Ligand-drug-composite containing single molecular weight polysarcosine

Abstract

To provide a ligand-drug-composite (LDC) containing a single molecular weight homopolymer particularly single molecular weight polysarcosine.SOLUTION: A ligand-drug-composite compound (LDC) has the following formula (XV). L denotes an orthogonal connector; HPSMW is generated from a covalent bond to the orthogonal connector L of a single molecular weight homopolymer having a specific, D denotes a cytotoxic drug; X denotes an arbitrary cleavable portion for releasing D; Z denotes arbitrary spacer; and a is 1 or more, b is 1 or more, and m is 1 or more.SELECTED DRAWING: None

Description

Detailed Description of the Invention

[0001] The present invention relates to single molecular weight homopolymers, methods for preparing such homopolymers, and in particular its use in composite technology.

[0002] The present invention also provides ligases comprising single molecular weight homopolymers, particularly single molecular weight polysarcosines. This relates to Ligand-Drug-Conjugates (LDCs).

[0003] A Ligand-Drug-Conjugate (LDC) comprises at least one Ligand unit, The Ligand unit is linked to at least one therapeutic molecule, diagnostic molecule or A polypeptide or a marker molecule (hereinafter referred to as a drug or D) covalently linked to the marker molecule. This synthetic linker is a protein that connects the Ligand unit and the Drug unit. It may contain one or several bivalent arms for the purpose of The linker may also be selected from a group consisting of a group of molecules that are suitable for storage stability, plasma stability, or pharmacokinetic properties, etc. The LDC may have any monovalent moiety that can improve the LDC performance of a protein or The polypeptide is usually a targeting unit, but may have its own therapeutic properties. The ligand unit of the complex is an antibody or an antibody fragment, and the cytotoxic or chemotherapeutic agent In connection with the subject matter, the term "antibody-drug-conjugate (ADC)" is commonly used.

[0004] The design of ADCs (antibody-drug-conjugates) depends on a number of diverse factors: (i) on the ligand (ii) the nature, number, overall hydrophobicity and location of the synthetic linkers used for attachment at the The nature and mechanism of action; (iii) Structural elements involved in drug release after cellular internalization and during intracellular trafficking; (iv) Considerations of the properties of monoclonal antibodies (mAbs) and selected antigen targets. This includes investigations of... Recent methodologies have addressed some of the drawbacks of available ADCs, such as heterogeneous drug payloads (ADC subspecies with different pharmacological properties), limited mAb-linker stability or drug-linker stability, and suboptimal pharmacokinetic properties (Beck et al. Nat. Rev. Drug. Disco., 2017, 16(5), 315-337).

[0005] Another important factor to consider when designing the conjugate is the drug ratio (or drug-antibody-ratio (DAR) for ADCs), which is the average number of drug units attached to the antibody. As a result of recent research findings, the current trend in the ADC field is to generate homogeneous conjugates with low to medium DARs (usually 2-4) and bring them to the clinic. Nevertheless, more recently, new linker-drug technologies have emerged with the aim of overcoming the drawbacks of highly loaded ADCs (forming aggregates and thus complicating conjugate formulations, unfavorable pharmacokinetic properties and tendencies). Such technologies have the potential to bring to the clinic next-generation ADCs with improved efficacy, improved pharmacokinetic properties, and improved therapeutic indices, and can target tumors with low target expression, slow internalization or inefficient intracellular processing. To achieve such high payload loading without sacrificing pharmacokinetic properties and formulation stability, new linker-drug design approaches have been developed aimed at masking the putative hydrophobicity of the cytotoxic payload. ... ... ... ... It is necessary to do so.

[0006] In WO2014 / 093394A1, protein-polymer-drug conjugates showing high drug loading and strong binding to the target antigen have been reported. This conjugate contains a biodegradable and biocompatible poly-[1-hydroxymethylethylene hydroxymethyl formal] polymer entity, which enables a complex of about 12 to 25 cytotoxic molecules per mAb, having good pharmacokinetic properties. The main drawbacks of this approach are (i) the polydispersity of the linker, (ii) the non-uniform number of cytotoxic molecules per polymer arm, and (iii) the non-uniform number of grafted polymer arms per mAb, resulting in extreme polydispersity of the final conjugate.

[0007] In WO2015 / 057699A2 and WO2016 / 059377A1, formulation of 8 - 36 drug-loaded ADCs has been reported by including a polyethylene glycol (PEG) moiety orthogonal to the linker design. PEG is well-known for improving the hydrophilicity, stability and circulation time of small drugs, proteins, bioconjugates and nanoparticles due to its hydrophilicity, biocompatibility and high hydration shell. However, PEG is not free from drawbacks such as non-biodegradability, possible complement activation leading to hypersensitivity and unclear pharmacokinetics due to anti-PEG antibodies expressed by some healthy individuals.

[0008] (i) High drug loading while maintaining favorable pharmacokinetic properties and stability, (ii) complete homogeneity of the conjugate at the drug-linker level (chemically monodisperse drug-linker) and at the conjugate level (homogeneously loaded conjugate), and (iii) as a hydrophobic masking moiety A ligand-drug complex that acts on and combines with a biodegradable hydrophilic homopolymer is required.

[0009] Polysarcosine (poly-N-methylglycine or PSAR) is an alternative to PEG and can be used to design novel protein complexes with improved properties. PSAR is a highly hydrophilic, biodegradable, non-immunogenic, water-soluble polymer and has been used in several drug delivery systems or diagnostics. So far, PSAR has only been available in a polydisperse form since it is obtained via a condensative ring-opening polymerization reaction of sarcosine N-carboxyanhydride (NCA) or sarcosine N-thiocarboxyanhydride (NTA). In certain applications that require the use of shorter homopolymer compounds with a defined length (a single and unique molecular weight) and thus absolute homogeneity, these polydisperse PSARs cannot be used. The use of discrete monodisperse PSARs for macromolecule modification is a prerequisite for developing complexes with absolute chemical homogeneity. Such homogeneous complexes have the advantage of sharing exactly the same pharmacological properties (pharmacokinetics and efficacy), making it easier to determine the properties, allowing for greater control of the reproducibility of the manufacturing process, and meeting the increasingly stringent regulatory requirements for biocomplexes. According to the present invention, a defined chain length is obtained using a stepwise resin-based submonomer approach. Although relatively well-defined with an acceptable dispersity (a Gaussian distribution of molecular weights with a polydispersity index > 1),

[0010] The use of discrete monodisperse PSARs for macromolecule modification is a prerequisite for developing complexes with absolute chemical homogeneity. Such homogeneous complexes have the advantage of sharing exactly the same pharmacological properties (pharmacokinetics and efficacy), making it easier to determine the properties, allowing for greater control of the reproducibility of the manufacturing process, and meeting the increasingly stringent regulatory requirements for biocomplexes.

[0011] According to the present invention, a defined chain length is obtained using a stepwise resin-based submonomer approach. ​​​​​​A discrete monodisperse PSAR homopolymer was obtained. This method is inexpensive and can be easily extended, giving a final product with an acceptable yield and excellent monomer purity These monodisperse PSAR homopolymers are used in protein binding technology to provide ligand-drug conjugates (LDCs) with improved drug loading capacity, pharmacokinetics and therapeutic effects.

[0012] Accordingly, the present invention provides a single molecular weight monofunctional homopolymer that meets the above requirements for use in conjugate technology, particularly LDCs.

[0013] This homopolymer has the following formula (I):

[0014] [Chemical formula]

[0015] In the formula, R1 and R2 are different, one of R1 and R2 is H or an inert group, and the other of R1 and R2 is a functionalized reactive group which is reactive to covalently bond with a bondable group under reaction conditions where the inert group does not react, Z1 and Z2 are the same or different and are any spacer, n is 1 or more and k is 2 or more.

[0016] Before explaining the present invention in detail, the definitions of the terms used in this specification are shown below.

[0017] [Definition] According to the present invention, any compound such as reactants, products, monomers, homopolymers, units, etc. is an acid addition salt, a base addition salt, a metal salt, ammonium and alkylated ammonium ​It may be in the form of a salt, including a salt. Such salts are well-known to those skilled in the art. For the intended use of the homopolymers of the present invention, they are preferably pharmaceutically acceptable in the form of salts.

[0018] A single molecular weight homopolymer has the same properties but is centered around the size distribution and average molecular weight as opposed to a mixture of homopolymers having a molecular weight distribution, and refers to a homopolymer having a unique and specific molecular weight. A single molecular weight homopolymer can be defined by one absolute molecular formula having an absolute number of atoms.

[0019] In contrast to the polydisperse homopolymers traditionally obtained by a one-pot polymerization process and having a PDI > 1, a single molecular weight homopolymer can also be called "monodisperse" having a polydispersity index (PDI: polydispersity index) equal to 1. Despite the fact that the term "monodisperse" does not accurately reflect the production procedure of the product, it is generally recognized herein that both the terms "monodisperse" and "discrete" which define a homopolymer having a unique and absolute molecular weight, molecular formula and molecular structure are interchangeable. An inert group or capping group refers to any chemical non-reactive group that terminates one end of the homopolymer, and said group is non-reactive when compared to a functionalized reactive group that terminates the other end of the homopolymer under certain reaction conditions. The resulting homopolymer

[0020] is, depending on the view, end-capped by this inert group and is specifically not intended to be covalently bonded when used in LDC technology. In one embodiment is end-capped by this inert group and is specifically not intended to be covalently bonded when used in LDC technology. In one embodiment ​​​In this case, the group only needs to be inactivated after covalent bonding to one end of the homopolymer. That's fine.

[0021] A non-exhaustive list of inert groups includes acyl groups, especially acetyl groups, amide groups, alkyl groups, especially C 1-20 alkyl groups, alkyl ether groups, alkyl ester groups, alkyl ortho ester groups, alkenyl groups, alkynyl groups, aryl groups, aryl ester groups, tertiary am ine groups, hydroxyl groups, aldehyde groups. The inert group may also be selected from the same list of groups that define the functionalized reactive group (see the definition of the following functionalized reactive group). (See the definition of the following functionalized reactive groups.)

[0022] A functionalized reactive group is any chemical moiety that is reactive to covalently bond with a bondable group Yes, it is reactive when compared to an inert group under certain reaction conditions. In particular, it can bind to the following groups: carboxylic acid; primary amine; secondary amine; tertiary amine; hydroxy l; halogen; N-hydroxysuccinimide ester, perfluoroester, nitro phenyl ester, aza-benzotriazole and benzotriazole activated ester, activated esters such as acyl urea; alkynyl; alkenyl; azide; isocyanate; i sothiocyanate; aldehyde; maleimide, halomaleimide, haloacetyl, pyridyl thiol-reactive moieties such as disulfide; thiol; acrylate; mesylate; tosylate tos; triflate, hydroxylamine; chlorosulfonyl; boronic acid - B(OR')2 derivatives (where R' is hydrogen or an alkyl group).

[0023] A non-exhaustive list of functionalized reactive groups includes the following: carboxylic acid; primary amine; Secondary amine; tertiary amine; hydroxyl; halogen; N-hydroxysuccinimide ester ter, perfluoroester, nitrophenyl ester, aza-benzotriazole and benzotriazole-activated ester, activated esters such as acylurea; alkynyl; al kenyl; azide; isocyanate; isothiocyanate; aldehyde; maleimide, halo maleimide, haloacetyl, pyridyldisulfide and other thiol-reactive moieties; thiol; acrylate; mesylate; tosylate; triflate, hydroxylamine; chloros ulfonyl; boronic acid -B(OR’)2 derivative (wherein R’ is hydrogen or an alkyl group ).

[0024] Regarding the terms "inert" and "functionalized reactive" for inert groups and functionalized reactive groups, it should be noted that they are interdependent. This means that under the predetermined reaction conditions of the homopolymers of the present invention defined in any one of formulas (I), (II) and (I II), the inert groups do not react, and the functionalized reactive groups react to form a covalent bond with the reactants. Therefore, the inert groups and the functionalized reactive groups in any one of the homopolymers of formulas (I), (II) and (III) are different, but they may be entirely selected from the same list of groups.

[0025] The term "group" in the functionalized reactive group or inert group according to the present invention should be understood as a group that can form a covalent bond with the reactant or does not show any function other than being inert under the predetermined reaction conditions.

[0026] Alkyl is used alone or as part of an alkyl ether or alkyl ester ​​​​and, for example, having 1 to 20 carbon atoms, preferably 1 to 12, more preferably 1 to 6 , and in particular having 1 to 4, a saturated straight-chain or branched hydrocarbon group.

[0027] Alkenyl and alkynyl have 2 to 20 carbon atoms, preferably 2 to 12, more preferably 2 to 6, and in particular 2 to 4, a straight-chain or branched hydrocarbon group that is at least partially unsaturated.

[0028] Aryl is used alone or as part of an aryl ester and has, for example, 6 to 14 ring carbon atoms, preferably 6 to 10, and in particular 6, an aromatic group having one or more rings.

[0029] Alkylene is used alone or as part of an alkylene glycol and has, for example, 1 to 20 carbon atoms, preferably 1 to 12, more preferably 1 to 6, and in particular 1 to 4 , a divalent saturated straight-chain or branched hydrocarbon group.

[0030] Arylene refers to the divalent aryl group defined above.

[0031] Heteroalkyl consists of 1 to 20 or 1 to 10 carbon atoms and 1 to 10, preferably 1 to 3, heteroatoms selected from the group consisting of O, N, Si, and S, a straight-chain or branched hydrocarbon chain, where the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatoms may optionally be quaternized. The heteroatoms O, N, and S are located at any internal position of the heteroalkyl group or at the position where the alkyl group is attached to the rest of the molecule.

[0032] Heteroalkylene refers to the divalent heteroalkyl defined above. The heteroalkylene group with respect to the heteroatom, may also occupy either or both of the ends of the chain.

[0033] A C3-C8 carbocyclic ring refers to a monocyclic or bicyclic, saturated or unsaturated, non-aromatic carbocyclic ring having 3, 4, 5, 6, 7 or 8 carbon atoms, which is monovalent and substituted or unsubstituted.

[0034] C3-C8 carbocylo refers to the divalent C3-C8 carbocyclic ring defined above.

[0035] A C3-C8 heterocyclic ring refers to a monovalent substituted or unsubstituted aromatic or non-aromatic monocyclic or bicyclic ring system having 3 to 8 carbon atoms (also called ring members) and 1 to 4 heteroatom ring members independently selected from N, O, P or S. One or more N, C or S atoms in the heterocyclic ring may be oxidized. The ring containing heteroatoms may be aromatic or non-aromatic. Unless otherwise specified, the heterocyclic ring is attached to its pendant group by any heteroatom or carbon atom that results in a stable structure.

[0036] C3-C8 heterocylo refers to the divalent C3-C8 heterocyclic ring defined above.

[0037] Furthermore, the terms alkyl, alkenyl, alkynyl, aryl, alkylene, arylene, he teroalkyl, heteroalkylene, C3-C8 carbocyclic ring, C3-C8 carbocylo, C3- C8 heterocyclic ring, C3-C8 heterocylo also include -X, -R’, -O - , -OR’, =O, -SR’, -S - , -NR’2, -NR’3, =NR’, -CX3, -CN, -O CN, -SCN, -N=C=O, -NCS, -NO, -NO2, =N2, -N3, -NR C(=O)R', -C(=O)R', -C(=O)NR'2, -SO3 - , -SO3H, -S(=O)2R', -OS(=O)2OR', -S(=O)2NR', -S(=O)R ', -OP(=O)(OR')2, -P(=O)(OR')2, -PO3 - , -POH 2, -C(=O)R', -C(=O)X, -C(=S)R', -CO2R', -CO2, -C(=S)OR', C(=O)SR', C(=S)SR', C(=O)NR'2, C( and C(=NR')NR'2. , an optionally substituted group, where each X is independently a halogen: -F, -CI, -B r, or -I, and each R' is independently -H, -C 20 Alkyl, -C6-C 20 Aryl, or -C3-C 14 It is a heterocycle.

[0038] An acyl group refers to a -CO-alkyl group, where alkyl is defined above.

[0039] Monofunctional homopolymers consist of a single type of monomer (e.g., N for polysarcosine). -methylglycine monomers), said single type of monomer being functionalized as defined above. One end has a reactive group and the other has H or an inert group as defined above. and an end thereof.

[0040] Support for solid-phase peptide synthesis (SPPS) sis) refers to supports commonly used in the well-known process SPPS, In the method, the peptide and insoluble polymer immobilized on the carrier are deprotected, washed, and then coupled. Assembled via repeated cycles of washing, through successive addition of Fmoc-protected amino acids or Boc-protected amino acids. Each amino acid addition consists of (i) cleavage of the Nα-protecting group, (ii) a washing step, (iii) coupling of a fluorenylmethoxycarbonyl-(Fmoc)-protected amino acid or a tert-butyloxycarbonyl-(Boc)-protected amino acid using a coupling reagent and a non-nucleophilic base, and (iv) a washing step, and is called a cycle. Since the growing chain is attached to the carrier, excess reagents and soluble by-products can be removed by simple filtration. Repeated coupling reactions with hindered Fmoc-protected N-methylated amino acids or hindered Boc-protected N-methylated amino acids are difficult and often sub-optimal, and thus this technique is expected to have low crude purity, difficult purification, and low yield. Examples of the carrier are Wang resin, Rink amide resin, trityl- and 2-chlorotrityl resin, PAM resin, PAL resin, Sieber amide resin, MBHA resin, HMPB resin, HMBA resin, which are commercially available and to which peptides are directly or indirectly attached. The term "orthogonal linker" refers to a branched linker unit component, and the branched linker unit component binds a ligand to a homopolymer unit and a drug unit, such that the homopolymer unit is arranged in parallel (as opposed to a serial arrangement) with respect to the drug unit. The orthogonal linker is a scaffold having attachment sites for the components of the ligand-drug complex, i.e., the ligand, the homopolymer, and the drug unit. The term "parallel" fluorenylmethoxycarbonyl-(Fmoc)-protected amino acid or tert-butyloxycarbonyl-(Boc)-protected amino acid Since the growing chain is attached to the carrier, excess reagents and soluble by-products can be removed by simple filtration. Repeated coupling reactions with hindered Fmoc-protected N-methylated amino acids or hindered Boc-protected N-methylated amino acids are difficult and often sub-optimal, and thus this technique is expected to have low crude purity, difficult purification, and low yield. Examples of the carrier are Wang resin, Rink amide resin, trityl- and 2-chlorotrityl resin, PAM resin, PAL resin, Sieber amide resin, MBHA resin, HMPB resin, HMBA resin, which are commercially available and to which peptides are directly or indirectly attached. hindered Fmoc-protected N-methylated amino acid or hindered Boc-protected N-methylated amino acid difficult and often sub-optimal, and thus this technique is expected to have low crude purity, difficult purification, and low yield. Examples of the carrier are Wang resin, Rink amide resin, trityl- and 2-chlorotrityl resin, PAM resin, PAL resin, Sieber amide resin, MBHA resin, HMPB resin, HMBA resin, which are commercially available and to which peptides are directly or indirectly attached. 2-chlorotrityl resin, PAM resin, PAL resin, Sieber amide resin, MBHA resin, HMPB resin, HMBA resin peptides are directly or indirectly attached.

[0041] The term "orthogonal linker" refers to a branched linker unit component, and the branched linker unit component binds a ligand to a homopolymer unit and a drug unit, such that the homopolymer unit is arranged in parallel (as opposed to a serial arrangement) with respect to the drug unit. The orthogonal linker is a scaffold having attachment sites for the components of the ligand-drug complex, i.e., the ligand, the homopolymer, and the drug unit. The term "parallel" linker unit component binds a ligand to a homopolymer unit and a drug unit, such that the homopolymer unit is arranged in parallel (as opposed to a serial arrangement) with respect to the drug unit. such that the homopolymer unit is arranged in parallel (as opposed to a serial arrangement) with respect to the drug unit. The orthogonal linker is a scaffold having attachment sites for the components of the ligand-drug complex, i.e., the ligand, the homopolymer, and the drug unit. The term "parallel" ​​​​​is used to show the branching of two components of a ligand-drug complex (LDC). is not used to show that the two components are necessarily very close in space or have the same distance between them.

[0042] An exemplary schematic presentation of an LDC having a homopolymer (e.g., , polysarcosine) unit in a parallel (i.e., branched) orientation with respect to the drug unit is as follows:

[0043]

Chemical formula

[0044] wherein (L) is an orthogonal connector unit, w is 1 or more, typically 1 to 5, preferably 1 to 4, more preferably 1 to 3, and still more preferably 1 to 2. This orthogonal structure should not be confused with the linear structure . An exemplary schematic presentation of an LDC having a homopolymer (e.g., polysarcosine) unit in a series (i.e., linear) orientation with respect to the drug unit is as follows: is as follows:

[0045]

Chemical formula

[0046] A non-exhaustive list of orthogonal connectors includes natural or non-natural amino acids, e.g., lysine, glu tamic acid, aspartic acid, serine, tyrosine, cysteine, selenocysteine, gly cine, homoalanine; amino alcohols; amino aldehydes; polyamines, or any combination thereof . From his knowledge, one skilled in the art can select an appropriate orthogonal connector for the expected LDC compound. Advantageously, L is one or more natural or non-natural amino acids It is an acid. In one embodiment, L is selected from glutamic acid, lysine, and glycine .

[0047] The spacer is a divalent linear arm that covalently binds to two components of a ligand-drug complex as follows: - A ligand unit and an orthogonal connector unit, - An orthogonal connector unit and a homopolymer unit, - An orthogonal connector and a cleavable moiety, - A cleavable moiety and a drug, or - An orthogonal connector and an agent.

[0048] For example, the spacer is a divalent linear alkylene group, preferably (CH2)4.

[0049] A non-exhaustive list of spacer units includes alkylene, heteroalkylene (alkylene interrupted by at least one heteroatom selected from Si, N, O, and S), ; alkoxy; polyalkylene glycol and polyethers such as typically polyethylene glycol, ; one or more natural or non-natural amino acids such as glycine, alanine, proline, valine, N-methylglycine, etc., ; C3-C8 heterocyclo; C3-C8 carbocyclo; arylene, and any combination thereof. When the spacer is present between a cleavable moiety and a drug unit, or between an orthogonal connector and a drug unit, it can be linked to one or more drug units. For example, the spacer can be linked to 1 to 4 drug units, preferably 1 to 2 drug units. In one embodiment, the spacer between the cleavable moiety and the drug unit is (4-amino-1,3-phenylene) dimethanol.

[0050] ​​​​​In one embodiment, the spacer unit is of formula (XVII), formula (XVIII), formula ( XIX), formula (XX), formula (XXI) or formula (XXII):

[0051]

Chemical formula

[0052] The waveform coupling represents a coupling point, and R6 is -C1-C 10 alkylene-, -C1-C 10 he teroalkylene-, -C3-C8 carbocyclo-, -O-(C1-C8 alkyl)-, - arylene-, -C1-C 10 alkylene-arylene-, -arylene-C1-C 10 alkylene-, -C1-C 10 alkylene-(C3-C8 carbocyclo)-, -(C3- C8 carbocyclo)-C1-C 10 alkylene-, -C3-C8 heterocyclo-, -C1 -C 10 alkylene-(C3-C8 heterocyclo)-, -(C3-C8 heterocyclo)- C1-C 10 alkylene-, -C1-C 10 alkylene-C(=O)-, -C1-C 10 heteroalkylene-C(=O)-, -C3-C8 carbocyclo-C(=O)-, -O-( C1-C8 alkyl)-C(=O)-, -arylene-C(=O)-, -C1-C 10 al kylene-arylene-C(=O)-, -arylene-C1-C 10 alkylene-C(= O)-, -C1-C 10 alkylene-(C3-C8 carbocyclo)-C(=O)-, -( C3-C8 carbocyclo)-C1-C 10 alkylene-C(=O)-, -C3-C8 het cyclo-C(=O)-, -C1-C 10 alkylene-(C3-C8 heterocycle)-C (=O)-, -(C3-C8 heterocycle)-C1-C 10 alkylene-C(=O)-, -C1-C 10 alkylene-NH-, -C1-C 10 heteroalkylene-NH-, -C3 -C8 carbocycle-NH-, -O-(C1-C8 alkyl)-NH-, -arylene- NH-, -C1-C 10 alkylene-arylene-NH-, -arylene-C1-C 10 alkylene-NH-, -C1-C 10 alkylene-(C3-C8 carbocycle)-NH- , -(C3-C8 carbocycle)-C1-C 10 alkylene-NH-, -C3-C8 hetero cyclo-NH-, -C1-C 10 alkylene-(C3-C8 heterocycle)-NH-, -(C3-C8 heterocycle)-C1-C 10 alkylene-NH-, -C1-C 10 alk ylene-S-, -C1-C 10 heteroalkylene-S-, -C3-C8 carbocycle-S -, -O-(C1-C8 alkyl)-)-S-, -arylene-S-, -C1-C 10 alk ylene-arylene-S-, -arylene-C1-C 10 alkylene-S-, -C1- C 10 alkylene-(C3-C8 carbocycle)-S-, -(C3-C8 carbocycle) -C1-C 10 alkylene-S-, -C3-C8 heterocycle-S-, -C1-C 10 alk ylene-(C3-C8 heterocycle)-S-, -(C3-C8 heterocycle)-C1- C10 alkylene-S-, -C1-C 10 alkylene-O-C(=O)-, -C3-C8 carbocyclo-O-C(=O)-, -O-(C1-C8 alkyl)-O-C(=O)-, -arylene-O-C(=O)-, -C1-C 10 alkylene-arylene-O-C( O)-, -arylene-C1-C 10 alkylene-O-C(=O)-, -C1-C 10 a lkylene-(C3-C8 carbocyclo)-O-C(=O)-, -(C3-C8 carbocycl o)-C1-C 10 alkylene-O-C(=O)-, -C3-C8 heterocyclo-O-C (=O)-, -C1-C 10 alkylene-(C3-C8 heterocyclo)-O-C(=O) -, -(C3-C8 heterocyclo)-C1-C 10 is alkylene-O-C(=O)- 。

[0053] Any of the R6 groups is -X, -R’, -O - , -OR’, =O, -SR’, -S - , - NR’2, -NR’3 + , =NR’, -CX3, -CN, -OCN, -SCN, -N=C =O, -NCS, -NO, -NO2, =N2, -N3, -NR’C(=O)R’, -C( =O)R’, -C(=O)NR’2, -SO3 - , -SO3H, -S(=O)2R’, - OS(=O)2OR’, -S(=O)2NR’, -S(=O)R’, -OP(=O)(O R’)2, -P(=O)(OR’)2, -PO3 - , -PO3H2, -C(=O)X, - C(=S)R’, -CO2R’, -CO2, -C(=S)OR’, C(=O)SR’, C (=S)SR’, C(=O)NR’2, C(=S)NR’2, and C(=NR’)NR’ Optionally substituted by one or more substituents selected from 2, wherein each X is independently a halogen: -F, -Cl, -Br, or -I; and each R’ is independently -H, -C1-C2 alkyl, -C6-C aryl, or -C3-C 20 heterocycle. 14 Preferably, the spacer unit is of the formula (XVII), formula (XVIII), formula (XIX),

[0054] formula (XX), formula (XXI), or formula (XXII):

[0055]

Chemical Structure

[0056] The waveform bond represents the bonding point, and R6 is -C1-C 10 alkylene-, -C1-C 10 he teroalkylene-, -C1-C 10 alkylene-C(=O)-, -C1-C 10 hetero alkylene-C(=O)-, -arylene-C1-C 10 alkylene-C(=O)-, -a rylene-C1-C 10 alkylene-O-C(=O)-.

[0057] Any of the R6 groups is optionally substituted by one or more =O.

[0058] The ligand refers to any macromolecule (polypeptide, protein, peptide, typically an antibody) as commonly used in LDC (e.g., antibody-drug conjugate) technology, or a small molecule such as folic acid or an aptamer. Using bioconjugate technology, the synthetic phosphorus in this study ​​​It may be covalently linked to a car or drug linker (see Greg T. Hermanson, Bioconjugate Tech niques, 3rd Edition, 2013, Academic Press). Ligands are traditionally compounds selected for their targeting ability. A non-exhaustive list of ligands includes proteins, polypeptides, peptides, antibodies, full-length antibodies and antigen-binding fragments thereof, interleukins, lymphokines, hormones, growth factors, vitamins, transferrin, or any other cell-binding molecule or substance. The main classification of ligands used to prepare the complex is antibodies. As used herein, the term "antibody" is used in the broadest sense and includes monoclonal antibodies, polyclonal antibodies, modified monoclonal antibodies and modified polyclonal antibodies, multispecific antibodies such as bispecific antibodies, single specificity antibodies, antibody fragments, and antibody mimetics (Affibody®, Affilin®, Affimer®, Nanofitin®, Cell Penetrating Alphabody®, Anticalin®, Avimer® Fynomer®, Monobodies or nanoCLAM P®). An example of an antibody is trastuzumab. An example of a protein is human serum albumin.

[0059] As referred to herein, the term "antibody" includes whole antibodies and any antigen-binding fragment (i.e., "antigen-binding portion"), or a single chain thereof.

[0060] Naturally occurring "antibodies" are composed of at least two It is a glycoprotein containing a heavy (H) chain and two light (L) chains. Each heavy chain is composed of a heavy chain variable region ( abbreviated as V H in this specification) and a heavy chain constant region. The heavy chain constant region is composed of three domains, CH1, CH2, and CH3. Each light chain is composed of a light chain variable region (in this specification abbreviated as V L ) and a light chain constant region. The light chain constant region is composed of one domain C L . V H and V L regions can be further subdivided into hypervariable regions named complementarity determining regions (CDRs) in which more conserved regions named framework regions (FRs) are interspersed. Each V and V H and V L is composed of three CDRs and four FRs arranged in the following order from the amino terminus to the carboxy terminus: FR 1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant region of the antibody may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system ( e.g., effector cells) and the first component of the classical complement system (Clq).

[0061] The term "antigen-binding portion" (or simply "antigen portion") of an antibody, as used herein, refers to the full-length antibody or one or more fragments thereof that retain the ability to specifically bind to an antigen. It has been shown that the antigen-binding function of an antibody can be carried out by fragments of the full-length antibody. Examples of binding fragments included in the term "antigen-binding portion" of an antibody are V , V L , V H , C L and CH1 dom A Fab fragment, which is a monovalent fragment consisting of the main part; two Fab fragments linked by a disulfide bridge in the hinge region An F(ab)2 fragment, which is a divalent fragment containing two Fab fragments linked by a disulfide bridge in the hinge region; V H and CH An Fd fragment consisting of the V L and V H domains of a single arm of the antibody; an Fv fragment consisting of the V and V H domains (Ward et al., 1989 Nature 341:544-546); and Isolated complementarity-determining regions (CDRs), or any fusion protein containing such antigen-binding portions are included.

[0062] Furthermore, the two domains of the Fv fragment, V L and V H are encoded by separate genes, but they can be linked using recombinant methods by a synthetic linker that enables them to be made as a single-chain protein, in which the V and V regions pair to form a monovalent molecule (known as a single-chain Fv (scFv); see, for example, Bird et al., 1988 Science 242:423-426; and Huston et al., 1988 Proc Natl Acad. Sci L i. 85:5879-5883). Such single-chain antibodies are also intended to be encompassed by the term H "antigen-binding portion" of the antibody. These antibody fragments can be obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as the complete antibody. In certain embodiments, the ligand of the LDC is a chimeric antibody, a humanized antibody, or a human anti body. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as the complete antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as the complete antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as the complete antibody.

[0063] In certain embodiments, the ligand of the LDC is a chimeric antibody, a humanized antibody, or a human anti It is a body.

[0064] As used herein, the term "human antibody" is intended to include antibodies having a variable region in which both the framework region and the CDR region are derived from sequences of human origin. Further, when the antibody includes a constant region, the constant region is also derived from such human sequences, such as human germline sequences, or mutant versions of human germline sequences, or consensus framework sequences derived from human framework sequence analysis (e.g., as described in Knappik et al. (2000. J Mol Biol 296, 57-86)). Moreover, a human antibody may contain amino acid residues not encoded by human sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, are grafted onto human framework sequences. As used herein, "isotype" refers to the antibody class provided by the heavy chain constant region gene (e.g., IgG such as IgM, IgE, IgG1 or IgG4). The phrases "antibody that recognizes an antigen" and "antibody specific for an antigen" are synonymous with the term "antibody specific for an antigen".

[0065] A human antibody may contain amino acid residues not encoded by human sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, are grafted onto human framework sequences.

[0066] As used herein, the term "human monoclonal antibody" refers to an antibody that exhibits a single binding specificity and has a variable region in which both the framework region and the CDR region are derived from human sequences.

[0067] As used herein, "isotype" refers to the antibody class provided by the heavy chain constant region gene (e.g., IgG such as IgM, IgE, IgG1 or IgG4).

[0068] The phrases "antibody that recognizes an antigen" and "antibody specific for an antigen" are synonymous with the term "antibody specific for an antigen". ​​​​​​​​​"an antibody that binds to", which is used interchangeably herein.

[0069] A cleavable group (also referred to as a "releasable assembly unit") links the drug unit to the remainder of the ligand-drug complex. The function of the cleavable group is to release the drug at the site targeted by the ligand. This unit can thus form a cleavable bond for release of the drug unit, for example, by enzymatic processing or disulfide removal mechanisms. Recognition sites for enzymatic processing are usually dipeptide cleavage sites (e.g., Val-Cit, Val-Ala, or Phe-Lys), or sugar cleavage sites (e.g., glucuronide cleavage sites). For example, the cleavable group is a glucuronide group. This technology is well known to those skilled in the art, and from his knowledge, he can select a suitable cleavable group for the drug of an LDC (e.g., ADC) compound. For example, the cleavable group includes a disulfide-containing linker cleavable by disulfide exchange, an acid-labile linker cleavable at acidic pH, and a linker cleavable by hydrolytic enzymes (e.g., peptidases, esterases, and glucuronidases). The cleavable group can be in a form selected from: - one or more natural or unnatural amino acids, e.g., a cleavable peptide containing 2 to 12 amino acids, - a sugar moiety linked to a self-destructive group via an oxygen glycosidic bond, - a disulfide linker, and - an acid-labile linker hydrolyzable in lysosomes. - one or more natural or unnatural amino acids, e.g., a cleavable peptide containing 2 to 12 amino acids, - a sugar moiety linked to a self-destructive group via an oxygen glycosidic bond, - a disulfide linker, and - an acid-labile linker hydrolyzable in lysosomes.

[0070] Advantageously, the cleavable group can be in a form selected from: ​- One or more natural or non-natural amino acids, e.g., a cleavable peptide containing 2 to 12 amino acids, and - A sugar moiety linked to a self-destructive group via an oxygen glycosidic bond. When the sugar moiety is used, the self-destructive group is considered to be part of a cleavable group. The "self-destructive group" is a trifunctional chemical moiety that can covalently link three separated chemical moieties together: a sugar moiety (via a glycosidic bond), a drug D (directly or indirectly via a spacer Z), and an orthogonal linker L (directly or indirectly via a spacer Z). The glycosidic bond can be cleaved at the target site to initiate a self-destruction reaction sequence that results in the release of the drug.

[0071] When the sugar moiety is used, the self-destructive group is considered to be part of a cleavable group. The "self-destructive group" is a trifunctional chemical moiety that can covalently link three separated chemical moieties together: a sugar moiety (via a glycosidic bond), a drug D (directly or indirectly via a spacer Z), and an orthogonal linker L (directly or indirectly via a spacer Z). The glycosidic bond can be cleaved at the target site to initiate a self-destruction reaction sequence that results in the release of the drug. The "self-destructive group" is a trifunctional chemical moiety that can covalently link three separated chemical moieties together: a sugar moiety (via a glycosidic bond), a drug D (directly or indirectly via a spacer Z), and an orthogonal linker L (directly or indirectly via a spacer Z). The glycosidic bond can be cleaved at the target site to initiate a self-destruction reaction sequence that results in the release of the drug. When the sugar moiety is used, the self-destructive group is considered to be part of a cleavable group. The "self-destructive group" is a trifunctional chemical moiety that can covalently link three separated chemical moieties together: a sugar moiety (via a glycosidic bond), a drug D (directly or indirectly via a spacer Z), and an orthogonal linker L (directly or indirectly via a spacer Z). The glycosidic bond can be cleaved at the target site to initiate a self-destruction reaction sequence that results in the release of the drug. The "self-destructive group" is a trifunctional chemical moiety that can covalently link three separated chemical moieties together: a sugar moiety (via a glycosidic bond), a drug D (directly or indirectly via a spacer Z), and an orthogonal linker L (directly or indirectly via a spacer Z). The glycosidic bond can be cleaved at the target site to initiate a self-destruction reaction sequence that results in the release of the drug. The glycosidic bond can be cleaved at the target site to initiate a self-destruction reaction sequence that results in the release of the drug. The glycosidic bond can be cleaved at the target site to initiate a self-destruction reaction sequence that results in the release of the drug.

[0072] When a disulfide linker is used, cleavage occurs between the two sulfur atoms of the disulfide. A variety of disulfide linkers are known in the art and can be adapted for use in the present disclosure, including those that can be formed using SATA (N-succinimidyl-S-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), SMPT (N-succinimidyl-oxycarbonyl-α-methyl-α-(2-pyridyl-dithio)toluene), and SPP (N-succinimidyl 4-(2-pyridyldithio)penta-noate). See, e.g., U.S. Patent No. 4,880,935. When a disulfide linker is used, cleavage occurs between the two sulfur atoms of the disulfide. A variety of disulfide linkers are known in the art and can be adapted for use in the present disclosure, including those that can be formed using SATA (N-succinimidyl-S-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), SMPT (N-succinimidyl-oxycarbonyl-α-methyl-α-(2-pyridyl-dithio)toluene), and SPP (N-succinimidyl 4-(2-pyridyldithio)penta-noate). See, e.g., U.S. Patent No. 4,880,935. When a disulfide linker is used, cleavage occurs between the two sulfur atoms of the disulfide. A variety of disulfide linkers are known in the art and can be adapted for use in the present disclosure, including those that can be formed using SATA (N-succinimidyl-S-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), SMPT (N-succinimidyl-oxycarbonyl-α-methyl-α-(2-pyridyl-dithio)toluene), and SPP (N-succinimidyl 4-(2-pyridyldithio)penta-noate). See, e.g., U.S. Patent No. 4,880,935. When a disulfide linker is used, cleavage occurs between the two sulfur atoms of the disulfide. A variety of disulfide linkers are known in the art and can be adapted for use in the present disclosure, including those that can be formed using SATA (N-succinimidyl-S-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), SMPT (N-succinimidyl-oxycarbonyl-α-methyl-α-(2-pyridyl-dithio)toluene), and SPP (N-succinimidyl 4-(2-pyridyldithio)penta-noate). See, e.g., U.S. Patent No. 4,880,935. When a disulfide linker is used, cleavage occurs between the two sulfur atoms of the disulfide. A variety of disulfide linkers are known in the art and can be adapted for use in the present disclosure, including those that can be formed using SATA (N-succinimidyl-S-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), SMPT (N-succinimidyl-oxycarbonyl-α-methyl-α-(2-pyridyl-dithio)toluene), and SPP (N-succinimidyl 4-(2-pyridyldithio)penta-noate). See, e.g., U.S. Patent No. 4,880,935. When a disulfide linker is used, cleavage occurs between the two sulfur atoms of the disulfide. A variety of disulfide linkers are known in the art and can be adapted for use in the present disclosure, including those that can be formed using SATA (N-succinimidyl-S-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), SMPT (N-succinimidyl-oxycarbonyl-α-methyl-α-(2-pyridyl-dithio)toluene), and SPP (N-succinimidyl 4-(2-pyridyldithio)penta-noate). See, e.g., U.S. Patent No. 4,880,935. When a disulfide linker is used, cleavage occurs between the two sulfur atoms of the disulfide. A variety of disulfide linkers are known in the art and can be adapted for use in the present disclosure, including those that can be formed using SATA (N-succinimidyl-S-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), SMPT (N-succinimidyl-oxycarbonyl-α-methyl-α-(2-pyridyl-dithio)toluene), and SPP (N-succinimidyl 4-(2-pyridyldithio)penta-noate). See, e.g., U.S. Patent No. 4,880,935. When a disulfide linker is used, cleavage occurs between the two sulfur atoms of the disulfide. A variety of disulfide linkers are known in the art and can be adapted for use in the present disclosure, including those that can be formed using SATA (N-succinimidyl-S-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), SMPT (N-succinimidyl-oxycarbonyl-α-methyl-α-(2-pyridyl-dithio)toluene), and SPP (N-succinimidyl 4-(2-pyridyldithio)penta-noate). See, e.g., U.S. Patent No. 4,880,935. See, e.g., U.S. Patent No. 4,880,935.

[0073] In some embodiments, the cleavable unit is pH-sensitive, e.g., A linker labile to hydrolyzable acids in lysosomes (e.g., hydrazone, semicarbazone, thiosemicarbazone, cis-aconitic amide, orthoester, acetal, or ketal group) may be used (e.g., see U.S. Patents No. 5,122,368; No. 5,824,805; No. 5,622,929). Such linkers are relatively stable under neutral pH conditions (e.g., pH conditions in blood), but are unstable at pH 5.5 or 5.0 (approximate pH of lysosomes). A ligand-drug conjugate (LDC) binds to a ligand and a drug as defined above, includes any

[0074] mean as defined above, and refers to any conjugate described in the examples herein. When the ligand is an antibody, it may refer to an antibody-drug conjugate (ADC), which is a preferred embodiment of the present disclosure. A bondable group refers to a group that can react with a functionalized reactive group to form

[0075] a covalent bond. A bondable group thus includes a reactive group that reacts with a functionalized reactive group under predetermined reaction conditions. Specifically, a bondable group can include one of the following groups: carboxylic acid; primary amine; secondary amine; tertiary amine; hydroxyl; halogen; N-hydroxysuccinimide ester, perfluoroester, nitrophenyl ester, aza-benzotriazole and benzotriazole activated esters, activated esters such as acylurea; alkynyl; alkenyl; azide; isocyanate; isothiocyanate; aldehyde; maleimide, halomaleimide, haloacetyl, pyridyl disulfide and other thiol-reactive groups. Responsive moiety; thiol; acrylate; mesylate; tosylate; triflate, hydroxy silylamine; chlorosulfonyl; boronic acid - B(OR’)2 derivative (wherein R’ is hydrogen or an alkyl group).

[0076] The drug refers to any type of drug or compound, for example, cytotoxic, cytostatic, immunosuppressive, anti - inflammatory or anti - infectious compounds. Among the cytotoxic compounds, calicheamicin ; uncysalamycin; auristatin (such as monomethyl auristatin E known as MMAE); tubulysin analogs; maytansine; cryptophycin; benzodiazepine dimers (including pyrrolo[2,1 - c][1,4]benzodiazepines known as PDB’s); indolinobenzodiazepine pseudodimers (IGNs); duocarmycin; anthracyclines (such as doxorubicin or PNU159682 etc.); camptothecin analogs (such as 7 - ethyl - 10 - hydroxy - camptothecin known as SN38 or exatecan etc.); Bcl2 and Bcl - xl inhibitors; tyranstatin; amatoxin (including α - amanitin); kinesin spindle protein (KSP) inhibitors; vinorelbine; cyclin - dependent kinase (CDK) inhibitors; bleomycin; dactinomycin or radioactive nuclides and their complexing agents (such as DOTA / Lu etc.) can be mentioned. Among the anti - inflammatory drugs, corticosteroids such as dexamethasone or fluticasone etc. can be mentioned. Among the anti - infectious drugs, antibiotics such as rifampicin or vancomycin etc. can be mentioned. Next, the present invention will be described in more detail. It is the single molecular weight of polysarcosine homopolymer ; kinematic spindle protein (KSP) inhibitor; vinorelbine; cyclin - dependent kinase (CDK) inhibitor; bleomycin; dactinomycin or radioactive nuclides and their complexing agents (such as DOTA / Lu etc.) can be mentioned. Among the anti - inflammatory 177 drugs, corticosteroids such as dexamethasone or fluticasone etc. can be mentioned. Among the anti - infectious drugs, antibiotics such as rifampicin or vancomycin etc. can be mentioned. Next, the present invention will be described in more detail. It is the single molecular weight of polysarcosine homopolymer ; kinematic spindle protein (KSP) inhibitor; vinorelbine;

[0077] Next, the present invention will be described in more detail. It is the single molecular weight of polysarcosine homopolymer Although described more specifically with respect to it, the scope should be recognized to extend to any single molecular weight encompassed within the formula (I). Further, the benefits of the present invention are specifically demonstrated in the LDC technology. Of course, the advantages are not limited to such technology and can exhibit similar or better performance in any area where a single molecular weight, biocompatible, biodegradable homopolymer is required. Accordingly, the present invention more particularly relates to a single molecular weight homopolymer of sarcosine having the formula (II): Of course, the advantages are not limited to such technology and can exhibit similar or better performance in any area where a single molecular weight, biocompatible, biodegradable homopolymer is required. In the formula, R1 and R2 are different, one of R1 and R2 is H or an inert group, and the other of R1 and R2 is a functionalized reactive group, which is reactive to covalently bond to a bondable group under reaction conditions where the inert group does not react. Z1 and Z2 are the same or different and are each an optional spacer, and k is 2 or more.

[0078] Accordingly, the present invention more particularly relates to a single molecular weight homopolymer of sarcosine having the formula (II): :

[0079]

Chemical formula

[0080] (In the formula, R1 and R2 are different, one of R1 and R2 is H or an inert group, and the other of R1 and R2 is a functionalized reactive group, which is reactive to covalently bond to a bondable group under reaction conditions where the inert group does not react. One of R1 and R2 is H or an inert group, and the other of R1 and R2 is a functionalized reactive group, which is reactive to covalently bond to a bondable group under reaction conditions where the inert group does not react. Z1 and Z2 are the same or different and are each an optional spacer, and k is 2 or more. Z1 and Z2 are the same or different and are each an optional spacer, and k is 2 or more. Z1 and Z2 are the same or different and are each an optional spacer, and k is 2 or more. k is an integer of 2 or more.)

[0081] Further features of the homopolymer of formula (I), particularly the homopolymer of formula (II), are given below, alone or in any combination. k is an integer of at least 2, preferably at most 100, more preferably at most 50, specifically 2 to 30, more specifically 2 to 24, 6 to 24, or 12 to 24.

[0082] k is an integer of at least 2, preferably at most 100, more preferably at most 50, specifically 2 to 30, more specifically 2 to 24, 6 to 24, or 12 to 24. k is an integer of at least 2, preferably at most 100, more preferably at most 50, specifically 2 to 30, more specifically 2 to 24, 6 to 24, or 12 to 24. k is an integer of at least 2, preferably at most 100, more preferably at most 50, specifically 2 to 30, more specifically 2 to 24, 6 to 24, or 12 to 24.

[0083] In any of formula (I) or formula (II), the functionalized reactive group R1 or R2 may be selected from the following groups: - A carboxylic acid group, - An amino group NRR'' (wherein R and R'' are independently selected from H and (C1-C6)alkyl optionally interrupted by at least one heteroatom selected from O, N, and S), - A hydroxyl group, - A halogen atom, - A hydrazine (-NH2-NH2) group, - A nitro group, - A hydroxylamine group, - An azide group, - A (C2-C6) alkynyl group, - A (C2-C6) alkenyl group, - A thiol group, - An N-hydroxysuccinimide ester, perfluoroester, nitrophenyl ester, aza-benzotriazole and benzotriazole activated ester, acylurea and other activated ester groups, - A boronic acid -B(OR'''')2 group (wherein R'''' is a hydrogen atom or a C1-C6 alkyl group), - A thiol reactive group such as maleimide, halomaleimide, haloacetyl, pyridyl disulfide, etc., - A mesylate group, - A tosylate group, - A triflate group, - An aldehyde group, - An isocyanate group or an isothiocyanate group, - A chlorosulfonyl group, - An acrylate group.

[0084] As described above, the spacer Z is optional, both Z1 and Z2 may be present, and Z ​​​​​​Only one of 1 and Z2 may be present, or they may not be present either. In the latter case, when the homopolymer of the present invention is a homopolymer of sarcosine, it has the formula (III):

[0085] [Chemical formula]

[0086] (wherein R1, R2 and k are as defined above).

[0087] In formula (I), formula (II) or formula (III), R1 is H or an inert group, and R2 may be a functionalized reactive group, or R1 may be a functionalized reactive group and R2 may be H or an inert group.

[0088] According to a preferred embodiment, the functionalized reactive group R1 or R2 is a secondary amine, and the inert group R1 or R2 is a carboxylic acid that remains unreacted and is not attached to the final LDC structure.

[0089] In a preferred embodiment, in formula (I), formula (II) or formula (III), R1 is selected from OH and NH2, when R1 is OH, R2 is COCH3, when R1 is NH2, R2 is CO-G-COOH, and G is CH2CH2, CH2 CH2CH2, CH2CH2CH2CH2, CH2OCH2, CH2SCH2, CH2C H(CH3)CH2, CH2C(CH3)2CH2 or CH2N(CH3)CH2 .

[0090] The present invention also relates to a single molecular weight homopolymer of any one of formula (I), formula (II) or formula (III) ​Relates to a method for preparing a limmer. Generally, each N-methylglycine monomer is assembled on a solid support from two sub-monomers, namely haloacetic acid and methylamine. Each monomer addition is a cycle called (i) acylation of the resin-bound secondary amine with haloacetic acid and carbodiimide, or acylation by other suitable carboxylic acid salt activation methods, (ii) a washing step, (iii) nucleophilic substitution of the resin-bound halogen with methylamine, (iv) a washing step.

[0091] A method according to the invention, comprising the following steps: a) A compound of formula (IV)

[0092]

Chemical formula

[0093] (wherein R3 is a peptide synthesis solid support, m is 1 or more and less than k) is reacted with an acid of formula (V)

[0094]

Chemical formula

[0095] (wherein Hal is a halogen) to obtain a compound of formula (VI)

[0096]

Chemical formula

[0097] (wherein R3, m and Hal are as defined above) , b) Reacting the compound of formula (VI) with methylamine to obtain a compound of formula (VII)

[0098] [Chemical formula]

[0099] (wherein, R3 and m are as defined above) to obtain a compound of c) Formula (VIII)

[0100] [Chemical formula]

[0101] (wherein, R3 is as defined above and k is as defined above) Repeat steps a) and b) until a compound of d) React the compound (VIII) to obtain a compound of formula (IX)

[0102] [Chemical formula]

[0103] (wherein, R2 is an inert group, R3 is as defined above, and k is as defined above ) to obtain a compound of e) A cleavage reaction to obtain a single molecular weight homopolymer of formula (III) as defined above.

[0104] According to an embodiment of this method, in step a), reacting a compound of formula (IV) (wherein R3 is a peptide synthesis solid support and m is 3), said compound is obtained by the Fmoc-solid phase peptide synthesis method. They are well known to those skilled in the art, and from his knowledge, he can use any suitable coupling reagent, such as N-[(dimethylamino)- 1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-me Chirumethanium hexafluorophosphate N-oxide (HATU) can be selected. It is possible.

[0105] In an alternative embodiment of the method of the present invention, preparing a single molecular weight homopolymer comprises the following steps: a) A compound of formula (X)

[0106]

Chemical formula

[0107] (wherein R3 is a peptide synthesis solid phase, m is 1 or more, and is less than k) is reacted with an acid of formula (V)

[0108]

Chemical formula

[0109] (wherein Hal is a halogen) to obtain a compound of formula (XI)

[0110]

Chemical formula

[0111] (wherein R3, m and Hal are as defined above) to obtain a compound of b) Reacting the compound of formula (XI) with methylamine to obtain a compound of formula (XII)

[0112]

Chemical formula

[0113] (wherein R3 and m are as defined above) to obtain a compound of c) A compound of formula (XIII)

[0114]

Chem.

[0115] (wherein R3 and k are as defined above) Repeat steps a) and b) until the compound of is obtained, d) Formula (XIV)

[0116]

Chem.

[0117] (wherein k is as defined above) A cleavage reaction to obtain the compound of e) React the compound (XIV) with at least one of succinic anhydride, glutaric anhydride, adipic anhydride , diglycolic anhydride, thiodiglycolic anhydride, 3-methylglutaric anhydride, 3 -3-dimethylglutaric anhydride or 4-methylmorpholine-2,6-dione to obtain a single molecular weight homopolymer of formula (III) as defined above . .

[0118] As described above, the homopolymer of the present invention is useful in LDC technology without being limited to this technology .

[0119] Accordingly, the present invention also relates to a ligand-drug-complex compound (L DC) having the following formula (XV):

[0120]

Chem.

[0121] (wherein L allows (HP SMW ) to be in an orthogonal orientation with respect to (X-D) is an orthogonal connector, HP SMW is from the covalent bond of the single molecular weight homopolymer of the present invention as described above to the orthogonal connector resulting from the covalent bond to L, D is a drug, in particular, a cytotoxic drug such as monomethyl auristatin E (MMAE) or SN38, etc., is a cytotoxic drug, X is any cleavable moiety for releasing D, Z is any spacer, a is 1 or more, b is 1 or more, and m is 1 or more).

[0122] The single molecular weight homopolymer, particularly single molecular weight polysarcosine, when grafted in a parallel (i.e., orthogonal) orientation with respect to the drug unit, compared to a ligand-drug-complex that does not contain the single molecular weight homopolymer grafted in parallel, results in efficient hydrophobic masking properties of the complex, reduced apparent hydrophobicity, better pharmacokinetic properties, and improved in vivo activity. activity. activity.

[0123] In an alternative embodiment, D is selected from the group consisting of bioactive molecules, therapeutic molecules such as anti-cancer agents, contrast agents, and fluorophores.

[0124] According to an alternative embodiment of the present invention, a is an integer, at least 1, preferably at most 6, more preferably at most 3, particularly 2, more particularly 1, and / or, b is an integer, at least 1, preferably at most 6, more preferably at most 3, particularly 2, more particularly 1, and / or, m is an integer, at least 1, preferably at most 30, more preferably at most 1 5, particularly 8, more particularly 4.

[0125] Advantageously, the single molecular weight homopolymer is polysarcosine.

[0126] In one embodiment, there is a spacer Z between L and the ligand, and / or between L and HP SMW and / or between L and X, and / or between X and D.

[0127] Typically, the orthogonal connector is through one or more linker unit configuration elements so that the (X-D) or (D) unit is in a parallel configuration (not in a series configuration) with respect to the homopolymer unit, connecting the releasable assembly-drug unit (X-D) or drug unit (D).

[0128] The present invention also relates to an intermediate compound having formula (XVI):

[0129]

Chemical formula

[0130] (wherein L is an orthogonal connector, HP SMW results from the covalent bond of the single molecular weight homopolymer of the present invention to the orthogonal connector L, D is a cytotoxic drug, X is any cleavable moiety for releasing D, Z is any spacer that can bind to the ligand, a is 1 or more, and b is 0 or 1 or more).

[0131] The present disclosure also relates to a compound having formula (XXIII):

[0132]

Chemical formula

[0133] In the formula, R6 is -C1-C 10 alkylene-, -C1-C 10 heteroalkylene-, -C3-C8 carbocyclo-, -O-(C1-C8 alkyl)-, -arylene-, -C 1-C 10 alkylene-arylene-, -arylene-C1-C 10 alkylene-, -C 1-C 10 alkylene-(C3-C8 carbocyclo)-, -(C3-C8 carbocyclo) -C1-C 10 alkylene-, -C3-C8 heterocyclo-, -C1-C 10 alkylene -(C3-C8 heterocyclo)-, -(C3-C8 heterocyclo)-C1-C 10 alk ylene-, -C1-C 10 alkylene-C(=O)-, -C1-C 10 heteroalkylene- C(=O)-, -C3-C8 carbocyclo-C(=O)-, -O-(C1-C8 alkyl )-C(=O)-, -arylene-C(=O)-, -C1-C 10 alkylene-aryle ne-C(=O)-, -arylene-C1-C 10 alkylene-C(=O)-, -C1-C 10 alkylene-(C3-C8 carbocyclo)-C(=O)-, -(C3-C8 carbosi clo)-C1-C 10 alkylene-C(=O)-, -C3-C8 heterocyclo-C(=O )-, -C1-C 10 alkylene-(C3-C8 heterocyclo)-C(=O)-, -(C 3-C8 heterocyclo)-C1-C 10 alkylene-C(=O)-, -C1-C 10 alk ylene-NH-, -C1-C 10heteroalkylene-NH-, -C3-C8carbocyclo -NH-, -O-(C1-C8alkyl)-NH-, -arylene-NH-, -C1-C 10 alkylene-arylene-NH-, -arylene-C1-C 10 alkylene-NH- , -C1-C 10 alkylene-(C3-C8carbocyclo)-NH-, -(C3-C8ca rbocyclo)-C1-C 10 alkylene-NH-, -C3-C8heterocyclo-NH-, , -C1-C 10 alkylene-(C3-C8heterocyclo)-NH-, -(C3-C8het erocyclo)-C1-C 10 alkylene-NH-, -C1-C 10 alkylene-S-, -C 1-C 10 heteroalkylene-S-, -C3-C8carbocyclo-S-, -O-(C1- C8alkyl)-)-S-, -arylene-S-, -C1-C 10 alkylene-aryle n-S-, -arylene-C1-C 10 alkylene-S-, -C1-C 10 alkylene- (C3-C8carbocyclo)-S-, -(C3-C8carbocyclo)-C1-C 10 al kylene-S-, -C3-C8heterocyclo-S-, -C1-C 10 alkylene-(C3- C8heterocyclo)-S-, -(C3-C8heterocyclo)-C1-C 10 alkylene- S-, -C1-C 10 alkylene-O-C(=O)-, -C3-C8carbocyclo-O- C(=O)-, -O-(C1-C8alkyl)-O-C(=O)-, -arylene-O- C(=O)-, -C1-C 10 alkylene-arylene-O-C(=O)-, -aryle -C1-C 10 alkylene-O-C(=O)-, -C1-C 10 alkylene-(C3- C8 carbocyclo)-O-C(=O)-, -(C3-C8 carbocyclo)-C1-C 10 alkylene-O-C(=O)-, -C3-C8 heterocyclo-O-C(=O)-, -C1 -C 10 alkylene-(C3-C8 heterocyclo)-O-C(=O)-, -(C3-C8 heterocyclo)-C1-C 10 is alkylene-O-C(=O)-, any of the R6 groups is -X, -R', -O - , -OR', =O, -SR', -S - , -NR'2, -NR'3 + , =NR', -CX3, -CN, -OCN, -SCN, -N= C=O, -NCS, -NO, -NO2, =N2, -N3, -NR'C(=O)R', -C (=O)R', -C(=O)NR'2, -SO3 - , -SO3H, -S(=O)2R', -OS(=O)2OR', -S(=O)2NR', -S(=O)R', -OP(=O)( OR')2, -P(=O)(OR')2, -PO3 - , -PO3H2, -C(=O)X, -C(=S)R', -CO2R', -CO2, -C(=S)OR', C(=O)SR', C(=S)SR', C(=O)NR'2, C(=S)NR'2, and C(=NR')NR '2 is optionally substituted with one or more substituents selected from, where each X is independently a halo gen (-F, -CI, -Br, or -I), and each R' is independently -H, -C1- C 20 alkyl, -C6-C 20 aryl, or -C3-C 14 heterocycle, Z is an arbitrary spacer, L is an orthogonal connector, X is an arbitrary cleavable moiety for releasing D, D is a cytotoxic drug, a is 1 or greater, b is 0 or 1 or greater; HP SMW results from the covalent attachment of the single molecular weight homopolymer of the present invention to the orthogonal connector L as defined above.

[0134] According to a preferred embodiment, HP SMW results from the covalent attachment of the polysarcosine homopolymer of the present invention to the orthogonal connector L. In this case, in formula (XV), formula (XVI) and formula (XXIII), HP represents the following: SMW

[0135]

Chemical formula

[0136] (wherein the wavy bond represents the bonding point to L or spacer Z if present, k is 2 or greater, preferably, k is 2 to 50, R4 represents a capping group).

[0137] Advantageously, R4 is -R’, -O - -, -OR’, -SR’, -S - -, -NR’2, -NR ’3 + =NR’, -CX3, -CN, -NRC(=O)R’, -C(=O)R’, -C (=O)NR’2, -SO3 - -, -SO3H, -S(=O)2R’, -OS(=O)2O R’, -S(=O)2NR’, -S(=O)R’, -OP(=O)(OR’)2, -P( =O)(OR’)2, -PO3 - ​​, -PO3H2, -C(=O)X, -C(=S)R’, , -CO2R’, -CO2, -C(=S)OR’, C(=O)SR’, C(=S)SR’, , C(=O)NR’2, C(=S)NR’2, or C(=NR’)NR’2, where , each X is independently halogen: -F, -CI, -Br, or -I, and each R’ is independently , -H, -C1-C 20 alkyl, -C6-C 20 aryl, or -C3-C 14 heterocycle . Typically, R4 is -OR’, -NR’2, or -C(=O)R’.

[0138] In one embodiment, the present disclosure also relates to a ligand-drug-complex compound (LDC) having the following formula (XV):

[0139]

Chemical formula

[0140] wherein the ligand is an antibody, L is an orthogonal connector that enables HP SMW to be in an orthogonal orientation with respect to (X-D), and is selected from natural or unnatural amino acids, amino alcohols, amino aldehydes, polyamines, and combinations thereof, HP represents the following, SMW where,

[0141]

Chemical formula

[0142] (wherein the wavy bond represents a bonding point to L or spacer Z when present, k is 2 or more, preferably, k is 2-50, and R4 represents a capping group), D is a drug, in particular, a cytotoxic drug such as monomethyl auristatin E (MMAE) or SN38, etc. X is any cleavable moiety for releasing D, · One or more natural or unnatural amino acids, for example, containing 2 to 12 amino acids , a cleavable peptide, · A sugar moiety bonded to a self-destructive group via an oxygen glycoside bond, · A disulfide linker, and · An acid-labile linker that is hydrolyzable in lysosomes and is in a form selected from Z is any spacer and can be present between L and X, and / or between X and D, and / or between L and HP SMW and is selected from alkylene, heteroalkylene; alkoxy yl; polyether; one or more natural or unnatural amino acids; C3-C8 heterocycle; C 3-C8 carbocycle; arylene, and any combination thereof, a is 1 or more, b is 1 or more, and m is 1 or more.

[0143] The present invention also relates to a pharmaceutical composition comprising at least one LDC compound of the present invention and a pharmaceutically acceptable carrier.

[0144] The present disclosure also relates to an LDC compound as described above for use as a medicament.

[0145] The compound of formula (XXIII) can react with a protein such as serum albumin in vivo and then serve as a ligand, so it can be used as it is without a ligand. Accordingly, the present disclosure also relates to a compound of formula (XXIII) as described above for use as a medicament.

[0146] BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 represents a hydrophobic interaction chromatogram according to Example 12.

[0147] FIG. 2 shows the hydrophobic interaction chromatogram according to Example 13.

[0148] FIG. 3 shows the pharmacokinetic profile in mice according to Example 14.

[0149] FIG. 4A shows the tumor volume as a function of time according to Example 15. FIG. The figures show the survival rates of mice treated with the antibody.

[0150] FIG. 5 shows the pharmacokinetic profile in mice according to Example 16.

[0151] FIG. 6 shows the tumor volume as a function of time according to Example 17.

[0152] [Example] Materials and General Methods All solvents and reagents were obtained from commercial suppliers (Sigma-Aldrich, Alfa Aesar, Fluorochem, The rmo was obtained from Fisher Scientific, Carbosynth, and used without further purification unless otherwise noted. Anhydrous DMF and DCM were purchased from Sigma-Aldrich. Fmoc-amino acids, 2 -Chlorotrityl and Rink amide resins were purchased from Novabiochem. Uristatin E (MMAE) and 7-ethyl-10-hydroxycamptothecin (SN3 8) was purchased from DC Chemicals. PNU159682 was purchased from Kerui Biotechnology Co. Ltd. Exatecan mesylate was purchased from Angene Chemical. Min (cat# A3782) was purchased from Sigma-Aldrich. Anti-CD19 and anti-CD 22 antibodies were purchased from Euromedex. Trastuzumab (Herceptin®) was purchased from Roch e. Synthesis on resin was carried out in an empty SPE plastic tube equipped with a 20 μm polyethylene frit (Sigma-Aldrich). A Titramax 101 plate form shaker (Heidolph) was used for stirring. Unless otherwise specified, all chemical reactions were carried out at room temperature under an inert argon atmosphere.

[0153] Liquid nuclear magnetic resonance spectra were recorded on a Bruker Fourier 300HD spectrometer using the residual solvent peak for calibration. Mass spectrometry was performed at the Centre Commun de Spectrometrie de Ma sse (CCSM) of the UMR5246 CNRS Institute of the University Claude Bern ard Lyon 1.

[0154] Normal-phase flash chromatography was performed on a Teledyne Isco CombiFlash® Compan ion® device or a Teledyne Isco CombiFlash® Rf200 device, using either Int erchim (spherical HP 50 μm) or Biotage® ZIP® (50 μm) silica cartridges. Reverse-phase chromatography was performed using a Biotage® SNAP Ultra C18 (25 μm) cartridge or an Interchim PuriF lash RP-AQ (30 μm) cartridge. Chemical reactions and compound characterization were , respectively, pre-coated 40 - 63 μm silica gel (Macherey - Nagel), HPLC -UV (Agilent 1050) or UHPLC - UV / MS (Bruker Impact I I (trademark) Q - ToF mass spectrometer equipped Thermo UltiMate 3000 UHPLC system, or Agilent 1260 HPLC system equipped with Bruker MicrOTOF - QII mass spectrometer was used for monitoring and analysis by thin - layer chromatography using (Agilent 1260 HPLC system).

[0155] HPLC method 1: Agilent 1050 with DAD detection. Mobile phase A was water, and mobile phase B was acetonitrile. The column was Agilent Zorbax SB - Aq 4.6×150 mm 5 μm (room temperature). The gradient was 5%B - 95%B in 20 minutes and then held at 95%B for 5 minutes. The flow rate was 1.5 mL / min. UV detection was monitored at 214 nm.

[0156] HPLC method 2: Agilent 1050 with DAD detection. Mobile phase A was water and mobile phase B was acetonitrile. The column was Agilent Zorbax SB -Aq 4.6×150 mm 5 μm (room temperature). The gradient was 0%B - 50% B in 30 minutes and then held at 50%B for 5 minutes. The flow rate was 1.0 mL / min. UV detection was monitored at 214 nm.

[0157] HPLC method 3: The same as HPLC method 1, but containing 0.1% TFA in mobile phase A .

[0158] HPLC method 4: The same as HPLC method 2, but containing 0.1% TFA in mobile phase A It is.

[0159] UHPLC method 5: Thermo UltiMate 3000 UHPLC system + Bruker impact II (trademark) Q-ToF mass spectrometer. Mobile phase A was water + 0 .1% formic acid, and mobile phase B was acetonitrile + 0.1% formic acid. The column was Ag ilent PLRP-S 1000Å 2.1×150mm 8μm (80 °C) It was. The gradient was 10% B to 50% B in 25 minutes. The flow rate was 0.4 mL / min. UV detection was monitored at 280 nm. The Q-ToF mass spectrometer was used in the m / z range of 500 to 350 0 (ESI + )). The data was deconvoluted using the MaxEnt algorithm included in Bruker Compass (registered trademark) software.

[0160] HPLC method 6: Agilent 1050 equipped with DAD detection. Mobile phase A was water + 5m M ammonium formate, and mobile phase B was acetonitrile. The column was Agile nt Poroshell 120EC-C18 3.0×50mm 2.7μm (room temperature ). The gradient was 5% B to 90% B in 10 minutes, followed by holding at 90% B for 2 minutes . The flow rate was 0.8 mL / min. UV detection was monitored at 214 nm.

[0161] Examples 1 to 4 below illustrate the synthesis of single molecular weight polysarcosine of the present invention, and this synthesis is part of the present invention including various solid-phase synthesis methods.

[0162] Example 1: Synthesis of polysarcosine compound (synthesis method 1 on resin) The reaction scheme is described below.

[0163]

Number

[0164] 1.1) General method Resin-on synthesis was carried out in an empty SPE plastic tube equipped with a 20-μm polyethylene frit (Sigma-Aldrich). A Titramax 101 platform shaker (Heidolph) was used for stirring. All reported synthesis yields are based on an initial theoretical resin loading of 0.63 mmol / g (the degree of labeling indicated by the manufacturer). Unless otherwise specified, all reactions were carried out at room temperature. (Heidolph) was used for stirring. All reported synthesis yields are based on an initial theoretical resin loading of 0.63 mmol / g (the degree of labeling indicated by the manufacturer). Unless otherwise specified, all reactions were carried out at room temperature. Unless otherwise specified, all reactions were carried out at room temperature.

[0165] 1.2) Resin loading Typically, 500 mg of NovaGEL™ Rink amide beads (0.63 mmol / g, Novabiochem) were swollen in 5 mL of DMF for 15 minutes. The first monomer was added by reacting 10 equivalents of bromoacetic acid and 13 equivalents of diisopropylcarbodiimide (Sigma-Aldrich) in 5 mL of DMF at room temperature for 60 minutes, followed by thorough washing with DMF (5 times 5 mL). The bromoacetylated resin was incubated on a shaker platform with 5 mL of a 40% (wt) aqueous methylamine solution (Sigma-Aldrich) for 30 minutes, followed by thorough washing with DMF (5 times 5 mL) and DCM (5 times 5 mL). The resulting resin was ready for elongation. The resulting resin was ready for elongation.

[0166] 1.3) Elongation of polysarcosine compounds The polysarcosine oligomers were elongated by alternately performing the bromoacetylation step and the amine substitution step until the desired length was obtained. The bromoacetylation step was carried out with 5 ​​​​​​​​​10 equivalents of bromoacetic acid and 13 equivalents of diisopropylcarbodiimide were added to mL of DMF This was carried out by addition. The mixture was stirred for 30 minutes, drained, and washed with DMF (5 mL, 4 times) For the amine substitution step, 5 mL of a 40% (by weight) methylamine (Sigma - Aldrich) aqueous solution was added, the vessel was shaken for 30 minutes, drained, and washed with DMF (5 mL, 4 times) and DCM (5 mL, 4 times).

[0167] 1.4) Cleavage from the resin Cleavage of the polysarcosine oligomer was carried out at room temperature with stirring using a 5 mL solution of TFA / triisopropylsilane (95 :5). The resin was filtered, and the resulting solution was evaporated under reduced pressure to obtain an oily transparent substance.

[0168] At this stage, PSARn - N(CH3)H was either dissolved in water for purification (see below ) or involved in the final functionalization.

[0169] 1.5) Final functionalization To obtain the PSARn - CH2 - CH2 - COOH compound, the N - terminal of the oligomer was functionalized using 2.5 equivalents of succinic anhydride and 10 equivalents of DIPEA in anhydrous acetonitrile The mixture was stirred at room temperature for 1 hour, and the volatile substances were removed under reduced pressure.

[0170] 1.6) Purification The PSAR compound was purified on an Interchim® RP - AQ (30 μm) cartridge The mobile phase A was water + 0.05% TFA, and the mobile phase B was acetonitrile + 0.05% TFA. The gradient was in the range of 0 - 30% B.

[0171] 1.7) Single - molecular - weight polysarcosine compound The obtained PSAR compounds are listed in Table 1 below.

[0172] [Table 1]

[0173] Example 2: Synthesis of polysarcosine compound (synthesis method 2 on resin) The reaction scheme is described below.

[0174] [Number]

[0175] 2.1) General method Solid-phase synthesis was carried out in an empty SPE plastic tube equipped with a 20-μm polyethylene frit (Sigma-Aldrich). A Titramax 101 platform shaker (Heidolph) was used for stirring. All reported synthesis yields are based on an initial resin loading of 1.1 mmol / g (the degree of labeling indicated by the manufacturer). Unless otherwise specified, all reactions were carried out at room temperature. Unless otherwise specified, all reactions were carried out at room temperature. Unless otherwise specified, all reactions were carried out at room temperature.

[0176] 2.2) Synthesis of Fmoc-Sar-Sar-OH

[0177] [Number]

[0178] 2.2.1) Synthesis of Fmoc-Sar-Sar-OtBu Fmoc-Sar-OH (2000 mg / 6.42 mmol) and HATU (2443 mg / 6.42 mmol) were dissolved in 28 mL of anhydrous DMF in a round-bottom flask. DIPEA (2491 mg / 19.27 mmol) was added, and the mixture was stirred at room temperature for 3 minutes. Then Then, sarcosine tert-butyl ester hydrochloride (1167 mg / 6.42 mmol) was added, and the reaction mixture was stirred at room temperature for 90 minutes. The volatile substances were removed under vacuum, and the residue was diluted with water and extracted three times with EtOAc. The organic phase was dried over MgSO4, filtered, and evaporated under vacuum to obtain a solid crude product. The crude product was dissolved in EtOAc / DCM 80:20 (v / v) and the white insoluble material was removed by filtration. The filtrate was purified by silica gel chromatography (petroleum ether / EtOAc, gradient 60:40~20:80) to obtain Fmoc-S ar-Sar-OtBu (2310 mg / 82%) as a white solid. HRMS m / z (ESI ): Calc [M+H] + = 439.2227; Exp [M+H] + = 4 + = 4 39.2234; Error = -1.5 ppm. HPLC method 1 retention time = 13.3 minutes. TLC eluted with 1 00% EtOAc: Rf = 0.8.

[0179] 2.2.2) Removal of tert-butyl ester Fmoc-Sar-Sar-OtBu (2310 mg / 5.27 mmol) was dissolved in 20 mL of DCM, and 8.5 mL of TFA was slowly added. The aqueous solution was stirred at room temperature until complete tert-butyl ester deprotection was observed by HPLC (about 2 hours). Then, the volatile substances were removed under vacuum, and the residue was triturated with diethyl ether to obtain Fmo c-Sar-Sar-OH (1690 mg / 84%) as a white solid. 1 H NM R (500 MHz, DMSO-d6, 100 °C) δ (ppm) 2.84 (s, 3H), 2 .93 (s, 3H), 4.01 (s, 2H), 4.05 (s, 2H), 4.25 (t, J = 4.3 Hz, 1H), 4.34 (d, J = 6.4 Hz, 2H), 7.33 (t, J = 7 .4 Hz, 2H), 7.41 (t, J = 7.4 Hz, 2H), 7.63 (d, J = 7.4 Hz, 2H), 7.85 (d, J = 7.5 Hz, 2H). HRMS m / z (ESI + ) : Calc [M + H] + = 383.1601; Exp [M + H] + = 383.1602; Error = 0.0 ppm. HPLC method 1 retention time = 6.2 min. Eluted with DCM / MeOH 85 :15 (v / v) TLC: Rf = 0.65.

[0180] 2.3) Resin loading Typically, 1000 mg of 2-chlorotrityl chloride resin beads (100 - 20 0 mesh, 1% DVB, 1.1 mmol / g, Novabiochem) were swollen in 10 mL of DCM for 10 minutes. Fmoc-Sar-OH (1. 2 equivalents), previously dissolved in 10 m of dry DCM, was added onto the resin. DIPEA (5 equivalents) was added and the reaction vessel was stirred at room temperature for 2 hours . After draining, the resin was washed with DCM (3 times), DMF (2 times), DCM (3 times), and MeOH (2 times). The resin was dried under high vacuum overnight. The substitution level was evaluated from the weight gain of the resin and / or Fmoc cleavage test (absorbance measurement at 301 nm) and found to be semi - quantitative (usually , 0.95 - 1.1 mmol / g). The resin was stored at -20 °C until further use .

[0181] 2.4) Fmoc-Sar-Sar-OH coupling method The resin was treated with 20% piperidine in DMF (1 mL per 100 mg of resin) at room temperature for 15 minutes twice. Then, the resin was washed with DMF (4 times) and DCM (4 times). The resin In DMF, a solution of Fmoc-Sar-Sar-OH (3 equivalents), HATU (2.85 equivalents), and DIPEA (6 equivalents) was added (1 mL per 100 mg of resin). The reaction vessel was stirred for 2 hours, and the resin was thoroughly washed with DMF (5 times) and DCM (5 times). The resin was dried under vacuum and stored at -20 °C until further use.

[0182] 2.5) Elongation of polysarcosine compound The resin was treated twice with 20% piperidine in DMF (1 mL per 100 mg of resin) at room temperature for 15 minutes each. Subsequently, the resin was washed with DMF (4 times) and DCM (4 times).

[0183] The bromoacetylation step and the amine substitution step were alternately performed until the desired length was obtained to elongate the polysarcosine oligomer. The bromoacetylation step was carried out by adding 10 equivalents of bromoacetic acid and 13 equivalents of diisopropylcarbodiimide to DMF (2 mL per 100 mg of resin) . The mixture was stirred for 30 minutes, drained, and washed with DMF (4 times). For the amine substitution step, 40% (by weight) aqueous methylamine solution was added (1.5 mL per 100 mg of resin), the vessel was shaken for 30 minutes, drained, and washed with DMF ( 4 times) and DCM (4 times).

[0184] 2.6) Final acetylation When the desired oligomer length was obtained, the N-terminus was acetylated using a capping solution consisting of acetic anhydride / DIPEA / DMF (1:2: 3 v / v) (the vessel was shaken for 30 minutes). The solution was drained, and the reaction was repeated once with fresh capping solution. The resin was washed with DMF ( 4 times) and DCM (4 times).

[0185] 2.7) Resin cleavage Cleavage of polysarcosine oligomers from the resin was carried out using a HFIP / DCM (20:80 v / v ) solution with stirring for 30 minutes. The resin was filtered and the volatiles were removed under reduced pressure to obtain a solid crude product.

[0186] 2.8) Purification The PSAR compound was purified on an Interchim® RP-AQ (30 μm) cartridge edge. Mobile phase A was water + 0.1% TFA and mobile phase B was acetonitrile + 0.1% TFA.

[0187] 2.9) Single molecular weight polysarcosine compound The obtained PSAR compounds are listed in Table 2 below.

[0188]

Table 2

[0189] Example 3: Synthesis of polysarcosine compounds having one or several azide-functionalized orthogonal connectors (on-resin synthesis method 3) (on-resin synthesis method 3) The reaction scheme is described below.

[0190]

Number

[0191] 3.1) General method On-resin synthesis was carried out in an empty SPE plastic tube equipped with a 20 μm polyethylene frit (Sigma-Aldrich). A Titramax 101 platform shaker (Heidolph) was used for stirring. All reported synthetic yields were 1.1 mmol / g (manufactured (Heidolph) was used for stirring. All reported synthetic yields were 1.1 mmol / g (manufactured Based on the initial resin loading (degree of labeling shown by the manufacturer). Unless otherwise specified, all reactions were carried out at room temperature. The starting materials were obtained as described in Example 2 above .

[0192] 3.2) Step (1) A 3 M solution of 2-azidoethane-1-amine in DMF was added (1 mL per 100 mg of resin), the vessel was shaken for 45 minutes, drained, and washed with DMF (4 times) and DCM (4 times) .

[0193] 3.3) Step (2) To the resin, a solution of commercially available 2-[4-(2,5-dioxo-2,5-dihydro-1H- pyrrol-1-yl)phenyl]acetic acid (5 equivalents), COMU (4.9 equivalents), and DIP EA (4.9 equivalents) in DMF was added (1 mL per 100 mg of resin). The reaction vessel was stirred for 90 minutes , and the resin was washed with DMF (3 times) and DCM (3 times).

[0194] 3.4) Step (3) Cleavage of the desired compound from the resin was carried out using 1% TFA in DCM (v / v) solution with stirring for 5 minutes (repeated 2 times). The resin was filtered, and the volatiles were removed under reduced pressure to obtain a solid crude product, which was purified using the protocol described in Example 2 above.

[0195] 3.5) Step (4) The bromoacetylation step was carried out by adding 10 equivalents of bromoacetic acid and 13 equivalents of diisopropylcarbodi imide in DMF (2 mL per 100 mg of resin). The mixture was stirred for 30 minutes, drained, and washed with DMF (4 times). For the amine substitution step, 3 0% (wt) aqueous ammonia solution was added (2 mL per 100 mg of resin), and the vessel was stirred for 30 minutes . It was shaken intermittently, discharged, and washed with DMF (4 times) and DCM (4 times). At this stage, the compound was cleaved from the resin (as described in step (3)), and purified using the protocol described in Example 2 .

[0196] 3.6) Step (5) The final bromoacetylation step was carried out by adding 10 equivalents of bromoacetic acid and 13 equivalents of di isopropylcarbodiimide in DMF (2 mL per 100 mg of resin ). The mixture was stirred for 30 minutes, discharged, and washed with DMF (4 times) and DCM (4 times). At this stage, the compound was cleaved from the resin (as described in step (3)), and purified using the protocol described in Example 2 .

[0197] 3.7) Step (6) The bromoacetylation step was carried out by adding 10 equivalents of bromoacetic acid and 13 equivalents of diisopropylcarb odiimide in DMF (2 mL per 100 mg of resin). The mixture was stirred for 30 minutes, discharged, and washed with DMF (4 times). For the amine substitution step, 4 0% (by weight) aqueous methylamine solution was added (1.5 mL per 100 mg of resin), and the vessel was shaken for 3 0 minutes, discharged, and washed with DMF (4 times) and DCM (4 times).

[0198] 3.8) Step (7) The bromoacetylation step was carried out by adding 10 equivalents of bromoacetic acid and 13 equivalents of diisopro pylcarbodiimide in DMF (2 mL per 100 mg of resin). The mixture was stirred for 30 minutes, discharged, and washed with DMF (4 times). For the amine substitution step, in D MF, 3 M 2-azidoethane-1-amine solution was added (1 mL per 100 mg of resin ) The container was shaken for 45 minutes, drained, and washed with DMF (4 times) and DCM (4 times).

[0199] 3.9) Step (8) To the resin, a solution of commercially available 2-[4-(2,5-dioxo-2,5-dihydro-1H- pyrrol-1-yl)phenyl]acetic acid (5 equivalents), COMU (4.9 equivalents), and DIP EA (4.9 equivalents) was added (1 mL per 100 mg of resin). The reaction vessel was stirred for 90 minutes and the resin was washed with DMF (3 times) and DCM (3 times). At this stage, the compound was cleaved from the resin (as described in step (3)) and purified using the protocol described in Example 2 above.

[0200] 3.10) Single molecular weight polysarcosine compound The obtained PSAR compounds are listed in Table 3 below.

[0201]

Table 3

[0202] Example 4: Synthesis of a polysarcosine compound having a terminal non-orthogonal azide functionalized linker (on-resin synthesis method 4) The reaction scheme is described below.

[0203]

Number

[0204] 4.1) General method On-resin synthesis was carried out in an empty SPE plastic tube equipped with a 20 μm polyethylene frit (Sigma-Aldrich). A Titramax 101 platform shaker (Heidolph) was used for stirring. All reported synthesis yields were 0.47 mmol / g ( ​ is based on the initial theoretical resin loading (degree of labeling indicated by the manufacturer). Unless otherwise specified, all reactions were carried out at room temperature. Unless otherwise specified, all reactions were carried out at room temperature.

[0205] 4.2) Resin Loading Typically, 500 mg of Ramage ChemMatrix® beads ( 0.47 mmol / g, Sigma - Aldrich) were swollen in 5 mL of DCM for 15 minutes. The resin was treated with 20% piperidine in DMF (1 mL per 100 mg of resin) at room temperature for 15 minutes twice. Then the resin was washed with DMF (4 times) and DCM (4 times). To the resin, a solution of Fmoc - L - γ - azidohomoalanine OH (3 equivalents), HATU (2.9 equivalents ), and DIPEA (6 equivalents) in DMF was added (1 ml per 100 mg of resin). The reaction vessel was stirred for 1.5 hours and the resin was thoroughly washed with DMF (5 times) and DCM (5 times). The unreacted sites were acetylated using a capping solution consisting of acetic anhydride / DIPEA / DMF (1:2:3 v / v) (the vessel was shaken for 30 minutes). The solution was drained and the resin was washed with DMF (4 times) and DCM (4 times). The resin was treated with 20% piperidine in DMF (1 mL per 100 mg of resin) at room temperature for 15 minutes twice. Then the resin was washed with DMF (4 times) and DCM (4 times). mg of resin) at room temperature for 15 minutes twice. Then the resin was washed with DMF (4 times) and DCM (4 times).

[0206] 4.3) Fmoc - Sar - Sar - OH Coupling Method To the resin, a solution of Fmoc - Sar - Sar - OH (4 equivalents), HATU (3.9 equivalents ), and DIPEA (8 equivalents) in DMF was added (1 ml per 100 mg of resin). The reaction vessel was stirred for 2 hours and the resin was thoroughly washed with DMF (4 times) and DCM (4 times). The resin was In DMF, with 20% piperidine (1 mL per 100 mg of resin), the treatment was carried out twice at room temperature for 15 minutes each. Subsequently, the resin was washed with DMF (4 times) and DCM (4 times).

[0207] 4.4) Elongation of polysarcosine compound The bromoacetylation step and the amine substitution step were alternately performed until the desired length was obtained to elongate the polysarcosine oligomer. The bromoacetylation step was carried out by adding 10 equivalents of bromoacetic acid and 13 equivalents of diisopropylcarbodiimide in DMF (2 mL per 100 mg of resin). The mixture was stirred for 30 minutes, discharged, and washed with DMF (4 times). For the amine substitution step, 40% (by weight) methylamine in aqueous solution was added (1.5 mL per 100 mg of resin), the container was shaken for 30 minutes, discharged, and washed with DMF (4 times) and DCM (4 times). In DMF, By adding In an aqueous solution, 40% (by weight) methylamine was added (1.5 mL per 100 mg of resin), the container was shaken for 30 minutes, discharged, and washed with DMF (4 times) and DCM (4 times). (4 times) and DCM (4 times).

[0208] 4.5) Step (6) To the resin, a solution of commercially available 2-[4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)phenyl]acetic acid (5 equivalents), COMU (4.9 equivalents), and DIPEA (4.9 equivalents) in DMF was added (1 mL per 100 mg of resin). The reaction vessel was stirred for 90 minutes, and the resin was washed with DMF (3 times) and DCM (3 times). (3 times) and DCM (3 times).

[0209] 4.5) Cleavage and purification from resin The cleavage of the oligomer from the resin was carried out using a 5 mL solution of TFA / DCM (50:50) at room temperature with stirring for 30 minutes. This step was repeated once, and the pooled filtrate was evaporated under reduced pressure to obtain a crude solid product, which was purified as described in Example 2 above.

[0210] ​​​​​​4.6) Single molecular weight polysarcosine compound The obtained PSAR compounds are listed in Table 4 below.

[0211]

Table 4

[0212] Example 5: Synthesis of a polyethylene glycol (PEG) compound with an azide-functionalized orthogonal connector (Synthesis method 5 on resin) The reaction scheme is described below. The reaction scheme is described below.

[0213]

Number

[0214] 5.1) General method Solid-phase synthesis was carried out in an empty SPE plastic tube equipped with a 20 μm polyethylene frit (Sigma-Aldrich). A Titramax 101 platform shaker (Heidolph) was used for stirring. All reported synthesis yields are based on an initial resin loading of 1.1 mmol / g (the degree of labeling indicated by the manufacturer). Unless otherwise specified, all reactions were carried out at room temperature. Solid-phase synthesis was carried out in an empty SPE plastic tube equipped with a 20 μm polyethylene frit (Sigma-Aldrich). A Titramax 101 platform shaker (Heidolph) was used for stirring. All reported synthesis yields are based on an initial resin loading of 1.1 mmol / g (the degree of labeling indicated by the manufacturer). Unless otherwise specified, all reactions were carried out at room temperature. (Heidolph) was used for stirring. All reported synthesis yields are based on an initial resin loading of 1.1 mmol / g (the degree of labeling indicated by the manufacturer). Unless otherwise specified, all reactions were carried out at room temperature. All reported synthesis yields are based on an initial resin loading of 1.1 mmol / g (the degree of labeling indicated by the manufacturer). Unless otherwise specified, all reactions were carried out at room temperature. Unless otherwise specified, all reactions were carried out at room temperature.

[0215] 5.2) Resin loading Typically, 200 mg of 2-chlorotrityl chloride resin beads (100 - 200 mesh, 1% DVB, 1.1 mmol / g, Novabiochem) were swollen in 4 mL of DCM for 10 minutes. Fmoc-PEG -CH2CH2COOH (PurePEG™ 、1.2 equivalents) was pre-dissolved in 2 mL of dry DCM and added to the resin. DI 12 -CH2CH2COOH (PurePEG™ (1.2 equivalents) was pre-dissolved in 2 mL of dry DCM and added to the resin. DI PEA (3 equivalents) was added, and the reaction vessel was stirred at room temperature for 1 hour. 300 μL of MeOH was added to quench the unreacted resin. After shaking for 10 minutes, the solution was drained, and the resin was washed with DMF (3 times) and DCM (3 times). The resin was dried under high vacuum.

[0216] 5.2) Step (1) The resin was treated twice with 20% piperidine in DMF (1 mL per 100 mg of resin) at room temperature for 15 minutes each. Subsequently, the resin was washed with DMF (4 times) and DCM (4 times). The bromo acetylation step was carried out by adding 10 equivalents of bromoacetic acid and 13 equivalents of diisopropylcarbodiimide in DMF (2 mL per 100 mg of resin). The mixture was stirred for 30 minutes, drained, and washed with DMF (4 times). For the amine substitution step, a solution of 3 M 2-azidoethane-1-amine in DMF was added (1 mL per 100 mg of resin), the vessel was shaken for 45 minutes, drained, and washed with DMF (4 times) and DCM (4 times).

[0217] 5.3) Step (2) The coupling method of 2-[4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)phenyl]acetic acid and cleavage from the resin were carried out as described in Example 3 above, and the purification of the compound was carried out as described in Example 2 above.

[0218] 5.4) Single molecular weight PEG compound The obtained PEG compounds are listed in Table 5 below.

[0219]

Table 5

[0220] Example 6: Intermediates Based on MMAE, SN38, Exatecan, and PNU159682 Synthesis of Compounds 6.1) Synthesis of Compound Alkyne-Glucuronide-MMAE

[0221]

Number

[0222] 110.8 mg (0.087 mmol) of the starting material (synthesized as described in Renoux et al., Chem. Sci., 2017, 8(5), 3427-3433) was dissolved in MeOH (10 mL) at 0 °C. LiOH monohydrate (36.7 mg / 0.87 mmol) was dissolved in water (1 mL) and slowly added to the reaction vessel. After stirring at 0 °C for 70 minutes, the mixture was neutralized with acetic acid (68.2 mg / 1. 14 mmol) and concentrated under reduced pressure. The resulting material was dissolved in a water / MeOH / DMF aqueous solution (1:1:1 v / v) and purified on a 30 g Biotage® SNAP Ultra C18 (25 μm) cartridge. Mobile phase A was water + 0.05% TFA and mobile phase B was acetonitrile + 0.05% TFA. The gradient was in the range of 10 - 60% B . Compound Alkyne-Glucuronide-MMAE (a mixture of two diastereoisomers) was obtained as a white solid (95 mg / 96%). LC-HRMS m / z (ESI

[0223] ): Cal c[M+H] + = 1127.5758; Exp [M+H] = 1127.5757; Er + ror = 0.1 ppm. HPLC method 3 retention time = 10.3 minutes. +

[0224] 6.2) Synthesis of Compound Alkyne-r-Glucuronide-b(MMAE)2

[0225] [Number]

[0226] 6.2.1) Synthesis of Compound Alkyne-Glucuronide-(PNP)2 165 mg (0.240 mmol) of the starting material (Renoux et al., Chem. Sci., 2017, 8 (5), 3427 - 3433), 64.2 mg (0.419 mmol) of commercially available 4-amino-3-( hydroxymethyl)phenylmethanol and 40.7 mg (0.300 mmol) of HOB t were dissolved in anhydrous DMF. After stirring at 50 °C for 3 hours, the volatile substances were evaporated, and the residue was purified by silica gel chromatography (petroleum ether / EtOAc, gradient 40:60~0:10 0) to obtain the intermediate diol compound as a yellow foamy substance.

[0227] 4 molar equivalents of anhydrous pyridine were added dropwise to a cooled solution (0 °C) of 4-nitrophenyl chloroformate (4 molar equivalents) in anhydrous DCM. The mixture was stirred at 0 °C for 15 minutes. An aqueous solution of the previous intermediate diol compound (1 molar equivalent) in DCM was added, and the mixture was stirred at room temperature for 1 hour . The reaction was quenched with a saturated solution of NaCl and extracted 3 times with DCM. The organic phase was dried over MgSO4 and filtered, and evaporated under reduced pressure to obtain a solid crude product, which was purified by silica gel chroma tography (petroleum ether / EtOAc, gradient 60:40~30:70) to obtain Compound Alkyne-Glucuronide-(PNP)2 (52 mg / 21%, in two steps) as a white solid 11H NMR (300 MHz, CDCl3) δ (ppm) 2.04 (s, 3H), 2.06 (s, 3H), 2.11 (d, J = 2.0 Hz, 3H), 2.71 - 2 .92 (m, 2H), 3.72 (s, 3H), 4.11 (q, J = 7.1 Hz, 1H), 4.22 (d, J = 8.6 Hz, 1H), 5.18 - 5.41 (m, 8H), 5.87 ( t, J = 6.5 Hz, 1H), 7.31 - 7.41 (m, 5H), 7.45 - 7.55 ( m, 2H), 7.56 - 7.71 (m, 2H), 7.83 (d, J = 8.2 Hz, 1H) 、7.89 (s, 1H), 8.21 - 8.32 (m, 4H). HRMS m / z (ESI + ): Calc [M + Na] + = 1055.1925; Exp [M + Na] + = 1055 .1955; Error = -2.9 ppm.

[0228] 6.2.2) Synthesis of Compound Alkyne - Glucuronide - (MMAE)2 52 mg (0.050 mmol) of the previous compound Alkyne - Glucuronide - (PNP)2 、13.7 mg (0.100 mmol) of HOBt and 74.1 mg (0.103 mmo l) of monomethyl auristatin E (MMAE) were dissolved in 1 mL of an 8:2 (v / v) mixture of anhydrous DMF / pyridine. The reaction mixture was stirred at room temperature for 24 hours, and the volatile substances were evaporated under reduced pressure . The crude product residue was purified by silica gel chromatography (DCM / MeOH gradient 9 7:3 ~ 90:10) to obtain 77 mg (70%) of the intermediate compound, which was directly involved in the deprotection step without extensive characterization. 77 mg (0.035 mmol) of this compound was dissolved in MeOH (7 mL) at 0 °C. Lithium hydroxide monohydrate (14.7 mg / 0.350 mmol) was dissolved in water (0.7 mL) and slowly added to the reaction vessel. At 0 °C After stirring for 60 minutes, the mixture was neutralized with acetic acid (27.4 mg / 0.457 mmol), and concentrated under reduced pressure. The resulting material was dissolved in water / MeOH / DMF solution (1:1:1 v / v), and purified on a 30 g Biotage® SNAP Ultra C18 (25 μm) cartridge. Mobile phase A was water + 0.05% TFA, and mobile phase B was acetonitrile + 0.05% TFA.

[0229] Compound alkyne-glucuronide-(MMAE)2 (a mixture of two diastereoisomers ) was obtained as a white solid (40 mg / 56%). LC-HRMS m / z (ESI + ): Calc[M+2H] 2+ = 1025.5623; Exp[M+2H] 2+ = 1025. 5599; Error = 2.4 ppm. HPLC method 3 retention time = 13.0 minutes.

[0230] 6.3) Synthesis of compound alkyne-val-cit-PAB-MMAE

[0231]

Number

[0232] 58 mg (0.052 mmol) of the starting material (synthesized as described in Tang et al., Org. Biomol. Chem., 201 6, 14(40), 9501-9518) and 15 mg (0.077 mmol) of 4-pentynoic acid succinimidyl ester were dissolved in 3 mL of anhydrous DCM. 16 .7 mg (0.129 mmol) of DIPEA was added, and the reaction was carried out at room temperature under an argon atmosphere . It was stirred at room temperature for 16 hours. Subsequently, volatile substances were removed under reduced pressure, and the resulting substance was dissolved in DMF solution, and purified on a 30 g Biotage (registered trademark) SNAP Ultra C18 (25 μm) cartridge. Mobile phase A was water + 0.1% TFA, and mobile phase B was ace tonitrile + 0.1% TFA. The gradient was in the range of 25 - 70% B.

[0233] Compound alkyne-val-cit-PAB-MMAE was obtained as a white solid (21 mg / 34%). ESI + [M+Na] + = 1225.7. HPLC method 3 retention time = 9.0 minutes .

[0234] 6.4) Synthesis of compound alkyne-SN38

[0235]

Number

[0236] 156 mg (0.308 mmol) of the starting material TBDMS-SN38 (synthesized as described in Moon et al., J Med. Chem., 2008, 51(21), 6916 - 6926), 113 mg (0.9 24 mmol) of 4-(dimethylamino)pyridine, and 75 mg (0.369 mmol ) of 4-nitrophenyl chloroformate were dissolved in 8 mL of anhydrous DCM. The solution was stirred at room temperature for 90 minutes, diluted with 5% acetic acid in water, and extracted 3 times with DCM. The organic phase was dried over MgSO4 , filtered, and evaporated under reduced pressure to obtain a yellow solid, which was used in the next step without further purification .

[0237] 175 mg (0.261 mmol) of this yellow solid was dissolved in 4 mL of anhydrous DMF, and prop 43 mg (0.782 mmol) of rugylamine was slowly added. The reaction mixture was stirred at room temperature for 16 h under an argon atmosphere. The volatiles were removed under vacuum, and the residue was purified by silica gel chromatography (petroleum ether / EtOAc, gradient 40:60–0:100) to afford compound alkyne-SN38 (71 mg / 56%) as a pale yellow solid. HRMS m / z (ESI ): Calc [M+H] = 474.1660; Exp [M+H] = 474.1664; Error = -0.9 ppm. HPLC method 3 retention time = 9.7 min S + + + . TLC eluted with 100% EtOAc: Rf = 0.15

[0238] 6.5) Synthesis of compound alkyne glucuronide-SN38

[0239]

No.

[0240] 50 mg (0.074 mmol) of the starting material TBDMS-SN38-OPNP (synthesized as described in the previous section 6.4) and 5 mg (0.037 mmol) of HOBt were weighed into a reaction vessel. 58 .5 (0.223 mmol) of tert-butyl (2-((2-(2-hydroxyethoxy)ethyl)amino)ethyl)(methyl)carbamate (synthesized as described in WO2011 / 133039), which had been pre-dissolved in 1 mL of an 8:2 (v / v) mixture of anhydrous DMF / pyridine, was added to the reaction vessel. The reaction mixture was stirred at room temperature for 16 h, and the volatiles were evaporated under reduced pressure. The crude product residue was purified by silica gel chromatography (DCM / MeOH gradient 98:2–90:10) to afford 48 mg (95%) of the intermediate compound . The volatiles were removed under vacuum, and the residue was purified by silica gel chromatography (DCM / MeOH gradient 98:2–90:10) to afford 48 mg (95%) of the intermediate compound ​​​​A substance (yellow solid) was obtained and used directly in the deprotection step. HRMS m / z (ESI + ): Calc [M+H] + = 681.3130; Exp [M+H] + = 681.3113 ; Error = 2.5 ppm.

[0241] 48 mg (0.071 mmol) of this compound was dissolved in 2 mL of DCM, and 500 μL of TFA was added. The solution was stirred at room temperature for 90 minutes, and volatile substances were removed under reduced pressure. The crude product residue was purified by silica gel chromatography (DCM / MeOH gradient 94:6 - 80:20 ) to give 39.8 mg (96%) of the compound SN38-methylamine as a yellow solid HRMS m / z (ESI + ): Calc [M+H] + = 581.2606 ; Exp [M+H] + = 581.2601; Error = 0.8 ppm. HPLC retention time = 7.7 minutes.

[0242] 48 mg (0.069 mmol) of the starting material (synthesized as described in Renoux et al., Chem. Sci., 2017, 8(5 ), 3427 - 3433), 39.8 mg (0.069 mmol) of the previous compound SN38-methylamine and 9.3 mg (0.069 mmol) of HOBt were dissolved in 1 . 5 mL of an 8:2 (v / v) mixture of anhydrous DMF / pyridine. The reaction mixture was stirred at room temperature for 16 hours, and volatile substances were evaporated under reduced pressure. The crude product residue was purified by silica gel chromatography (DCM / MeOH gradient 98:2 - 95:5) to give 57 mg( 74%) of the intermediate compound (yellow solid), which was used directly in the deprotection step. ESI + ​​[M+H] + = 1130.4。

[0243] 57 mg (0.050 mmol) of this compound was dissolved in MeOH (6 mL) at 0 °C. LiOH monohydrate (21.2 mg / 0.504 mmol) was dissolved in water (0.6 mL), and slowly added to the reaction vessel. After stirring at 0 °C for 70 minutes, the mixture was neutralized with acetic acid (39.4 mg / 0.656 mmol) and concentrated under reduced pressure. The resulting material was dissolved in water / MeOH / D MF solution (1:1:1 v / v) and purified on a 30 g Biotage® SNAP Ultra C18 (25 μm) cartridge. Mobile phase A was water + 0.05% TFA and mobile phase B was acetonitrile + 0.05% TFA. The gradient was in the range of 10 - 6 0% B.

[0244] Compound alkyne - glucuronide - SN38 was obtained as a yellow solid (20.5 mg / 42 %). LC - HRMS m / z (ESI + ): Calc [M + H] + = 990.3251 ; Exp [M + H] + = 990.3210; Error = 4.1 ppm. HPLC method 3 retention time = 8.2 minutes.

[0245] 6.6) Synthesis of compound alkyne - glucuronide - exatecan

[0246]

Number

[0247] 132.1 mg (0.192 mmol) of the starting material (Renoux et al., Chem. Sci., 201 Synthesized as described in 7, 8(5), 3427-3433), 102 mg (0.192 mmol) of e Xatecan mesylate and 26 mg (0.192 mmol) of HOBt were dissolved in 1 mL of an 8:2 (v / v) mixture of anhydrous DMF / pyridine. The reaction mixture was stirred at room temperature for 16 hours , and the volatiles were evaporated under reduced pressure. The crude product residue was purified by silica gel chromatography (D CM / MeOH gradient 98:2~90:10) to obtain 165 mg (87%) of the intermediate compound (yellow solid), which was used directly in the deprotection step. ESI [M+H] + + = 985.3.

[0248] 165 mg (0.168 mmol) of this compound was dissolved in MeOH / THF 1:1 v / v (16 mL) at 0 °C. Lithium hydroxide monohydrate (70.3 mg / 1.675 mmol) was dissolved in water (1.6 mL) and slowly added to the reaction vessel. After stirring at 0 °C for 70 minutes, the mixture was neutralized with acetic acid (131 mg / 2.18 mmol) and concentrated under reduced pressure. The resulting substance was dissolved in a water / MeOH / DMF solution (1:1:1 v / v) and purified on a 30 g Biotage (registered trademark) SNAP Ultra C18 (25 μm) cartridge. The mobile phase A was water + 0.05% TFA, and the mobile phase B was acetonitrile + 0.05% TFA . The gradient was in the range of 10~50% B.

[0249] Compound alkyne-glucuronide-xatecan was obtained as a yellow solid (98 mg / 69 %). LC-HRMS m / z (ESI + ): Calc [M+H] + = 845.2312 ; Exp [M+H] +=845.2360; Error = -4.8 ppm. HPLC method 3 Retention time = 8.0 minutes.

[0250] 6.7) Synthesis of compound alkyne-PNU159682

[0251]

Number

[0252] 6.7.1) N-(2-((2-Aminoethyl)amino)-2-oxoethyl)prop anol amide synthesis 762 mg (4.54 mmol) of glycine tert-butyl ester hydrochloride, 318.3 mg (4.54 mmol) of propiolic acid, and 61.4 mg (0.454 mmol) of HOBt were dissolved in 5 mL of anhydrous DMF. 1103 mg (10.9 mmol) of DIEA was added, and the solution was stirred on ice (0 °C). 871 mg (4.54 mmol) of EDC hydrochloride was suspended in 12 mL of anhydrous DMF and added to the reaction vessel. The mixture was stirred in the dark at room temperature for 1 6 hours. Then, the volatile substances were removed under reduced pressure. A saturated solution of NH4Cl was added , and the mixture was extracted 3 times with DCM. The organic phase was dried over MgSO4, filtered, and evaporated. The crude product residue was purified by silica gel chromatography (petroleum ether / EtOAc gradient 80:20~5 0:50) to obtain 255 mg (31%) of tert-butyl propioloyl glycinate as a clear oil. MS (ESI + ): [M+H] + =184.0; Eluted with petroleum ether / EtOAc (40:60 v / v) and stained with KMnO4: Rf = 0. 75. 75.

[0253] 255 mg (1.39 mmol) of tert-butyl propioloyl glycinate in 5 mL The deprotection reaction was evaluated by TLC analysis. The reaction was complete after stirring at room temperature for 1 hour as determined by the reaction mixture. The volatiles were removed under reduced pressure to give As an oily residue, 188 mg (105%) of propioloyl glycine was obtained, which was not purified. It was used in the next step without any reaction.

[0254] 178 mg (1.40 mmol) of propioloylglycine and 504.8 mg (1. 33 mmol) of HATU was dissolved in 3 mL of anhydrous DMF. 1 mmol) of DIPEA was added and the reaction was stirred at room temperature for 5 min. Add 291 mg (1.82 mmol) of N-Boc-ethylenediamine dissolved in F1m. The reaction mixture was stirred in the dark at room temperature for 30 min. The volatiles were then removed under reduced pressure. A saturated solution of NH4Cl was added and extracted three times with DCM. The organic phase was dried over MgSO4. The crude residue was purified by silica gel chromatography (petroleum ether Purification by ethanol / EtOAc gradient 20:80 to 0:100 yielded 204 mg (54%). tert-Butyl (2-(2-propiolamidoacetamido)ethyl)carbamate Obtained as a slightly yellow oil. MS(ESI + ):[M+H] + =270.1, 100% TLC eluted with EtOAc and stained with KMnO4: Rf=0.35.

[0255] 204 mg (0.758 mmol) of tert-butyl (2-(2-propiolamido) Dissolve cetoamidoethyl carbamate in 5 mL of DCM / TFA (7:3 v / v) solution. The deprotection reaction was stirred at room temperature for 45 minutes as evaluated by TLC analysis and then was completed. Volatiles were removed under high vacuum overnight to afford 196 mg (92%) of N-(2- ((2-aminoethyl)amino)-2-oxoethyl)prop-2-ynamide TFA salt as a slightly yellowish thick wax. 1 1H NMR (300 MHz, DMSO-d6) δ (ppm) 2.84 (q, J = 6.2 Hz, 2H), 3.29 (q, J = 6.3 Hz, 2H), 3.72 (d, J = 6.0 Hz, 2H), 4.20 (s, 1H), 7.78 (br s, 3H), 8.12 (t, J = 5.6 Hz, 1H), 8.94 (t, J = 5.9 H z, 1H). MS (ESI + ): [M + H] + = 170.0.

[0256] 6.7.2) Synthesis of alkyne-PNU159682 25 mg (0.040 mmol) of the PNU159692 carboxylic acid derivative (a purple solid synthesized as described in WO / 20 16 / 040825, see the chemical structure above) and 15.1 mg (0.040 mmol) of HATU were dissolved in 1 mL of anhydrous DMF in a round-bottom flask. 10.2 mg (0.080 mmol) of DIPEA was added and the mixture was stirred at room temperature for 2 minutes. 13.4 mg (0.047 mmol) of N-(2-((2-aminoethyl)amino)-2-oxoethyl)prop-2-ynamide TFA salt (previously dissolved in 500 μ L of anhydrous DMF) was added and the reaction mixture was stirred at room temperature for 5 minutes. Volatiles were removed under high vacuum and the residue was purified by silica gel chromatography (DCM / MeOH, gradient 99 :1 to 90:10) to afford 14.7 mg (49%) of alkyne-PNU15 9682 as a yellow solid. 9682 was obtained as a red solid. HRMS m / z (ESI + ): Calc[M+H] + =779.2770; Exp[M+H] + =779.2758; Error = 1.5 pp m. HPLC retention time = 5.4 min.

[0257] Example 7: Synthesis of a polysarcosine-based drug conjugate linker using glutamic acid as the orthogonal moiety The reaction scheme is described below.

[0258]

Number

[0259] 7.1) Resin loading of ethylenediamine 500 mg of 2-chlorotrityl chloride resin beads (100 - 200 mesh, 1% DVB, 1.1 mmol / g, 0.55 mmol scale, Novabiochem) were swollen in 5 mL of D CM for 10 minutes, and 5 equivalents (2.75 mmol, 165.3 mg) of ethylenedi amine (Sigma - Aldrich) were added. The mixture was shaken at room temperature for 4 hours and then washed thoroughly with DCM (5 times 5 mL). The unreacted sites on the resin were capped using a DCM / MeOH / DIPEA (17 :2:1 v / v) solution (treated for 20 minutes). The resin was washed thoroughly with DCM (5 times 5 m L) and MeOH (5 times 5 mL), dried under vacuum, and stored at -20 °C until further use.

[0260] 7.2) Fmoc - Glu(OAll)-OH coupling To the resin containing the deprotected N-terminus (1 equivalent), in DMF, Fmoc - Glu(OA ​A solution of ll)-OH (3 equivalents), HATU (2.85 equivalents), and DIPEA (6 equivalents) was added (1 mL per 100 mg of resin). The reaction vessel was stirred for 2 hours, and the resin was washed thoroughly with DMF (5 times 3 mL) and DCM (5 times 3 mL). The completeness of the reaction was confirmed by a negative Kais er test. The resin was dried under vacuum and stored at -20 °C until further use.

[0261] 7.3) Removal of the Alloc protecting group The resin was suspended in DCM (4 mL per 100 mg of resin), and the mixture was gently stirred by an argon stream introduced from below a fritted disk. Phenylsilane (20 equivalents) was added, and after stirring was continued for 5 minutes, Pd(PPh3)4 (0.25 equivalents) was added. Under an argon flow, stirring of the mixture at room temperature was continued for 30 minutes under protection from light, after which the solution was drained. Treatment with phenylsilane and Pd(PPh3)4 was repeated once, and the resin was washed thoroughly with DCM (5 times 5 mL), DMF (5 times 5 mL), and MeOH (5 times 5 mL). The resin was dried under vacuum and stored at -20 °C until further use. The resin loading was evaluated (Fmoc cleavage test, absorbance measurement at 301 nm), and was usually 0.70 - 0.80 mmol / g.

[0262] 7.4) (2R,3R,4R,5S,6R)-6-(2-(3-Aminopropanamide )-4-((5S,8S,11S,12R)-11-((S)-sec-Butyl)-12 -(2-(2-((1R,2R)-3-(((1S,2R)-1-Hydroxy-1-phen ylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl) ​(Pyrrolidin-1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-di methyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetra decyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2- carboxylic acid (NH2-glucuronide-MMAE) conjugation method To a resin containing a deprotected carboxylic acid group (1 equivalent), a solution of HATU (4 equivalents ) and DIPEA (4.2 equivalents) in DMF was added. The reaction vessel was stirred for 25 minutes and drained , and the resin was washed with DMF (4 times 5 mL). Then, 1.5 equivalents of the compound (2R,3R,4R,5S,6R)-6-(2-(3-aminopropanamide)-4- ((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2- (2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpro pan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolid in-1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl- 3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl) phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (“NH2-glucuronide-MMAE”; synthesized as described in Jeffrey SC et al., Bioconjug. Chem., 200 6, 17(3), 831-840) and DIPEA (3.2 equivalents) were added. The reaction vessel was stirred for 3 hours, drained, and washed with DMF (5 times 3 mL) and DCM (5 times 3 m L). The resin was dried under vacuum and stored at -20 °C until further use.

[0263] 7.5) Fmoc Deprotection Method The resin containing Fmoc-protected amino acid was treated twice with 20% piperidine (1 mL per 100 mg of resin) in DMF for 15 minutes each at room temperature. Then, the resin was washed with DMF (5 times, 5 mL each) and DCM (5 times, 5 mL each). The resin was dried under vacuum and stored at -20 °C until further use.

[0264] 7.6) Polysarcosine Coupling Method To the resin containing a deprotected primary amine group (1 equivalent), a solution of polysarcosine-CH2-CH2-COOH (2.2 equivalents), HATU (2 equivalents), and DIPEA (6 equivalents) in DMF was added. The reaction vessel was stirred for 2.5 hours, drained, and the resin was washed with DMF (3 times, 5 mL each) and DCM (3 times, 5 mL each). The resin was dried under vacuum and stored at -20 °C until further use.

[0265] 7.7) Cleavage from Resin Using 20% (v / v) HFIP in DCM solution (2 mL per 100 mg of resin), the final cleavage from the 2-chlorotrityl resin was carried out with stirring at room temperature for 60 minutes. The resin was filtered, and the resulting solution was evaporated under a stream of argon gas. The final residue was dried under high vacuum and used as such in the following steps.

[0266] 7.8) 3-(Maleimido)propionic Acid N-Hydroxysuccinimide Ester Coupling Method To the residue dissolved in anhydrous DMF, 3-(maleimido)propionic acid N-hydroxysuccinimide ester (8 equivalents) was added. DIPEA (10 equivalents) was added, and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was quenched with water / TFA (99.5:0.5 v / v). ​​​​​​​​30 g Biotage (registered trademark) SNAP Ultra C18 (25 μm) cartridge was purified on a cartridge. Mobile phase A was water + 0.05% TFA, and mobile phase B was acetonitrile + 0.05% TFA. The gradient was in the range of 10 - 60% B.

[0267] Compound MAL-Glu (glucuronide MMAE)-CH2-CH2-PSAR6 was obtained as a clear oil (yield based on 5.8 mg / initial resin loading was 14%). LC-HRMS m / z (ESI ): Calc [M+2H] + = 989.5064; 2+ Exp [M+2H] 989.5023; Error = 4.2 ppm. HPLC method 1 retention 2+ time = 6.7 minutes.

[0268] Compound MAL-Glu (glucuronide MMAE)-CH2-CH2-PSAR12 was obtained as a clear oil (yield 20% based on 4.6 mg / initial resin loading). LC-HRMS m / z (ESI ): Calc [M+2H] + = 1202.6178; 2+ Exp [M+2H] 2+ 1202.6178; Error = 0.0 ppm. HPLC method 1 retention time = 6.9 minutes.

[0269] Compound MAL-Glu (glucuronide MMAE)-CH2-CH2-PSAR18 was obtained as a clear oil (yield based on 2.4 mg / initial resin loading was 14%). LC-HRMS m / z (ESI ): Calc [M+2H] + = 1415.7291 2+ ; Exp [M+2H] 2+ 1415.7282; Error = 0.7 ppm. HPLC method​​​ Retention time = 6.8 minutes.

[0270] Example 8: Synthesis of a polysarcosine-based drug conjugate linker using lysine as the orthogonal moiety Synthesis The reaction scheme is described below.

[0271]

Number

[0272] 8.1) Synthesis of Fmoc-D-Lys (glucuronide MMAE)-NH2

[0273]

Chemical formula

[0274] Compound (2R,3R,4R,5S,6R)-6-(2-(3-aminopropanamide) -4-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12- (2-(2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenyl propyl)amino)-1-methoxy-2-methyl-3-oxopropyl)pi rolidin-1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-dime thyl-3,6,9-trioXO-2,13-dioxa-4,7,10-triazatetrade syl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-car boxylic acid (“NH2-glucuronide-MMAE”; synthesized as described in Jeffrey SC et al., Bioconjug. Chem ., 2006, 17(3), 831-840) (69.9 mg / 0.06 2 mmol) and Fmoc-D-Lys(Boc)-OSu (35 mg / 0.062 mm (ol) was dissolved in 1.2 mL of anhydrous DMF. DIPEA (24.0 mg / 0.186 m mol) was added and the mixture was stirred at room temperature for 20 hours. Volatiles were removed under vacuum. The flask containing the slightly yellowish crude product was placed on an ice bath (0 °C) and 7 mL of DCM / TFA (7:3 v / v) solution was added slowly. The solution was stirred on ice until complete Boc deprotection was observed by HPLC (about 2 hours). Then, volatiles were removed under reduced pressure and the residue was dissolved in DMF for purification on a 30 g Biotage® SNAP Ultra C18 (25 μm) cartridge. Mobile phase A was water + 0.05% TFA and mobile phase B was acetonitrile + 0.05% TFA. The gradient was 10 - 60% B and the compound Fmoc-D-Lys(glucuronide MMAE)-NH2 was obtained as a white solid (59 mg / 65%). LC-HRMS m / z (ESI ): Calc[M+H] = 1480.7862; Exp[M+H] + ): Calc[M+H] + = 1480.7862; Exp[M+H] + = 1480.7890; Error = -1. 9 ppm. HPLC method 1 retention time = 10.5 minutes.

[0275] 8.2) Synthesis of Fmoc-D-Lys(glucuronide MMAE)-PSARn

[0276]

Chemical Structure

[0277] Compound PSARn-COOH (2 equivalents; obtained as described in Example 2 and pre-dissolved as a 0.15 M stock solution in anhydrous DMF) was added to HATU (1.8 equivalents) in a vial. DIPEA (5 equivalents) was added and the mixture was stirred at room temperature for 3 minutes. Then, in anhydrous DMF) was added to HATU (1.8 equivalents) in a vial. DIPEA (5 equivalents) was added and the mixture was stirred at room temperature for 3 minutes. Then, in anhydrous DMF) was added to HATU (1.8 equivalents) in a vial. DIPEA (5 equivalents) was added and the mixture was stirred at room temperature for 3 minutes. Then, Compound Fmoc-D-Lys(glucuronide MMAE)-NH2 (1 equivalent; pre-dissolved as a 0.05 M stock solution in anhydrous DMF) was added. Stirred at room temperature for 1 hour and loaded onto a 30 g B iotage (registered trademark) SNAP Ultra C18 (25 μm) cartridge for purification. Mobile phase A was water + 0.05% TFA and mobile phase B was acetonitrile + 0.05% TFA. The gradient was in the range of 10 - 60% B.

[0278] Compound Fmoc-D-Lys(glucuronide MMAE)-PSAR6 was obtained as a white solid (9.8 mg / 38%). LC-HRMS m / z (ESI ): Calc[M + 2 + H] 2+ = 975.0133; Exp[M + 2H] 2+ = 975.0088; Error = 4.6 ppm. HPLC method 1 retention time = 7.5 minutes.

[0279] Compound Fmoc-D-Lys(glucuronide MMAE)-PSAR12 was obtained as a white solid (3.6 mg / 28%). LC-HRMS m / z (ESI ): Calc[M + + 2H] 2+ = 1188.1247; Exp[M + 2H] 2+ = 1188.1233; Er ror = 1.1 ppm. HPLC method 1 retention time = 7.6 minutes.

[0280] 8.3) 6-(Maleimide)hexanoic acid N-hydroxysuccinimide ester coupling ring method The compound Fmoc-D-Lys(glucuronide MMAE)-PSARn from the previous step was treated with 20% piperidine in DMF at room temperature for 5 minutes. Volatiles were removed under high vacuum ​​​​​, the dry residue was dissolved in anhydrous DMF. Then, N-hydroxysuccinimide ester of 6-(maleimido)hexanoic acid (8 equivalents) and DIPEA (10 equivalents) were added, and the mixture was stirred at room temperature for 30 minutes. It was quenched with water / TFA (99.5:0.5 v / v) and purified on a 30 g B iotage (registered trademark) SNAP Ultra C18 (25 μm) cartridge. Mobile phase A was water + 0.05% TFA, and mobile phase B was acetonitrile + 0.0 5% TFA. After 10 minutes of isocratic hold (5% B), the target compound was eluted isocratically at 40% B.

[0281]

Chemical formula

[0282] Compound MAL-Lys (glucuronide MMAE)-PSAR6 was obtained as a clear oil ( 5.0 mg / 52%). LC-HRMS m / z (ESI + ): Calc [M + 2H] 2 + = 960.5163; Exp [M + 2H] 2+ = 960.5167; Error = -0 .5 ppm. HPLC method 1 retention time = 7.1 minutes.

[0283] Compound MAL-Lys (glucuronide MMAE)-PSAR12 was obtained as a clear oil (1.8 mg / 51%). LC-HRMS m / z (ESI + ): Calc [M + 2H] 2+ = 1173.6276; Exp [M + 2H] 2+ 1173.6229; Error = 4.0 ppm. HPLC method 1 retention time = 7.0 minutes.

[0284] ​​​Example 9: Polyalkosine-based or polyethylene glycol-based using glycine as the orthogonal moiety Synthesis of a recall-based drug conjugate linker 9.1) Compound bromoacetamide-Ngly(triazole-)glucuronide MMA E)-PSARn

[0285]

Number

[0286] Alkyne-glucuronide-MMAE (1 equivalent; obtained as described in Example 6), P SARn-N3-bromoacetamide (1.1 equivalents; obtained as described in Example 3) , and tetrakis(acetonitrile)copper(I) hexafluorophosphate (3 equivalents) were combined in a reaction vessel. A DCM / acetonitrile 1:1 (v / v) solution was added to reach a final alkyne-glucuronide-MMAE concentration of 12 μ mol / mL. The reaction mixture was stirred at room temperature under argon in the dark for 16 - 20 hours. After removal of the volatiles under reduced pressure, the residue was dissolved in DMF and purified on a 30 g Biotage® SNAP Ultra C18 (25 μm) cartridge. Mobile phase A was water + 0.1% TFA, and mobile phase B was acetonitrile + 0.1% TFA. The gradient was in the range of 10 - 50% B.

[0287] Compound bromoacetamide-Ngly(triazole-glucuronide MMAE)-PS AR12 was obtained as a white solid (8.5 mg / 51%). LC-HRMS m / z (ES I + ): Calc[M + 2H] 2+ = 1151.0179; Exp[M + 2H] 2+ = 1 151.0188; Error = -0.8 ppm. HPLC method 3 retention time = 8.5 minutes.

[0288] 9.2) Compound MAL-phenyl-Ngly(triazole-glucuronide MMAE) -PSARn

[0289]

Number

[0290] Alkyne-glucuronide-MMAE (obtained as described in Example 6) and PSAR n-N3-phenyl-MAL (obtained as described in Example 3) was reacted and purified as described above in Section 9.1 using DCM as the reaction solvent.

[0291] Compound MAL-phenyl-Ngly(triazole-glucuronide MMAE)-PSA R6 was obtained as a white solid (3.3 mg / 20%). LC-HRMS m / z (ESI + ): Calc [M+2H] 2+ = 955.9566; Exp [M+2H] 2+ = 955. 9533; Error = 3.4 ppm. HPLC method 3 retention time = 9.2 minutes.

[0292] Compound MAL-phenyl-Ngly(triazole-glucuronide MMAE)-PSA R12 was obtained as a white solid (9.0 mg / 33%). LC-HRMS m / z (ESI + ): Calc [M+2H] 2+ = 1169.0679; Exp [M+2H] 2+ = 11 69.0621; Error = 4.9 ppm. HPLC method 3 retention time = 8.7 minutes.

[0293] ​Compound MAL-Phenyl-Ngly(Triazole-Glucuronide MMAE)-PSA R18 was obtained as a white solid (11.5 mg / 40%). LC-HRMS m / z (ES I + ): Calc [M+2H] 2+ = 1382.1792; Exp [M+2H] 2+ = 1 382.1803; Error = -0.7 ppm. HPLC method 3 retention time = 8.6 minutes.

[0294] Compound MAL-Phenyl-Ngly(Triazole-Glucuronide MMAE)-PSA R24 was obtained as a white solid (15 mg / 44%). LC-HRMS m / z (ESI + ): Calc [M+4Na] 4+ = 820.1309; Exp [M+4Na] 4+ = 82 0.1324; Error = -1.8 ppm. HPLC method 3 retention time = 8.4 minutes.

[0295] 9.3) Compound MAL-Phenyl-Ngly(Triazole-Glucuronide MMAE) -PEGn

[0296]

Number

[0297] Alkyne-Glucuronide-MMAE (obtained as described in Example 6) and PEGn-N 3-Phenyl-MAL (obtained as described in Example 5) were reacted as described above in Section 9.1 using NMP / DC M2:1 (v / v) as the reaction solvent, and purified.

[0298] Compound MAL-Phenyl-Ngly(Triazole-Glucuronide MMAE)-PEG 12 was obtained as a slightly yellow oil (10.4 mg / 45%). LC-HRMS m / z (ESI + ): Calc[M+2H] 2+ = 1042.5211; Exp[M+2H] 2+ = 1042.5218; Error = -0.7 ppm. HPLC method 3 retention time = 8. 0 minutes.

[0299] 9.4) Compound MAL-phenyl-Ngly(triazole-glucuronide MMAE) -Ngly(triazole-glucuronide MMAE)-PSARn

[0300]

Number

[0301] Alkyne-glucuronide-MMAE (3 equivalents; obtained as described in Example 6), PSA Rn-N3-N3-phenyl-MAL (1 equivalent; obtained as described in Example 3), and te trakis(acetonitrile)copper(I) hexafluorophosphate (5 equivalents) were combined in a reaction vessel and stirred in DCM in the dark under argon at room temperature for 16 - 20 hours . After removal of volatiles under reduced pressure, the residue was dissolved in DMF and purified on a 30 g Biotage (registered trademark) SNAP Ultra C18 (25 μm) cartridge. The mobile phase A was water + 0.1% TFA and the mobile phase B was acetonitrile + 0.1% TFA. The gradient was in the range of 10 - 50% B.

[0302] Compound MAL-phenyl-Ngly(triazole-glucuronide MMAE)-Ngl y(triazole-glucuronide MMAE)-PSAR18 was obtained as a white solid (10 .1 mg / 44%). LC-HRMS m / z (ESI + ): Calc [M+4H] 4+ = 1022.5082; Exp [M+4H] 4+ = 1022.5093; Error = - 1.0 ppm. HPLC method 3 retention time = 9.5 min.

[0303] 9.5) Compound MAL-phenyl-Ngly[triazole-glucuronide (MMAE )2]-PSARn

[0304]

Number

[0305] Alkyne-glucuronide-(MMAE)2 (obtained as described in Example 6) and P SARn-N3-phenyl-MAL (obtained as described in Example 3) were used as the reaction solvent and reacted as described above in Section 9.1 using DCM and purified.

[0306] Compound MAL-phenyl-Ngly[triazole-glucuronide (MMAE)2]- PSAR24 was obtained as a white solid (12.5 mg / 39%). LC-HRMS m / z (ESI + ): Calc [M+3H] 3+ = 1371.3767; Exp [M+3H] 3 + = 1371.3818; Error = -3.8 ppm. HPLC method 3 retention time = 10. 0 min.

[0307] 9.6) Compound MAL-phenyl-Ngly[triazole-galactoside (MMAE )2]-PSARn

[0308]

Number

[0309] Alkyne-galactoside-(MMAE)2 (synthesized as described in Alsarraf et al., Chem. Commun., 2015, 51(87), 15792 - 15795) and PSARn-N3-phenyl -MAL (obtained as described in Example 3) were reacted using DCM as the reaction solvent as described in Section 9.1 and purified.

[0310] Compound MAL-phenyl-Ngly[triazole-galactoside-(MMAE)2]- PSAR24 was obtained as a white solid (6.5 mg / 55%). LC-HRMS m / z( ESI + ): Calc [M + 4H] 4+ = 1022.7895; Exp [M + 4H] 4+ = 1022.7903; Error = 0.8 ppm. HPLC method 3 retention time = 9.2 minutes.

[0311] 9.7) Compound MAL-phenyl-Ngly(triazole-val-cit-PAB -MMAE)-PSARn

[0312]

No.

[0313] Alkyne-val-cit-PAB-MMAE (obtained as described in Example 6) and PSARn-N3-phenyl-MAL (obtained as described in Example 3) were reacted using NMP as the reaction solvent as described above in Section 9.1 and purified.

[0314] ​Compound MAL-phenyl-Ngly(triazole-val-cit-PAB-MMA E)-PSAR12 was obtained as a white solid (4.5 mg / 12%). LC-HRMS m / z(ESI + ): Calc[M+2H] 2+ =1207.1494; Exp[M+2H 2+ =1207.1535; Error=-3.5 ppm. HPLC method 3 retention time = 8 .6 minutes.

[0315] 9.8) Compound MAL-phenyl-Ngly(triazole-SN38)-PSARn

[0316]

Number

[0317] Alkyne-SN38 (obtained as described in Example 6) and PSARn-N3-phe nyl-MAL (obtained as described in Example 3) were reacted and purified as described in Section 9.1 using DCM / DMF 2 :1 (v / v) as the reaction solvent.

[0318] Compound MAL-phenyl-Ngly(triazole-SN38)-PSAR18 was obtained as a light yellow solid (4.0 mg / 20%). LC-HRMS m / z(ESI + ): Ca lc[M+2Na] 2+ =1077.4562; Exp[M+2Na] 2+ =1077. 4588; Error=-2.4 ppm. HPLC method 3 retention time = 7.5 minutes.

[0319] 9.9) Compound MAL-phenyl-Ngly(triazole-SN38)-Ngly( triazole-SN38)-PSARn ​

[0320]

Number

[0321] Alkyne-SN38 (obtained as described in Example 6) and PSARn-N3-N3 -Phenyl-MAL (obtained as described in Example 3) were reacted as described in Section 9.4 and purified.

[0322] Compound MAL-Phenyl-Ngly (Triazole-SN38)-Ngly (Triazole -SN38)-PSAR18 was obtained as a yellow solid (5.1 mg / 43%). LC- HRMS m / z (ESI + ): Calc [M+2Na] 2+ = 1412.5812; E xp [M+2Na] 2+ = 1412.5852; Error = -3.8 ppm. HPLC method 3 retention time = 8.4 minutes.

[0323] 9.10) Compound MAL-Phenyl-Ngly (Triazole-glucuronide SN38 )-Ngly (Triazole-glucuronide SN38)-PSARn

[0324]

Number

[0325] Alkyne-glucuronide-SN38 (obtained as described in Example 6) and PSAR n-N3-N3-Phenyl-MAL (obtained as described in Example 3) were reacted with the reaction solvent using DCM / MeOH 8:2 (v / v) and reacted as described in Section 9.4 and purified.

[0326] Compound MAL-phenyl-Ngly(triazole-glucuronide SN38)-Ngl y(triazole-glucuronide SN38)-PSAR18 was obtained as a yellow solid (7. 0 mg / 30%). LC-HRMS m / z (ESI + ): Calc [M+2H] 2+ = 1271.5080; Exp [M+2H] 2+ = 1271.5103; Error = -1 .8 ppm. HPLC method 3 retention time = 7.8 minutes.

[0327] 9.11) Compound MAL-phenyl-Ngly(triazole-glucuronide-exa tecane)-PSARn

[0328]

Number

[0329] Alkyne-glucuronide-exatecan (obtained as described in Example 6) and PS ARn-N3-phenyl-MAL (obtained as described in Example 3) were used as the reaction solvent and reacted using DCM as described above in Section 9.1 and purified.

[0330] Compound MAL-phenyl-Ngly(triazole-glucuronide-exatecan)- PSAR18 was obtained as a yellow solid (13.2 mg / 64%). LC-HRMS m / z (ESI + ): Calc [M+2H] 2+ = 1241.0069; Exp [M+2H] 2 + = 1241.0088; Error = -1.1 ppm. HPLC method 3 retention time = 7.1 minutes.

[0331] 9.12) Compound MAL-Phenyl-Ngly(Triazole-PNU159682) -PSARn

[0332]

Number

[0333] Alkyne-PNU159682 (obtained as described in Example 6) and PSARn- N3-Phenyl-MAL (obtained as described in Example 3) were used with DCM as the reaction solvent During reverse-phase purification, the 0.1% TFA additive in the mobile phase was replaced with 0.1% formic acid and reacted and purified as described above in Section 9.1

[0334] Compound MAL-Phenyl-Ngly(Triazole-PNU159682)-PSAR 12 was obtained as a red solid (4.8 mg / 40%). LC-HRMS m / z (ESI + ): Calc[M+2H] 2+ = 994.9185; Exp[M+2H] 2+ = 994. 9184; Error = 0.1 ppm. HPLC method 6 retention time = 5.0 minutes

[0335] Compound MAL-Phenyl-Ngly(Triazole-PNU159682)-PSAR 18 was obtained as a red solid (7.2 mg / 33%). LC-HRMS m / z (ESI + ): Calc[M+2H] 2+ = 1208.0298; Exp[M+2H] 2+ = 120 8.0295; Error = 0.3 ppm. HPLC method 6 retention time = 4.9 minutes

[0336] Example 10: Negative control drug conjugate linker MAL-Glucuronide MMAE, MAL-F Enil-triazole-glucuronide MMAE, and MAL-phenyl-PSARn-t Synthesis of Triazole-glucuronide MMAE 10.1) Synthesis of Compound MAL-glucuronide MMAE

[0337]

Number

[0338] Starting compound (2R,3R,4R,5S,6R)-6-(2-(3-aminopropanami do)-4-((5S,8S,11S,12R)-11-((S)-sec-butyl)-1 2-(2-(2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-f enylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl )pyrrolidin-1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10- dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatet radecyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2 -carboxylic acid (“NH2-glucuronide-MMAE”; synthesized as described in Jeffrey SC et al., Bioconjug. Chem., 2006, 17(3), 831-840) (6.2 mg / 5 μm ol) and N-hydroxysuccinimide ester of 3-(maleimide)propionic acid (1 4.6 mg / 55 μmol) were weighed and dissolved in 200 μL of anhydrous DMF. DIPE A (8.5 mg / 66 μmol) was added and the mixture was stirred at room temperature for 30 minutes. 1.5 mL of water / TFA (99:1 v / v) was used to quench, and purification was carried out on a 30 g Biotage® SN AP Ultra C18 (25 μm) cartridge. Mobile phase A is water + 0. It was 0.5% TFA, and mobile phase B was acetonitrile + 0.05% TFA. The gradient was 1 in the range of 0 - 70% B.

[0339] The titled compound MAL - glucuronide MMAE was obtained as a white solid (4.1 mg / 59 %). LC - HRMS m / z (ESI + ): Calc [M + H] + = 1281.650 1; Exp [M + H] + = 1281.6489; Error = 0.9 ppm. HPLC method Retention time = 7.1 minutes.

[0340] 10.2) Synthesis of compound MAL - phenyl - triazole - glucuronide MMAE 10.2.1) Synthesis of perfluorophenyl 2 - (4 - (2,5 - dioxo - 2,5 - dihydro - 1H - pyrrol - 1 - yl)phenyl)acetate

[0341]

Chemical Structure

[0342] Commercially available 2 - [4 - (2,5 - dioxo - 2,5 - dihydro - 1H - pyrrol - 1 - yl )phenyl]acetic acid (299 mg / 1.29 mmol), N,N’ - dicyclohexylcarb odiimide (267 mg / 1.29 mmol) and pentafluorophenol (238 m g / 1.29 mmol) were dissolved in 15 mL of anhydrous 1,2 - dimethoxyethane in a reaction vessel . After stirring at room temperature for 2 hours, the insoluble substances were removed by filtration, and the filtrate was purified by silica gel chromatography (petroleum ether / EtOAc, gradient 80:20 - 20:80) to obtain the titled compound (400 mg / 78%) as a white solid. 1 H NMR (30 0 MHz, CDCl3) δ (ppm) 4.01 (s, 2H), 6.87 (s, 2H), 7 .40 (d, J = 8.7 Hz, 2H), 7.47 (d, J = 8.7 Hz, 2H). HRM S m / z (ESI + ): Calc [M + H] + = 398.0446; Exp [M + H] + = 398.0448; Error = -0.4 ppm.

[0343] 10.2.2) Synthesis of N-(2-azidoethyl)-2-(4-(2,5-dioxo-2,5- dihydro-1H-pyrrol-1-yl)phenyl)acetamide

[0344]

Chemical Structure

[0345] The previous compound perfluorophenyl 2-(4-(2,5-dioxo-2,5-dihydro- 1H-pyrrol-1-yl)phenyl)acetate (78 mg / 0.20 mmol) was dissolved in 1 mL of anhydrous DCM in a reaction vessel. 2-Azidoethan-1-amine (33.8 mg / 0.40 mmol) was added and the reaction mixture was stirred at room temperature for 1 hour. Then, 1N HCl l solution was added and the mixture was extracted three times with DCM. The organic phase was dried over MgSO4, filtered , and evaporated under reduced pressure to obtain a crude solid product, which was purified by silica gel chromatography (petroleum ether / EtOAc, gradient 60:40~0:100) to give the title compound (18 mg / 31%) as a white solid. MS (ESI ): [M + H] + = 300.1; + = 300.1; HPLC method 1 retention time = 8.4 minutes. TLC eluted with 100% EtOAc: Rf = 0.6 5.

[0346] 10.2.3) Synthesis of Compound MAL-Phenyl-Triazole-Glucuronide MMAE

[0347]

No.

[0348] Compound Alkyne-Glucuronide MMAE from Example 6 (17 mg / 15.1 μmol ), tetrakis(acetonitrile)copper(I) hexafluorophosphate (11.2 mg / 30 μmol), and N-(2-azidoethyl)-2-(4-(2,5- dioxo-2,5-dihydro-1H-pyrrol-1-yl)phenyl)acetamide (6 .3 mg / 21 μmol) from the previous step were combined in an HPLC vial. 800 μL of anhydrous DCM / acetonitrile / NMP 1:1:1 (v / v / v) solution was added, and the reaction mixture was stirred under argon at room temperature for 16 h. After removal of the volatiles under reduced pressure, the residue was dissolved in DMF and purified on a 30 g Biotage® SNAP Ultra C18 (25 μm) cartridge . Mobile phase A was water + 0.1% TFA, and mobile phase B was acetonitrile + 0.1 % TFA. The gradient was in the range of 10 - 60% B.

[0349] The title compound MAL-Phenyl-Triazole-Glucuronide MMAE was obtained as an off-white solid (10.3 mg / 48%). LC-HRMS m / z (ESI + ): C alc[M+2H] 2+ = 713.8425; Exp[M+2H] 2+ = 713.841 5; Error = 1.3 ppm. HPLC method 3 retention time = 10.2 min.

[0350] 10.3) Compound MAL-Phenyl-PSARn-Triazole-Glucuronide MMA Synthesis of E

[0351] [Number]

[0352] Compound Alkyne-Glucuronide MMAE from Example 6 (20 mg / 17.7 μmol ), Tetrakis(acetonitrile)copper(I) Hexafluorophosphate (13.2 mg / 35 μmol), and N3-PSARn-Phenyl-MAL from Example 4 (34.3 mg / 28 μmol) were combined in an HPLC vial. 900 μL of NMP / DCM2 :1 (v / v) solution was added and the reaction was stirred under argon at room temperature for 16 h. After removal of volatiles under reduced pressure, the residue was dissolved in DMF and purified on a 30 g Biotage® SNA P Ultra C18 (25 μm) cartridge. Mobile phase A was water + 0.1 % TFA and mobile phase B was acetonitrile + 0.1% TFA. The gradient was in the range of 10 - 6 0% B.

[0353] The title compound MAL-Phenyl-PSAR n-Triazole-Glucuronide MMAE was obtained as a white solid (16.0 mg / 39%). LC-HRMS m / z (ESI + ) : Calc [M+2H] 2+ = 1168.5759; Exp [M+2H] 2+ = 1168 .5792; Error = -2.8 ppm. HPLC method 3 retention time = 6.8 min.

[0354] Example 11: Preparation of the LDC Compounds of the Invention The following LDC compounds were prepared and characterized: Trastuzumab-Glu (glucuronide MMAE)-CH2-CH2-PSAR6 Trastuzumab-Glu (glucuronide MMAE)-CH2-CH2-PSAR12 Trastuzumab-Glu (glucuronide MMAE)-CH2-CH2-PSAR18 Trastuzumab-Lys (glucuronide MMAE)-PSAR6 Trastuzumab-Lys (glucuronide MMAE)-PSAR12 Trastuzumab-BAC-Ngly (triazole-glucuronide MMAE)-PSA R12 Trastuzumab-MAL-phenyl-Ngly (triazole-glucuronide MMAE )-PSAR6 Trastuzumab-MAL-phenyl-Ngly (triazole-glucuronide MMAE )-PSAR12 Trastuzumab-MAL-phenyl-Ngly (triazole-val-cit-PA B-MMAE)-PSAR12 Trastuzumab-MAL-phenyl-Ngly (triazole-glucuronide MMAE )-PSAR18 Trastuzumab-MAL-phenyl-Ngly (triazole-SN38)-PSAR 18 Trastuzumab-MAL-phenyl-Ngly (triazole-glucuronide-exa tecane)-PSAR18 Trastuzumab-MAL-phenyl-Ngly (triazole-PNU159682) -PSAR12 Trastuzumab-MAL-phenyl-Ngly (triazole-PNU159682) -PSAR18 Trastuzumab-MAL-phenyl-Ngly (triazole-glucuronide MMAE )-PSAR24 CD19-MAL-phenyl-Ngly(triazole-glucuronide MMAE)-P SAR24 CD22-MAL-phenyl-Ngly(triazole-glucuronide MMAE)-P SAR24 Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide MMAE )-Ngly(triazole-glucuronide MMAE)-PSAR18 Trastuzumab-MAL-phenyl-Ngly[triazole-glucuronide(MMA E)2]-PSAR24 Trastuzumab-MAL-phenyl-Ngly[triazole-galactoside(MMA E)2]-PSAR24 Trastuzumab-MAL-phenyl-Ngly(triazole-SN38)-Ngly (triazole-SN38)-PSAR18 Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide SN38 )-Ngly(triazole-glucuronide SN38)-PSAR18 Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide MMAE )-PEG12 Trastuzumab-glucuronide MMAE Trastuzumab-MAL-phenyl-triazole-glucuronide MMAE Trastuzumab-MAL-phenyl-PSAR12-triazole-glucuronide MM AE Human albumin-MAL-phenyl-Ngly(triazole-glucuronide MMAE )-PSAR24 Their structures are described in Table 6 below.

[0355]

Table 6

[0356] 11.1) Preparation of the conjugate 11.1.1) Preparation of the antibody-drug conjugate The antibody solution (10 mg / mL in PBS 7.4 + 1 mM EDTA) was treated with 14 molar equivalents of tris(2-carboxyethyl)phosphine (TCEP) at 37 °C for 2 hours. For maleimide-based coupling, the fully reduced antibody was buffer-exchanged into 100 mM potassium phosphate pH 7.4 + 1 mM EDTA by 3 rounds of dilution / centrifugation using an Amicon 30K centrifugal filter device (Merck Millipore). 10 - 12 equivalents of the drug linker (from a 12 mM DMSO stock solution) were added to the antibody (residual DMSO < 10% v / v). The solution was incubated at room temperature for 30 minutes. For bromoacetamide-based coupling, the fully reduced antibody was buffer-exchanged into 50 mM borate buffer pH 8.1 + 1 mM EDTA and conjugated with 16 molar equivalents of drug-linker in the dark at 37 °C for 24 hours. The conjugate was buffer-exchanged / purified into PBS 7.4 by 4 rounds of dilution / centrifugation using an Amicon 30K centrifugal filter device. Alternatively, the conjugate was buffer-exchanged / purified using a PD MiniTra p G-25 column (GE Healthcare) and sterilized ​ Filtered (0.20 μm PES filter). The complex incorporating the self-hydrolyzable maleimide (MAL-phenyl) group was incubated at 5 mg / mL in PBS 7.4 at 37 °C for 48 h to ensure complete hydrolysis of the succinimidyl moiety. The final protein concentration was spectrophotometrically evaluated at 280 nm using a Colibri microvolume spectrometer device (Titertek Berthold).

[0357] 11.1.2) Preparation of human albumin-drug complex To a solution of human albumin (10 mg / mL in 100 mM potassium phosphate pH 7.4 + 1 mM EDTA), 2 molar equivalents of drug-linker (from a 12 mM DMSO stock aqueous solution) were added. The residual DMSO was <10% (v / v). The solution was incubated at room temperature for 4 h. The complex was buffer-exchanged / purified with PBS 7.4 by 4 rounds of dilution / centrifugation using an Amicon 30K centrifugal filter device. Alternatively, the complex was buffer-exchanged / purified using a PD MiniTrap G-25 column (GE Healthcare) and sterile filtered (0.20 μm PES filter). The complex was incubated at 5 mg / mL in PBS 7.4 at 37 °C for 48 h to ensure complete hydrolysis of the succinimidyl moiety. The final protein concentration was spectrophotometrically evaluated at 280 nm using a Colibri microvolume spectrometer device (Titertek Berthold).

[0358] 11.2) Characterization of the complex The obtained complex was characterized as follows: Reverse-phase liquid chromatography-mass spectrometry (RPLC-MS):​​​​​​​​​​​​​​ Denaturing RPLC-QToF analysis was performed using the UHPLC method 5 described above. Briefly, the complex was eluted using a gradient of mobile phase of water / acetonitrile + 0.1% formic acid (0.4 mL / min) on an Agilent PLRP-S 1000 Å 2.1×150 mm 8 μm ( 80 °C), and detected using a Bruke + ) to scan the range of 500 - 3500 m / z (ESI r Impact II (trademark) Q-ToF mass spectrometer. The data was deconvoluted using the MaxEnt algorithm included in Bruker Compass (registered trademark) software.

[0359] Size exclusion chromatography (SEC): SEC was performed using an Agilent 1050 HPLC system having a column dead volume of less than 15 μL (equipped with a short section of a peek tube with an inner diameter of 0.12 mm and a microvolume UV flow cell). The column was either a Waters Acquity UPLC (registered trademark) Protein BEH SEC 200 Å 4.6×150 mm 1.7 μm ( maintained at room temperature) or an Agilent AdvanceBio SEC 300 Å 4.6× 150 mm 2.7 μm (maintained at room temperature). The mobile phase was 100 mM sodium phosphate and 200 mM sodium chloride (pH 6.8). 10% acetonitrile (v / v) was added to the mobile phase to minimize secondary hydrophobic interactions with the stationary phase and prevent bacterial growth while maintaining a flow rate of 0.35 mL / min. UV detection was monitored at 280 nm.

[0360] Hydrophobic interaction chromatography (HIC): Hydrophobic interaction chromatography (HIC) was performed using an Agilent 1050 HPLC Performed on a system. The column was Tosoh TSK-GEL BUTYL-NPR 4. 6×35 mm 2.5 μm (25 °C). Mobile phase A was 1.5 M (NH4)2SO 4 + 25 mM potassium phosphate pH 7.0. Mobile phase B was 25 mM potassium phosphate pH 7.0 + 15% isopropanol (v / v). The linear gradient was 0% B to 100% B in 10 minutes, followed by holding at 100% B for 3 minutes. The flow rate was 0.75 mL / min . UV detection was monitored at 220 and 280 nm.

[0361] 11.3) Outline of Characterization of Complexes The complexes showed one LC- 1d (light chain with one drug linker attached) and one HC-3d (heavy chain with three drug-linker s attached) absorbance peak on their denaturing RPLC chromatograms (DAR8 complexes). For the mass spectrometry of the heavy chain, the major glycoform was reported (G0F for trastuzumab). The complexes showed a single absorbance peak on their HIC chromatograms.

[0362] Trastuzumab-Glu (glucuronide MMAE)-CH2-CH2-PSAR6 (D AR8): Deconvoluted LC-1d Calc: 25416; Obs: 25417 / De convoluted HC-3d Calc: 56529; Obs: 56528 Monomer purity: 97.2% HIC retention time: 8.8 minutes Trastuzumab-Glu (glucuronide MMAE)-CH2-CH2-PSAR12( DAR8): Deconvoluted LC-1d Calc: 25844; Obs: 25844 / De ​Convoluted HC-3d Calc:57805;Obs:57805 Monomer purity: 99.0% HIC retention time: 8.8 minutes Trastuzumab-Glu (glucuronide MMAE)-CH2-CH2-PSAR18( DAR8): Deconvoluted LC-1d Calc:26270;Obs:26270 / de Convoluted HC-3d Calc:59086;Obs:59086 Monomer purity: 96.5% HIC retention time: 8.8 minutes Trastuzumab-Lys (glucuronide MMAE)-PSAR6 (DAR8): Deconvoluted LC-1d Calc:25360;Obs:25360 / de Convoluted HC-3d Calc:56352;Obs:56353 Monomer purity: 99+% HIC retention time: 7.6 minutes Trastuzumab-Lys (glucuronide MMAE)-PSAR12 (DAR8): Deconvoluted LC-1d Calc:25786;Obs:25786 / de Convoluted HC-3d Calc:57634;Obs:57632 Monomer purity: 99+% HIC retention time: 7.5 minutes Trastuzumab-BAC-Ngly (triazole-glucuronide MMAE)-PSA R12 (DAR8) Deconvoluted LC-1d Calc:25661;Obs:25662 / de Convoluted HC-3d Calc:57264;Obs:57262 Monomer purity: 99+% HIC retention time: 7.0 minutes (DAR8 conjugate). This ADC is in the HIC chromatogram As observed, ~20% DAR6; ~20% DAR7, and ~60% DAR8 is a heterogeneous mixture containing the complex.

[0363] Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide MMAE )-PSAR6(DAR8)(=ADC-PSAR6) Deconvoluted LC-1d Calc: 25368; Obs: 25368 / de Convoluted HC-3d Calc: 56382; Obs: 56380 Monomer purity: 99+% HIC retention time: 7.5 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide MMAE )-PSAR12(DAR8)(=ADC-PSAR12) Deconvoluted LC-1d Calc: 25794; Obs: 25794 / de Convoluted HC-3d Calc: 57660; Obs: 57660 Monomer purity: 99+% HIC retention time: 7.1 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-val-cit-PA B-MMAE)-PSAR12(DAR8) Deconvoluted LC-1d Calc: 25871; Obs: 25870 / de Convoluted HC-3d Calc: 57889; Obs: 57888 Monomer purity: 99+% HIC retention time: 9.2 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide MMAE )-PSAR18(DAR8)(=ADC-PSAR18) Deconvoluted LC-1d Calc: 26221; Obs: 26221 / de Convoluted HC-3d Calc:58939;Obs:58939 Monomer purity: 99+% HIC retention time: 6.8 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-SN38)-PSAR 18 (DAR8) Deconvoluted LC-1d Calc:25567;Obs:25567 / de Convoluted HC-3d Calc:56979;Obs:56977 Monomer purity: 99+% HIC retention time: 5.1 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide-exa tecan)-PSAR18(DAR8) Deconvoluted LC-1d Calc:25938;Obs:25937 / de Convoluted HC-3d Calc:58092;Obs:58089 Monomer purity: 99+% HIC retention time: 5.7 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-PNU159682) -PSAR12(DAR8) Deconvoluted LC-1d Calc:25446;Obs:25446 / de Convoluted HC-3d Calc:56614;Obs:56612 Monomer purity: 99+% HIC retention time: 5.2 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-PNU159682) -PSAR18(DAR8) Deconvoluted LC-1d Calc:25872;Obs:25872 / de Convoluted HC-3d Calc:57894;Obs:57892 Monomer purity: 99+% HIC retention time: 5.1 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide MMAE )-PSAR24(DAR8)(=ADC-PSAR24) Deconvoluted LC-1d Calc: 26647; Obs: 26674 / De Convoluted HC-3d Calc: 60218; Obs: 60218 Monomer purity: 99+% HIC retention time: 6.7 minutes CD19-MAL-phenyl-Ngly(triazole-glucuronide MMAE)-P SAR24(DAR8) Deconvoluted LC-1d Calc: 27347; Obs: 27347 / De Convoluted HC-3d Calc: 60137; Obs: 60132 Monomer purity: 92.6% HIC retention time: 6.8 minutes CD22-MAL-phenyl-Ngly(triazole-glucuronide MMAE)-P SAR24(DAR8) Deconvoluted LC-1d Calc: 27341; Obs: 27341 / De Convoluted HC-3d Calc: 60314; Obs: 60311 Monomer purity: 97.4% HIC retention time: 6.8 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide MMAE )Ngly(triazole-glucuronide MMAE)-PSAR18(DAR16) Deconvoluted LC-1d Calc: 27544; Obs: 27545 / De Convoluted HC-3d Calc: 62910; Obs: 62907 Monomer purity: 98.5% HIC retention time: 8.7 minutes Trastuzumab-MAL-phenyl-Ngly[triazole-glucuronide(MMA E)2]-PSAR24(DAR16) Deconvoluted LC-1d Calc: 27569; Obs: 27570 / De Convoluted HC-3d Calc: 62985; Obs: 62983 Monomer purity: 98.2% HIC retention time: 9.9 minutes Trastuzumab-MAL-phenyl-Ngly[triazole-galactoside (MMA E)2]-PSAR24(DAR16) Deconvoluted LC-1d Calc: 27555; Obs: 27554 / De Convoluted HC-3d Calc: 62943; Obs: 62940 Monomer purity: 99+% HIC retention time: 10.2 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-SN38)-Ngly (triazole-SN38)-PSAR18(DAR16) Deconvoluted LC-1d Calc: 26237; Obs: 26237 / De Convoluted HC-3d Calc: 58989; Obs: 58987 Monomer purity: 99+% HIC retention time: 6.6 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide SN38 )-Ngly(triazole-glucuronide SN38)-PSAR18(DAR16) Deconvoluted LC-1d Calc: 27270; Obs: 27270 / De Convoluted HC-3d Calc: 62086; Obs: 62087 Monomer purity: 99+% HIC retention time: 5.7 minutes Trastuzumab-MAL-phenyl-Ngly(triazole-glucuronide MMAE )-PEG12(DAR8)(=ADC-PEG12) Deconvoluted LC-1d Calc: 25541; Obs: 25541 / De Convoluted HC-3d Calc: 56901; Obs: 56900 Monomer purity: 99+% HIC retention time: 7.4 minutes Trastuzumab-glucuronide MMAE (DAR8): Deconvoluted LC-1d Calc: 24721; Obs: 24720 / De Convoluted HC-3d Calc: 54439; Obs: 54438 Monomer purity: 95.2% HIC retention time: 9.2 minutes Trastuzumab-MAL-phenyl-triazole-glucuronide MMAE (DAR8 )(=ADC-PSAR0): Deconvoluted LC-1d Calc: 24884; Obs: 24884 / De Convoluted HC-3d Calc: 54926; Obs: 54926 Monomer purity: 98.5% HIC retention time: 8.4 minutes Trastuzumab-MAL-phenyl-PSAR12-triazole-glucuronide MM AE (DAR8)(=ADC-PSAR12L): Deconvoluted LC-1d Calc: 25793; Obs: 25793 / De Convoluted HC-3d Calc: 57657; Obs: 57657 Monomer purity: 99+% HIC retention time: 8.5 minutes Human albumin-MAL-phenyl-Ngly (triazole-glucuronide MMAE )-PSAR24 (DAR1): Deconvoluted Calc: 69765; Obs: 69645 Monomer purity: 90.8% HIC retention time: 3.9 minutes Trastuzumab: Deconvoluted LC Obs: 23439 / Deconvoluted HC O bs: 50595 Monomer purity: 99+% HIC retention time: 4.7 minutes Anti-CD19 antibody: Deconvoluted LC Obs: 24139 / Deconvoluted HC O bs: 50517 Monomer purity: 93.2% HIC retention time: 4.7 minutes Anti-CD22 antibody: Deconvoluted LC Obs: 24133 / Deconvoluted HC O bs: 50692 Monomer purity: 99+% HIC retention time: 4.8 minutes Human albumin: Deconvoluted Obs: 66556 Monomer purity: 92.4% HIC retention time: 2.5 minutes.

[0364] Example 12: Non-polysarcosine-based antibody-drug conjugate (ADC-PSAR0), Polysarcosine-based antibody-drug conjugate with orthogonal configuration (ADC-PSAR12) and polysarcosine-based antibody-drug conjugate with linear configuration (ADC-PSAR 12L) hydrophobic interaction chromatography (HIC) characteristics Relative exposure of the payload bound to the bulk solvent and trastuzumab-based DAR8 The apparent hydrophobicity of the ADC was evaluated by hydrophobic interaction chromatography (HIC) on a Tosoh TSK- GEL BUTYL-NPR column according to the method described in Example 11. The results are shown in Figure 1. When polysarcosine is grafted in a parallel (i.e., orthogonal) orientation with respect to the drug unit, efficient hydrophobic masking properties are obtained when grafted in a parallel (i.e., orthogonal) orientation with respect to the drug unit, efficient hydrophobic masking properties are obtained , the apparent hydrophobicity of the conjugate (ADC-PSAR12) decreases. However, when the polysialic acid is in a linear (i.e., continuous) arrangement, a decrease in the apparent hydrophobicity of the conjugate is not observed (ADC-PSAR12L).

[0365] Example 13: Hydrophobic interaction chromatography (HIC) properties of polysialic acid- and polyethylene glycol-based antibody-drug-conjugates The relative exposure of the payload bound to the bulk solvent and the apparent hydrophobicity of trastuzumab-based DAR8 ADCs were evaluated by hydrophobic interaction chromatography (HIC) on a Tosoh TSK- GEL BUTYL-NPR column according to the method described in Example 11. The results are shown in Figure 2. At equal length (n = 12 monomer units), polysialic acid gives better hydrophobic masking properties than polyethylene glycol (lower retention time).

[0366] Example 14: Pharmacokinetic properties (total antibody concentration over time) in mice after single intravenous administration of 3 mg / kg dose of non-polysialic acid-based antibody-drug-conjugate (AD C-PSAR0) and polysialic acid-based antibody-drug-conjugate (ADC-PSAR1 2) Male SCID mice (4-6 weeks old) were injected with ADC at 3 mg / kg via the tail vein (5 mice per treatment group, randomly assigned). Blood was collected into citrate tubes via retro-orbital bleeding at various time points and processed into plasma. Total ADC concentration was evaluated using a human IgG ELISA kit (Stemcell (trademark ) Technologies) according to the manufacturer's protocol. A standard curve of trastuzumab was used for quantification. Pharmacokinetic parameters (clearance and AUC ) ​​​​) was calculated by non-compartmental analysis using Microsoft (R) Excel (R) software incorporating PK function (add-in developed by Usansky et al., Department of Pharmacokinetics and Drug Metabolism, Allergan, Irvine, USA). The results are shown in Figure 3. ADCs containing polysarcosine show favorable pharmacokinetics when compared to ADCs not containing polysarcosine.

[0367] Example 15: Tumor volume (mm ) and survival curves of non-polysarcosine-based ADC (ADC-PSAR0) and polysarcosine-based ADC (ADC-PSAR12) at a single intravenous dose of 3 mg / kg in a BT-474 breast cancer xenograft model 3 BT-474 breast cancer cells were subcutaneously transplanted into female SCID mice (4 weeks old). The ADCs from Example 14 were intravenously administered at a dose of 3 mg / kg when the tumors had grown to approximately 150 mm 3 (day 20, 5 animals per group, assigned to minimize the difference in initial tumor volume between groups). The results are shown in Figures 4A and 4B. Tumor volume was measured every 3 - 5 days using a caliper device and calculated using the formula (L × W ) / 2. Mice were sacrificed when the tumor volume exceeded 1000 mm 2 3 . No significant weight change was observed in the treated mice.

[0368] Example 16: Polysarcosine-based antibody-drug conjugate (ADC) at a dose of 3 mg / kg -PSAR12) and polyethylene glycol-based antibody-drug conjugates (ADC -PEG12) after single intravenous administration in mice (total antibody concentration over time ) The experiment was conducted in male CD-1 mice (4-6 weeks old) according to the procedure described in Example 14 as shown in Figure 5. The ADC containing polysarcosine has improved pharmacokinetic parameters compared to the ADC containing polyethylene glycol .

[0369] Example 17: 2.5 mg / kg of polysarcosine-based antibody-drug conjugates (ADC-PSAR6, ADC-PSAR12, ADC -PSAR18, ADC-PSAR24) with different PSAR lengths in orthogonal orientations; orthogonal polyethylene glycol-based antibody -drug conjugates (ADC-PEG12) and linear polysarcosine-based antibody-drug conjugates (ADC-PSAR12L) were intravenously administered once to a BT-474 breast cancer xenograft model for tumor volume (mm ) 3 ) The experiment was conducted as described in Example 15. BT-474 breast cancer cells were subcutaneously transplanted into female SCID mice (4 weeks old). When the tumors grew to approximately 150 mm 3 , the ADC was intravenously administered once at a dose of 2. 5 mg / kg (on day 13, 6 animals per group, assigned to minimize the initial tumor volume difference between groups ). The results are shown in Figure 6. No significant weight changes were observed in the treated mice .

Brief Description of the Drawings

[0370]

Figure 1

Figure 2

Figure 3

Figure 4A

Figure 4B

Figure 5

Figure 6

Claims

1. A ligand-drug-conjugate compound (LDC) having the formula (XV): 【Chemistry 1】 In the formula, L is (HP ＳＭＷ ) is orthogonal to (X-D) an orthogonal connector; HP ＳＭＷ to said orthogonal connector L of a single molecular weight homopolymer having the formula (I): resulting from the covalent bonding of 【Chemistry 2】 (In the formula, R １ and R ２ Unlike, and R １ and R ２ is H or an inert group, and R １ and R ２ The other is functionalization reaction The inert group is a reactive group, which reacts with a bondable group under reaction conditions in which the inert group does not react. reactive to form bonds with Z １ and Z ２ are, either together or separately, any spacer, and n is 1 or more and k is 2 or more; D is a cytotoxic drug; X is any cleavable moiety to release D; Z is an optional spacer; and a is 1 or more, b is 1 or more, and m is 1 or more.

2. 2. The LDC compound of claim 1, wherein the single molecular weight homopolymer is polysarcosine. 。

3. HP ＳＭＷ to the orthogonal connector L of a single molecular weight homopolymer having the formula (II): The LDC compound according to claim 1 or 2, which is derived from the covalent bond of: 【Chemistry 3】 In the formula, R １ , R ２ , Z １ and Z ２ is as defined in claim 1, and k is an integer of 2 to 100, preferably 2 to 50.

4. R １ Or R ２ is a functionalized reactive group selected from the group consisting of: The LDC compound according to any one of claims 1 to 5, - a carboxylic acid group, - Amino group NRR'' wherein R and R″ are independently H and at least one selected from O, N and S. Optionally interrupted by one heteroatom (C １ -C ６ ) alkyl and ( - hydroxyl group, - halogen atoms, - Hydrazine (-NH ２ -NH ２ ) group, - nitro group, - hydroxylamine groups, - azido group, - (C ２ -C ６ ) an alkynyl group, - (C ２ -C ６ ) an alkenyl group, - thiol group, - N-hydroxysuccinimide ester, perfluoroester, nitrophenyl esters, aza-benzotriazoles and benzotriazole activated esters, acyl Activated ester groups such as urea, - boronic acid -B (OR'''') ２ base (wherein R"" is a hydrogen atom or C １ -C ６ is an alkyl group), - Thio groups such as maleimide, halomaleimide, haloacetyl, and pyridyl disulfide reactive groups, - mesylate group, - tosylate group, - triflate group, - aldehyde group, - isocyanate or isothiocyanate groups, - chlorosulfonyl group, - acrylate groups.

5. The ligand is selected from the group consisting of a polypeptide, a protein, an antibody, and an antibody fragment. The LDC compound according to any one of claims 1 to 4.

6. D is selected from the group consisting of biologically active molecules, therapeutic molecules such as anticancer drugs, imaging agents and fluorophores.

6. The LDC compound according to claim 1 , wherein the LDC compound is selected from the group consisting of:

7. 7. The method according to claim 1, wherein L is one or more natural or unnatural amino acids. LDC compounds.

8. 8. Any one of claims 1 to 7, wherein L is selected from glutamic acid, lysine and glycine. The LDC compound according to claim 1.

9. X is, one or more natural or unnatural amino acids, - a sugar moiety linked to a self-immolative group via an oxygen glycosidic bond, - a disulfide linker, and - Acid-labile linker that is hydrolyzable in the lysosome 9. The LDC compound according to claim 1, selected from:

10. X is, one or more natural or unnatural amino acids, and - A sugar moiety linked to a self-immolative group via an oxygen glycosidic bond 10. The LDC compound according to claim 1, selected from:

11. Z is an alkylene, heteroalkylene; an alkoxy; a polyether; one or more natural or is an unnatural amino acid; ３ -C ８ Heterocyclo; C ３ -C ８ Carbocyclo;Arylene; and any combination thereof. DC compound.

12. Z is a group represented by the formula (XVII), the formula (XVIII), the formula (XIX), the formula (XX), or the formula (XXI). or formula (XXII): 【Chemistry 4】 where the wavy bond represents the point of attachment, and R ６ is -C １ -C １０ Alkylene-, -C １ -C １０ Heteroalkylene-, -C １ -C １０ Alkylene-C(=O)-, -C １ -C １０ heteroalkylene-C(=O)-, -Arylene-C １ -C １０ Alkylene-C(=O)-, -arylene-C １ -C １０ a alkylene-O-C(=O)-, R ６ Any of the groups is optionally substituted with one or more =O will be done.

13. An intermediate compound having the formula (XVI): 【Chemistry 5】 where L is an orthogonal connector; HP ＳＭＷ to said orthogonal connector L of a single molecular weight homopolymer having the formula (I): resulting from the covalent bonding of 【Chemistry 6】 (In the formula, R １ and R ２ Unlike, and R １ and R ２ is H or an inert group, and R １ and R ２ The other is functionalization reaction The inert group is a reactive group, which reacts with a bondable group under reaction conditions in which the inert group does not react. reactive to form bonds with Z １ and Z ２ are, either together or separately, any spacer, and n is 1 or more and k is 2 or more; D is a cytotoxic drug; X is any cleavable moiety to release D; Z is an optional spacer; and a is 1 or more, and b is 0 or 1 or more.

14. 14. The intermediate of claim 13, wherein the single molecular weight homopolymer is polysarcosine. Compound.

15. 13. A LDC according to any one of claims 1 to 12 for use as a medicament.

16. Compounds having formula (XXIII): 【Chemistry 7】 In the formula, R ６ is -C １ -C １０ Alkylene-, -C １ -C １０ Heteroalkylene-, -C ３ -C ８ Carbocyclo-, -O-(C １ -C ８ Alkyl)-, -arylene-, -C １ -C １０ Alkylene-arylene-, -arylene-C １ -C １０ Alkylene-, -C １ -C １０ Alkylene-(C ３ -C ８ Carbocyclo)-, -(C ３ -C ８ Carbocyclo) -C １ -C １０ Alkylene-, -C ３ -C ８ Heterocyclo-, -C １ -C １０ Alkylene - (C ３ -C ８ Heterocyclo)-, -(C ３ -C ８ Heterocyclo)-C １ -C １０ Alki Ren-, -C １ -C １０ Alkylene-C(=O)-, -C １ -C １０ Heteroalkylene - C(=O)-, -C ３ -C ８ Carbocyclo-C(=O)-, -O-(C １ -C ８ Alkyl )-C(=O)-, -arylene-C(=O)-, -C １ -C １０ Alkylene-Aryl Arylene-C(=O)-, -arylene-C １ -C １０ Alkylene-C(=O)-, -C １ -C １０ Alkylene-(C ３ -C ８ Carbocyclo)-C(=O)-, -(C ３ -C ８ Carbosi Black) - C １ -C １０ Alkylene-C(=O)-, -C ３ -C ８ Heterocyclo-C(=O ) -, -C １ -C １０ Alkylene-(C ３ -C ８ Heterocyclo)-C(=O)-, -(C ３ -C ８ Heterocyclo)-C １ -C １０ Alkylene-C(=O)-, -C １ -C １０ Al Kiren-NH-, -C １ -C １０ Heteroalkylene -NH-, -C ３ -C ８ Carbocyclo -NH-, -O-(C １ -C ８ alkyl)-NH-, -arylene-NH-, -C １ -C １０ Alkylene-arylene-NH-, -arylene-C １ -C １０ Alkylene -NH- , -C １ -C １０ Alkylene-(C ３ -C ８ Carbocyclo)-NH-, -(C ３ -C ８ mosquito Rubocyclo)-C １ -C １０ Alkylene -NH-, -C ３ -C ８ heterocyclo-NH-, -C １ -C １０ Alkylene-(C ３ -C ８ Heterocyclo)-NH-, -(C ３ -C ８ Haeta Rocyclo)-C １ -C １０ Alkylene -NH-, -C １ -C １０ Alkylene-S-, -C １ -C １０ Heteroalkylene -S-, -C ３ -C ８ Carbocyclo-S-, -O-(C １ - C ８ Alkyl)-S-, -arylene-S-, -C １ -C １０ Alkylene-Aryl Arylene-S-, -Arylene-C １ -C １０ Alkylene-S-, -C １ -C １０ Alkylene - (C ３ -C ８ Carbocyclo)-S-, -(C ３ -C ８ Carbocyclo)-C １ -C １０ Al Kiren-S-, -C ３ -C ８ Heterocyclo-S-, -C １ -C １０ Alkylene-(C ３ - C ８ Heterocyclo)-S-, -(C ３ -C ８ Heterocyclo)-C １ -C １０ Alkylene - S-, -C １ -C １０ Alkylene -O-C(=O)-, -C ３ -C ８ Carbocyclo-O- C(=O)-, -O-(C １ -C ８ Alkyl)-O-C(=O)-, -arylene-O- C(=O)-, -C １ -C １０ Alkylene-arylene-O-C(=O)-, -Arylene N-C １ -C １０ Alkylene -O-C(=O)-, -C １ -C １０ Alkylene-(C ３ - C ８ Carbocyclo)-O-C(=O)-, -(C ３ -C ８ Carbocyclo)-C １ -C １０ Alkylene -O-C(=O)-, -C ３ -C ８ Heterocyclo-O-C(=O)-, -C １ -C １０ Alkylene-(C ３ -C ８ Heterocyclo)-O-C(=O)-, -(C ３ -C ８ Heterocyclo)-C １ -C １０ alkylene -O-C(=O)-; R ６ Any of the groups may be -X, -R', -O － , -OR', =O, -SR', -S － , -NR’ ２ 、-NR’ ３ ＋ 、=NR’、-CX ３ 、-CN、-OCN、-SCN、-N= C=O、-NCS、-NO、-NO ２ 、=N ２ 、-N ３ 、-NR’C(=O)R’、-C (=O)R’、-C(=O)NR’ ２ 、-SO ３ － 、-SO ３ H、-S(=O) ２ R’、 -OS(=O) ２ OR’、-S(=O) ２ NR’、-S(=O)R’、-OP(=O)( OR’) ２ 、-P(=O)(OR’) ２ 、-PO ３ － 、-PO ３ H ２ 、-C(=O)X、 -C(=S)R'、-CO ２ R'、-CO ２ 、-C==OOR、C==OO)SR'、 C(=S)SR', C(=O)NR' ２ , C(=S)NR' ２ and C(=NR')NR ' ２ wherein each X is independently selected from halogen: -F, -CI, -Br, or -I, and each R' is independently -H, -C １ -C ２０ Alkyl, -C ６ -C ２０ Aryl, or -C ３ -C １４ heterocyclic ring); Z is an optional spacer; L is an orthogonal connector; X is any cleavable moiety to release D; D is a cytotoxic drug; a is 1 or more, and b is 0 or 1 or more; and HP ＳＭＷ to said orthogonal connector L of a single molecular weight homopolymer having the formula (I): Arising from the covalent bonding of: 【Chemistry 8】 (In the formula, R １ and R ２ Unlike, and R １ and R ２ is H or an inert group, and R １ and R ２ The other is functionalization reaction The inert group is a reactive group, which reacts with a bondable group under reaction conditions in which the inert group does not react. reactive to form bonds with Z １ and Z ２ are, either together or separately, any spacer, and n is 1 or more and k is 2 or more).

17. 17. The compound of claim 16, wherein the single molecular weight homopolymer is polysarcosine.

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