (90%, 2x TFA salt) MS: m / z 1081.42 = [M+H] + , (calculated = 1081.42).

[0510] [Example 2] Synthesis of ubiquitin-linker-PEG20k conjugate 2b:

[0511] [Chemical formula]

[0512] Ubiquitin (157 μL, 12.75 mg / ml) from bovine erythrocytes in 60 mM sodium phosphate buffer pH 7.4 was mixed with solution 1i (4.86 μL, 0.1 M) in DMSO. The reaction was incubated at rt for 10 min and quenched by adding 39.3 μL of pH 3 sodium succinate buffer (0.5 M) to shift the pH of the reaction to approximately 4. To remove the unconjugated linker from the protein conjugate mixture, an Akta pure system equipped with a GE HiTrap column was used at a flow rate of 2 ml / min for buffer exchange to 5 mM sodium succinate pH 4. 1.5 ml of the product fraction was collected, which contained ubiquitin and ubiquitin linker conjugate 2a.

[0513] 9.3 mg of mPEG thiol 20 kDa (Sunbright ME-200SH) was dissolved in the product fraction, and 225 μL of 0.5 M sodium phosphate buffer at pH 7.8 containing 200 mM TriMED was added to the reaction to promote the cleavage of the protecting group and the rearrangement of ester 2a to amide 2b. The reaction mixture was combined and incubated at 25°C for 20 hours.

[0514] Ubiquitin-linker-PEG20k monoconjugate was purified by ion-exchange chromatography using a GE Healthcare 15S 4.6 / 100 PE column connected to an Akta pure system. The reaction product was diluted 10-fold with water to a volume of 20 ml, and the pH was adjusted to approximately 4 by adding 50 μL of 10% TFA in water. For chromatography, 20 mM sodium acetate and 10 mM methionine (pH 4) were used as buffer A, and 20 mM sodium acetate, 10 mM methionine, and 0.5 M NaCl (pH 4.5) were used as buffer B, creating a gradient of 0-50% buffer B across 13 column volumes. The total reaction volume was loaded into the column in four 5 ml injections and eluted in a single operation. Pure PEG monoconjugate was found in a 2 ml fraction. The fraction was buffered with PBS (pH 7.4) using three HiTrap columns in succession. The resulting 3 ml fraction was concentrated to 0.4 ml using a Vivaspin Turbo 4 MWCO 5000 and stored at -20°C.

[0515] [Example 3] Synthesis of Protected Diamino Alcohols 3a-c 3a was synthesized according to the following scheme:

[0516] [ka]

[0517] A dimethylamine solution (536 μL, 2M, 1.07 mmol) and dichloromethane (5 mL) in THF were stirred at rt. DIPEA (204 μL, 1.17 mmol), Boc-Ser-OH (200 mg, 0.97 mmol), HOBt (198 mg, 1.46 mmol), and EDC (224 mg, 1.17 mmol) were added to the reaction mixture. The reaction mixture was stirred for 23 hours. The reaction mixture was diluted with 5 ml of dichloromethane and washed with saturated bicarbonate, 5% citric acid, water, and brine (once each). Each washing solution was back-extracted with 5 ml of DCM. The combined organic phase was dried (MgSO4), filtered, and concentrated under vacuum. The resulting Boc-Ser-NMe2 was used in the next step without further purification. Yield: 140 mg (62%) MS: m / z 232.96 = [M+H] + (Calculated value = 233.15).

[0518] The crude product from the final step (74 mg, 0.32 mmol) was dissolved in THF (0.4 mL) and toluene (0.2 mL) and stirred in an ice bath under a nitrogen atmosphere. Bis(2-methoxyethoxy)aluminum sodium hydride (70% w / w in toluene, 445 μL, 1.6 mmol) was added dropwise, accompanied by gas release. After 4 hours, the reaction mixture was quenched with 1 M NaOH aqueous solution (0.65 mL). The reaction mixture was stirred at rt for 30 minutes. The reaction mixture was acidified with TFA (pH < 2). The white precipitate was filtered off and washed with acetonitrile / water 1:1 (approximately 1.5 ml) containing 0.1% TFA. The filtrate was purified by RP-HPLC to obtain 3a. Yield: 61 mg (57%, TFA salt) MS: m / z 219.17 = [M+H] + (Calculated value = 219.17).

[0519] 3b was synthesized according to the following scheme:

[0520] [ka]

[0521] A dimethylamine solution (536 μL, 2 M, 1.07 mmol) and dichloromethane (5 mL) in THF were stirred at rt. DIPEA (204 μL, 1.17 mmol), (S)-3-((tert-butoxycarbonyl)amino)-2-hydroxypropanoic acid (200 mg, 0.97 mmol), HOBt (198 mg, 1.46 mmol), and EDC (224 mg, 1.17 mmol) were added to the reaction mixture. The reaction mixture was stirred for 19 hours. The reaction mixture was diluted with 5 ml of DCM and washed with saturated bicarbonate, 5% citric acid, water, and brine (1 × 5 ml each). Each washing solution was back-extracted with 5 ml of DCM. The organic phase was dried with (MgSO4), filtered, and concentrated under vacuum. The resulting intermediate was used in the next step without further purification. Yield: 204 mg (90%) MS: m / z 233.15 = [M+H] + (Calculated value = 233.15).

[0522] The crude product from the final step (204 mg, 0.88 mmol) was dissolved in THF (1 mL) and toluene (0.5 mL) and stirred in an ice bath under a nitrogen atmosphere. Bis(2-methoxyethoxy)aluminum sodium hydride (70% w / w in toluene, 1.22 mL, 4.39 mmol) was added dropwise, accompanied by gas release. After 3 hours, the reaction mixture was quenched with 1 M NaOH aqueous solution (1.63 mL). The reaction mixture was stirred at rt for 30 minutes. The aqueous layer was extracted three times with 5 ml of diethyl ether. The ether layer was extracted with 10 ml of 0.64 M HCl aqueous solution, and the product was isolated from the aqueous acidic phase by RP-HPLC to obtain 3b. Yield: 173 mg (59%, TFA salt) MS: m / z 219.17 = [M+H] + (Calculated value = 219.17).

[0523] 3c was synthesized according to the following scheme:

[0524] [ka]

[0525] H-Dpr(Boc)-OMe HCl salt (200 mg, 0.79 mmol) was dissolved in MeOH (9.4 mL), and aqueous formaldehyde solution (37% w / w, 175 μL) was added to this solution. The reaction mixture was stirred for 10 minutes, and sodium borohydride cyanohydride (148 mg, 2.36 mmol) was added all at once. The reaction mixture was stirred for 2.5 hours. Aqueous formaldehyde solution (88 μL) and sodium borohydride cyanohydride (74 mg, 1.18 mmol) were added to the reaction mixture. After 5 hours, the reaction mixture was quenched with a solution prepared from 5 ml of saturated bicarbonate solution and 5 ml of water. The mixture was stirred overnight in an open flask in a fume hood. The aqueous phase was extracted by DCM (4 × 20 mL). The combined organic phase was dried (MgSO4), filtered, and concentrated under vacuum. The resulting intermediate was used in the next step without further purification. Yield: 161 mg (83%) MS: m / z 247.17 = [M+H] + (Calculated value = 247.17).

[0526] The crude product from the final step (160 mg, 0.65 mmol) was dissolved in THF (1.5 mL), and the reaction mixture was cooled in an ice bath. 1.3 mL, 1.3 mmol of lithium aluminum hydride solution (1 M in THF) was added dropwise while stirring (gas formation). After 1 hour, the reaction mixture was diluted with 5 mL of diethyl ether, and 50 μL of water was added dropwise. After gas formation subsided, 50 μL of 4 M NaOH was added, followed by 150 μL of water. The reaction mixture was stirred at rt for 30 minutes. MgSO4 was added, and the reaction mixture was stirred for 15 minutes. The solid was filtered off and washed with 5 mL of diethyl ether. Volatile substances were removed under a nitrogen stream. The product was purified by RP-HPLC to obtain 3c. Yield: 158 mg (73%, TFA salt) MS: m / z 219.17 = [M+H] + (Calculated value = 219.17).

[0527] [Example 4] Synthesis of Ac-N-Me-Asp(OBzl)-OH Fmoc-N-Me-Asp(OBzl)-OH (500 mg, 1.09 mmol) was dissolved in THF (5 mL). DBU (325 μL, 2.18 mmol) was added dropwise with stirring. A suspension was formed. After 20 min, a solution of N-acetoxysuccinimide (342 mg, 2.18 mmol) in THF (5 mL) was added to the reaction. The suspension slowly changed to a solution. After 50 min, the reaction solution was diluted with DCM (20 ml) and extracted with saturated sodium bicarbonate (30 ml). The aqueous phase was washed again with DCM (20 ml). The organic phase was discarded. Ethyl acetate (30 ml) was added to the bicarbonate phase, which was acidified to <pH 2 with concentrated HCl.

[0528] The organic phase was further washed with 0.1 M HCl (2 × 30 ml). The organic phase was dried (MgSO4), filtered, and concentrated in vacuo to give Ac-N-Me-Asp(OBzl)-OH (4). Yield: 299 mg (99%) MS: m / z 280.12 = [M+H] + , (calculated value = 280.12).

[0529] [Example 5] Synthesis of linker reagents 5e~h 5e~h were synthesized according to the following scheme:

[0530]

Chemical formula

[0531] Synthesis of 5e: Fmoc-N-Me-Asp(OBzl)-OH (138 mg, 0.30 mmol) was dissolved in DCM (1 mL), and N,N'-bis-Boc-2-hydroxypropylenediamine (105 mg, 0.36 mmol) and EDC (86 mg, 0.45 mmol) were added with stirring. A catalytic amount of DMAP was added. After 2.5 hours, the reaction mixture was diluted with 10 ml of DCM and washed twice with citric acid, twice with saturated bicarbonate solution, and once with brine (10 ml each). The organic phase was dried with (MgSO4), filtered, and reduced to a volume of 2 ml under vacuum.

[0532] DBU (90 μL, 0.60 mmol) was added while stirring. After 15 minutes, the reaction was complete, and acetic anhydride (142 μL, 1.50 mmol) was added while stirring. After 30 minutes, the reaction product was acidified with 100 μL of acetic acid, and the solvent was removed under a nitrogen stream. Intermediate 5a was purified by RP-HPLC. Yield: 107 mg (65%) MS: m / z 552.29 = [M+H] + (Calculated value = 552.29).

[0533] 5a (107 mg, 0.19 mmol) was dissolved in THF (2 mL), and palladium 10% Pd standard (41 mg, 0.04 mmol) on activated carbon was added. The reaction mixture was vigorously stirred under a hydrogen atmosphere. The reaction mixture was diluted with THF (8 mL) and filtered through a 0.2 μm PTFE syringe filter. TSTU (117 mg, 0.39 mmol) and DIPEA (67 μL, 0.39 mmol) were added. The suspension was stirred at rt. After 16 h, the suspension was filtered. The filtrate was concentrated under vacuum. The residue was partitioned between ethyl acetate and pH 7 sodium phosphate buffer (100 mM). The organic phase was washed twice with pH 7 buffer and once with brine. The organic phase was dried (MgSO4), filtered, and concentrated under vacuum. The oily residue was dissolved in DCM and concentrated under vacuum to obtain 5e as a white foam. Yield: 106 mg (98%) MS: m / z 559.26 = [M+H] + (Calculated value = 559.26).

[0534] Synthesis of 5b~d: 5b:4 (47 mg, 167 μmol) was dissolved in DCM (1 ml). The solution was added to 3a (61 mg, 183 μmol). The reaction mixture was stirred, and EDC (35 mg, 183 μmol) was added. DMAP (2 mg, 17 μmol) was added. After 1 hour, EDC (35 mg, 183 μmol) and DMAP (2 mg, 17 μmol) were added. After 2 hours, volatile substances were removed under a nitrogen stream. 5b was purified by RP-HPLC. Yield: 48 mg (48%, TFA salt) MS: m / z 480.27 = [M+H] + (Calculated value = 480.27).

[0535] According to 5c:5b, 5c was synthesized using 4 (50 mg, 0.18 mmol) and 3b (65 mg, 0.2 mmol). EDC (76 mg, 0.39 mmol) and DMAP (2 mg, 0.02 mmol) were added at once, and the reaction time was 18 hours. Yield: 39 mg (37%, TFA salt) MS: m / z 480.27 = [M+H] + (Calculated value = 480.27).

[0536] 5d was synthesized according to the procedure, similar to 5c, but using 3c instead of 3b. Yield: 30 mg (28%, TFA salt) MS: m / z 480.27 = [M+H] + (Calculated value = 480.27).

[0537] Synthesis of 5f~h: 5f:5b (48 mg, 100 μmol) and palladium on activated carbon (21 mg, based on 10% Pd) were suspended in THF (2 mL) under a nitrogen atmosphere. The container was filled with hydrogen and held under a hydrogen atmosphere for 2.5 hours. The reaction mixture was filtered through a 0.2 μm PTFE syringe filter and rinsed with THF (4 mL). TSTU (60 mg, 200 μmol) and DIPEA (35 μl, 200 μmol) were added to the filtrate. The suspension was stirred at rt for 16 hours. After removing volatile substances, the product was purified by RP-HPLC to obtain 5f. Yield: 36 mg (60%, TFA salt) MS: m / z 487.24 = [M+H] + (Calculated value = 487.24).

[0538] Following the 5g:5f ratio, 5g was synthesized using 5c ​​(39mg, 36μmol), Pd on carbon (4mg), and THF (2ml) for a reaction time of 1h. The reaction mixture was filtered through a 1μm PTFE syringe filter and rinsed with DCM (2ml). TSTU (40mg, 132μmol) and DIPEA (46μL, 265μmol) were added to the filtrate. The suspension was stirred for 1h. After removing volatile substances, the product was purified by RP-HPLC to obtain 5g. Yield: 25 mg (63%, TFA salt) MS: m / z 487.24 = [M+H] + (Calculated value = 487.24).

[0539] 5h was synthesized following the procedure for 5g, but 5d (30mg, 51μmol) was used instead of 5c, and the reagents were adjusted accordingly. Yield: 19 mg (61%, TFA salt) MS: m / z 487.24 = [M+H] + (Calculated value = 487.24).

[0540] [Example 6] Synthesis of AcAKF-OH linker conjugates 6a-d from linker reagents 5e-h

[0541] [ka]

[0542] AcAKF tripeptides were synthesized on 2-chlorotrityl chloride resin using Fmoc amino acids Fmoc-Ala-OH, Fmoc-Lys(ivDDe)-OH, and Fmoc-Phe-OH. The N-terminus of the peptides was acetylated on the resin using acetic anhydride / DIPEA, and ivDDE was cleaved using hydrazine. Linkers 5e-h were linked to the free side-chain amine of Lys on the resin in DMF using 8-9 mg of peptide-loaded resin, 2 equivalents of each linker, and 3 eq of DIPEA. The resin was stirred for 30 minutes. The resin was washed 5 times with DMF and 5 times with DCM, and the Boc-protected peptide was cleaved from the resin using 20% ​​HFIP in DCM. The Boc-protected peptide was purified by RP-HPLC. The Boc protecting group was cleaved with TFA / DCM 1:1. Volatile substances were removed; the residue was dissolved in acetonitrile, and the peptide linker conjugate was isolated after ether precipitation as the TFA salt. 6a: Yield: 3 mg (37%, 2x TFA salt) MS: m / z 650.30 = [M+H] + (Calculated value = 650.35). 6b Yield: 2 mg (25%, 2x TFA salt) MS: m / z 678.40 = [M+H] + (Calculated value = 678.38). 6c yield: 2.4 mg (34%, 2x TFA salt) MS: m / z 678.40 = [M+H] + (Calculated value = 678.38). 6d yield: 2.4 mg (34%, 2x TFA salt) MS: m / z 678.39 = [M+H] + (Calculated value = 678.38).

[0543] [Example 7] In vitro release kinetics

[0544] [ka]

[0545] The cleavage rate of tripeptide AcAKF from conjugates 6a-d was monitored by LCMS (UV detection) in aqueous buffer (pH 7.4, 60 mM sodium phosphate) at pH 7.4 and 37°C. In the first step, the rearrangement of the linker portion occurred within a few minutes (as exemplified for conjugate 6b above), followed by the time-dependent disappearance of conjugate 7b, which was then fitted into curve fitting software to obtain the half-life for slow release. compound t 1 / 2 (pH 7.4) 6a 16d 6b 27d 6c 15d 6d 3.0d

[0546] [Example 8] In vitro release kinetics The cleavage rate of the reversible bond from conjugate 2b was monitored by LC-MS (UV detection) in aqueous buffer (PBS pH 7.4) at pH 7.4 and 37°C. The percentage of the increasing free ubiquitin peak area over time was determined and fitted to curve fitting software to obtain the release half-life. compound t 1 / 2 (pH 7.4) 2b 30d

[0547] [Example 9] Synthesis of 9g of linker reagent

[0548] [ka]

[0549] 9a (1.5 g, 5.7 mmol) was dissolved in THF (37.5 mL). TSTU (2.6 g, 8.6 mmol) and DIPEA (3.97 mL, 22.8 mmol) were added. A dense suspension was formed upon stirring. The mixture was stirred for 22 hours, and TSTU (1.7 g, 5.5 mmol), DIPEA (2 mL, 11.5 mmol), and DMF (13 mL) were added, causing the reaction product to change color to dark brown. After a total of 26 hours, the reaction mixture was diluted with 350 mL of ethyl acetate and washed with 2 × 200 mL of 0.1 N HCl and 1 × 100 mL of brine. The organic phase was dried over Na₂SO₄ and evaporated. The residue was dried overnight under high vacuum. The product was purified using flash chromatography, and 9b was obtained as a colorless oil. Yield: 1.65 g (81%) MS: m / z 361.17 = [M+H] + (Calculated value = 361.16).

[0550] 9b (1.65 g, 4.58 mmol) was dissolved in DCM (11.6 mL), and N-Me-L-Asp(tBu)-OH (932 mg, 4.59 mmol) and DIPEA (1.6 mL, 9.2 mmol) were added. The white suspension was stirred at RT. The mixture slowly turned into a pale yellow solution over time.

[0551] After 1 hour, acetic acid (786 μL, 13.7 mmol) was added. The solvent was evaporated, and the product was purified by RP-LPLC to obtain 9c. Yield: 1.77 g (86%) MS: m / z 449.15 = [M+H] + (Calculated value = 449.25).

[0552] 9c (1.23 g, 2.74 mmol) and 1c (1.99 g, 2.28 mmol) were dissolved in acetonitrile (53 mL). DMAP (557 mg, 4.56 mmol) was added under stirring, and DIC (1.41 mL, 9.12 mmol) was added to the resulting solution. After 1 hour, 0.7 mL of TFA was added, and the solvent was removed under vacuum. The product was purified by RP-LPLC to obtain 9d. Yield: 2.33 g (78%, 2x TFA salt) MS: m / z 1077.65 = [M+H] + (Calculated value = 1077.57).

[0553] 9d (2.33 g, 1.78 mmol) was dissolved in DCM (10 mL). TFA (10 mL, 131 mmol) was added under stirring. After 45 min, the solvent was evaporated, and the residue was co-evaporated with 50 mL of DCM. The residue was dried overnight under high vacuum to obtain 2.90 g of 9e, which was used without further purification. 9e was dissolved in acetonitrile (68 mL), and 3-maleimidopropionic acid N-hydroxysuccinimide (1.19 g, 4.45 mmol) was added under stirring. DIPEA (3.1 mL, 17.8 mmol) was added. After 80 min, the reaction mixture was quenched by adding TFA (1.36 mL, 17.8 mmol). The reaction mixture was concentrated under vacuum to a volume of 40 mL, and the product was purified by RP-LPLC to obtain 9f. Yield: 1.73 g (75% in two stages, 2x TFA salt) MS: m / z 1072.60 = [M+H] + (Calculated value = 1072.49).

[0554] 9f (1.73 g, 1.33 mmol) was dissolved in acetonitrile (17 mL), and EDC (767 mg, 4 mmol), HOSu (462 mg, 4 mmol), and DMAP (19 mg, 0.15 mmol) were added under stirring. After 1 hour and 30 minutes, the reaction mixture was quenched by adding TFA (100 μL, 1.3 mmol), and the reaction mixture was concentrated under vacuum to a volume of 8.5 mL. The product was purified by RP-LPLC to obtain 9 g. Yield: 1.36 g (73%, 2x TFA salt) MS: m / z 1169.71 = [M+H] + (Calculated value = 1169.50).

[0555] [Example 10] Preparation of amine-HA 10a and 10b Sodium hyaluronate (90-130 kDa, 504 mg, 1.25 mmol COOH, 1.00 equivalent) was dissolved in 100 mM MES and 400 mM 1,3-diaminopropane buffer pH 5.5 (62.5 mL) under vigorous stirring. HOBt (573 mg; 3.74 mmol, 3.00 equivalent) and EDC (223 mg; 1.17 mmol, 0.93 equivalent) were added. The suspension was stirred overnight at ambient temperature. Sodium acetate trihydrate (8.48 g) was added, and the suspension was converted into a solution. The crude amine-modified HA was precipitated by adding anhydrous ethanol, washed with 80% (v / v) ethanol and anhydrous ethanol, and dried under high vacuum for 1 hour. The precipitate was dissolved in water (40 mL) to form a clear solution. 4M NaOH (13.3 mL) was added, and the solution was stirred at ambient temperature for 2 hours, after which acetic acid (3.05 mL) was added. The product was precipitated by adding anhydrous ethanol, washed with 80% (v / v) ethanol and anhydrous ethanol, and dried under high vacuum to obtain amine-functionalized HA 10a as acetate. The amine content of this material was determined by OPA assay. Yield: 432 mg (acetate, amine content: 0.253 mmol / g, 10.4% DS)

[0556] Amine-HA 10b was prepared simply by using different amounts of EDC (95.8 mg; 0.50 mmol, 0.404 equivalents) according to the procedure described above. Yield: 449 mg (acetate, amine content: 0.114 mmol / g, 4.6% DS)

[0557] [Example 11] Preparation of thiol-HA11 from amine-HA10a Amine-functionalized HA 10a (400 mg, 0.101 mmol amine, 1.0 equivalent) was dissolved in 100 mM HEPES buffer pH 8.4 (33.25 mL). A freshly prepared solution of SPDP in acetonitrile (18 mL) (318 mg, 1.02 mmol, 10.1 equivalents) was added to the mixture with stirring. The mixture was stirred at ambient temperature for 120 minutes, and then a freshly prepared solution of TCEP in water (5.13 mL) (582 mg, 2.03 mmol, 20.1 equivalents) was added to the reaction mixture. This solution was stirred at ambient temperature for 1 hour, and then 1 M sodium acetate buffer pH 5.5 (56.4 mL) was added. The product was collected by adding anhydrous ethanol and centrifugation. After washing with 80% (v / v) ethanol and anhydrous ethanol, and drying under high vacuum for 5 hours, crude thiol-HA was obtained as a white solid. The crude material was dissolved in 1% acetic acid (40 mL) by vigorous stirring under an argon atmosphere. 1 M sodium acetate buffer pH 5.5 (40 mL) was added to this solution, and the resulting mixture was filtered through a 0.22 μm PES bottle-top filter. The product was precipitated from the filtrate by adding anhydrous ethanol and centrifugation. After washing with 80% (v / v) ethanol and anhydrous ethanol, the material was dried under high vacuum for 6 hours to obtain thiol-HA 11 as a grayish-white precipitate. The thiol content was determined by the Ellman assay. Yield: 366 mg (Thiol content: 0.209 mmol / g)

[0558] [Example 12] Preparation of maleimide-HA12 from amine-HA10b Amine-functionalized HA 10b (443 mg, 0.05 mmol amine, 1.0 equivalent) was dissolved in 100 mM HEPES buffer pH 7.4 (44.25 mL). A freshly prepared solution of 3-maleimidepropionic acid NHS ester (134 mg, 0.49 mmol, 10.0 equivalent) in acetonitrile (9.7 mL) was added to the mixture with stirring. The mixture was stirred at ambient temperature for 60 minutes, after which 1 M sodium acetate buffer pH 5.5 (54 mL) was added. The product was collected by adding anhydrous ethanol and centrifugation. After washing with 80% (v / v) ethanol and then anhydrous ethanol, the material was stored overnight at -20°C and dried under high vacuum for 2 hours the following day to obtain crude maleimide-HA as a white solid. The crude material was dissolved in 1% acetic acid (44.25 mL) with vigorous stirring. 1M sodium acetate buffer pH 5.5 (54 mL) was added to the solution, and the resulting mixture was filtered through a 0.22 μm PES bottle-top filter. The product was precipitated from the filtrate by adding anhydrous ethanol and centrifugation. After washing with 80% (v / v) ethanol and anhydrous ethanol, the material was dried under high vacuum for 6 hours to obtain maleimide-HA12 as a white precipitate. The maleimide content was determined by reverse Ellman assay. Yield: 376 mg (Maleimide content: 0.109 mmol / g)

[0559] [Example 13] Preparation of crosslinked HA microparticles 13 containing free thiols Thiol-HA 11 (90.5 mg) was dissolved in 200 mM MES and 3 mM EDTA buffer pH 5.5 (3015 μL) by vigorous shaking under an argon atmosphere to produce a 30 mg / mL solution of the compound in the buffer (Solution A). Maleimide-HA 12 (70.7 mg) was dissolved in 200 mM MES and 3 mM EDTA buffer pH 5.5 (2355 μL) by vigorous shaking to produce a 30 mg / mL solution of the compound in the buffer (Solution B). In a 2 mL Eppendorf tube equipped with a magnetic stirring bar, 200 mM MES and 3 mM EDTA buffer pH 5.5 (94.2 μL) were mixed with Solution A (717.7 μL) and Solution B (688.1 μL) under vigorous shaking. For gelation, the mixture was left to stand overnight under an argon atmosphere at rt. The gel was transferred to a 5 mL Luer lock syringe connected to a male / female Luer lock adapter, a 2 x 1 mm PTFE O-ring, a 144 μm stainless steel mesh (3.8 mm diameter), a 2 x 1 mm PTFE O-ring, a male / female Luer lock adapter, a 2 x 1 mm PTFE O-ring, a 144 μm stainless steel mesh (4 mm diameter), a 2 x 1 mm PTFE O-ring, and a male / female Luer lock adapter line. The gel portion in the syringe was passed through two 144 μm stainless steel meshes into a 15 mL Falcon tube containing 200 mM MES and 3 mM EDTA buffer at pH 5.50. The hydrogel was washed by shaking, centrifugation, and removal of the supernatant with 3 mM EDTA buffer pH 5.5, followed by 200 mM succinate, 3 mM EDTA buffer pH 4.0, and then 200 mM succinate, 3 mM EDTA, and 0.5% Tween 20 buffer pH 4.0. After the final washing step, the volume of the gel suspension was adjusted to 10 mL in a 15 mL Falcon tube using 3 mM EDTA and 0.5% Tween 20 buffer pH 4.0 to obtain a colorless, almost completely clear suspension of cross-linked HA containing free thiol groups. The thiol content of the hydrogel suspension was determined by the Ellman assay.

[0560] [Example 14] Preparation of CTLA-4 mAB-linker conjugate mixture 14 In this example, 204.13 mL of CTLA-4 mAB was used in a solution of 26 mM Tris-HCl, 100 mM NaCl, 55 mM mannitol, 0.1 mM pentetate (DTPA), 0.01% Tween 80, pH 7.0, at a concentration of 5.341 mg / mL. The mAB was buffered with 30 mM sodium phosphate, pH 7.4, concentrated, and the protein concentration was adjusted to 9.74 mg / mL. A 103.14 mL mAB solution was prepared.

[0561] 3 mol equivalents (218.6 μL) of 9 g of linker reagent (100 mM storage solution in DMSO) were added to the protein solution. The reaction mixture was carefully mixed and incubated at ambient temperature for 5 minutes to obtain a mixture 14 of unmodified CTLA-4 mAB and protected CTLA-4 mAB-linker conjugate (e.g., monoconjugate, bisconjugate).

[0562] To a volume of mAB solution (103.1 mL), 0.12 vol. equivalents (12.4 mL) of 0.5 M succinic acid, pH 3.0, were added to achieve a pH shift to approximately pH 4, and the solution was carefully mixed. 14 was purified by cation exchange chromatography at a flow rate of 4.0 mL / min using a linear salt gradient elution with a sodium chloride solution (0-60% 20 mM succinic acid, 1 M NaCl, pH 5.5 in 15 CV) with an Eshmuno CPX column (ID 8 mm × length 200 mm, CV = 10 mL), using 20 mM succinic acid, pH 5.5, as the mobile phase. Three operations were performed with an injection volume of approximately 39 mL (approximately 337 mg) per operation, and 119.27 mL of 14 was collected at a concentration of 7.32 mg / mL.

[0563] To determine the reactive maleimide content, 20 μL of 14 was PEGylated with 20 kDa PEG-thiol, followed by SE-HPLC analysis. After peak integration of the SE chromatogram, a total maleimide content of 41% was determined for 14.

[0564] After analysis and overnight storage at 4°C, 118.38 mL of solution 14 was adjusted to a final concentration of 5 mM EDTA and 0.01% Tween 20 using 1 / 19 vol. equivalent of 20 mM succinic acid, 100 mM EDTA, and 0.2% Tween 20, pH 5.5 (6.2 mL) relative to the 118.38 mL volume, and the solution was carefully shaken. The sample was filtered using a single qpore Plastic vacuum filter (PVDF membrane) with a pore size of 0.22 μm.

[0565] 122.67 mL of solution 14 was obtained at a concentration of 7.82 mg / mL.

[0566] [Example 15] Synthesis of temporary CTLA-4 mAB-linker-hydrogel prodrug 15b Thiol functionalization and conjugation of the CTLA-4 mAB / protected CTLA-4 mAB-linker conjugate mixture 14 to the crosslinked HA hydrogel 13 were performed by adding 1.5 mol of mixture 14 to the hydrogel 13 in proportion to the determined total maleimide content of 41% (4 μM).

[0567] A 7.5 mL hydrogel suspension, prepared in 20 mM succinic acid, 150 mM NaCl, 3 mM EDTA, 0.1% Tween 20, pH 4.0 according to Example 13 (nominal gel content of 4.22 mg / mL with 200.8 μM thiol content), was transferred to a 15 mL Falcon tube. A total of four 15 mL Falcon tubes were prepared in this manner. The hydrogel particles were allowed to settle by centrifugation at 4000 rcf for 1 minute, and the supernatant was removed by pipetting. Washing of the particles was achieved via five cycles of washing steps, including the addition of 10 mL of 20 mM succinic acid, 5 mM EDTA, 0.01% Tween 20, pH 5.5 buffer, centrifugation at 1000 rcf for 1 minute, and careful removal of the supernatant by pipetting. After the final washing step, each of the four Falcon tubes was filled with the buffer described above to a nominal total volume of 4 mL of suspension. From each Falcon tube, 2.6 mL of hydrogel suspension was transferred to a new 50 mL Falcon tube, resulting in four Falcon tubes, each containing 2.6 mL of washed hydrogel suspension.

[0568] 122.62 mL of 14 (c=7.82 mg / mL, 958.3 mg) at pH 5.5 was divided into four portions, and approximately 33 mL of each portion was added to each of the four 50 mL Falcon tubes containing the hydrogel suspension described above. The resulting suspensions were mixed by rotation and incubated overnight at ambient temperature under gentle stirring to obtain protective temporary CTLA-4 mAB-linker hydrogel prodrug 15a.

[0569] The hydrogel suspension was centrifuged at 1000 rcf for 1 minute and allowed to stand for 3 minutes. The supernatant after hydrogel loading was pipetteed into a 250 mL Corning bottle. The hydrogels were then combined into a single 50 mL Falcon tube.

[0570] The hydrogel was initially washed seven times with 30 mL of 10 mM IAA in 30 mM sodium phosphate, 50 mM TriMED, 0.01% Tween 20, pH 7.4. Then, 30 mL of 10 mM IAA in 30 mM sodium phosphate, 50 mM TriMED, 0.01% Tween 20, pH 7.4 was added to the hydrogel and incubated at ambient temperature for 1 hour with gentle stirring. Removal of the IAA blocking solution was achieved via 10 cycles of washing, including the addition of 30 mL of 30 mM sodium phosphate, 50 mM TriMED, 0.01% Tween 20, pH 7.4 buffer, centrifugation at 1000 rcf for 1 minute, and careful removal of the supernatant by pipetting after a 3-minute pause. Subsequently, to deprotect the protected transient CTLA-4 mAB-linker hydrogel prodrug 15a, 30 mL of 30 mM sodium phosphate, 50 mM TriMED, 0.01% Tween 20, pH 7.4 buffer was added to the precipitated hydrogel, and the resulting suspension was incubated overnight at 25°C to obtain transient CTLA-4 mAB-linker hydrogel prodrug 15b.

[0571] The final formulation of 15b was prepared by washing the hydrogel 10 times with 20 mM succinic acid, 10 wt% α-α-D-trehalose, 0.01% Tween20, and pH 5.5.

[0572] [Example 16] In vitro release kinetics for 15b 25 mg of 15b (equivalent to approximately 0.45 mg of protein) was transferred to a sterile 1.5 mL Eppendorf tube. A total of 8 tubes were prepared. 1 mL of 60 mM sodium phosphate, 3 mM EDTA, 0.01% Tween 20, pH 7.4 was added to each tube, and the contents were mixed by rotating and incubated for 5 minutes without agitation. The supernatant was removed to obtain a suspension of 0.5 mL in each vial. The suspension was incubated in a water bath at 37°C. After different time intervals, one vial was removed from 37°C, centrifuged, and the supernatant was analyzed by A280 measurement and SE-HPLC at 215 nm. The relative amount of CTLA-4 mAB released was recorded based on the concentration of the supernatant relative to the total amount of CTLA-4 mAB. Release kinetics from 15b:

[0573] [Table 1]

[0574] [Example 17] Synthesis of linker reagent 17f

[0575] [ka] TIFF0007851853000111.tif123162

[0576] 17a (880 mg, 2.68 mmol) was dissolved in DCM (4 mL) and added to a suspension of N-Me-L-Asp(tBu)-OH (519 mg, 2.55 mmol) in DCM (3 mL) and DIPEA (892 μL, 5.10 mmol). DCM (2 mL) and DMF (1.5 mL) were added to the colorless suspension and heated to 40°C to obtain a colorless solution. Acetic acid (438 μL, 7.66 mmol) was added after 7 hours and 30 minutes. The solvent was evaporated, and the product was purified by RP-LPLC to obtain 17b. Yield: 480 mg (45%) MS: m / z 417.25 = [M+H] +(Calculated value = 417.51).

[0577] 17b (480 mg, 1.15 mmol) and 1c (760 mg, 0.87 mmol) were dissolved in acetonitrile (20 mL). DMAP (212 mg, 1.74 mmol) was added under stirring, and DIC (541 μL, 3.48 mmol) was added to the resulting solution. After 1 hour and 55 minutes, TFA (332 μL, 4.35 mmol) was added, and the solvent was removed under vacuum. The product was purified by RP-LPLC to obtain 17c. Yield: 810 mg (73%, 2x TFA salt) MS: m / z 1045.74 = [M+H] + (Calculated value = 1045.57).

[0578] 17c (810 mg, 0.64 mmol) was dissolved in DCM (3.5 mL). TFA (3.5 mL, 45.7 mmol) was added under stirring. After 2 hours and 15 minutes, the solvent was evaporated, and the residue was co-evaporated with 25 mL of DCM. The residue was dried overnight under high vacuum to obtain 945 mg of 17d, which was used without further purification. 17d (56 mg, 45.2 μmol) was dissolved in acetonitrile (500 μL), and DBCO-C4-NHS ester (20 mg, 49.7 μmol) was added. DIPEA (79 μL, 0.452 mmol) was added. After 45 minutes, the reaction product was quenched by adding TFA (35 μL, 0.452 mmol). Water (400 μL) was added, and the product was purified by preparative HPLC to obtain 17e. Yield: 54 mg (84% in two stages, 2x TFA salt) MS: m / z 1176.77 = [M+H] + (Calculated value = 1176.55).

[0579] 17e (35 mg, 24.9 μmol) was dissolved in acetonitrile (500 μL), and EDC (14 mg, 74.8 μmol) and HOSu (8.6 mg, 74.8 μmol) were added. After 4 hours and 15 minutes, the reaction product was quenched by adding TFA (1.9 μL, 24.9 μmol). The reaction mixture was diluted with water (500 μL), and the product was purified by preparative HPLC to obtain 17f. Yield: 31.6 mg (84%, 2x TFA salt) MS: m / z 1274.86 = [M+H] + (Calculated value = 1273.57).

[0580] [Example 18] Preparation of PEG-based aminohydrogel PEG-based aminohydrogels were synthesized using different degrees of crosslinking, as described in Example 3 of WO2011 / 012715A1, to obtain different levels of amine content. All crosslinking agents were based on 3.3 kDa PEG and synthesized using azelaic acid, as described in Example 2 of WO2011 / 012715A1. The hydrogels were characterized by their free amine content: HG-1: 0.191 mmol / g, HG-3: 0.215 mmol / g.

[0581] [Example 19] Preparation of PEG-based aminohydrogel HG-2 containing free azides

[0582] [ka]

[0583] HG-1 (50 mg, 9.55 μmol) was placed in a 5 mL frit syringe, swollen in 3 mL of 1% DIPEA in NMP, and washed with 1% DIPEA in NMP (10 × 3 mL). Azide-PEG8-NHS ester (16.2 mg, 28.7 μmol) was dissolved in 1% DIPEA in NMP and added to a hydrogel. The syringe was shaken overnight at room temperature. After 2 hours and 45 minutes, shaking was stopped, and the hydrogel was washed with NMP (10 × 3 mL). HG-2 was suspended in 1.1 mL of NMP, transferred to an Eppendorf tube, and this suspension was used without further characterization.

[0584] [Example 20] Preparation of ubiquitin-linker conjugate mixture 20b

[0585] [ka]

[0586] Ubiquitin (50 mg, 5.86 μmol) from bovine erythrocytes was dissolved in 3.84 mL of 60 mM sodium phosphate buffer at pH 7.4, and a solution of 17f in DMSO (87.9 μL, 0.1 M, 8.79 μmol) was added. The reaction mixture was incubated at rt for 10 min, and 653 μL of 60 mM phosphate buffer at pH 7.4 containing 200 mM TriMED was added to the reaction mixture to promote the cleavage of the protecting group and the rearrangement of ester 20a to amide 20b. The reaction mixture was mixed and incubated at 25°C for 23 h 20 min. The concentrated reaction mixture was filtered through a PES membrane filter. To remove unconjugated linkers from the protein conjugate mixture, buffer exchange to PBST pH 7.4 was performed using an Akta pure system with GE HiTrap columns (3 in a row) at a flow rate of 1.5 ml / min. A 10.5 ml product fraction was collected, containing ubiquitin and ubiquitin linker conjugate 20b. A 6.5 ml product fraction was concentrated to 4 ml using an Amicon Ultra-15 centrifugal filtration unit (MWCO: 3 kDa). The concentrated solution, with an estimated protein concentration of 0.89 mM, was used directly for loading onto hydrogel HG-2.

[0587] [Example 21] Preparation of Ubiquitin-Linker-Hydrogel Conjugate 21

[0588] [ka]

[0589] Azide functionalization and conjugation of ubiquitin / ubiquitin-linker conjugate mixture 20b to cross-linked PEG hydrogel HG-2 were carried out by adding solution 20b to hydrogel HG-2. 75 μL of hydrogel suspension HG-2 was transferred to a 5 mL frit syringe and washed with water (10 × 2 mL) and PBST buffer pH 7.4 (10 × 2 mL). The hydrogel was incubated with 4 mL of protein / protein-linker conjugate solution 20b and shaken at room temperature for 18 h 30 min. The hydrogel was washed with PBST buffer pH 7.4 (10 × 2 mL), transferred to a 1.5 mL Eppendorf tube, and a 7.2 wt% solution in PBST buffer pH 7.4 was obtained.

[0590] [Example 22] In vitro release kinetics related to 21 A 40 μL suspension 21 (corresponding to approximately 0.74 mg of ubiquitin) was transferred to a sterile 1.5 mL Eppendorf tube. 1 mL of 60 mM sodium phosphate, 3 mM EDTA, and pH 7.4 buffer were added to the tube, mixed, and incubated in a water bath at 37°C. After different time intervals, small samples of the supernatant were analyzed by HPLC at 215 nm, and the released ubiquitin was quantified by comparison with a ubiquitin standard. The obtained values ​​were corrected for ubiquitin removal based on the previous sampling time. Release from 21:

[0591] [Table 2]

[0592] [Example 23] Synthesis of MTS-PEG12-NHS ester 16c

[0593] [ka]

[0594] 6-bromohexanoic acid (5.89 g, 30.2 mmol, 1.0 equivalent) and sodium methanethiosulfonate (4.05 g, 30.2 mmol, 1.0 equivalent) were dissolved in anhydrous DMF (47.1 mL) under an argon atmosphere and stirred at 80°C for 3 hours. After cooling to rt, the mixture was diluted with water (116 mL) and extracted with diethyl ether (3 × 233 mL). The combined organic layer was washed with brine (350 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to a volume of 40 mL. The solution was divided and added to two portions of cold n-heptane (2 × 1150 mL), and the mixture was cooled overnight to -18°C. The supernatant solution was decanted, and the precipitate was dissolved in diethyl ether (80 mL in total). This solution was divided and added to two portions of cold n-heptane (2 × 1000 mL), and the mixture was cooled to -18°C for 2 hours. The precipitate was collected by filtration and dried overnight under high vacuum to obtain intermediate 16a (5.62 g, 24.8 mmol, 82%). MS: m / z 249.02 = [M+Na] + (Calculated monoisotopic mass: [M] = 226.03)

[0595] DIPEA (2.76 mL, 15.9 mmol, 3.28 equivalents) was added to a stirring solution of 16a (1.15 g, 5.08 mmol, 1.05 equivalents) and PyBOP (2.64 g, 5.08 mmol, 1.05 equivalents) in anhydrous DCM (54.8 mL). After stirring for 30 minutes, 1-amino-3,6,9,12,15,18,21,24,27,30,33,36-dodecaoxanonateriacontan-39-acid (2.99 g, 4.84 mmol, 1.00 equivalent) was added, and the mixture was stirred at room temperature for another 30 minutes. Cold MTBE (55 mL) was added to the slightly yellow reaction mixture, and the mixture was cooled overnight to -20°C. No precipitate formed. All volatile substances were removed under vacuum, and the residue was dissolved in DCM. After adding TFA (1.2 mL), the solution was concentrated to 10 mL. Cold MTBE (55 mL) was added to the slightly yellow solution, and the mixture was cooled overnight to -20°C. The supernatant was decanted, and the yellow precipitate was washed with cold MTBE (55 mL). The white residue was then dried using a rota-vaporizer. After further purification by preparative RP-HPLC, intermediate 16b (2.81 g, 3.40 mmol, 70%) was obtained as a white solid. MS: m / z 826.35 = [M+H] + (Calculated monoisotopic mass: [M] = 825.39)

[0596] 16b (2.81 g, 3.40 mmol, 1.0 equivalent), HOSu (470 mg, 4.08 mmol, 1.2 equivalents), DMAP (41.6 mg, 0.34 mmol; 0.1 equivalent), and DCC (842 mg, 4.08 mmol, 1.2 equivalents) were dissolved in anhydrous DCM (32.6 mL), and the mixture was stirred at room temperature for 30 minutes. The precipitate was removed by filtration, and the solvent was evaporated from the filtrate. The residue was purified by preparative RP-HPLC to obtain pure water reagent 16c (1.74 g; 1.88 mmol, 55%). MS: m / z 923.45 = [M+H] + (Calculated monoisotopic mass: [M] = 922.40)

[0597] [Example 24] Synthesis of MTS-functionalized hydrogel HG-4

[0598] [ka]

[0599] PEG-hydrogel HG-3 (500 mg, amine content: 0.215 mmol / g, 1.0 equivalent), present as a suspension in a mixture of NMP / n-propylamine (99:1 v / v), was divided into equal aliquots between five 20 mL syringe reactors equipped with PE frit. Each hydrogel portion was successively washed with anhydrous NMP (10 × 8 mL) and NMP / DIPEA (99:1 v / v, 6 × 8 mL) until completely washed, after which all solvent was completely drained. Each hydrogel portion was then drawn into an aliquot of 5 mL of freshly prepared solution of 16c (309 mg, 3.0 equivalents) in anhydrous NMP (22 mL) and NMP / DIPEA (99:1 v / v, 3 mL). The syringe reactors were stirred at 500 rpm for 180 minutes. The reaction mixture was drained from all syringes, and each hydrogel portion was successively washed with anhydrous NMP (10 × 8 mL), water containing 0.1% AcOH and 0.01% Tween 20 (10 × 8 mL), and 20 mM succinate 0.01% Tween 20 pH 4.0 buffer (10 × 8 mL). The hydrogel aliquots were combined in a 50 mL Falcon tube with an additional 20 mM succinate 0.01% Tween 20 pH 4.0 buffer. After a short centrifugation, the suspension volume was adjusted to 24 mL by removing an appropriate volume of clear supernatant, obtaining a suspension of MTS-hydrogel HG-4 in 20 mM succinate 0.01% Tween 20 pH 4.0 buffer with a volume of 24 mL and a hydrogel content of 23.7 mg / mL. The MTS load of the dried hydrogel was determined to be 0.189 mmol / g.

[0600] [Example 25] HHC MET - Preparation of linker conjugate mixture 31

[0601] [ka]

[0602] 154 mL of HHC at pH 7.4 and 5.94 mg / mL in PBS MET HHC was used in this embodiment. MET The protein was concentrated using Amicon Ultra 15 and MWCO 3kDa (Merck), and the protein concentration was determined. 28.18 mL of HHC at a concentration of 30.3 mg / mL in PBS at pH 7.4 was measured. MET We prepared it.

[0603] HHC MET For the amount of protein solution, 9 g of linker reagent (1.5 mol equivalent (508 μL), corrected for NHS content and stored in 100 mM DMSO), was added. The reaction mixture was carefully mixed and incubated at ambient temperature for 5 minutes to obtain unmodified HHC. MET and protection HHC MET A mixture 31 of conjugates (e.g., monoconjugate, bisconjugate) was obtained.

[0604] The linker-conjugation reaction immediately follows a pH shift to approximately pH 4, followed by buffer exchange and HHC. MET / HHC MET - Excess linker species were removed from the linker conjugate mixture 31. HHC MET A buffer shift was achieved by adding 0.047 vol. equivalents (1.324 mL) of 0.4 M succinic acid, pH 3.0, to the solution volume (28.18 mL), and the solution was carefully mixed by rotation. Akta purifier 100 system equipped with a GE HiPrep column was used at a flow rate of 8.0 mL / min to exchange the buffer to 20 mM succinic acid, pH 4.0. Six operations were performed with an injection volume of approximately 5 mL per operation.

[0605] HHC MET / protected HHC MET - Protective HHC in linker conjugate mixture 31 METTo determine the content of -linker mono-, bis-, and tris-conjugates, PEGylation using 20 kDa PEGthiols and subsequent SE-HPLC analysis were performed.

[0606] 24.4 μL of unmodified HHC MET / protected HHC MET - Linker conjugate mixture 31 (c=10.89 mg / mL) MET The pH was adjusted to 5.5 by adding 0.154 vol. equivalents (3.8 μL) of 0.5 M succinic acid, pH 6.2, to a volume of 24.4 μL of solution. Next, to the resulting solution, 1.5 μL of 20 mM succinic acid, 100 mM EDTA, and 0.2% Tween 20, pH 5.5, were added to a volume of 28.2 μL. Unmodified HHC MET / protected HHC MET - The protein concentration of linker conjugate mixture 31 at pH 5.5 was adjusted to 4 mg / mL by mixing 15.8 μL of the solution with 20.2 μL of 20 mM succinic acid, 5 mM EDTA, 0.01% Tween 20, and pH 5.5. The PEGylation reaction was initiated by adding 4 μL of 15 mM PEG20-SH solution in water. After incubation at ambient temperature for 15 minutes, SE-HPLC was performed using an Agilent 1200 system connected to a Superdex 200 Increase 10 / 300 GL column with PBS-T, pH 7.4 as the mobile phase. Maleimide content was calculated by multiplying the peak area of ​​the conjugate by the number of bound PEG reagents. HHC MET / protected HHC MET -Regarding the linker conjugate mixture 31, a total maleimide content of 47.7% was determined.

[0607] After analysis and overnight storage at 4°C, 71.98 mL of HHC was found. MET / protected HHC MET- The pH of linker conjugate mixture 31 (c=10.89 mg / mL) was adjusted to pH 5.5 by adding 0.154 vol. equivalents (11.08 mL) of 0.5 M succinic acid pH 6.2. To the resulting solution, 1 / 19 vol. equivalents of 20 mM succinic acid, 100 mM EDTA, and 0.2% Tween 20 pH 5.5 (4.37 mL) were added, and the solution was mixed by rotation. The sample was filtered using one qpore Plastic vacuum filter (PVDF membrane) with a pore size of 0.22 μm.

[0608] [Example 26] temporary HHC MET Synthesis of Linker-Hydrogel Prodrug 32

[0609] [ka]

[0610] HHC MET / HHC MET -Conjugation of linker conjugate mixture 31 to reduced thiol-functionalized hydrogel HG-5, HHC MET / HHC MET - Linker conjugate mixture 31, HHC MET / HHC MET - This was performed by adding 1.75 mol. equivalents of thiol groups in hydrogel HG-5 to the determined total maleimide content of 47.7% (19.13 μmol) in linker conjugate mixture 31.

[0611] 8.5 mL of MTS-functionalized hydrogel HG-4 (nominal gel content of 23.7 mg / mL with 0.183 mmol / g thiol content) in 20 mM succinic acid, 0.01% Tween 20, pH 4.0 was transferred to a 20 mL syringe equipped with frit. The thiol-functionalized hydrogel was reduced by replacing the storage solution with 20 mL of 50 mM TCEP solution in PBS-T and incubating at ambient temperature for 15 minutes. Subsequently, the 50 mM TCEP solution was removed from the syringe, and the hydrogel was washed 10 times in the syringe with 20 mL of 20 mM succinic acid, 5 mM EDTA, 0.01% Tween 20, pH 5.5. It was then resuspended in approximately 6.7 mL of 20 mM succinic acid, 5 mM EDTA, 0.01% Tween 20, pH 5.5 to obtain HG-5.

[0612] 3.06 mL of hydrogel HG-5 was transferred to two 50 mL Falcon tubes. Then, 43.2 mL of HHC at pH 5.5 was added. MET / protected HHC MET -Linker conjugate mixture 31 (c=9.26 mg / mL) was added to each Falcon tube containing hydrogel HG-5. The resulting suspension was thoroughly mixed and incubated overnight at ambient temperature under gentle rotation to protect temporary HHC. MET - A linker hydrogel prodrug was obtained.

[0613] After overnight incubation, protective temporary HHC MET - The Linker Hydrogel Prodrug was transferred to a 20 mL syringe equipped with frit, and washed once in the syringe with 20 mL of 20 mM succinic acid, 5 mM EDTA, 0.01% Tween20, pH 5.5, and twice with 20 mL of 10 mM iodoacetamide in 30 mM sodium phosphate, 50 mM TriMED, 0.01% Tween20, pH 7.4. Protective temporary HHC MET- The linker hydrogel prodrug was incubated in a syringe at ambient temperature for 60 minutes with gentle rotation in 30 mM sodium phosphate, 10 mM iodoacetamide, 50 mM TriMED, 0.01% Tween20, pH 7.4 buffer. The hydrogel was then washed 10 times in the syringe with 20 mL of 30 mM sodium phosphate, 200 mM TriMED, 0.01% Tween20, pH 7.4. The solvent was discarded after each wash.

[0614] 20 mL of 30 mM sodium phosphate, 200 mM TriMED, 0.01% Tween20, pH 7.4 buffer was drawn into a syringe, and the resulting suspension was incubated at 25°C for 26 hours under gentle rotation to obtain transient HHC. MET -Linker Hydrogel Prodrug 32 was obtained. The hydrogel was washed 10 times in a syringe with 20 mL of 20 mM succinic acid, 8.5% α-α-D-trehalose, 1% carboxymethylcellulose, 0.01% Tween 20, pH 5.0 to obtain temporary HHC MET - We have successfully formulated Linker Hydrogel Prodrug 32.

[0615] [Example 27] In vitro release kinetics related to 32 25 μL of each suspension 32 (approximately 0.76 mg of HHC) MET The sample (equivalent to ) was transferred to 16 sterile 1.5 mL Eppendorf tubes. 60 mM sodium phosphate, 3 mM EDTA, 0.01% Tween-20, pH=7.40 were added to the tubes (filling to 0.5 mL or 1 mL). The samples were then mixed and incubated in a water bath at 37°C. After different time intervals, two samples were sacrificed, and the supernatant was analyzed for protein concentration using absorbance at 280 nm. Protein release over time was fitted using curve fitting software to obtain a half-life of 27 d.

[0616] [Example 28] temporary HHC MET- After subcutaneous (SC) and intramuscular (IM) injection of Linker Hydrogel Prodrug 32, and free HHC MET HHC in Wistar rats after intravenous (IV) and subcutaneous (SC) injection. MET Plasma pharmacokinetics temporary HHC MET - After subcutaneous and intramuscular administration of Linker Hydrogel Prodrug 32, or free HHC MET HHC in Wistar rats after subcutaneous or intravenous administration MET This study was conducted to investigate the plasma pharmacokinetics of the drug. Animals (n=3 per group) were given formulation 32 (10 mg / kg HHC). MET A single SC injection into the cervical region (equivalent to) or a single IM injection into the thigh muscle, or HHC MET Preparation (10 mg / kg of HHC MET The patient received either a single SC injection into the cervical region or an IV injection into the tail vein. At a selected time point, 200 μL of blood was collected in a Li-heparin tube and processed by centrifugation at 3,000 g at 4°C for 10 minutes to obtain plasma.

[0617] Using our proprietary sandwich ELISA mechanism, we analyze HHC in rat plasma. MET The concentration was determined. HHC MET To capture the target, human CTLA-4 (AA Ala37-Ser160)-Fc tag fusion protein (supplier AcroBiosystem, Newark, DE;USA, catalog number CT4-H5255) was coated onto ELISA plate wells and read out via rabbit anti-camelid VHH antibody conjugated with horseradish peroxidase (supplier Genscript, Piscataway, NJ, USA, catalog number A01861-200).

[0618] HHC in blank plasma MET The calibration standard was prepared as follows: Homogenized thawed Li-heparin Wistar rat plasma. Free HHC METThe formulations were added to blank plasma at concentrations ranging from 96.0 ng / mL to 3.00 ng / mL, with additional higher and lower anchor points. These solutions were used to create calibration curves. The calibration curves were analyzed via 4-parameter log-fitting and 1 / Y weighting. The calibration curves were verified against quality control standards prepared separately at 10, 40, and 80 ng / mL. HHC detected MET Plasma concentrations are shown in Table 3.

[0619] [Table 3]

[0620] For details, transient HHC MET - Linker Hydrogel Prodrug 32 (10 mg / kg HHC MET HHC levels 72 hours after intratissue (subcutaneous or intramuscular) injection (equivalent to) MET The concentration is temporary HHC MET - Linker Hydrogel Prodrug 32 (10 mg / kg HHC MET HHC 1 hour after intratissue (subcutaneous or intramuscular) injection (equivalent to) MET The concentration is at least 80%.

[0621] Abbreviation A. Alanine Acetyl Ala Alanine aq. Water-based Asp (Aspartate) Bzl (benzyl) Boc tert-butyloxycarbonyl CBZ Benzyloxycarbonyl DCC N,N'-Dicyclohexylcarbodiimide DBCO (Dibenzoazacyclooctin) DBU 1,8-Diazabicyclo(5.4.0)Undeca-7-Ene DCM Dichloromethane DIC N,N'-Diisopropylcarbodiimide DIPEA Diisopropylethylamine DMAP dimethylaminopyridine DMF Dimethylformamide DMSO (Dimethyl Sulfoxide) Dpr 2,3-diaminopropionic acid DS substitution degree DTNB 5,5'-Dithiobis-(2-nitrobenzoic acid) Eq equivalent EDC 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride EDTA (Ethylenediaminetetraacetic acid) F Phenylenine Fmoc Fluorenylmethyloxycarbonyl HA Hyaluronic Acid HG Hydrogel HEPES 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid HFIP 1,1,1,3,3,3-Hexafluoroisopropanol HOBt 1-hydroxybenzotriazole HOSu N-hydroxysuccinimide HPLC (High-Performance Liquid Chromatography) IAA Iodoacetamide ivDDe 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-3-methylbutyl K Lysine LC (Reset Chromatography) LCMS (Liquid Chromatography Mass Spectrometry) LLOQ Lower limit of quantification LPLC Low-Pressure Liquid Chromatography Lysine mAB monoclonal antibody MeOH methanol Me methyl MES 4-Morpholine Ethanesulfonic Acid MTBE methyl tert-butyl ether MTS Methanethiosulfonyl MWCO Fractionated Molecular Weight NHS N-hydroxysuccinimide NMP N-methyl-2-pyrrolidone OPA o-phthalaldehyde PBS (Phosphate-buffered saline) pH 7.4 Phosphate-buffered saline containing PBST Tween-20, pH 7.4 PE (Polyethylene) PEG polyethylene glycol PES Polyethersulfone Phe phenylalanine PTFE (Polytetrafluoroethylene) PVDF (Polyvinylidene Fluoride) PyBOP Benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate RP inverse phase rt / rt room temperature sat. saturation SE size exclusion Serine SPDP N-succinimidyl 3-(2-pyridyldithio)propionate Su succinimide tBu and t-Bu tert-butyl TCEP Tris(2-carboxyethyl)phosphine hydrochloride TFA (Trifluoroacetic Acid) THF (Tetrahydrofuran) TriMED N,N,N'-trimethylethylenediamine TSTU O-(N-succinimidyl)-N,N,N',N'-tetramethyluroniumtetrafluoroborate UPLC Ultra-High-Speed ​​Liquid Chromatography (Note) (Note 1) at least one -L 1 -L 2 - A conjugate comprising at least one -D moiety conjugated to at least one Z moiety via a moiety, or a pharmaceutically acceptable salt thereof, -L 1-The - portion is conjugated to the nitrogen of the primary or secondary amine of the -D portion, and -D and -L 1 The concatenation of - is reversible, and -L 2 -The part is conjugated to Z, Each -D is independently a primary or secondary amine-containing portion of drug DH. Each-L 2 - is an independent single bond or spacer portion, Each Z independently forms a polymer portion or C 8~24 It is alkyl, Each-L 1 - is independent of equation (I): [ka] [In the formula, The dashed line indicates the bonding of a primary or secondary amine of -D to the nitrogen. v is selected from the group consisting of 0 or 1. -X 1 - is -C(R 8 )(R 8a )-,-N(R 9 Selected from the group consisting of )- and -O-, =X 2 is =O and =N(R 10 Selected from the group consisting of, -X 3 - is selected from the group consisting of -O-, -S-, and -Se-, Each p is independently selected from the group consisting of 0 or 1, provided that at most one p is 0. -R 6 ,-R 6a ,-R 10 is -H, -C(R 11 )(R 11a )(R 11b Independently selected from the group consisting of ) and -T, -R 9 -C(R 11 )(R 11a )(R 11b Selected from the group consisting of ) and -T, -R 1 ,-R 1a ,-R2 ,-R 2a ,-R 3 ,-R 3a ,-R 4 ,-R 4a ,-R 5 ,-R 5a ,-R 7 ,-R 8 ,-R 8a ,-R 11 ,-R 11a and -R 11b -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkynnyls, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is one or more identical or different -R 13 In some cases, it is replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R) 14 )-,-S(O)2N(R 14 )-,-S(O)N(R14 )-, -S(O)2-, -S(O)-, -N(R 14 )S(O)2N(R 14a )-, -S-, -N(R 14 )-, -OC(OR 14 )(R 14a )-,-N(R 14 )C(O)N(R 14a )- and -OC(O)N(R 14 )- is optionally interrupted by one or more groups selected from the group consisting of, -R 12 ,-R 12a ,-R 12b -H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkynyls, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is one or more identical or different -R 13 In some cases, it is replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R) 14 )-,-S(O)2N(R 14 )-,-S(O)N(R 14 )-, -S(O)2-, -S(O)-, -N(R 14 )S(O)2N(R 14a )-, -S-, -N(R 14 )-, -OC(OR 14 )(R 14a )-,-N(R 14 )C(O)N(R 14a )- and -OC(O)N(R 14 )- is optionally interrupted by one or more groups selected from the group consisting of, Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Independently selected from the group consisting of cycloalkyls, 3-10 membered heterocyclyls, and 8-11 membered heterobicyclyls, each T is one or more identical or different -R groups.13 It is independently substituted depending on the case, -R 13 These are halogen, -CN, oxo, and -C(O)OR 15 , -OR 15 , -C(O)R 15 ,-C(O)N(R 15 )(R 15a ), -S(O)2N(R 15 )(R 15a ), -S(O)N(R 15 )(R 15a ), -S(O)2R 15 ,-S(O)R 15 , -N(R 15 )S(O)2N(R 15a )(R 15b ), -SR 15 , -N(R 15 )(R 15a ), -NO2, -OC(O)R 15 , -N(R 15 )C(O)R 15a , -N(R 15 )S(O)2R 15a , -N(R 15 )S(O)R 15a , -N(R 15 )C(O)OR 15a , -N(R 15 )C(O)N(R 15a )(R 15b ), -OC(O)N(R 15 )(R 15a ) and C 1~6 Selected from the group consisting of alkyl groups, C 1~6 Alkyl is optionally substituted with one or more halogens, which are the same or different. -R 14 ,-R 14a ,-R 15 ,-R 15a and -R 15b -H and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 Alkyl is optionally substituted with one or more halogens, which are the same or different. Depending on the case, -R 1 / -R 1a ,-R2 / -R 2a ,-R 3 / -R 3a ,-R 4 / -R 4a ,-R 5 / -R 5a or -R 8 / -R 8a One or more of the pairs, together with the atom they are bonded to, become C 3~10 Forming cycloalkyl, 3-10 membered heterocyclyl, or 8-11 membered heterobicyclyl, Depending on the case, -R 1 / -R 2 ,-R 1 / -R 8 ,-R 1 / -R 9 ,-R 2 / -R 9 or -R 2 / -R 10 One or more of the pairs, together with the atom to which they are bonded, form a ring-A-. -A- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl, Depending on the case, -R 3 / -R 6 ,-R 4 / -R 6 ,-R 5 / -R 6 ,-R 6 / -R 6a or -R 6 / -R 7 One or more of these pairs can form a ring -A'- together with the atom to which they are bonded. -A'- is selected from the group consisting of 3- to 10-membered heterocyclyls and 8- to 11-membered heterobicyclyls. This is the linker part, Each-L 1 - but at least one -L 2 Substituted with -, and possibly further substituted, provided that the hydrogens indicated by the asterisk in formula (I) are not replaced by substituents. Conjugate or a pharmaceutically acceptable salt thereof. (Note 2) -D is selected from the group consisting of small molecules, medium molecules, oligonucleotides, peptide nucleic acids, peptides, and protein drug moieties, as described in Appendix 1, or a pharmaceutically acceptable salt thereof. (Note 3) -D is selected from the group consisting of small molecules, medium molecules, peptides, and protein drug moieties, and is a conjugate as described in Appendix 1 or 2 or a pharmaceutically acceptable salt thereof. (Note 4) - A conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 3, wherein D is the protein drug portion. (Note 5) A conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 4, wherein v is 0. (Note 6) =X 2 A conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 5, wherein ≠ O. (Note 7) -X 3 A conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 6, wherein - is -O-. (Note 8) -R 3 and -R 3a A conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 7, wherein both atoms are -H. (Note 9) -L 1 -but, equation (Ia): [ka] [In the formula, The dashed line indicates the bonding of the primary or secondary amine of -D to the nitrogen as described in Appendix 1. -R 1 ,-R 1a ,-R 2 ,-R 2a ,-R 5 ,-R 5a,-R 6 ,-R 6a , -L 2 - and Z are used as defined in Appendix 1. The conjugate described in any one of the appendices 1 to 8, or a pharmaceutically acceptable salt thereof. (Note 10) -R 6 and -R 6a A conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 9, wherein both atoms are -H. (Note 11) -R 1 and -R 1a A conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 10, wherein both atoms are -H. (Note 12) -L 1 -but, formula (Id): [ka] [In the formula, A dashed line marked with an asterisk indicates the bonding of a primary or secondary amine of -D to the nitrogen as described in Appendix 1, and an unmarked dashed line indicates -L 2 - indicates binding to -L 2 - and Z are used as defined in Appendix 1. The conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 11. (Note 13) A conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 12, wherein Z is the polymer portion. (Note 14) A conjugate or a pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 13, wherein Z is a water-insoluble polymer portion. (Note 15) Z is 2-methacryloyloxyethyl phosphorylcholine, poly(acrylic acid), poly(acrylate), poly(acrylamide), poly(alkyloxy)polymer, poly(amide), poly(amideamine), poly(amino acid), poly(anhydride), poly(aspartamide), poly(butyric acid), poly(glycolic acid), polybutylene terephthalate, poly(caprolactone), poly(carbonate), poly(cyanoacrylate), poly(dimethylacrylamide), poly(ester), poly(ethylene), poly(ethylene glycol), poly(ethylene oxide), poly(ethyl phosphate), poly(ethyl oxazoline), poly(glycolic acid), poly(hydroxyethyl acrylate), poly(hydroxyethyl oxazoline), poly(hydroxymethacrylate), poly(hydroxypropyl methacrylamide), poly(hydroxypropyl methacrylate), poly(hydroxypropyl oxazoline), poly(iminocarbonate), poly(milk A water-insoluble polymer portion comprising a polymer selected from the group consisting of acids, poly(lactic acid-coglycolic acid), poly(methacrylamide), poly(methacrylate), poly(methyloxazoline), poly(organophosphazene), poly(orthoester), poly(oxazoline), poly(propylene glycol), poly(siloxane), poly(urethane), poly(vinyl alcohol), poly(vinylamine), poly(vinyl methyl ether), poly(vinylpyrrolidone), silicone, cellulose, carbomethylcellulose, hydroxypropylmethylcellulose, chitin, chitosan, dextran, dextrin, gelatin, hyaluronic acid and derivatives, functionalized hyaluronic acid, mannan, pectin, rhamnogalacturonan, starch, hydroxyalkyl starch, hydroxyethyl starch and other carbohydrate-based polymers, xylan, and copolymers thereof, the conjugate described in any one of the appendices 1 to 14 or a pharmaceutically acceptable salt thereof. (Note 16) A conjugate or a pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 13, wherein Z is a water-soluble polymer portion. (Note 17) -L 2- is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-,-S(O)2N(R y1 )-,-S(O)N(R y1 )-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-,-N(R y1 )C(O)N(R y1a )-,-OC(O)N(R y1 )-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 The spacer portion is selected from the group consisting of alkynnyls, -T'-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is one or more identical or different -R y2 In some cases, it is replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-,-S(O)2N(R y3 )-,-S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-,-N(R y3 )C(O)N(R y3a )- and -OC(O)N(R y3 )- is optionally interrupted by one or more groups selected from the group consisting of, -R y1 and -R y1a However, -H, -T', C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Independently selected from the group consisting of alkynnyls, -T', C 1~50Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is one or more identical or different -R y2 In some cases, it is replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkinyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 )-,-S(O)2N(R y4 )-,-S(O)N(R y4 )-, -S(O)2-, -S(O)-, -N(R y4 )S(O)2N(R y4a )-, -S-, -N(R y4 )-, -OC(OR y4 )(R y4a )-,-N(R y4 )C(O)N(R y4a )- and -OC(O)N(R y4 )- is optionally interrupted by one or more groups selected from the group consisting of, Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl, each T' is one or more identical or different -R y2 It is independently substituted depending on the case, Each-R y2 However, halogen, -CN, oxo (=O), -COOR y5 , -OR y5 , -C(O)R y5 ,-C(O)N(R y5 R y5a ), -S(O)2N(R y5 R y5a ), -S(O)N(R y5 R y5a ), -S(O)2R y5 ,-S(O)R y5 , -N(R y5 )S(O)2N(R y5a R y5b ), -SR y5 , -N(Ry5 R y5a ), -NO2, -OC(O)R y5 , -N(R y5 )C(O)R y5a , -N(R y5 )S(O)2R y5a , -N(R y5 )S(O)R y5a , -N(R y5 )C(O)OR y5a , -N(R y5 )C(O)N(R y5a R y5b ), -OC(O)N(R y5 R y5a ) and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 The alkyl group is optionally substituted with one or more halogens, which are either the same or different. Each-R y3 ,-R y3a ,-R y4 ,-R y4a ,-R y5 ,-R y5a and -R y5b However, -H and C 1~6 Independently selected from the group consisting of alkyls, C 1~6 The alkyl group is optionally substituted with one or more halogens, which are the same or different. The conjugate or pharmaceutically acceptable salt thereof as described in any one of the appendices 1 to 16. (Note 18) -L 2 - A conjugate or pharmaceutically acceptable salt thereof as described in any one of Appendix 1 to 17, having a molecular weight in the range of 14 g / mol to 750 g / mol. (Note 19) A reagent containing an -L*- portion, wherein the -L*- portion is conjugated to -Q. -Q is -OH or -LG, and -LG is the leaving group portion. -L*- is equation (II): [ka] [In the ceremony The dashed line indicates a connection to -Q. v is selected from the group consisting of 0 or 1. -X 1 - is -C(R 8 )(R 8a )-,-N(R 9 Selected from the group consisting of )- and -O-, =X 2 is =O and =N(R 10 Selected from the group consisting of, -X 3 - is selected from the group consisting of -O-, -S-, and -Se-, Each p is independently selected from the group consisting of 0 or 1, provided that at most one p is 0. -R 6 -PG and -R 6a is -H, -C(R 11 )(R 11a )(R 11b ), selected from the group consisting of -T and -PG, or -R 6 and -R 6a -C(R 11 )(R 11a )(R 11b Independently selected from the group consisting of ) and -T, -R A and -R B It is independently selected from the group consisting of -H and -PG, except for -R A or -R B The condition is that one or less of them can be -H, -PG is the amine protecting group moiety, -R 9 -C(R 11 )(R 11a )(R 11b Selected from the group consisting of ) and -T, -R 10 is -H, -C(R 11 )(R 11a )(R 11b Selected from the group consisting of ) and -T, -R 1 ,-R 1a ,-R 2 ,-R 2a ,-R3 ,-R 3a ,-R 4 ,-R 4a ,-R 5 ,-R 5a ,-R 7 ,-R 8 ,-R 8a ,-R 11 ,-R 11a and -R 11b -H, halogen, -CN, -C(O)OR 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)2R 12 ,-S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkynnyls, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkinyl is one or more identical or different -R 13 In some cases, it is replaced by C 1~6 Alkyl, C 2~6 Alke...

Claims

1. -L 1 -L 2 A conjugate comprising a -D portion conjugated to a Z portion via a - portion, or a pharmaceutically acceptable salt thereof, wherein the -L 1 -The portion is conjugated to the nitrogen of the primary or secondary amine of the -D portion, and -D and -L 1 - The connection is reversible, and -L 2 -The part is conjugated to Z, -D is the primary or secondary amine-containing portion of the protein drug DH. -L 2 - indicates a single bond or spacer portion. Z is the polymer part or C 8～24 It is alkyl, -L 1 - is equation (Ia): 【Chemistry 1】 [In the formula, The dashed line indicates the bonding of a primary or secondary amine of -D to the nitrogen. -R 6 and -R 6a is independently selected from the group consisting of -H, -C(R 11 )(R 11a )(R 11b ), and -T; -R 1 ,-R 1a ,-R 2 ,-R 2a ,-R 5 ,-R 5a ,-R 11 ,-R 11a and -R 11b -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 ,-C(O)N(R 12 )(R 12a ), -S(O) 2 R 12 ,-S(O)R 12 , -SR 12 , -NO 2 , -T, and C 1～6 Independently selected from the group consisting of alkyls, C 1～6 Alkyl is one or more identical or different -R 13 In some cases, it is replaced by C 1～6 Alkyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R) 14 )-,-S(O) 2 N(R 14 )-,-S(O)N(R 14 )-,-S(O) 2 -, -S(O)-, -S-, and -OC(OR 14 )(R 14a )- is optionally interrupted by one or more groups selected from the group consisting of, -R 12 , and -R 12a -H, -T, C 1～6 Independently selected from the group consisting of alkyls, -T, C 1～6 Alkyl is one or more identical or different -R 13 In some cases, it is replaced by C 1～6 Alkyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R) 14 )-,-S(O) 2 N(R 14 )-,-S(O)N(R 14 )-,-S(O) 2 -, -S(O)-, -S-, and -OC(OR 14 )(R 14a )- is optionally interrupted by one or more groups selected from the group consisting of, Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3～10 Independently selected from the group consisting of cycloalkyls and 3- to 10-membered heterocyclines, each T is one or more identical or different -R groups. 13 It is independently substituted depending on the case, -R 13 These are halogen, -CN, oxo, and -C(O)OR 15 , -OR 15 , -C(O)R 15 ,-C(O)N(R 15 )(R 15a ), -S(O) 2 R 15 ,-S(O)R 15 , -SR 15 , -NO 2 -OC(O)R 15 , -N(R 15 )C(O)R 15a and C 1～6 Selected from the group consisting of alkyl groups, C 1～6 Alkyl is optionally substituted with one or more halogens, which are the same or different. -R 14 ,-R 14a ,-R 15 , and -R 15a -H and C 1～6 Independently selected from the group consisting of alkyls, C 1～6 Alkyl is optionally substituted with one or more halogens, which are the same or different. Depending on the case, -R 1 / -R 1a ,-R 2 / -R 2a , or -R 5 / -R 5a One or more of the pairs, together with the atom they are bonded to, become C 3～10 Forming cycloalkyl or 3-10 membered heterocyclines, Depending on the case, -R 1 / -R 2 The pairs, together with the atoms they are bonded to, form a ring-A-. -A- is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3～10 cycloalkyl, and 3- to 10-membered heterocyclyl, Depending on the case, -R 5 / -R 6 , or -R 6 / -R 6a One or more of these pairs can form a ring -A'- together with the atom to which they are bonded. -A'- is selected from the group consisting of 3- to 10-membered heterocyclines. This is the linker part, -R of formula (I-a) 1 、-R 1a 、-R 2 、-R 2a 、-R 5 、-R 5a 、-R 6 、or -R 6a one of them is -H, and the -H is replaced by -L 2 ​ Conjugate or a pharmaceutically acceptable salt thereof.

2. -R in equation (Ia) 1 ,-R 1a ,-R 2 , or -R 2a One of them is -H, and this H is -L 2 - The conjugate or pharmaceutically acceptable salt thereof according to claim 1, which is replaced by -.

3. -R in equation (Ia) 2 , or -R 2a One of them is -H, and this H is -L 2 -A conjugate or pharmaceutically acceptable salt thereof according to claim 1 or 2, which is replaced by -.

4. -R 6 and -R 6a A conjugate or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein both are -H.

5. -R 1 and -R 1a A conjugate or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein both are -H.

6. -L 1 -but, formula (Id): 【Chemistry 2】 [In the formula, A dashed line marked with an asterisk indicates the bonding of a primary or secondary amine to the nitrogen of -D as described in claim 1, and an unmarked dashed line indicates -L 2 - indicates binding to -L 2 - and Z are used as defined in claim 1. A conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5.

7. A conjugate or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein Z is a polymer portion.

8. A conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Z is a water-insoluble polymer portion.

9. Z is poly(2-methacryloyl-oxyethyl phosphorylcholine), poly(acrylic acid), poly(acrylate), poly(acrylamide), poly(alkyloxy)polymer, poly(amide), poly(amideamine), poly(amino acid), poly(anhydride), poly(aspartamide), poly(butyric acid), poly(glycolic acid), polybutylene terephthalate, poly(caprolactone), poly(carbonate), poly(cyanoacrylate), poly(dimethylacrylamide), poly(ester), poly(ethylene), poly(ethylene glycol), poly(ethylene oxide), poly(ethyl phosphate), poly(ethyl oxazoline), poly(glycolic acid), poly(hydroxyethyl acrylate), poly(hydroxyethyl oxazoline), poly(hydroxymethacrylate), poly(hydroxypropyl methacrylamide), poly(hydroxypropyl methacrylate), poly(hydroxypropyl oxazoline), poly(iminocarbonate), poly A water-insoluble polymer portion comprising a polymer selected from the group consisting of (lactic acid), poly(lactic acid-coglycolic acid), poly(methacrylamide), poly(methacrylate), poly(methyloxazoline), poly(organophosphazene), poly(orthoester), poly(oxazoline), poly(propylene glycol), poly(siloxane), poly(urethane), poly(vinyl alcohol), poly(vinylamine), poly(vinyl methyl ether), poly(vinylpyrrolidone), silicone, cellulose, carbomethylcellulose, hydroxypropylmethylcellulose, chitin, chitosan, dextran, dextrin, gelatin, hyaluronic acid and derivatives, functionalized hyaluronic acid, mannan, pectin, rhamnogalacturonan, starch, hydroxyalkyl starch, hydroxyethyl starch and other carbohydrate-based polymers, xylan, and copolymers thereof, the conjugate according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof.

10. A conjugate or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Z is a water-soluble polymer portion.

11. -L 2 - is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-,-S(O) 2 N(R y1 )-,-S(O)N(R y1 )-,-S(O) 2 -, -S(O)-, -N(R y1 )S(O) 2 N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-,-N(R y1 )C(O)N(R y1a )-,-OC(O)N(R y1 )-, C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 The spacer portion is selected from the group consisting of alkynnyls, -T'-, C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 Alkinyl is one or more identical or different -R y2 In some cases, it is replaced by C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 Alkinyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-,-S(O) 2 N(R y3 )-,-S(O)N(R y3 )-,-S(O) 2 -, -S(O)-, -N(R y3 )S(O) 2 N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-,-N(R y3 )C(O)N(R y3a )- and -OC(O)N(R y3 )- is optionally interrupted by one or more groups selected from the group consisting of, -R y1 and -R y1a However, -H, -T', C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 Independently selected from the group consisting of alkynnyls, -T', C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 Alkinyl is one or more identical or different -R y2 In some cases, it is replaced by C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 Alkinyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 )-,-S(O) 2 N(R y4 )-,-S(O)N(R y4 )-,-S(O) 2 -, -S(O)-, -N(R y4 )S(O) 2 N(R y4a )-, -S-, -N(R y4 )-, -OC(OR y4 )(R y4a )-,-N(R y4 )C(O)N(R y4a )- and -OC(O)N(R y4 )- is optionally interrupted by one or more groups selected from the group consisting of, Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3～10 Independently selected from the group consisting of cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl, each T' is either identical or different from one or more -R y2 It is independently substituted depending on the case, Each-R y2 However, halogen, -CN, oxo (=O), -COOR y5 , -OR y5 , -C(O)R y5 ,-C(O)N(R y5 R y5a ), -S(O) 2 N(R y5 R y5a ), -S(O)N(R y5 R y5a ), -S(O) 2 R y5 ,-S(O)R y5 , -N(R y5 )S(O) 2 N(R y5a R y5b ), -SR y5 , -N(R y5 R y5a ), -NO 2 -OC(O)R y5 , -N(R y5 )C(O)R y5a , -N(R y5 )S(O) 2 R y5a , -N(R y5 )S(O)R y5a , -N(R y5 )C(O)OR y5a , -N(R y5 )C(O)N(R y5a R y5b ), -OC(O)N(R y5 R y5a ) and C 1～6 Independently selected from the group consisting of alkyls, C 1～6 The alkyl group is optionally substituted with one or more halogens, which are either the same or different. Each-R y3 ,-R y3a ,-R y4 ,-R y4a ,-R y5 ,-R y5a and -R y5b However, -H and C 1～6 Independently selected from the group consisting of alkyls, C 1～6 The alkyl group is optionally substituted with one or more halogens, which are the same or different. A conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 10.

12. -L 2 - The conjugate or pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein - has a molecular weight in the range of 14 g / mol to 750 g / mol.

13. A pharmaceutical composition comprising a conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12.

14. Use of a conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12 in the manufacture of a pharmaceutical product.

15. A pharmaceutical composition for use in a method of treating a disease treatable with DH, comprising the conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12.

Images