Compound comprising fc-binding unit, and conjugate prepared using same

Abstract

Some aspects of the present application provide a compound comprising an Fc-binding unit. The compound comprising an Fc-binding unit, of the present application, can be used for site-specific delivery of a target group to an antibody. In addition, some aspects of the present application provide a method for preparing an antibody conjugate comprising a target group (for example, a reactive group) by using a compound comprising an Fc-binding unit.

Description

Compounds comprising FC binding units and conjugates prepared using the same

[0001] The present application relates to compounds comprising an Fc-binding unit, conjugates prepared using the same, and methods for preparing conjugates using compounds comprising an Fc-binding unit. Compounds comprising an Fc-binding unit provided according to some embodiments of the present application enable delivery of a target substance (e.g., a reactive group) to a target location on an antibody. That is, a target substance can be site-specifically delivered to an antibody via a compound comprising an Fc-binding unit.

[0002] Furthermore, the present application provides an antibody-functional group conjugate (e.g., an antibody-drug conjugate) characterized by having a branched linker.

[0003]

[0004]

[0005] Antibodies are biomolecules that recognize specific molecules and are used in a variety of industrial applications. For example, antibodies can be used to detect or screen specific substances, identify the pathways through which specific substances move within the body or cells, and induce immune responses to specific substances for therapeutic purposes.

[0006] To expand the functionality of these antibodies, attempts have been made to improve them. Representative examples include attempts to label or conjugate various substances (e.g., drugs or radioactive moieties) to complement or expand their functions. For example, antibodies can be labeled with fluorescent substances for use in fluorescence assays, or agents for treating specific diseases can be labeled or conjugated to antibodies to maximize their therapeutic efficacy. These attempts and technologies may be referred to as antibody labeling or antibody-target moiety conjugation, and the present application relates to antibody labeling or antibody-target moiety conjugation.

[0007] In past studies, antibody-target conjugates were prepared using highly reactive amino acid residues (e.g., amine groups or thiol groups) among the amino acid residues that constitute antibodies. Specifically, reactive groups capable of reacting with the above residues were introduced into a target moiety, and then a target moiety capable of reacting with the reactive residues of the antibody (more precisely, a modified target moiety into which the above reactive groups were introduced) was prepared. An antibody-target conjugate was prepared through a reaction between the modified target moiety and the antibody.

[0008] These past studies were conducted by randomly attaching target substances to antibodies, and this past method caused many problems.

[0009] Essentially, past methods could not precisely control the "location" where the target moiety binds to the antibody, nor could they precisely control the "number" of targets bound to the antibody. In other words, antibody-target moiety conjugates manufactured using conventional methods suffer from the problem of drug structure inhomogeneity.

[0010] This heterogeneity in drug structure inevitably leads to heterogeneity in drug efficacy, arising from the "differences" in drug structure. This problem has significantly hindered the development of antibody-drug conjugate (ADC) technology, which demands high safety and reproducibility.

[0011] Furthermore, the heterogeneity of drug structures can lead to problems that hinder antibody function. Antibodies comprise a Fab region, which contains an antigen-binding domain that recognizes antigens, and an Fc region, which is involved in antibody crystallization. Non-site-specific conjugation / labeling makes it impossible to precisely control the binding site of the target moiety on the antibody. Consequently, it is impossible to prevent the target moiety from binding to the antigen-binding domain of the antibody or a site adjacent to it, thereby disrupting the antibody's recognition function.

[0012] This has led to a demand in this field for techniques for site-specific antibody labeling to ensure structural homogeneity of antibody-target conjugates. While some techniques have been developed, most have lacked technical and economic benefits, such as requiring genetic manipulation or modification of antibodies.

[0013] Under the circumstances described above, active development of technologies for site-specifically delivering a group of interest to an antibody has been underway.

[0014] In particular, the document [Korean Patent Application No. 10-2020-0091826 (Application No. 10-2020-0009162)] discloses a technique for delivering a target substance, a click chemistry functional group, to an antibody using a compound comprising an Fc-binding unit. Specifically, the document discloses a technique for position-specifically delivering a target substance to an antibody, either with or together with the Fc-binding unit, using a compound comprising an Fc-binding unit. Meanwhile, with respect to a compound comprising an Fc-binding unit for delivering a target substance to an antibody with the release of the Fc-binding unit, according to the research results disclosed in the present application by the inventors of the present application, the compound comprising an Fc-binding unit disclosed in Patent Application No. 10-2020-0091826 has several problems and is therefore not suitable for use. Accordingly, the inventors of the present application developed a compound comprising a novel and improved Fc binding unit based on patent application No. 10-2020-0091826.

[0015]

[0016]

[0017] A method for position-specifically delivering a target substance (e.g., a reactive group or a functional group) to an antibody via a compound comprising an Fc-binding unit has been developed. However, when using the compound disclosed in Patent Application No. 10-2020-0091826, the yield of the desired product, i.e., a conjugate, is low, the reaction time is too long, or the conjugate cannot be obtained. Accordingly, the present application provides a compound comprising an Fc-binding unit having an improved or enhanced effect (e.g., enhanced reaction efficiency).

[0018]

[0019]

[0020] The present application provides a compound comprising an Fc-binding unit. The compound comprising the Fc-binding unit of the present application can be used in the preparation of an antibody conjugate.

[0021] Furthermore, the present application provides a method for preparing an antibody conjugate (e.g., an antibody conjugate comprising a desired group or an antibody conjugate comprising a reactive group) using a compound comprising the Fc binding unit of the present application.

[0022] Furthermore, the present application provides a method of preparing an antibody-payload conjugate using a compound, antibody, and payload comprising the Fc binding unit of the present application; or an antibody conjugate and payload comprising the reactive group of the present application.

[0023] Furthermore, the present application provides antibody-payload conjugates comprising a branched linker and payloads for preparing the same, which have improved or enhanced efficacy (e.g., improved plasma stability).

[0024]

[0025]

[0026] The present application provides compounds comprising an Fc binding unit having improved or enhanced effects (e.g., improved reaction efficiency and / or improved stability).

[0027] Furthermore, the present application discloses antibody-payload conjugates comprising a branched linker having improved or enhanced efficacy (e.g., improved plasma stability).

[0028]

[0029]

[0030] Figure 01 illustrates the positions of lysine residues on the Fc region, including lysine 246 and lysine 248.

[0031] Figures 02 and 03 show the positional relationship between the Fc binding peptide and the Fc region.

[0032] Figure 04 is Xa of Fc binding peptide 1 Lysine 246 and lysine 248 in the Fc region are depicted.

[0033] Figure 05 shows the amine group of lysine 246 in the Fc region and Xa of the Fc-binding peptide. 1 It is about the distance between the beta carbons.

[0034] Figure 06 shows the amine group of lysine 248 in the Fc region and Xa of the Fc-binding peptide. 1 It is about the distance between the beta carbons.

[0035] Figure 07 illustrates the reaction of a compound comprising the Fc binding unit of the present application with an antibody, and an antibody conjugate comprising the desired group produced by the reaction.

[0036] Figure 08 illustrates an antibody conjugate comprising two target groups, wherein the target groups are linked to K246 of the Fc region of the antibody.

[0037] Figure 09 illustrates an antibody conjugate comprising two target groups, wherein the target groups are linked to K248 of the Fc region of the antibody.

[0038] Figure 10 illustrates an antibody conjugate comprising two target groups, one of which is linked to K246 and the other is linked to K248.

[0039] FIG. 11 relates to one embodiment of a method for preparing an antibody conjugate comprising a desired group.

[0040] Figure 12 illustrates an antibody and an antibody unit.

[0041] FIG. 13 relates to one embodiment of a method for preparing an antibody-payload conjugate.

[0042] Figure 14 shows the results of confirming the conjugation efficiency of compound 1.

[0043] Figure 15 shows the results of confirming the conjugation efficiency of compound 3.

[0044] Figure 16 shows the results of confirming the conjugation efficiency of compound 4.

[0045] Figure 17 shows the results of confirming the conjugation efficiency of compound 5.

[0046] Figure 18 shows the results of confirming the conjugation efficiency of each of compounds 6 and 7.

[0047] Figures 19 and 20 show the results of confirming the conjugation efficiency of compound 8.

[0048] Figure 21 shows the results of confirming the conjugation efficiency of each of compounds 6 to 8 according to a long-term reaction.

[0049] Figure 22 shows the results of confirming the conjugation efficiency of each compound when the reaction time between the compound (compound 6 to compound 8) and the antibody is 48 hours.

[0050] Figures 23 to 24 illustrate a schematic diagram of the preparation of an antibody-payload conjugate using compound 9, anti-CLDN18.2 mAb, and payload 1.

[0051] Figure 25 specifically illustrates the (PEG8)2-BG-MMAE portion of the antibody-payload conjugate (DAR2) of Figure 24.

[0052] Figures 26 and 27 show the results of confirming the conjugation efficiency of compound 9.

[0053] Figure 28 shows the results of confirming the conjugation efficiency of compound 9.

[0054] Figure 29 shows the results of confirming the conjugation efficiency of compound 10.

[0055] Figure 30 shows the results of confirming the conjugation efficiency of compound 11.

[0056] Figures 31 and 32 show the results of confirming the conjugation efficiency of compound 12.

[0057] Figure 33 shows the results of stability verification of compound 14.

[0058] Figure 34 is a graph showing the measured absorbance after treatment with antibody-A and ADC-A, respectively, for a CHO-K1 cell line (MOCK CHO-K1) transiently delivered with only the NOCK vector (empty vector).

[0059] Figure 35 is a graph showing the measured absorbance after treatment with antibody-A and ADC-A, respectively, for CHO-K1 cell lines (Claudin 18.1 CHO-K1) transiently transfected with a gene (SEQ ID NO: 29) encoding claudin 18.1 protein (CLDN18.1).

[0060] Figure 36 is a graph showing the measured absorbance after treatment with antibody-A and ADC-A, respectively, for a CHO-K1 cell line (Claudin 18.2 CHO-K1) transiently transfected with a gene (SEQ ID NO: 30) encoding claudin 18.2 protein (CLDN18.2).

[0061] Figure 37 is a graph showing the measured absorbance after antibody-A and ADC-A were respectively treated on CLDN18.2~VLP (Virus-Like Particle) expressing claudin 18.2 protein (CLDN18.2).

[0062] Figure 38 is a graph showing the measured absorbance after MIA PaCa-2~CLDN18.2 cell lines were treated with antibody-A, ADC-A, antibody-B, and ADC-B, respectively.

[0063] Figure 39 is a graph showing the measured absorbance after each treatment of the SNU601 cell line with antibody-A, ADC-A, antibody-B, and ADC-B.

[0064] Figure 40 is a graph showing the measured absorbance after the PATU8988S cell line was treated with antibody-A, ADC-A, antibody-B, and ADC-B, respectively.

[0065] Figure 41 is a graph showing the measured absorbance after treatment of MIA PaCa-2 (CLDN18.2-) cell line with antibody-A, ADC-A, antibody-B, and ADC-B, respectively.

[0066] Figure 42 is a graph showing the internalization levels measured every hour after treatment of MIA PaCa-2~CLDN18.2 cell lines with antibody-A and ADC-A, respectively. Here, the red area ratio of the cell axis refers to the ratio of the area of ​​antibody-A or ADC-A that has penetrated into the cell (area measured by red dots) to the total area occupied by the cell.

[0067] Figure 43 is a graph showing the internalization levels measured every hour after treatment of MIA PaCa-2 (CLDN18.2-) cells with antibody-A and ADC-A, respectively. Here, the red area ratio of the cell axis refers to the ratio of the area of ​​antibody-A or ADC-A that has penetrated into the cell (area measured by red dots) to the total area occupied by the cell.

[0068] Figure 44 is a graph depicting the internalization levels measured every hour after treatment of SNU601 cells with antibody-A and ADC-A, respectively. Here, the red area ratio of the cell axis refers to the ratio of the area of ​​antibody-A or ADC-A that has penetrated into the cell (the area measured by the red dots) to the total area occupied by the cell.

[0069] Figure 45 is a graph showing the changes in cell viability measured after treatment of MIA PaCa-2~CLDN18.2 cell lines with various concentrations of antibody-A, ADC-A, antibody-B, ADC-B, and a combination of antibody-A and MMAE.

[0070] Figure 46 is a graph showing the changes in cell viability measured after treating the PATU8988S cell line with various concentrations of antibody-A, ADC-A, antibody-B, ADC-B, and a combination of antibody-A and MMAE.

[0071] Figure 47 is a graph showing the changes in cell viability measured after treating SNU601 cell line with various concentrations of antibody-A, ADC-A, antibody-B, ADC-B, and a combination of antibody-A and MMAE.

[0072] Figure 48 is a graph showing the changes in cell viability measured after treating NUGC4 cell line with various concentrations of antibody-A, ADC-A, antibody-B, ADC-B, and a combination of antibody-A and MMAE.

[0073] Figure 49 is a graph showing the changes in cell viability measured after treatment of MIA PaCa-2 (CLDN18.2-) cell line with various concentrations of antibody-A, ADC-A, antibody-B, ADC-B, and a combination of antibody-A and MMAE.

[0074] Figure 50 is a graph showing the changes in cell viability measured after treating AGS cell lines with various concentrations of antibody-A, ADC-A, antibody-B, ADC-B, and a combination of antibody-A and MMAE.

[0075] Figure 51 is a graph showing the relative proportions of total antibodies or total ADC measured after supernatants were extracted from samples cultured for various periods of time after treatment with antibody-A, ADC-A and ADC-B in human plasma, respectively.

[0076] Figure 52 is a graph showing the relative proportions of total antibodies or total ADC measured after supernatants were extracted from samples cultured for various periods of time after monkey plasma was treated with antibody-A, ADC-A and ADC-B, respectively.

[0077] Figure 53 is a graph showing the relative proportions of total antibodies or total ADC measured after supernatants were extracted from samples cultured for various periods of time after treatment with antibody-A, ADC-A and ADC-B in rat plasma, respectively.

[0078] Figure 54 is a graph showing the relative proportions of total antibodies or total ADC measured after supernatants were extracted from samples cultured for various periods of time after treatment with antibody-A, ADC-A and ADC-B in mouse plasma, respectively.

[0079] Figure 55 is a graph showing the measured tumor volumes after intravenous injection of antibody-A, ADC-A, and ADC-C into tumor model mice under various conditions (groups G1 to G8).

[0080] Figure 56 is a graph showing the measured body weights after intravenous injection of antibody-A, ADC-A, and ADC-C into tumor model mice under various conditions (groups G1 to G8).

[0081] Figure 57 shows comparative photographs of tumors that were intravenously injected into tumor model mice with antibody-A, ADC-A, and ADC-C under various conditions (groups G1 to G8), and autopsied 28 days later.

[0082] Figure 58 is a graph showing the weight of tumors 28 days after intravenous injection of antibody-A, ADC-A, and ADC-C into tumor model mice under various conditions (groups G1 to G8).

[0083] Figure 59 is a graph showing the concentrations of total antibody and total ADC measured after intravenous administration of ADC-A at various concentrations to rats over a certain period of time.

[0084] Figure 60 shows the stability test results of compounds 5 to 9.

[0085]

[0086] Some embodiments of the present application provide compounds comprising an Fc binding unit.

[0087] Some embodiments of the present application provide a compound comprising an Fc binding unit having the structure of Formula 2-2:

[0088] [Chemical Formula 2-2]

[0089] ,

[0090] At this time,

[0091] D a is a spacer A, wherein the spacer A is a bond, substituted or unsubstituted C 1-20 Alkylene, substituted or unsubstituted C 1-20 Heteroalkylene, substituted or unsubstituted C 2-20Alkenylene, substituted or unsubstituted C 2-20 Heteroalkenylene, substituted or unsubstituted C 2-20 Alkynylene, or substituted or unsubstituted C 2-20 Heteroalkynylene, wherein substituted indicates that one or more hydrogen atoms in the group described by the term substituted is replaced by one or more substituents, wherein the substituents are each independently selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH, wherein the substituent is not H, and wherein the heteroalkylene, heteroalkenylene, heteroalkynylene, heterocycloalkyl, or heteroaryl comprises one or more heteroatoms, wherein each heteroatom is independently selected from N, O, and S,

[0092] L a is a linker A, wherein the linker A is a bond, substituted or unsubstituted C 1-100 Alkylene, substituted or unsubstituted C 1-100 Heteroalkylene, substituted or unsubstituted C 2-100 Alkenylene, substituted or unsubstituted C 2-100 Heteroalkenylene, substituted or unsubstituted C 2-100 Alkynylene, or substituted or unsubstituted C 2-100Heteroalkynylene, wherein substituted means that one or more hydrogen atoms in the group described by the term substituted are substituted with one or more substituents, wherein the substituents are each independently selected from -R, =O, =S, -NO2, -CR3, -NR2, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH, wherein the substituent is not H, and wherein the heteroalkylene, heteroalkenylene, heteroalkynylene, heterocycloalkyl, or heteroaryl comprises one or more heteroatoms, wherein each heteroatom is independently selected from N, O, and S,

[0093] X is -CH2-, -O-, or -NH-,

[0094] R a1 is H or C 1-6 It is alkyl,

[0095] R a2 is H or C 1-6 It is alkyl,

[0096] R a3 is H or C 1-6 It is alkyl,

[0097] J a is -C(=O)-, -S-, -NH-, or -C(=NH)-,

[0098] RG is a reactive group, wherein the reactive group comprises a reactive moiety,

[0099] FcBU is an Fc binding unit, wherein the Fc binding unit has a structure Have,

[0100] At this time,

[0101] Each Xaa is independently any amino acid residue,

[0102] Xa 2 is a glutamic acid residue or an asparagine residue,

[0103] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue,

[0104] The cysteine ​​residue adjacent to the N terminus and the cysteine ​​residue adjacent to the C terminus are optionally covalently linked,

[0105] Xa 1 'silver and,

[0106] Here, m is an integer greater than or equal to 1 and less than or equal to 5,

[0107] J f is -NH-, -S-, or -C(=O)-,

[0108] * and ** are each Xa 1 ' represents the attachment point with the adjacent amino acid residue,

[0109] *** is Xa 1 ', and represents an attachment site with a non-Fc-binding unit portion of a compound including an Fc-binding unit.

[0110] In certain embodiments, X can be -O- or -CH2-.

[0111] In certain embodiments, X can be -O-.

[0112] In certain embodiments, R a1 Silver C 1-3 It could be an alkyl.

[0113] In certain embodiments, R a1 can be methyl.

[0114] In certain embodiments, R a2 and R a3 are each independently H and C 1-3 It can be any one selected from alkyl.

[0115] In certain embodiments, R a2 and R a3 can all be H.

[0116] In certain embodiments, J a can be -C(=O)-.

[0117] In certain embodiments, D a is unsubstituted C 1-10 alkylene, unsubstituted C 1-10 Heteroalkylene, unsubstituted C 2-10 Alkenylene, or unsubstituted C 2-10 It may be a heteroalkenylene.

[0118] In certain embodiments, D a is unsubstituted C 1-10 It may be alkylene.

[0119] In certain embodiments, the Fc binding unit can have the structure:

[0120] .

[0121] In certain embodiments, J f can be -NH-.

[0122] In certain embodiments, m can be an integer greater than or equal to 1 and less than or equal to 4.

[0123] In certain embodiments, m may be 3.

[0124] In certain embodiments, L a is a bond, unsubstituted C 1-60 alkylene, unsubstituted C 1-60 Heteroalkylene, unsubstituted C 2-60 Alkenylene, or unsubstituted C 2-60 It may be a heteroalkenylene.

[0125] In certain embodiments, L a is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60Heteroalkylene, wherein the unsubstituted heteroalkylene may contain 0 to 20 ethylene glycol units.

[0126] In certain embodiments, L a is a bond, unsubstituted C 1-30 Alkylene, or unsubstituted C 1-30 Heteroalkylene, wherein the unsubstituted heteroalkylene may contain 0 to 10 ethylene glycol units.

[0127] In certain embodiments, L a is a bond, unsubstituted C 1-24 Alkylene, or unsubstituted C 1-24 Heteroalkylene, wherein the unsubstituted heteroalkylene may contain 0 to 8 ethylene glycol units.

[0128] In certain embodiments, L a Is and, at this time, sf may be an integer from 0 to 8, sg may be an integer from 0 to 15, and sh may be an integer from 0 to 8. In a specific embodiment, at this time, sf may be an integer from 0 to 3, sg may be an integer from 0 to 10, and sh may be an integer from 0 to 3.

[0129] In certain embodiments, RG can be represented by the following structure:

[0130] ,

[0131] At this time, D RG is the spacer of the reactive group (spacer RG),

[0132] At this time, the spacer of the above reactive group is bonded, substituted or unsubstituted C 1-6 Alkylene, substituted or unsubstituted C 1-6 Heteroalkylene, substituted or unsubstituted C 2-6 Alkenylene, or substituted or unsubstituted C 2-6Heteroalkenylene, wherein substituted means that one or more hydrogen atoms in the group modified by the term substituted are replaced by one or more substituents, wherein the substituents are each independently -C 1-4 alkyl, -C(=O)H, -C(=O)CH3, -C(=O)OH, -C(=O)NH2, -NH2, =NH, =O, =S, -OH, -NO2, and -SH, wherein the heteroalkylene or heteroalkenylene contains one or more heteroatoms, wherein the heteroatoms are each independently selected from O, N, and S,

[0133] H RG is a reactive moiety.

[0134] In certain embodiments, the reactive moiety may be a bioorthogonal functional group.

[0135] In certain embodiments, the reactive moiety may have any of the following structures:

[0136]

[0137]

[0138] and

[0139] ,

[0140] At this time,

[0141] hn is an integer from 1 to 3,

[0142] R H are each independently H or -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10Selected from heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH.

[0143] In a particular embodiment, the reactive moiety is selected from an azide group, a terminal alkyne group, a terminal alkene group, a cyclooctyne group, a tetrazine group, a norbornene group, a cyclooctene group, an oxime group, and an isocyanide group, wherein the cyclooctyne group is any one selected from OCT cyclooctyne, BCN (Bicyclononyne), DBCO (Dibenzocyclooctyne), DIBAC (aza-dibenzocyclooctynes), DIBO (dibenzocyclooctynol), DIFO (difluorinated cyclooctynes), BARAC (biarylazacyclooctynone), DIMAC (dimethoxyazacyclooctyne), and DIFBO (difluorobenzocyclooctyne), and wherein the cyclooctene group may be any one selected from a cis-cyclooctene group and a trans-cyclooctene group.

[0144]

[0145] Some embodiments of the present application provide compounds comprising an Fc binding unit having the structure of Formula 2-15:

[0146] [Chemical Formula 2-15]

[0147] ,

[0148] At this time,

[0149] aa is an integer greater than or equal to 1 and less than or equal to 10,

[0150] X is -O- or -CH2-,

[0151] L a is a linker A, wherein the linker A is a bond, unsubstituted C 1-30alkylene, or unsubstituted C containing 0 to 10 ethylene glycol units 1-30 It is heteroalkylene,

[0152] is a reactive group,

[0153] D RG is a spacer of a reactive group (spacer RG), wherein the spacer of the reactive group is a bond, substituted or unsubstituted C 1-6 Alkylene, substituted or unsubstituted C 1-6 Heteroalkylene, substituted or unsubstituted C 2-6 Alkenylene, or substituted or unsubstituted C 2-6 Heteroalkenylene, wherein substituted means that one or more hydrogen atoms in the group modified by the term substituted are replaced by one or more substituents, wherein the substituents are each independently -C 1-4 alkyl, -C(=O)H, -C(=O)CH3, -C(=O)OH, -C(=O)NH2, -NH2, =NH, =O, =S, -OH, -NO2, and -SH, wherein the heteroalkylene or heteroalkenylene contains one or more heteroatoms, wherein the heteroatoms are each independently selected from O, N, and S,

[0154] H RG is a reactive moiety,

[0155] FcBU is an Fc binding unit, wherein the Fc binding unit has a structure

[0156] Have,

[0157] At this time,

[0158] Each Xaa is independently any amino acid,

[0159] Xa 2 is a glutamic acid residue or an asparagine residue,

[0160] Xa 3is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue,

[0161] The cysteine ​​residue adjacent to the N terminus and the cysteine ​​residue adjacent to the C terminus are optionally covalently linked,

[0162]

[0163] Xa 1 'silver and,

[0164] Here, m is an integer greater than or equal to 1 and less than or equal to 4,

[0165] * and ** are each Xa 1 ' represents the attachment point with the adjacent amino acid residue,

[0166] *** is Xa 1 ', and represents an attachment site with a non-Fc-binding unit portion of a compound including an Fc-binding unit.

[0167] In certain embodiments, aa can be an integer greater than or equal to 1 and less than or equal to 6.

[0168] In certain embodiments, aa can be 3.

[0169] In certain embodiments, the Fc binding unit can have the structure:

[0170] .

[0171] In certain embodiments, m may be 3.

[0172] In certain embodiments, the reactive moiety may be a bioorthogonal functional group.

[0173] In certain embodiments, the reactive moiety may have any of the following structures:

[0174]

[0175]

[0176] and

[0177] ,

[0178] At this time,

[0179] hn is an integer from 1 to 3,

[0180] R H are each independently H or -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Selected from heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH.

[0181] In a particular embodiment, the reactive moiety is selected from an azide group, a terminal alkyne group, a terminal alkene group, a cyclooctyne group, a tetrazine group, a norbornene group, a cyclooctene group, an oxime group, and an isocyanide group, wherein the cyclooctyne group is any one selected from OCT cyclooctyne, BCN (Bicyclononyne), DBCO (Dibenzocyclooctyne), DIBAC (aza-dibenzocyclooctynes), DIBO (dibenzocyclooctynol), DIFO (difluorinated cyclooctynes), BARAC (biarylazacyclooctynone), DIMAC (dimethoxyazacyclooctyne), and DIFBO (difluorobenzocyclooctyne), and wherein the cyclooctene group may be any one selected from a cis-cyclooctene group and a trans-cyclooctene group.

[0182]

[0183] Some embodiments of the present application provide methods for preparing an antibody conjugate comprising a reactive group.

[0184] Some embodiments of the present application provide a method of preparing an antibody conjugate comprising a reactive group, comprising:

[0185] Contacting a compound comprising the Fc binding unit of the present application with an antibody, wherein the compound comprising the Fc binding unit comprises a reactive group.

[0186] In certain embodiments, the antibody may be an IgG antibody.

[0187] In certain embodiments, the antibody is an IgG antibody, and the IgG antibody can be a human IgG antibody, a humanized IgG antibody, or a chimeric IgG antibody.

[0188] In a particular embodiment, the Fc region of the antibody comprises the amino acid sequence of SEQ ID NO: 14 or an amino acid sequence having at least 80% sequence identity thereto, wherein the Fc region of the antibody may comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and the amino acid sequence of MHEALHNHY (SEQ ID NO: 12).

[0189] In certain embodiments, contacting an antibody with a compound comprising an Fc binding unit results in transfer of a reactive group to a targeting region of the antibody, wherein the targeting region consists of five consecutive amino acid residues and may include K246 and K248 of the Fc region of the antibody.

[0190] In certain embodiments, contacting an antibody with a compound comprising an Fc binding unit can result in transfer of a reactive group to one or more of K246 and K248 of the Fc region of the antibody.

[0191] In certain embodiments, the method of preparing an antibody conjugate comprising the reactive group may further comprise obtaining an antibody conjugate comprising the reactive group.

[0192] In certain embodiments, an antibody conjugate comprising a reactive group is prepared by contacting an antibody with a compound comprising an Fc binding unit, wherein the antibody conjugate comprising a reactive group may comprise from one to four reactive groups.

[0193] In certain embodiments, an antibody conjugate comprising a reactive group is prepared by contacting an antibody with a compound comprising an Fc binding unit,

[0194] At this time, the antibody conjugate containing the above reactive group contains two reactive groups,

[0195] In the antibody conjugate containing the reactive group at this time, one of the two reactive groups (the first reactive group) is linked to one of K246 and K248 of one of the two heavy chains of the antibody (the first heavy chain),

[0196] In the antibody conjugate containing the reactive group at this time, the other reactive group (the second reactive group) of the two reactive groups can be linked to either K246 or K248 of the other heavy chain (the second heavy chain) of the two heavy chains of the antibody.

[0197] In certain embodiments, an antibody conjugate comprising a reactive group is prepared by contacting an antibody with a compound comprising an Fc binding unit,

[0198] At this time, the antibody conjugate containing the above reactive group contains two reactive groups,

[0199] At this time, each of the two reactive groups may be linked to K246 of one heavy chain of the antibody and K246 of the other heavy chain, or may be linked to K248 of one heavy chain of the antibody and K248 of the other heavy chain, respectively.

[0200] In certain embodiments, contacting a compound comprising an Fc-binding unit with an antibody can be accomplished by a method comprising: mixing a composition comprising a compound comprising an Fc-binding unit with a composition comprising an antibody. In certain embodiments, the mixing of the composition comprising a compound comprising an Fc-binding unit with a composition comprising an antibody can be performed under conditions of pH 6 to pH 8.5.

[0201]

[0202]

[0203] Below, the invention is described in more detail through embodiments and examples. The invention disclosed by this application can be implemented in various ways and is not limited to the specific embodiments described herein.

[0204] Those skilled in the art will readily recognize various modifications and alternative embodiments of the invention disclosed herein. Therefore, it should be understood that the invention disclosed herein is not limited to the specific embodiments or examples described herein, and that modifications and alternative embodiments are also encompassed within the scope of the invention disclosed herein.

[0205]

[0206] Explanation of Terms

[0207] Unless otherwise stated, all technical and scientific terms used in this application have the meanings commonly understood by those skilled in the art to which this application pertains. All publications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0208]

[0209] "Halogen" or "halo" refers to the group of elements in the halogen group of the periodic table, which includes fluorine, chlorine, bromine, and iodine.

[0210] The term "hetero" as used herein refers to a compound or group that contains one or more heteroatoms. That is, the term hetero can be used in conjunction with terms used to refer to the molecule itself or terms used to refer to a portion of the molecule. For example, heteroalkylene refers to an alkylene group that contains one or more heteroatoms in the main chain. As another example, heteroaryl refers to an aryl group that contains one or more heteroatoms on the ring (e.g., a C6 aryl group in which one or more carbons on the ring are each independently replaced by a heteroatom selected from among others). The term "heteroatom" refers to an atom other than carbon or hydrogen, such as B, Si, N, P, O, S, F, Cl, Br, I, and Se. Preferably, it includes multivalent elements such as N, O, and S. For example, when a structure contains one or more heteroatoms, each heteroatom can be independently selected from N, O, and S.

[0211] The term "alkyl" or "alkane", when used to refer to a molecule by itself or to a portion of a molecule, is used to mean a chain or branched hydrocarbon group that is fully saturated. Chain and branched alkyl groups can be, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, iso-butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl. Alkyl groups can include cyclic structures. "C x-y "The term "is intended to include residues containing x to y carbons in a chain or ring, for example, when used with the term alkyl. For example, "C x-yThe term "alkyl" may mean a substituted or unsubstituted, chain-like alkyl group, branched alkyl group, or cyclic alkyl group containing x to y carbons. C0 alkyl means hydrogen. C 1-4 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, and iso-butyl. For example, a chain or branched alkyl group can have from 1 to about 60, from 1 to 20, or from 1 to 10 carbon atoms.

[0212] The term "heteroalkyl" as used herein means an alkyl group containing one or more heteroatoms, wherein each heteroatom is independently selected.

[0213] The term "alkylene," when used to refer to a molecule by itself or to refer to a portion of a molecule, refers to a divalent radical derived from alkyl. The term "alkylene" may be used with the terms "substituted" or "unsubstituted," as appropriate. When the term "alkylene" is not used with the terms "substituted" or "unsubstituted," the term "alkylene" is intended to encompass both substituted and unsubstituted alkylenes. For example, alkylene can refer to a group having from 1 to 100 carbon atoms in the main chain. Alkylene can be exemplified by, but not limited to, -CH2-, -CH2CH2-, -CH2CH2CH2-, and -CH2CH2CH2CH2-. For example, alkylene can be used as C2alkylene, which refers to an alkylene group having two carbon atoms in the main chain. For example, in this specification, "C x-y "Alkylene" is used to mean substituted or unsubstituted alkylene having X to Y number of carbon atoms in the main chain.

[0214] The term "heteroalkylene," when used to refer to a molecule by itself or to refer to a portion of a molecule, refers to a divalent radical derived from heteroalkyl. The term "heteroalkylene" may be used with the terms "substituted" or "unsubstituted," as appropriate. When the term "heteroalkylene" is not used with the terms "substituted" or "unsubstituted," the term "heteroalkylene" is intended to encompass aspects of substituted and unsubstituted heteroalkylenes. For example, heteroalkylene can refer to a group having from 1 to 100 carbon atoms and a heteroatom in the main chain, wherein the sum of the number of carbon atoms in the main chain and the number of heteroatoms is from 1 to 100. By way of example, heteroalkylene groups include, but are not limited to, -CH2-CH2-O-CH2-CH2-, and -CH2-O-CH2-CH2-NH-CH2-. The heteroalkylene group may include one or more heteroatoms, and each heteroatom may be the same or different. For example, the heteroalkylene group may include one or more heteroatoms at a position other than the terminal of the chain or branch, and each heteroatom may be the same or different. For example, the heteroalkylene group may include one or more heteroatoms at each or all terminals of the chain or branch, and each heteroatom may be the same or different. By way of example, as used herein, "C x-y"Heteroalkylene" is used to refer to a substituted or unsubstituted heteroalkylene having a total of x to y carbon atoms and heteroatoms in the main chain (e.g., the sum of the number of carbon atoms and the number of heteroatoms located in the main chain is x to y). For example, C3 heteroalkylene can be used to mean a heteroalkylene having two carbon atoms and one heteroatom in the main chain. As another example, C5 heteroalkylene can be used to mean a heteroalkylene having three carbon atoms and two heteroatoms in the main chain. C5 heteroalkylene encompasses structures such as, for example, -CH2-CH2-O-CH2-CH2- and -CH2-O-CH2-NH-CH2-.

[0215] The term "cycloalkyl" is used to refer to a fully saturated cyclic hydrocarbon group. "Cycloalkyl" includes monocyclic and polycyclic groups. Unless otherwise defined, a monocyclic cycloalkyl group generally has from 3 to about 20 carbon atoms, preferably from 3 to 10 carbon atoms per ring. Rings other than the first ring of a polycyclic cycloalkyl may be selected from saturated, unsaturated, and aromatic rings. Cycloalkyl is a bicyclic molecule, including those in which one, two, three, or more atoms are shared between two rings. The term "fused cycloalkyl" refers to a polycyclic cycloalkyl group in which each ring shares two adjacent atoms with the other ring. Rings other than the first ring of a fused polycyclic cycloalkyl may be selected from saturated, unsaturated, and aromatic rings. Cycloalkyl can be used with the terms substituted or unsubstituted, wherein substituted cycloalkyl refers to a group in which one or more hydrogen atoms attached to a carbon atom on the ring are replaced by one or more independent substituents. Furthermore, cycloalkyl can be used with the term hetero, wherein heterocycloalkyl refers to a cycloalkyl group containing one or more heteroatoms on the ring.

[0216] The term "cycloalkylene" is used to mean a divalent radical derived from cycloalkyl. For example, the term cycloalkylene can be used with the terms substituted or unsubstituted. For example, the term cycloalkylene can be used with the term hetero.

[0217] The term "alkene" or "alkenyl," when used to refer to a molecule in its own right or to a portion of a molecule, refers to a chain or branched non-aromatic hydrocarbon group containing one or more double bonds. For example, a chain or branched alkenyl group can have from 2 to about 60 carbon atoms, from 2 to 20 carbon atoms, or from 2 to 10 carbon atoms.

[0218] The term "heteroalkene" or "heteroalkenyl" means an alkenyl containing one or more heteroatoms, each of which is independently selected.

[0219] The term "alkenylene", when used to refer to a molecule by itself or to refer to a portion of a molecule, refers to a divalent radical derived from alkenyl. The term "alkenylene" may be used with the terms "substituted" or "unsubstituted", as appropriate. When the term alkenylene is not used with the terms substituted or unsubstituted, the term alkenylene is intended to encompass aspects of substituted alkenylene and unsubstituted alkenylene. For example, alkenylene may refer to a group having 2 to 100 carbon atoms in the main chain. Alkenylene may be exemplified by, but is not limited to, -C=C-, -CCC=CC=C-, or -CCCC=C-. As used herein, "C x-y When used as "alkenylene", C x-y Alkenylene is used to mean a substituted or unsubstituted alkenylene having x to y number of carbon atoms in the main chain.

[0220] The term "heteroalkenylene", used to refer to a molecule in itself or to a portion of a molecule, refers to a divalent radical derived from heteroalkenyl. For example, the term "heteroalkenylene" can be used to refer to an alkenylene group that includes one or more heteroatoms in the main chain. For example, heteroalkenylene can refer to a group having 2 to 100 carbon atoms and a heteroatom in the main chain, wherein the sum of the number of carbon atoms and the number of heteroatoms is 2 to 100. The term "heteroalkenylene" can be used with the terms "substituted" or "unsubstituted", as appropriate. In the present specification, "C x-y When used as "heteroalkenylene", C x-y Heteroalkenylene is used to mean a substituted or unsubstituted heteroalkenylene having x to y numbers of carbon atoms and heteroatoms in the main chain (e.g., the sum of the number of carbon atoms and the number of heteroatoms is x to y).

[0221] The term "cycloalkene" or "cycloalkenyl" refers to a cyclic hydrocarbon group containing one or more double bonds in the ring. "Cycloalkenyl" includes monocyclic and polycyclic groups. Unless otherwise defined, a monocyclic cycloalkenyl generally has from 3 to about 20 carbon atoms, preferably from 3 to 10 carbon atoms, in the ring. Rings other than the first ring of a polycyclic cycloalkenyl may be selected from saturated, unsaturated, and aromatic rings. Cycloalkenyl is a bicyclic molecule containing one, two, three, or more atoms shared between two rings. The term "fused cycloalkenyl" refers to a polycyclic cycloalkenyl in which each ring shares two adjacent atoms with the other ring. Rings other than the first ring of a fused polycyclic cycloalkenyl may be selected from saturated, unsaturated, and aromatic rings. The term "cycloalkenyl" may be used with the terms "substituted" or "unsubstituted", wherein a substituted cycloalkenyl refers to a group in which one or more hydrogen atoms attached to a carbon atom on the ring are replaced by one or more independent substituents. Furthermore, the term "cycloalkenyl" may be used with the term "hetero", wherein heterocycloalkenyl refers to a cycloalkenyl group that includes one or more heteroatoms on the ring.

[0222] The term "cycloalkenylene" is used to mean a divalent radical derived from cycloalkenyl. For example, the term cycloalkenylene can be used with the terms substituted or unsubstituted. For example, the term cycloalkenylene can be used with the term hetero.

[0223] The terms "alkyne" or "alkynyl," when used to refer to a molecule in its own right or to a portion of a molecule, refer to a chain or branched non-aromatic hydrocarbon group containing one or more triple bonds. For example, a chain or branched alkynyl group can have from 2 to about 60 carbon atoms, from 2 to 20 carbon atoms, or from 2 to 10 carbon atoms.

[0224] The term "heteroalkynyl" or "heteroalkyne" means an alkynyl containing one or more heteroatoms, each of which is independently selected.

[0225] The term "alkynylene", when used to refer to a molecule by itself or to refer to a portion of a molecule, means a divalent radical derived from alkynyl. The term "alkynylene" may be used with the terms "substituted" or "unsubstituted", as appropriate. When the term alkynylene is not used with the terms substituted or unsubstituted, the term alkynylene is intended to encompass both substituted and unsubstituted alkynylene embodiments. For example, alkynylene may refer to a group having from 2 to 100 carbon atoms in the main chain. As used herein, "C x-y When used as "alkynylene", C x-y Alkynylene is used to mean substituted or unsubstituted alkynylene having x to y number of carbon atoms in the main chain.

[0226] The term "heteroalkynylene", used to refer to a molecule in itself or to a part of a molecule, means a divalent radical derived from heteroalkynyl. For example, the term "heteroalkynylene" can be used to refer to an alkynylene group containing one or more heteroatoms in the main chain. For example, heteroalkynylene can refer to a group having 2 to 100 carbon atoms and a heteroatom in the main chain, wherein the sum of the number of carbon atoms in the main chain and the number of heteroatoms is 2 to 100. The term "heteroalkynylene" can be used with the terms "substituted" or "unsubstituted", as appropriate. In the present specification, "C x-y When used as "heteroalkynylene", C x-y Heteroalkynylene is used to mean a substituted or unsubstituted heteroalkynylene having x to y numbers of carbon atoms and heteroatoms in the main chain (e.g., the sum of the number of carbon atoms and the number of heteroatoms is x to y).

[0227] The term "cycloalkyne" or "cycloalkynyl" refers to a cyclic hydrocarbon group containing one or more triple bonds on the ring, also referred to as a "strained alkyne." "Cycloalkynyl" includes monocyclic and polycyclic groups. Generally, unless otherwise defined, a monocyclic cycloalkynyl has from 3 to about 10 carbon atoms on the ring. Rings other than the first ring of a polycyclic cycloalkynyl can be selected from saturated, unsaturated, and aromatic rings. Cycloalkynyl is a bicyclic molecule that includes one, two, three, or more atoms shared between two rings. The term "fused cycloalkynyl" refers to a polycyclic cycloalkynyl in which each ring shares two adjacent atoms with the other ring. Rings other than the first ring of a fused polycyclic cycloalkynyl can be selected from saturated, unsaturated, and aromatic rings. The term "cycloalkynyl" may be used with the terms "substituted" or "unsubstituted", wherein substituted cycloalkynyl refers to a group in which one or more hydrogen atoms attached to a carbon atom on the ring are replaced by one or more independent substituents. Furthermore, the term "cycloalkynyl" may be used with the term "hetero", wherein heterocycloalkynyl refers to a cycloalkynyl group that includes one or more heteroatoms on the ring.

[0228] The term "cycloalkynylene" is used to mean a divalent radical derived from cycloalkynyl. For example, the term cycloalkynylene can be used with the terms substituted or unsubstituted. For example, the term cycloalkynylene can be used with the term hetero.

[0229] The term "aryl" is used to refer to a group containing an aromatic ring, and refers to a group derived from the aromatic compound arene. The term aryl includes monocyclic groups and polycyclic groups. The term "aryl" may be used with the term "hetero," where heteroaryl is used to refer to an aryl group having one or more heteroatoms on the ring. The term "aryl" may be used with the terms "substituted" or "unsubstituted," where substituted aryl is used to refer to an aryl group where one or more hydrogen atoms attached to an atom on the ring are replaced with one or more substituents. The term "aryl" may be used to encompass both substituted and unsubstituted aryls, and substituted and unsubstituted heteroaryls. Examples of aryls include, but are not limited to, phenyl, pyridyl, naphthyl, and biphenyl.

[0230] The term "arylene" is used to mean a divalent radical derived from aryl. For example, the term arylene can be used with the terms substituted or unsubstituted. For example, the term arylene can be used with the term hetero. The term arylene can be used to encompass both substituted or unsubstituted arylenes and substituted or unsubstituted heteroarylenes.

[0231] The term "substituted" as used herein means that one or more hydrogen atoms on an atom are replaced by a substituent including deuterium and hydrogen variants, wherein the valence of the atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), this means that two hydrogen atoms are replaced. When one substituent is a halogen (e.g., Cl, F, Br, and I, etc.), this means that one hydrogen atom is replaced by the halogen. When two or more substituents are present in a group, the substituents present in the group may be the same or different. Unless otherwise specified, the type and number of substituents may be arbitrary, as long as it is chemically achievable. By way of example, the substituents include -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH can be selected from, but are not limited to, (provided that the substituent is not -H). Representative examples of the substituent are -C 1-4 alkyl, -C(=O)H, -C(=O)CH3, -C(=O)OH, -C(=O)NH2, -NH2, =NH, =O, =S, -OH, -NO2, and -SH, but are not limited thereto. The term substituted or unsubstituted may be used with the term used to refer to the molecule itself or with the term used to refer to a part of the molecule. For example, a substituted C 10-20 Alkylene may mean that one or more hydrogen atoms connected to the main chain are substituted with a substituent, and each substituent may be independently selected.

[0232] In this specification, when expressing the structure (partial structure) of a compound, when indicating a part where a group of the partial structure and another group are connected, a wavy line drawn in a direction approximately perpendicular to the bond (e.g., ) is used. For example, the structure When expressed as , it indicates that group X within a substance, molecule, or compound is connected to other parts through bonds. For example, structure When expressed as such, it indicates that the group X in the substance, molecule, or compound is connected to other parts through bonds. For example, in a compound having the structure of "AX", if only the structure of group X is shown, the structure can be expressed as. For example, in a compound having the structure of "AXB", if only the X structure is shown, the structure can be expressed as. If necessary, the wavy line drawn almost perpendicular to the bond can be expressed through additional notation. For example, in a compound having the structure of AXB, if only the structure of the X group is shown, the structure of the X group, if necessary, It can be depicted as follows, and by stating something like "* is the part that connects to A, and ** is the part that connects to B", information can be provided about which part each wavy line represents.

[0233] Furthermore, the wavy lines depicted in a direction roughly perpendicular to the bond should not be construed as indicating that the "structure depicted with the wavy lines" is shared and directly connected with the "group other than the structure depicted with the wavy lines," and should not be construed as suggesting that additional elements may be included between the "structure depicted with the wavy lines" and the "group other than the structure depicted with the wavy lines." If additional elements may be included, they will be described in a separate, related description. The following structures are illustrative examples.

[0234] - Example A

[0235] [Example Chemical Formula A]

[0236] ,

[0237] At this time, FcBU is an Fc binding unit, and the Fc binding unit has the following structure:

[0238] .

[0239] Here, Xa 1 ' is J a Directly connected to (i.e. J a Wow Xa 1 ' Xa without any additional elements in between 1 ' and J a will be interpreted as being connected.

[0240] Based on the above description, the structure of the example chemical formula A is illustrated as follows:

[0241] [Example Chemical Formula A-1]

[0242] .

[0243] - Example B

[0244] [Example Chemical Formula B]

[0245] ,

[0246] At this time, GOI is the target group, the target group is the reactive group, and the reactive group has the following structure:

[0247] .

[0248] Here, D RG is L a Directly connected to (i.e. D RG Wow L a D without any additional elements in between RG Wow L a will be interpreted as being connected.

[0249] Based on the above explanation, the structure of the example chemical formula B is illustrated as follows:

[0250] [Example Chemical Formula B-1]

[0251] .

[0252] "A structure or structure used within a chemical formula disclosed herein" " is C x It is used to mean alkylene. For example, the structure can be used to represent C4 alkylene such as -CH2-CH2-CH2-CH2-. Here, when x is 0, it means a bond. That is, the structure is a structure can be expressed as

[0253] The compounds of the present disclosure may have specific geometric or stereoisomeric forms. Unless otherwise specified, when a compound is disclosed in the present application, isomers of the compound, such as cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemates, are included within the scope of the present application. That is, designations relating to isomers in chemical formulas or structures disclosed herein (e.g., In the absence of a chemical formula or structure, it is meant to include all possible isomers.

[0254] The term "amino acid" as used herein may refer to both amino acids that are not bound to other amino acids and amino acid residues bound to other amino acids contained in proteins or peptides, and may be appropriately interpreted depending on the content or context of the paragraph in which the term "amino acid" is used. The term "amino acid" as used herein may be used to encompass both natural amino acids and unnatural amino acids. As used herein, "natural amino acid" collectively refers to the 20 amino acids synthesized through the transcription and translation of genes in the human body. Specifically, the natural amino acids are alanine (Ala, A), arginine (Arg, R), asparagine (Asn, N), aspartic acid (Asp, D), cysteine ​​(Cysteine, C), glutamic acid (Glu, E), glutamine (Gln, Q), glycine (Gly, G), histidine (His, H), isoleucine (Ile, I), leucine (Leu, L), lysine (Lys, K), methionine (Met, M), phenylalanine (Phe, F), proline (Pro, P), serine (Serine, S), threonine (Thr, T), Includes tryptophan (Trp, W), tyrosine (Tyr, Y), and valine (Val, V). As used herein, an unnatural amino acid refers to an amino acid that is not synthesized through the transcription and translation process of genes in the human body, but is synthesized through a process other than transcription and translation, is synthesized artificially, or can be synthesized in an organism other than a human.The non-natural amino acids may include, for example, ornithine (orn), diaminopropionic acid (Dap), diaminobutyric acid (Dab), and naphthylalanine. As mentioned above, the term "amino acid" as used herein may be used to refer to both an amino acid that is not bound to another amino acid and an amino acid residue that is bound to another amino acid in a protein or peptide. For example, alanine may be used to refer to an alanine and / or an alanine residue. For example, arginine may be used to refer to an arginine and / or an arginine residue. The term "amino acid" as used herein may be used to include both L-form amino acids and D-form amino acids. In some embodiments, if there is no mention of the L-form or D-form, it may be interpreted as the L-form amino acid.

[0255] The term "amino acid residue" as used herein refers to a structure derived from an amino acid, which is covalently linked to another portion of a compound, peptide, and / or protein (e.g., an antibody, etc.) contained in the compound, peptide, and / or protein. For example, when alanine, arginine, and glutamic acid are linked via an amide bond to form a peptide having an ARE sequence, the peptide includes three amino acid residues, wherein A may be referred to as an alanine residue, R as an arginine residue, and E as a glutamic acid residue. Furthermore, as described above, in a peptide having an ARE sequence, the peptide may include three amino acids, wherein A may be referred to as alanine, R as arginine, and E as glutamic acid. As another example, when aspartic acid, phenylalanine, and lysine are linked via amide bonds to form a peptide having a DFK sequence, the peptide comprises three amino acid residues, wherein D may be referred to as an aspartic acid residue, F may be referred to as a phenylalanine residue, and K may be referred to as a lysine residue. Furthermore, as described above, in a peptide having a DFK sequence, the peptide may comprise three amino acids, wherein D may be referred to as aspartic acid, F may be referred to as phenylalanine, and K may be referred to as lysine.

[0256] Unless otherwise stated, amino acid sequences in this specification are written in the N-terminal to C-terminal direction using the amino acid single-letter notation or the three-letter notation. For example, when written as RNVP, it means a peptide in which arginine, asparagine, valine, and proline are sequentially connected from the N-terminal to the C-terminal. Another example, when written as Thr-Leu-Lys, it means a peptide in which threonine, leucine, and lysine are sequentially connected from the N-terminal to the C-terminal. In the case of amino acids that cannot be expressed in the single-letter notation or the three-letter notation, other letters are used and additional explanations are provided.

[0257] The term "click chemistry" used herein is a chemical concept introduced by K. Barry Sharpless of Scripps Research Institute to describe complementary chemical functional groups and chemical reactions designed to rapidly and stably form covalent bonds between two molecules. Click chemistry as used herein does not refer to a specific reaction, but rather to the concept of a fast and stable reaction. In one embodiment, several conditions must be met for the formation of bonds between molecules via click chemistry. These conditions include high yields, excellent selectivity for the reaction site, modular operation to organically bond molecules, and thermodynamically stable processes to rapidly and accurately produce products. Click chemistry in the present disclosure includes the reaction of reactive pairs of click chemistry functional groups (e.g., terminal alkynes, azides, strained alkynes, dienes (e.g., Diels-Alder dienes), dienophiles (e.g., Diels-Alder dienophiles), trans-cyclooctenes, alkenes, thiols, tetrazines, triazines, dibenzocyclooctynes ​​(DBCO), and bicyclononynes (bicyclo[6.1.0]non-4-yne)). Examples of click chemistry reactions include Huisgen 1,3-dipolar cycloaddition (Tornoe et al.Journal of Organic Chemistry (2002) 67: 3075-3064; Diels-Alder reaction; inverse-electron-demand Diels-Alder reaction; nucleophilic addition to small strained rings such as epoxides and aziridines; nucleophilic addition to activated carbonyl groups; Staudinger ligation and addition to carbon-carbon double or triple bonds.

[0258] The term "bio-orthogonal functional group" is used to refer to a biologically inert chemical reactive group. That is, the term "bio-orthogonal functional group" is used to refer to a chemical functional group that participates in bio-orthogonal chemistry or bio-orthogonal reactions, thereby performing a bio-orthogonal reaction. The term "bio-orthogonal functional group" can also be referred to as a bio-orthogonal chemical functional group or a bio-orthogonal chemical group. A bio-orthogonal functional group refers to a group that does not react with endogenous molecules or functional groups within living cells or organisms. These bio-orthogonal functional groups are designed to react with specific groups in complex biological systems without interfering with normal cellular processes. The term "bio-orthogonal chemistry" was first proposed by Carolyn R. Bertozzi in 2003, and bio-orthogonal chemistry remains widely used in organic chemistry and conjugation. The term "bio-orthogonal" refers to the ability of chemical reactions to occur without interfering with natural biological processes via bio-orthogonal functional groups. A key characteristic of bio-orthogonal functional groups is that they react specifically and efficiently with corresponding reactive groups in a biological environment. Bio-orthogonal functional groups, bio-orthogonal chemistry, or bio-orthogonal reactions are discussed in detail in the literature [Sletten, Ellen M., and Carolyn R. Bertozzi. "Bioorthogonal chemistry: fishing for selectivity in a sea of ​​functionality." Angewandte Chemie International Edition 48.38 (2009): 6974-6998.; Mbua, Ngalle Eric, et al."Strain-promoted alkyne-azide cycloadditions (SPAAC) reveal new features of glycoconjugate biosynthesis." ChemBioChem 12.12 (2011): 1912-1921.; Bird, Robert E., et al. "Bioorthogonal chemistry and its applications." Bioconjugate Chemistry 32.12 (2021): 2457-2479.; Devaraj, Neal K. "The future of bioorthogonal chemistry." ACS central science 4.8 (2018): 952-959.; and Scinto, Samuel L., et al. "Bioorthogonal chemistry." Nature Reviews Methods Primers 1.1 (2021): 30.], the entire contents of which are incorporated herein by reference. Bioorthogonal chemistry can overlap significantly with the broader field of click chemistry. In certain embodiments, a bioorthogonal functional group may be used to refer to a chemical group that is non-reactive with an antibody, but capable of a bioorthogonal reaction.Representative types of bioorthogonal reactions (or bioorthogonal chemistry) include, but are not limited to, Staudinger Ligation, Copper-Catalyzed Azide-Alkyne Cycloaddition (CuAAC), Copper-Free Azide-Alkyne Cycloaddition including strain-promoted azide-alkyne cycloaddition (SPAAC), Tetrazine Ligation, Tetrazole Ligation, Oxime Ligation, and Isocyanide Click Reaction. Bioorthogonal functional groups include, but are not limited to, azide, terminal alkyne, cyclic alkyne (e.g., cyclooctyne), tetrazine, norbornene, cycloalkene (e.g., cyclooctene), tetrazole, oxime, or isocyanide groups. For example, in a copper-free strain-promoted azide-alkyne cycloaddition (SPAAC), an azide group and a cyclooctyne group undergo a bioorthogonal reaction. The cyclooctyne group participating in SPAAC can be a monocyclic cyclooctyne or a polycyclic cyclooctyne, including a fused polycyclic group.Specific examples of cyclooctyne groups participating in SPAAC include, but are not limited to, BCN (Bicyclononyne), DBCO (Dibenzocyclooctyne), DIBAC (aza-dibenzocyclooctynes), DIBO (dibenzocyclooctynol), DIFO (difluorinated cyclooctynes), BARAC (biarylazacyclooctynone), DIMAC (dimethoxyazacyclooctyne), and DIFBO (difluorobenzocyclooctyne).

[0259] The term "antibody" as used herein refers to an immunoglobulin molecule or a fragment thereof. Immunoglobulins are commonly known and have the ability to specifically bind to one or more antigens. As used herein, the term "antibody" is also used to encompass fragments thereof, so it does not matter if the molecule does not have the ability to bind to a specific antigen, as in the case of an Fc fragment. Unless used in a specific limitation related to antibodies, the term "antibody" will be interpreted without special limitation to include all of monospecific antibodies, bispecific antibodies, trispecific antibodies, monoclonal antibodies, human antibodies, humanized antibodies, recombinant antibodies, and chimeric antibodies. For example, an antibody may comprise two heavy chains and two light chains. For example, it is known that an antibody can have a structure in which two heavy chains are linked through one or more bridges (e.g., disulfide bonds), one heavy chain and one light chain are linked through one or more bridges, and one other heavy chain and one other light chain are linked through one or more bridges. An antibody can be divided into an Fc region (or Fc domain) and a Fab region, the Fab region including a site capable of binding to an antigen, and the Fc region including a part of a constant region of a heavy chain. The term "antibody" as used herein can be used to include both conjugated antibodies and unconjugated antibodies (e.g., free antibodies).

[0260] As used herein, an active moiety is used to refer to a moiety having one or more functions or activities. An active moiety may include, but is not limited to, a drug (e.g., a toxin), an imaging moiety (e.g., a fluorescent moiety, a luminescent moiety including a luminescent material such as luciferin, etc.), a radioactive moiety, a protein having a specific function, a peptide having a specific function, an affinity agent (e.g., biotin, streptavidin, and aptamers, etc.), a stabilizing agent, a vitamin, a nucleic acid (DNA or RNA), or a PEG (polyethylene glycol) moiety. Here, one or more active moieties may each be independently selected. In some embodiments of the present application, the active moiety may be included in a functional group. Furthermore, in some embodiments of the present application, the active moiety may be included in a payload.

[0261] The term "radioactive moiety" as used herein refers to a ligand for a radioisotope and / or a moiety comprising a radioisotope (e.g., a radioactive metal nuclide) that is designed to bind to a radioisotope. A ligand for a radioisotope may be referred to, for example, as a chelator. The chelator may be selected from, but is not limited to, DOTA (tetraxetan), NOTA [2,2',2"-(1,4,7-triazacyclononane-1,4,7-triyl)triacetic acid], DTPA (Diethylenetriaminepentaacetic acid), and EDTA (Ethylenediaminetetraacetic acid), for example. Radiolabeling is useful in diagnostic imaging, radioimmunotherapy (RIT), and radiotherapy. The radioactive moiety may be, for example, 18 F, 11 C, 67 Cu, 90 Y, 125 I, 123 I, 124 I, 131 I, 177 Lu, 186 Re, 188 Re, 211 At, 213 Bi, 225 Ac or 99m May include, but is not limited to, Tc.

[0262] The term "fluorescent moiety" as used herein refers to a moiety comprising a dye, protein, or dye reagent for fluorescent purposes. Molecules that can be used as dyes and dye reagents are well known in the art. Fluorescent moieties can include, but are not limited to, green fluorescent protein (GFP), Cy3, Cy5, Texas Red, FITC, Rhodamine, or DAPI.

[0263] As used herein, the term "drug" or "drug moiety" is used to mean a molecule or portion of a molecule that has therapeutic efficacy against any disease. The drug according to the present application includes those known to those skilled in the art to be effective against any disease. Furthermore, the term "drug" as used herein can be used to encompass both conjugated drugs and unconjugated drugs (e.g., free drugs). Drugs include, for example, auristatin, eribulin, tubulysin, geldanamycin (Kerr et al., 1997, Bioconjugate Chem. 8(6):781-784), maytansinoids (EP 1391213, ACR 2008, 41, 98-107), calicheamicin (US Patent Publication No. 2009 / 0105461, Cancer Res. 1993, 53, 3336-3342), mertansine, daunomycin, doxorubicin, methotrexate, vindesine, SG2285 (Cancer Res. 2010, 70(17), 6849-6858), dolastatin, dolastatin analog auristatin (US Patent No.5,635,483), cryptophycin, camptothecin, camptothecin analogs (e.g., SN38, FL118, or exatecan), rhizoxin derivatives, CC 1065 analogs or derivatives, duocarmycin, enediyne antibiotics, esperamicin, epothilones, pyrrolobenzodiazepine (PBD) derivatives, α-amanitin, toxoids, TLR5 (toll-like receptor 5) agonists, TLR7 (toll-like receptor 7) agonists, TLR8 (toll-like receptor 8) agonists, NOTA (1,4,7-Triazacyclononane-1,4,7-triacetic acid), and Any one selected from DOTA (1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid), or an analog thereof, but is not otherwise limited thereto.

[0264] In this application, when there is a reference to the numbering of amino acid residues in the Fc domain (or Fc region) of an antibody, unless otherwise stated, the numbering of amino acid residues is according to the EU numbering system. The EU numbering system has been widely used as a sequence system for the Fc region since the sequence of IgG was studied in Edelman GM, et al., The covalent structure of an entire gammaG immunoglobulin molecule, Proc Natl Acad Sci U S A., 1969 May;63(1):78-85. For example, at lysine 246 in the Fc region, 246 is the number assigned according to the EU numbering system. As another example, at lysine 248 in the Fc region, 248 is the number assigned according to the EU numbering system.

[0265] The terms "connected" or "linked" as used herein mean that two or more elements within a single conceptual structure are directly or indirectly connected (e.g., via another element such as a linker), and does not imply that no other additional elements can exist between the two or more elements. For example, a description such as "element B connected to element A" is intended to encompass, but is not limited to, both instances where one or more other elements are present between element A and element B (i.e., element A is connected to element B via one or more other elements) and instances where one or more other elements are not present between element A and element B (i.e., element A and element B are directly connected).

[0266] The term "sequence identity" as used herein refers to the degree of similarity between two or more sequences. For example, the term "sequence identity" is used with terms referring to a reference sequence and terms indicating a ratio (e.g., a percentage). For example, the term "sequence identity" can be used to describe a sequence that is similar or substantially identical to a reference amino acid sequence. When described as "a sequence having at least 90% sequence identity to sequence A," the reference sequence is sequence A. For example, the percentage of sequence identity can be calculated by aligning the sequence being measured for the percentage of sequence identity with the reference sequence. The method for calculating and / or determining the percentage of sequence identity is not otherwise limited, and can be calculated and / or determined through a reasonable method or algorithm that can be utilized by a person skilled in the art.

[0267] As used herein, the term "unit" is used, in some embodiments, to distinguish a conjugated substance from a free substance. The term "unit," as used in some embodiments, is described by way of example, using an antibody unit and a free antibody. A free antibody refers to an antibody molecule that is not covalently bound to another molecule or group. An antibody unit refers to a group derived from a free antibody that is covalently bound to another molecule or group. For example, an antibody-functional group conjugate can be prepared when a free antibody and a functional substance are linked through a reaction between a primary amine group of a lysine residue of the free antibody and a reactive group of the functional group. In this case, the portion derived from the free antibody can be referred to as an antibody unit. In an antibody-functional group conjugate, the antibody unit can be understood to be structurally identical to the free antibody from which the antibody unit originated, except for the portion that is conjugated to a non-antibody portion of the antibody-functional group conjugate. For example, if the primary amine groups of the lysine residues participating in the reaction in a free antibody are depicted separately, the structure of the free antibody is " "can be expressed as follows. When the junction between the antibody unit and a non-antibody unit is depicted separately in the antibody unit, the structure of the antibody unit is " " can be expressed as such. In this way, except for the primary amine group of the lysine residue used in the reaction, the antibody unit and the free antibody can be understood to be structurally identical. In this relationship, in some embodiments, the antibody unit and the free antibody can be referred to as "antibody" without being otherwise distinguished, and these terms can be appropriately interpreted depending on the context. In some embodiments, when it is described as "the antibody unit is derived from an antibody," the antibody unit and the antibody can be understood to have the same relationship as described above. In some embodiments, when it is described as "the Fc-binding unit is derived from an Fc-binding substance," "the Fc-binding unit and the Fc-binding substance" can be understood to have a relationship similar to the above description. That is, except for the junction where the Fc-binding unit is conjugated to a part other than the Fc-binding unit, the Fc-binding unit can be understood to have the same structure as the Fc-binding substance from which it is derived. As in the case of the antibody unit, in some embodiments, the Fc-binding unit can be referred to as the Fc-binding substance from which it is derived. and these terms can be interpreted appropriately depending on the context.

[0268] In the present specification, when it is described as "A comprises B", the description of "A comprises B" should be interpreted as not excluding that A includes additional components other than B. That is, "A comprises B" is intended to encompass cases such as cases where A has additional components other than B (for example, cases where B and C exist in A), cases where A is B, and cases where A is composed of B. On the other hand, "A comprises B" can be modified and rewritten as "A is B", "A is composed of B", or "A is expressed by B" because it encompasses all of the above cases. The fact that the description "A includes B" can be rewritten as "A is B" or something like that means that the description "A is B" can be newly produced by using the description "A includes B" that already existed in the specification, and does not mean that the description "A includes B" should be limitedly interpreted as "A is B." When described as "A includes B," it will be interpreted that A may have additional elements in addition to B. In other words, when described as "A includes B" in the specification, it should be interpreted to include both cases where A is B and cases where A has additional elements in addition to B.

[0269] In the present specification, when it is described as "A has B", the description of "A has B" should be interpreted as not excluding that A has additional components other than B. That is, "A has B" is intended to encompass cases such as cases where A has additional components other than B (for example, cases where A has B and C), cases where A is B, and cases where A is composed of B. On the other hand, "A has B" can be rewritten as "A is B", "A is composed of B", or "A is expressed by B", as it encompasses all of the above cases. The fact that the description "A has B" can be rewritten as "A is B" or something like that means that the description "A is B" can be newly produced by using the description "A has B" that already existed in the specification, and does not mean that the description "A has B" should be limitedly interpreted as "A is B." When described as "A has B," it will be interpreted that A may have additional elements in addition to B. In other words, when described as "A has B" in this specification, it should be interpreted to encompass both cases where A is B and cases where A has additional elements in addition to B.

[0270] It will be understood that where a compound (e.g., a small compound, a peptide, an antibody, a conjugate, etc.) is disclosed herein, the salt form thereof is also disclosed. The ions forming the salt of the compound are, for example, ammonium, calcium, sodium, potassium, acetate (CH3COO- - ), carbonate (CO3 2- ), chloride (Cl - ), citrate, cyanide, fluoride (F - ), nitrate (NO3- ), nitrite (NO2 - ), phosphate (PO3 - ), and sulfate (SO4 2- ) and the like, and are not otherwise limited. Salt-forming ions commonly used in the art may be used, if desired, to form salts of the compounds. The salts may be, for example, pharmaceutically acceptable salts, wherein a pharmaceutically acceptable salt is a salt that retains the efficacy of the parent agent and is not biologically undesirable (e.g., has little or no toxicity). Suitable salts include, for example, salts that can be formed by mixing a solution of the parent agent with a solution of a pharmaceutically acceptable acid, such as hydrochloric acid, phosphoric acid, sulfuric acid, or acetic acid. For example, when the compound carries an acidic moiety, pharmaceutically acceptable salts may include salts formed with suitable organic ligands, such as alkali metal ions (sodium or potassium), alkaline earth metal ions (calcium or magnesium), and ammonium ions.

[0271]

[0272] Below, the structure of an antibody is specifically described based on information generally known in the art to aid understanding, and the scope of the present application is not limited by the description below.

[0273] The structure of antibodies is divided into a heavy chain region and a light chain region, depending on the chain type. The structure of antibodies is divided into a fragment antigen-binding region (Fab region) and a fragment crystallizable region (Fc region) depending on the function of binding to the antigen. The structure of antibodies is divided into a variable region and a constant region depending on the variability of the amino acid sequence. Other structures of antibodies include the hinge region and the tail region. The heavy chain region and the light chain region can be broadly explained as being functionally divided into a fragment antigen-binding region (Fab region) and a fragment crystallizable region (Fc region). The Fab region is a region that includes a portion that binds to an antigen (antigen-binding site). The Fc region is a portion that can bind to an Fc receptor. The heavy chain region can be described as having both a Fab region and an Fc region, and the light chain region can be described as having a Fab region.

[0274] The Fab region of the heavy chain region includes the heavy chain variable region (VH) and the heavy chain constant region 1 (CH1). For example, in IgG1, the Fc region is known to include the heavy chain constant region 2 (CH2) and the heavy chain constant region 3 (CH3). In this case, the entire heavy chain constant region of the antibody can be referred to as CH. For example, the entire region combining CH1, CH2, and CH3 of IgG1 can be expressed as CH.

[0275] The Fab region of the light chain region includes a light chain variable region (VL) and a light chain constant region (CL). The light chain region can be described as having no Fc region.

[0276] As mentioned above, VH, CH1, CH2, CH3, VL and CL can each be referred to as immunoglobulin domains.

[0277] The immunoglobulin domains included in the heavy chain region are known to be positioned in the order of VH, CH1, CH2, and CH3 or VH, CH1, CH2, CH3, and CH4 from the N-terminal to the C-terminal direction. The immunoglobulin domains included in the light chain region are known to be positioned in the order of VL and CL from the N-terminal to the C-terminal direction. In general, the heavy chain region and the light chain region are known to be connected by a disulfide bond, and the Fab region and the Fc region are known to be connected by a hinge region. Specifically, the C-terminal portion of CH1 and the N-terminal portion of CH2 of the heavy chain region are known to be connected by a hinge region.

[0278] The variable region (VH and VL) is a region including an antigen-binding portion, and in the variable region, there is a hypervariable region with the greatest variability, which is called a complementarity-determining region (CDR). The VH includes three CDRs, and the three CDRs included in the VH are generally referred to as CDRH1, CDRH2, or CDRH3, respectively. The CDRs in the VH can be understood to be positioned in the order of CDRH1, CDRH2, and CDRH3 from the N-terminal to the C-terminal. The VL includes three CDRs, and the three CDRs included in the VL are generally referred to as CDRL1, CDRL2, or CDRL3, respectively. The CDRs in the VH can be understood to be positioned in the order of CDRL1, CDRL2, and CDRL3, from the N-terminal to the C-terminal.

[0279] The constant region of an antibody is a region separated from the antigen-binding portion, and it is known that the constant region can interact with cells or molecules of the immune system. For example, the constant region can interact with (be bound to or linked to) the cell membrane of an immune cell (e.g., lymphocytes, neutrophils, dendritic cells, and / or macrophages). Specifically, the hinge region and / or CH2 portion of the constant region can bind to a receptor (e.g., FcεRIII) on the cell membrane of the immune cell. In another specific example, the constant region can bind to FcRn.

[0280] The constant region of the heavy chain region (hereinafter referred to as “heavy chain constant region”) is broadly divided into five types (classes or isotypes): alpha (α), gamma (γ), delta (δ), epsilon (ε), and mu (μ). At this time, the type of the heavy chain constant region is not determined individually for CH1, CH2, CH3, and CH4, but is determined by considering all heavy chain constant regions (CH1, CH2, and CH3; or CH1, CH2, CH3, and CH4) included in the antibody.

[0281] There are two types of constant regions of the above light chain region (hereinafter referred to as “light chain constant regions”), namely lambda (λ) and kappa (κ).

[0282] It is known that antibodies can be broadly classified into five types (classes or isotypes). These five types are determined by the type of heavy chain constant region.

[0283] The five types of antibodies mentioned above are Immunoglobulin M (IgM), Immunoglobulin D (IgD), Immunoglobulin G (IgG), Immunoglobulin A (IgA), and Immunoglobulin E (IgE). If the type of the heavy chain constant region of the antibody is classified as alpha, the type of antibody can be recognized as IgA. If the type of the heavy chain constant region of the antibody is classified as gamma, the type of antibody can be recognized as IgG. If the type of the heavy chain constant region of the antibody is classified as delta, the type of antibody can be recognized as IgD. If the type of the heavy chain constant region of the antibody is classified as epsilon, the type of antibody can be recognized as IgE. If the type of the heavy chain constant region of the antibody is classified as mu, the type of antibody can be recognized as IgM. For example, each heavy chain of IgG is known to contain four immunoglobulin domains (VH, CH1, CH2, and CH3).

[0284] Among the above five types of antibodies, it is known that IgG and IgA can be further classified into subtypes. For example, if the antibody is a human antibody, if the type of heavy chain constant region of the antibody is gamma 1 (γ1), the type of antibody is IgG1; if it is gamma 2 (γ2), the type of antibody is IgG2; if it is gamma 3 (γ3), the type of antibody is IgG3; and if it is gamma 4 (γ4), the type of antibody is IgG4. If the heavy chain constant region of the human antibody is alpha 1 (α1), the type of antibody is IgA1; and if it is alpha 2 (α2), the type of antibody is IgA2.

[0285]

[0286] Hereinafter, compounds comprising an Fc binding unit, which are provided as some embodiments of the present application, are described in detail.

[0287]

[0288]

[0289] Compounds comprising an Fc binding unit

[0290] Prior art and problems of prior art

[0291] As described above, research has been conducted on compounds comprising an Fc-binding unit for site-specific delivery of a target substance (e.g., a bioorthogonal functional group) to an antibody. Korean Patent Application No. 10-2020-0091826 (Application No. 10-2020-0009162) discloses a compound comprising an Fc-binding unit, the compound having the structure of the following chemical formula 1-1.

[0292] [Chemical Formula 1-1]

[0293] .

[0294] In the compound of the above chemical formula 1-1, the Fc binding unit (FcBU) induces a compound including the Fc binding unit near lysine residues 246 and 248 located in the Fc region of the antibody, and the carbonyl group directly connected to the norbornene group, which is a bioorthogonal functional group, reacts with the amino group of the lysine residue 246 (K246) or lysine residue 248 (K248) of the antibody. Through the reaction, the Fc binding unit is released, and the norbornene group is transferred to lysine 246 or lysine 248 of the Fc region of the antibody. In the compound of the above chemical formula 1-1, the group reacting with the antibody The structure (wherein * in the structure indicates a carbonyl group that reacts with the amino group of the lysine residue of the antibody) allows the reaction of a compound containing an antibody and an Fc-binding unit under physiological conditions (e.g., pH 7.4), and allows the norbornene group to be transferred to a desired position (e.g., K246 or K248). Meanwhile, in some cases, NHS ester is used as a group that reacts with the antibody, but when an NHS ester (N-Hydroxysuccinimide Ester) group is used, the reaction conditions need to be adjusted to acidic conditions. When the reaction conditions are acidic conditions, additional substances or additional processes are required to adjust the pH, and furthermore, since acidic conditions can affect the structure of the antibody, the reaction under acidic conditions is disadvantageous.

[0295] Meanwhile, the inventors of the present application confirmed that there was a problem with the reaction efficiency (i.e., reaction efficiency with an antibody) of the compound of chemical formula 1-1 disclosed in the prior literature [Korean Patent Application No. 10-2020-0091826 (Application No. 10-2020-0009162)].

[0296]

[0297] Development of compounds containing novel Fc-binding units

[0298] As described above, the inventors of the present application have identified problems with the reaction efficiency of the compound of formula 1-1 disclosed in the prior art. The results of a study on the reaction of the compound of formula 1-1 with an antibody are disclosed in Example 02. As disclosed in Example 02, the compound of formula 1-1 exhibited low reaction efficiency in the reaction with an antibody.

[0299] To improve the low reaction efficiency of the compound of Chemical Formula 1-1, the inventors of the present application first attempted to prepare a compound of Chemical Formula 1-2, a compound in which the norbornene group in Chemical Formula 1-1 was replaced with an azide group. However, the compound of Chemical Formula 1-2 was found to be extremely unstable. Accordingly, the inventors of the present application determined that the compound of Chemical Formula 1-2 could not be used to deliver a target substance to an antibody.

[0300] [Chemical Formula 1-2]

[0301] .

[0302]

[0303] In order to improve the low reaction efficiency of the compound of Chemical Formula 1-1, the inventors of the present application secondly designed a novel compound comprising an Fc-binding unit in which a peptide linker (Val-Gly or Ala-Gly) was added between the azide group and the carbonyl group. The newly designed compound with the added peptide linker has the structures of Chemical Formula 1-3 and Chemical Formula 1-4. The inventors of the present application prepared compounds having the structures of Chemical Formula 1-3 and Chemical Formula 1-4 and conducted conjugation experiments with these compounds and antibodies. However, as described in Examples 03 to 04, it was confirmed that compounds containing a peptide linker between the azide and carbonyl groups react irregularly with antibodies. Therefore, the inventors of the present application determined that the compounds of Chemical Formula 1-3 and Chemical Formula 1-4 are also difficult to use for delivering a target substance to an antibody. Chemical Formula 1-3 below is a compound in which a Val-Gly peptide linker is introduced. The following chemical formula 1-4 is a compound into which an Ala-Gly peptide linker is introduced.

[0304] [Chemical Formula 1-3]

[0305] ,

[0306] [Chemical Formula 1-4]

[0307] .

[0308] In this way, the inventors of the present application attempted various structural modifications to improve the low reaction efficiency of compounds containing conventional Fc-binding units, but could not easily find a compound containing an Fc-binding unit suitable for the reaction. Furthermore, because the positions where structural modifications can occur are diverse, and the resulting structures of the structural modifications are also diverse, it was difficult to develop a novel compound containing an Fc-binding unit with improved reaction efficiency. However, after much effort, the inventors of the present application eventually succeeded in developing a novel compound containing an Fc-binding unit with improved reaction efficiency.

[0309] Hereinafter, compounds comprising the Fc binding unit provided by the present application are described in detail.

[0310]

[0311] Overview of compounds comprising the Fc binding unit of the present application

[0312] Some embodiments of the present application provide a compound comprising an Fc binding unit. The compound comprising the Fc binding unit may be referred to as a compound for delivering a group of interest to an antibody. The compound comprising the Fc binding unit may be referred to as a compound for site-specifically delivering a group of interest to an antibody.

[0313] Compounds comprising an Fc binding unit can be used to transfer a desired group (e.g., a bioorthogonal functional group) to an antibody.

[0314] Compounds comprising an Fc binding unit can be used to deliver a desired group (e.g., a bioorthogonal functional group) to a target region of an antibody.

[0315] Compounds comprising an Fc-binding unit can be used to deliver a desired group (e.g., a bioorthogonal functional group) to a desired location on an antibody (e.g., K246 or K248 of the Fc region of an antibody).

[0316]

[0317] Below, the structure of a compound comprising an Fc binding unit provided according to some embodiments of the present application is described.

[0318]

[0319] A compound comprising an Fc binding unit according to some embodiments of the present application may have a structure represented by Formula 2.

[0320] Some embodiments of the present application provide a compound having the structure of Formula 2:

[0321] [Chemical Formula 2]

[0322] .

[0323] In chemical formula 2, D a is spacer A.

[0324] In chemical formula 2, L a is linker A.

[0325] In chemical formula 2, X is C, O, or N (i.e., -X- is -CH2-, -O-, or -NH-).

[0326] In chemical formula 2, GOI is the group of interest.

[0327] In chemical formula 2, FcBU is an Fc binding unit.

[0328] In chemical formula 2, R a1 is H, or C 1-6 It's alkyl.

[0329] In chemical formula 2, R a2 is H, or C 1-6 It's alkyl.

[0330] In chemical formula 2, R a3 is H, or C 1-6 It's alkyl.

[0331] In chemical formula 2, J a is -C(=O)-, -S-, -NH-, or -C(=NH)-.

[0332]

[0333] Below, each element of the compound of chemical formula 2 is described in detail.

[0334]

[0335] spacer A (spacer A; D"a)

[0336] In chemical formula 2, D a is spacer A. In the above table of contents "spacer A (spacer A; D"a)", "a of D"a represents superscript a. That is, D"a is D a will be understood as

[0337] In some embodiments, spacer A is a bond, substituted or unsubstituted C 1-20 Alkylene, substituted or unsubstituted C 1-20 Heteroalkylene, substituted or unsubstituted C 2-20 Alkenylene, substituted or unsubstituted C 2-20 Heteroalkenylene, substituted or unsubstituted C 2-20 Alkynylene, or substituted or unsubstituted C 2-20It may be heteroalkynylene. Here, "substituted" indicates that one or more hydrogen atoms in the group modified by the term "substituted" are replaced by one or more substituents. That is, the substituted alkylene, etc. may include one or more substituents. For example, the substituents may each independently be selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH. Here, heteroalkylene, heteroalkenylene, heteroalkynylene, heterocycloalkyl, and heteroaryl each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0338] In certain embodiments, spacer A is unsubstituted C 1-20 alkylene, unsubstituted C 1-20 Heteroalkylene, unsubstituted C 2-20 Alkenylene, unsubstituted C 2-20 Heteroalkenylene, unsubstituted C 2-20 Alkynylene, or unsubstituted C 2-20 It may be heteroalkynylene. Here, heteroalkylene, heteroalkenylene, and heteroalkynylene each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0339] In certain embodiments, spacer A is unsubstituted C 1-10 alkylene, unsubstituted C 1-10 Heteroalkylene, unsubstituted C 2-10 Alkenylene, unsubstituted C 2-10 Heteroalkenylene, unsubstituted C 2-10Alkynylene, or unsubstituted C 2-10 It may be heteroalkynylene. Here, heteroalkylene, heteroalkenylene, and heteroalkynylene each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0340] In certain embodiments, spacer A is unsubstituted C 1-6 Alkylene, or unsubstituted C 1-6 It may be a heteroalkylene. Here, the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, or S. In a particular embodiment, each heteroatom can be O.

[0341] In certain embodiments, spacer A can be an unsubstituted C3 alkylene, or an unsubstituted C3 heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, or S. In certain embodiments, each heteroatom can be O.

[0342] In certain embodiments, spacer A is unsubstituted C 1-6 It may be alkylene.

[0343] In certain embodiments, spacer A can be an unsubstituted C3 alkylene.

[0344] In some embodiments, the spacer can be designed to control the distance between groups within the molecule. For example, spacer A can be designed to control the distance between two adjacent carbonyl groups. For example, the length of spacer A can be 0 to 20 based on the number of atoms in the main chain (i.e., the number of atoms located in the main chain) (if the number of atoms in the main chain is 0, spacer A is a bond). For example, spacer A can include a cyclic group in the main chain, in which case the number of atoms in the main chain can be conveniently counted based on the two atoms forming a bond with a non-cyclic moiety on the ring, and in this case, the counting is done in the direction that can achieve the smallest number. For example, If the ring is present in the main chain of spacer A, the number of atoms of the main chain counted by the ring is 3. For example, When a ring is present in the main chain of spacer A, the number of atoms of the main chain counted by the ring is 5.

[0345]

[0346] Linker A (L"a)

[0347] In chemical formula 2, L a is linker A.

[0348] In some embodiments, the length of linker A can be from 0 to 100 based on the number of atoms in the main chain. For example, linker A can include a cyclic group in the main chain.

[0349] In some embodiments, linker A is a bond, substituted or unsubstituted C 1-100 Alkylene, substituted or unsubstituted C 1-100 Heteroalkylene, substituted or unsubstituted C 2-100 Alkenylene, substituted or unsubstituted C 2-100 Heteroalkenylene, substituted or unsubstituted C 2-100Alkynylene, or substituted or unsubstituted C 2-100 It may be heteroalkynylene. Here, "substituted" indicates that one or more hydrogen atoms in the group modified by the term "substituted" are replaced by one or more substituents. That is, the substituted alkylene, etc. may include one or more substituents. For example, the substituents may each independently be selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH. Here, heteroalkylene, heteroalkenylene, heteroalkynylene, heterocycloalkyl, and heteroaryl each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0350] In certain embodiments, linker A is a bond, unsubstituted C 1-60 alkylene, unsubstituted C 1-60 Heteroalkylene, unsubstituted C 2-60 Alkenylene, unsubstituted C 2-60 Heteroalkenylene, unsubstituted C 2-60 Alkynylene, or unsubstituted C 2-60 It may be heteroalkynylene. Here, heteroalkylene, heteroalkenylene, and heteroalkynylene each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0351] In certain embodiments, linker A is a bond, unsubstituted C 1-30 alkylene, unsubstituted C 1-30 Heteroalkylene, unsubstituted C 2-30Alkenylene, unsubstituted C 2-30 Heteroalkenylene, unsubstituted C 2-30 Alkynylene, or unsubstituted C 2-30 It may be heteroalkynylene. Here, heteroalkylene, heteroalkenylene, and heteroalkynylene each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S. In a particular embodiment, each heteroatom can be O.

[0352] In certain embodiments, linker A is a bond, unsubstituted C 1-30 Alkylene, or unsubstituted C 1-30 It may be a heteroalkylene. Wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be independently selected from N, O, and S. In a particular embodiment, each heteroatom may be O.

[0353] In certain embodiments, linker A is a bond, unsubstituted C 1-30 Alkylene, or unsubstituted C 1-30 It may be a heteroalkylene. Here, the heteroalkylene may include one or more heteroatoms, wherein each heteroatom may be independently selected from N, O, and S. In a particular embodiment, each heteroatom may be O. Here, the linker A may be an unsubstituted C 1-30 In the case of heteroalkylene, linker A is an unsubstituted C containing 0 to 10 ethylene glycol units. 1-30 It may be heteroalkylene.

[0354] The ethylene glycol unit can be represented as -[EG]-, where -[EG]- is -[CH2OCH2]-, -[OCH2CH2]-, or -[CH2CH2O]-.

[0355] In certain embodiments, linker A is a bond, unsubstituted C 1-25 Alkylene, or unsubstituted C 1-25It may be a heteroalkylene. Here, the heteroalkylene may include one or more heteroatoms, wherein each heteroatom may be independently selected from N, O, and S. In a particular embodiment, each heteroatom may be O. Here, the linker A may be an unsubstituted C 1-25 In the case of heteroalkylene, linker A is an unsubstituted C containing 0 to 8 ethylene glycol units. 1-25 It may be heteroalkylene.

[0356] In certain embodiments, linker A is a bond, unsubstituted C 1-20 Alkylene, or unsubstituted C 1-20 It may be a heteroalkylene. Here, the heteroalkylene may include one or more heteroatoms, wherein each heteroatom may be independently selected from N, O, and S. In a particular embodiment, each heteroatom may be O. Here, the linker A may be an unsubstituted C 1-20 In the case of heteroalkylene, linker A is an unsubstituted C containing 0 to 6 ethylene glycol units. 1-20 It may be heteroalkylene.

[0357] In certain embodiments, linker A is a bond, unsubstituted C 1-10 Alkylene, or unsubstituted C 1-10 It may be a heteroalkylene. Here, the heteroalkylene may include one or more heteroatoms, wherein each heteroatom may be independently selected from N, O, and S. In a particular embodiment, each heteroatom may be O. Here, the linker A may be an unsubstituted C 1-10 In the case of heteroalkylene, linker A is an unsubstituted C containing 0 to 3 ethylene glycol units. 1-10 It may be heteroalkylene.

[0358] In certain embodiments, linker A is a bond, unsubstituted C 1-5 Alkylene, or unsubstituted C 1-5It may be a heteroalkylene. Here, the heteroalkylene may include one or more heteroatoms, wherein each heteroatom may be independently selected from N, O, and S. In a particular embodiment, each heteroatom may be O. Here, the linker A may be an unsubstituted C 1-5 In the case of heteroalkylene, linker A is an unsubstituted C containing 0 to 1 ethylene glycol unit. 1-5 It may be heteroalkylene.

[0359]

[0360] In some embodiments, linker A may have the following structure:

[0361] -L a1 -L a2 -L a3 -.

[0362] At this time, L a1 can be connected to X of chemical formula 2. At this time, L a3 can be linked to the target group (GOI) of chemical formula 2.

[0363] In some embodiments, L a1 is a bond, or substituted or unsubstituted C 1-10 Alkylene, substituted or unsubstituted C 1-10 Heteroalkylene, substituted or unsubstituted C 2-10 Alkenylene, substituted or unsubstituted C 2-10 Heteroalkenylene, substituted or unsubstituted C 2-10 Alkynylene, or substituted or unsubstituted C 2-10It may be heteroalkynylene. Here, "substituted" indicates that one or more hydrogen atoms in the group modified by the term "substituted" are replaced by one or more substituents. That is, the substituted alkylene, etc. may include one or more substituents. For example, the substituents may each independently be selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH. Here, heteroalkylene, heteroalkenylene, heteroalkynylene, heterocycloalkyl, and heteroaryl each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0364] In some embodiments, L a2 is a combination, or -[EG] x- - may be. Here, x may be an integer from 1 to 20. Here, -[EG]- is an ethylene glycol unit, -[CH2OCH2]-, -[OCH2CH2]-, or -[CH2CH2O]-. -[EG] x- -is -[CH2OCH2] 1-20 -, -[OCH2CH2] 1-20 - or -[CH2CH2O] 1-20 - can be expressed as follows. That is, L a2 may be a combination or may consist of 1 to 20 ethylene glycol units.

[0365] In some embodiments, L a3 is a bond, or substituted or unsubstituted C 1-10 Alkylene, substituted or unsubstituted C 1-10Heteroalkylene, substituted or unsubstituted C 2-10 Alkenylene, substituted or unsubstituted C 2-10 Heteroalkenylene, substituted or unsubstituted C 2-10 Alkynylene, or substituted or unsubstituted C 2-10 It may be heteroalkynylene. Here, "substituted" indicates that one or more hydrogen atoms in the group modified by the term "substituted" are replaced by one or more substituents. That is, the substituted alkylene, etc. may include one or more substituents. For example, the substituents may each independently be selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH. Here, heteroalkylene, heteroalkenylene, heteroalkynylene, heterocycloalkyl, and heteroaryl each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0366] In certain embodiments, L a1 Silver bond, or unsubstituted C 1-3 Alkylene, or unsubstituted C 1-3 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S. In a particular embodiment, the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be O.

[0367] In certain embodiments, L a2 is a combination, or -[EG] x- may be. Wherein x may be an integer from 1 to 10. In a particular embodiment, x may be an integer from 1 to 7. In a particular embodiment, x may be an integer from 1 to 5. In a particular embodiment, x may be an integer from 1 to 3.

[0368] In certain embodiments, L a3 Silver bond, or unsubstituted C 1-3 Alkylene, or unsubstituted C 1-3 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S. In a particular embodiment, the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be O.

[0369] In some embodiments, linker A can have any of the following structures:

[0370] , and

[0371] ,

[0372] At this time,

[0373] se is an integer from 0 to 15,

[0374] sb is an integer from 0 to 3,

[0375] sc is an integer from 1 to 15, and

[0376] sd is an integer from 0 to 3.

[0377] In some embodiments, linker A may have the following structure:

[0378] ,

[0379] At this time,

[0380] sf is an integer from 0 to 8,

[0381] sg is an integer from 0 to 15, and

[0382] sh is an integer from 0 to 8.

[0383] In certain embodiments, sf can be an integer from 0 to 6. In certain embodiments, sf can be an integer from 0 to 3. In certain embodiments, sg can be an integer from 0 to 10. In certain embodiments, sg can be an integer from 0 to 7. In certain embodiments, sh can be an integer from 0 to 6. In certain embodiments, sh can be an integer from 0 to 3.

[0384] In some embodiments, X (L) of formula 2 a If X) connected to is O, the sum of sf, sg, and sh may not be 0 (i.e., at least one of sf, sg, and sh is an integer greater than or equal to 1).

[0385]

[0386] In some embodiments, linker A may have any one of the following structures:

[0387]

[0388] .

[0389]

[0390] In some embodiments, it may be desirable for linker A to be designed to have little or no reactivity. That is, linker A may be designed in an unsubstituted form, or, if it includes a substituent, it may be desirable for it to include a substituent that has little or no reactivity.

[0391] As described below, the reaction of a compound comprising an Fc-binding unit with an antibody is specifically directed to the carbonyl group (structure " " can be explained by the reaction between the *-marked carbonyl group) and the amino group of the 246th or 248th lysine residue of the antibody (i.e., the primary amine group of the lysine side chain). Meanwhile, the Fc-binding unit included in the compound including the Fc-binding unit guides the compound to the Fc region of the antibody, so that the *-marked carbonyl group and the amino group of the 246th or 248th lysine residue of the antibody come close. That is, the guide of the Fc-binding unit makes the *-marked carbonyl group and the amino group of the 246th or 248th lysine residue of the antibody reactable. Therefore, the length of the linker A, rather than the structure between the Fc-binding unit and the *-marked carbonyl group, can be designed more freely than other structures of the compound including the Fc-binding unit. Furthermore, the inventors of the present application conducted experiments on linkers A of various lengths, and found that the length of linker A Even though it was changed, it was confirmed that the compound containing the Fc binding unit had excellent reaction efficiency in the reaction with the antibody (see Examples 07 to 08).

[0392]

[0393] R"a1

[0394] As mentioned above, in chemical formula 2, R a1 is H or C 1-6 may be alkyl. In some embodiments, R a1 is H or C 1-4 It can be alkyl. In this case, C 1-4 Alkyl includes methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, and tert-butyl. In certain embodiments, R a1 is H or C 1-3 may be alkyl. In certain embodiments, R a1 is H or C 1-2may be alkyl. In certain embodiments, R a1 can be methyl.

[0395]

[0396] R"a2 and R"a3

[0397] As mentioned above, in chemical formula 2, R a2 is H or C 1-6 may be alkyl. In some embodiments, R a2 is H or C 1-4 It can be alkyl. In this case, C 1-4 Alkyl includes methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, and tert-butyl. In certain embodiments, R a2 is H or C 1-3 may be alkyl. In certain embodiments, R a2 is H or C 1-2 may be alkyl. In certain embodiments, R a2 may be H or methyl. In certain embodiments, R a2 can be H.

[0398] As mentioned above, in chemical formula 2, R a3 is H or C 1-6 may be alkyl. In some embodiments, R a3 is H or C 1-4 It can be alkyl. In this case, C 1-4 Alkyl includes methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, and tert-butyl. In certain embodiments, R a3 is H or C 1-3 may be alkyl. In certain embodiments, R a3 is H or C 1-2 may be alkyl. In certain embodiments, R a3 may be H or methyl. In certain embodiments, Ra3 can be H.

[0399] In certain embodiments, R a2 If not H, R a3 may be H. In certain embodiments, R a3 If not H, R a2 may be H. In certain embodiments, R a2 and R a3 can all be H.

[0400]

[0401] X

[0402] As described above, X can be C, O, or N (i.e., X can be -CH2-, -O-, or -NH-). In some embodiments, X can be C or O. In certain embodiments, X can be C. In certain embodiments, X can be O.

[0403]

[0404] J"a

[0405] As mentioned above, in chemical formula 2, J a is -C(=O)-, -S-, -NH-, or -C(=NH)-. J a In chemical formula 2, represents the bonding moiety of FcBU and other parts other than FcBU. J a- may be referred to, for example, as a junction moiety A or a junction moiety A. In certain embodiments, J a can be -C(=O)-.

[0406]

[0407] Group of interest (GOI)

[0408] Overview of the target group

[0409] In Formula 2, GOI is a group of interest. In some embodiments, the group of interest may comprise a reactive group. The group of interest may comprise one or more reactive groups. In some embodiments, the group of interest may comprise a functional group. The group of interest may comprise one or more functional groups. The group of interest may comprise one or more reactive groups and one or more functional groups. For example, the group of interest may comprise one or more reactive groups. For example, the group of interest may comprise one or more functional groups. For example, the group of interest may comprise one reactive group and one functional group. For example, the group of interest may comprise one reactive group and one functional group. For example, the group of interest may comprise two or more reactive groups and one functional group. For example, the group of interest may comprise two or more reactive groups and two or more functional groups. When the group of interest comprises multiple reactive groups, each reactive group is independently selected. When the group of interest comprises multiple functional groups, each functional group is independently selected. In some embodiments, the group of interest may be a reactive group or a functional group. In certain embodiments, the desired group may be a reactive group. For example, the desired group may be a group intended for delivery to an antibody.

[0410] In some embodiments, the sum of the atomic masses of all atoms comprising the desired group can be 5000 Daltons or less, 4000 Daltons or less, 3000 Daltons or less, 2000 Daltons or less, 1500 Daltons or less, 1000 Daltons or less, 900 Daltons or less, 800 Daltons or less, 700 Daltons or less, 600 Daltons or less, 500 Daltons or less, 400 Daltons or less, 300 Daltons or less, or 100 Daltons or less.

[0411]

[0412] Below, the reactive group and functional group are described in detail.

[0413]

[0414] reactive group (RG)

[0415] The target group may include a reactive group.

[0416] In some embodiments, the desired group may be a reactive group.

[0417] A reactive group may include a reactive moiety. A reactive moiety may refer to a moiety that is reactive with another molecule or a group within another molecule.

[0418] In some embodiments, the reactive moiety may be, but is not limited to, a click chemistry functional group or a bioorthogonal functional group, and may be selected from groups having reactivity known in the art.

[0419] In some embodiments, the sum of the atomic masses of all atoms comprising the reactive group can be 3000 Daltons or less, 2500 Daltons or less, 2000 Daltons or less, 1500 Daltons or less, 1000 Daltons or less, 900 Daltons or less, 800 Daltons or less, 700 Daltons or less, 600 Daltons or less, 500 Daltons or less, 400 Daltons or less, 300 Daltons or less, 200 Daltons or less, or 100 Daltons or less.

[0420]

[0421] In some embodiments, the reactive group can have the structure:

[0422] ,

[0423] At this time, D RG is the spacer of the reactive group (spacer RG), and H RG is a reactive moiety. In some embodiments, the reactive moiety may be a click chemistry functional group. In some embodiments, the reactive moiety may be a bioorthogonal functional group.

[0424] In some embodiments, the spacer of the reactive group (D RG- ) may be 0 to 6 based on the number of atoms in the main chain. In some embodiments, D RG is a bond, or substituted or unsubstituted C 1-6 Alkylene, substituted or unsubstituted C 1-6 Heteroalkylene, substituted or unsubstituted C 2-6 Alkenylene, substituted or unsubstituted C 2-6 Heteroalkenylene, substituted or unsubstituted C 2-6 Alkynylene, substituted or unsubstituted C 2-6 Heteroalkynylene, substituted or unsubstituted C 3-8 Cycloalkylene, substituted or unsubstituted C 3-8 Heterocycloalkylene, substituted or unsubstituted C 3-8 Cycloalkenylene (e.g., aryl), or substituted or unsubstituted C3-8 Heterocycloalkenylene (e.g., heteroaryl) may be substituted. Here, substituted indicates that one or more hydrogen atoms in the group modified by the term substituted is replaced by one or more substituents. That is, substituted alkylene, etc. may include one or more substituents. For example, the substituents may be each independently selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH. In certain embodiments, the substituents may be selected from, each independently, -C 1-4 Alkyl, -C(=O)H, -C(=O)CH3, -C(=O)OH, -C(=O)NH2, -NH2, =NH, =O, =S, -OH, -NO2, and -SH. Here, heteroalkylene, heteroalkenylene, and heteroalkynylene each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0425] In certain embodiments, D RG is a bond, or substituted or unsubstituted C 1-6 Alkylene, substituted or unsubstituted C 1-6 It may be heteroalkylene. In this case, the substituted alkylene or substituted heteroalkylene may include one or more substituents, and the substituents are each independently -C 1-4 Alkyl and =O may be selected from. Here, heteroalkylene contains one or more heteroatoms, and here, the heteroatoms may be independently selected from O, N, and S.

[0426] In certain embodiments, D RG is a bond, or substituted or unsubstituted C 1-3 Alkylene, substituted or unsubstituted C 1-3 It may be heteroalkylene. In this case, the substituted alkylene or substituted heteroalkylene may include one or more substituents, and the substituents are each independently -C 1-4 Alkyl and =O may be selected from. Here, heteroalkylene contains one or more heteroatoms, and here, the heteroatoms may be independently selected from O, N, and S.

[0427] In certain embodiments, D RG can be a combination.

[0428] In certain embodiments, the reactive group may have the structure:

[0429] .

[0430]

[0431] reactive moiety

[0432] A reactive moiety may refer to a moiety that is reactive with another group. A substance, molecule, or compound having a first reactive moiety (e.g., a compound including an Fc binding unit) may be bonded to a substance, molecule, or compound having a second reactive moiety by a reaction between the reactive moiety and a moiety capable of reacting with the reactive moiety (wherein the reactive moiety may be referred to as a first reactive moiety, and the moiety capable of reacting with the first reactive moiety may be referred to as a second reactive moiety). At this time, the substance, molecule, or compound having the first reactive moiety and the substance, molecule, or compound having the second reactive moiety may be covalently bonded.

[0433] In some embodiments, the reactive moiety may be a bio-orthogonal functional group. A bio-orthogonal functional group refers to a chemical functional group that participates in bio-orthogonal chemistry or a bio-orthogonal reaction, thereby performing a bio-orthogonal reaction. The bioorthogonal reaction may be, but is not limited to, for example, Staudinger Ligation, Copper-Catalyzed Azide-Alkyne Cycloaddition (CuAAC), Copper-Free Azide-Alkyne Cycloaddition including strain-promoted azide-alkyne cycloaddition (SPAAC), Tetrazine Ligation, Tetrazole Ligation, Oxime Ligation, or Isocyanide Click Reaction.

[0434] The bioorthogonal functional group can be selected from, but is not limited to, an azide group, a terminal alkyne group, a terminal alkene group, a cyclic alkyne (e.g., cyclooctyne) group, a tetrazine group, a norbornene group, a cycloalkene (e.g., cyclooctene) group, an oxime group, and an isocyanide group, for example. The cyclooctyne can be any one selected from, but is not limited to, OCT cyclooctyne, BCN (Bicyclononyne), DBCO (Dibenzocyclooctyne), DIBAC (aza-dibenzocyclooctynes), DIBO (dibenzocyclooctynol), DIFO (difluorinated cyclooctynes), BARAC (biarylazacyclooctynone), DIMAC (dimethoxyazacyclooctyne), and DIFBO (difluorobenzocyclooctyne). Cyclooctene can be, for example, any one selected from a cis-cyclooctene group and a trans-cyclooctene group. In some embodiments, the group exemplified as a bioorthogonal functional group can have one or more substituents. The substituents can each be independently selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein each R is independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 It can be selected from heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, =O, =S, and -SH.

[0435] In some embodiments, the bioorthogonal functional group can have any of the following structures:

[0436]

[0437]

[0438] In some embodiments, hn can be an integer from 1 to 3, and R H are each independently H or -R, =O, =S, -NO2, -CR3, -NR2, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 It can be selected from heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, =O, =S, and -SH. R H If =O or =S, then two R H It will be understood that it forms one =O or =S.

[0439]

[0440] In some embodiments, the reactive moiety may be a click chemistry functional group. A click chemistry functional group refers to a group that participates in a click chemistry reaction. The click chemistry reaction may be any one selected from the group consisting of Huisgen 1,3-dipolar cycloaddition; Diels-Alder reaction; inverse-electron-demand Diels-Alder (IEDDA) reaction; nucleophilic addition to small strained rings such as epoxides and aziridines; nucleophilic addition to activated carbonyl groups; and Staudinger ligation and addition to carbon-carbon double or triple bonds. In some embodiments, the click chemistry functional group can be selected from a diels-alder diene, a diels-alder dienophile, an IEDDA diene, and an IEDDA dienophile.

[0441]

[0442] In some embodiments, the reactive moiety can be selected from an azide group, a terminal alkyne group, a cyclic alkyne (e.g., cyclooctyne) group, a tetrazine group, a norbornene group, a cycloalkene (e.g., cyclooctene) group, a tetrazole group, a triazine group, an oxime group, and an isocyanide group, a halogen group, an aldehyde group, a nitrone group, a hydroxylamine group, a nitrile group, a hydrazine group, a ketone group, a bronic acid group, a cyanobenzothiazole group, an allyl group, a phosphine group, a maleimide group, a disulfide group, a thioester group, a halocarbonyl group, an isonitrile group, a cydnone group, a selenium group, a thiol group, and a protected thiol group.

[0443]

[0444] functional group (FG)

[0445] The target group may include a functional group.

[0446] In some embodiments, the desired group may be a functional group.

[0447] The functional group may comprise an active moiety. The active moiety may be, but is not limited to, a drug, an imaging moiety, a radioactive moiety, a protein having a specific function, a peptide having a specific function, an affinity agent (e.g., biotin, streptavidin, and aptamers), a stabilizer, a vitamin, a nucleic acid (e.g., DNA or RNA), or a PEG moiety. In some embodiments, the active moiety may be a drug moiety, an imaging moiety, a radioactive moiety, or an affinity agent.

[0448] In some embodiments, the molecular weight of the functional group (i.e., the sum of the atomic masses of all atoms comprising the functional group) can be 5000 Daltons or less, 4000 Daltons or less, 3000 Daltons or less, 2500 Daltons or less, 2000 Daltons or less, 1500 Daltons or less, or 1000 Daltons or less.

[0449] In some embodiments, the functional group can have the structure:

[0450] ,

[0451] At this time D FG is a spacer of a functional group (spacer FG), and AM is an active moiety. In some embodiments, the active moiety can be a drug (e.g., a toxin). In some embodiments, the active moiety can be a radioactive moiety. In some embodiments, the active moiety can be an imaging moiety.

[0452] In some embodiments, the spacer (D) of the functional group FG ) may be 0 to 6 based on the number of atoms in the main chain. In some embodiments, D FG is a bond, or substituted or unsubstituted C 1-6 Alkylene, substituted or unsubstituted C 1-6 Heteroalkylene, substituted or unsubstituted C 2-6 Alkenylene, substituted or unsubstituted C 2-6 Heteroalkenylene, substituted or unsubstituted C 2-6 Alkynylene, substituted or unsubstituted C 2-6 Heteroalkynylene, substituted or unsubstituted C 3-8 Cycloalkylene, substituted or unsubstituted C 3-8 Heterocycloalkylene, substituted or unsubstituted C 3-8 Cycloalkenylene (e.g., aryl), or substituted or unsubstituted C 3-8Heterocycloalkenylene (e.g., heteroaryl) may be substituted. Here, substituted indicates that one or more hydrogen atoms in the group modified by the term substituted is replaced by one or more substituents. That is, substituted alkylene, etc. may include one or more substituents. For example, the substituents may be each independently selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, =O, =S, and -SH. In certain embodiments, the substituents may be selected from, each independently, -C 1-4 Alkyl, -C(=O)H, -C(=O)CH3, -C(=O)OH, -C(=O)NH2, -NH2, =O, =S, -OH, -NO2, and -SH. Here, heteroalkylene, heteroalkenylene, and heteroalkynylene each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0453] In certain embodiments, D FG is a bond, or substituted or unsubstituted C 1-3 Alkylene, substituted or unsubstituted C 1-3 It may be heteroalkylene. In this case, the substituted alkylene or substituted heteroalkylene may include one or more substituents, and the substituents are each independently -C 1-4 Alkyl and =O may be selected from. Here, heteroalkylene contains one or more heteroatoms, and here, the heteroatoms may be independently selected from O, N, and S.

[0454] In certain embodiments, D FGcan be a combination.

[0455] In certain embodiments, the functional group may have the structure:

[0456] .

[0457]

[0458] In some embodiments, the active moiety can be a drug (e.g., a drug unit or a conjugated drug). Drugs include, for example, auristatins (e.g., monomethyl auristatin e), eribulin, tubulysin, geldanamycin (Kerr et al., 1997, Bioconjugate Chem. 8(6):781-784), maytansinoids (EP 1391213, ACR 2008, 41, 98-107), calicheamicin (US Patent Publication No. 2009 / 0105461, Cancer Res. 1993, 53, 3336-3342), mertansine, daunomycin, doxorubicin, methotrexate, Vindesine, SG2285 (Cancer Res. 2010, 70(17), 6849-6858), dolastatin, dolastatin analogs auristatin (US Patent No.5,635,483), cryptophycin, camptothecin, camptothecin analogs (e.g., SN38, FL118, or exatecan), rhizoxin derivatives, CC 1065 analogs or derivatives, duocarmycin, enediyne antibiotics, esperamicin, epothilones, pyrrolobenzodiazepine (PBD) derivatives, α-amanitin, toxoids, TLR5 (toll-like receptor 5) agonist TLR7 (toll-like receptor 7) agonist, TLR8 (toll-like receptor 8) agonist, NOTA (1,4,7-Triazacyclononane-1,4,7-triacetic acid), and DOTA (1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid), or an analog thereof, but is not otherwise limited thereto.

[0459] In some embodiments, the active moiety may comprise a radioactive moiety. The radioactive moiety may be a ligand (e.g., a chelator) designed to bind a radioisotope. In some embodiments, the radioactive moiety may comprise a complex (e.g., a chelate) of a chelator and a radioisotope.

[0460]

[0461] Fc binding unit (FcBU)

[0462] Overview of the Fc binding unit

[0463] As described above, in Chemical Formula 2, FcBU is an Fc-binding unit. The Fc-binding unit can be derived from an Fc-binding substance. The Fc-binding substance is described in detail below.

[0464]

[0465] Fc binding substance

[0466] Overview of Fc-binding substances

[0467] An Fc-binding material is a material that binds to the Fc region of an antibody, and is used as an element of a compound (e.g., a compound comprising an Fc-binding unit) for delivering a target group to a target position of an antibody. The technique of introducing a target material (e.g., a bioorthogonal functional group and a drug, etc.) to a specific position of an antibody using an Fc-binding material and a compound comprising the same is described in detail in the literature [European Patent Application No. 19818561.3, Publication No. EP 3811978; PCT Patent Application No. PCT / KR2020 / 003282, Publication No. WO2020 / 184944; and Korean Patent Application No. 10-2020-0009162, Publication No. 10-2020-0091826], the entire contents of each of which are incorporated herein by reference. As appropriate, the Fc binding agent may be referred to as, but is not limited to, an affinity agent for an antibody, an IgG binding agent, a site-specific antibody interactome, or a site-specific Fc interactome.

[0468]

[0469] Fc binding substance of the present application

[0470] Some embodiments of the present application provide an Fc-binding agent comprising an Fc-binding peptide having an amino acid sequence of SEQ ID NO: 01. Some embodiments of the present application provide an Fc-binding agent comprising an amino acid sequence of SEQ ID NO: 01.

[0471] The amino acid sequence of sequence number 01 is as follows:

[0472] (Xaa) 1-3 -C-(Xaa)2-H-Xa 1 -G-Xa 2 -LV-Xa 3 -C-(Xaa) 1-3 (Sequence number 01),

[0473] At this time,

[0474] Each Xaa is independently any amino acid,

[0475] Xa 1 silver , and here, m is an integer from 1 to 10, and X f is -NH2, -SH, or -C(=O)OH,

[0476] Xa 2 is a glutamic acid residue or an asparagine residue,

[0477] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue.

[0478] In some embodiments, in the amino acid sequence of SEQ ID NO: 01, the cysteine ​​residue adjacent to the N-terminus (i.e., the cysteine ​​residue located 2 to 4 from the N-terminus) and the cysteine ​​residue adjacent to the C-terminus (i.e., the cysteine ​​residue located 2 to 4 from the C-terminus) may optionally be covalently linked. For example, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus may optionally be covalently linked via a disulfide bond. For example, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus may optionally have a structure comprising a carbonyl group. can be shared through .

[0479] In some embodiments, each Xaa can independently be any amino acid other than cysteine.

[0480] In some embodiments, Xa 1 The residue may be a diaminopropionic acid (Dap) residue, a diaminobytyric acid (Dab) residue, an ornithine (Orn) residue, a lysine (Lys) residue, a 2,7-diaminoheptanoic acid residue, a 2,8-diaminooctanoic acid residue, a 2,9-diaminononanoic acid residue, a cysteine ​​residue, or a 2-aminosuberic acid residue.

[0481] For example, Xa 1 The structure of In , m is 1 and X f If -NH2, Xa 1 is a Dap residue. m is 2 and X f If -NH2, Xa 1 is a Dab residue. m is 3 and X f If -NH2, Xa 1 is an Orn residue. m is 4 and X f If -NH2, Xa 1 is a Lys residue. m is 5 and X f If -NH2, Xa 1 is a 2,7-diaminoheptanoic acid residue. m is 1 and X f If -SH, Xa 1 is a Cys residue. m is 5 and X f If it is -COOH, Xa 1 is a 2-aminosuberic acid residue.

[0482] In some embodiments, Xa 1 The structure of In , m can be an integer from 1 to 5. In some embodiments, m can be an integer from 1 to 4.

[0483] Xa 1 The structure of Silver structure Xa 1 (structure Xa1) and is not otherwise limited. For example, Xa 1 may be referred to as an amino acid residue having the structure Xa1, but is not otherwise limited.

[0484]

[0485] In certain embodiments, Xa 1 can be expressed in the following structure:

[0486] ,

[0487] At this time, m can be an integer from 1 to 4.

[0488] In the above structure, when m is 1, Xa 1 is a Dap residue. If m is 2, Xa 1 is a Dab residue. If m is 3, Xa 1 is an Orn residue. If m is 4, Xa 1 is a Lys residue.

[0489]

[0490] An Fc-binding peptide (e.g., the amino acid sequence of SEQ ID NO: 01) may include modifications commonly used in the art. In some embodiments, an Fc-binding peptide may include modifications that do not impair the inherent function of the Fc-binding peptide. For example, an Fc-binding peptide or Fc-binding substance that has binding affinity for the Fc region of an antibody may include modifications that do not impair this function. For example, the modifications may be modifications that control the stability or hydrophilicity of the Fc-binding peptide. For example, the modification can be addition of a hydrophilic moiety, addition of a hydrophobic moiety, addition of a PEG moiety (e.g., pegylation), addition of an amide group (e.g., amidation of the C-terminus), addition of a carbohydrate group, addition of a hydroxyl group, addition of a phosphate group, addition of a prenyl group (e.g., prenylation), and / or addition of a farnesyl group (e.g., farnesylation).

[0491] For example, the Fc-binding agent can further comprise one or more PEG moieties in addition to the Fc-binding peptide having the amino acid sequence of SEQ ID NO: 01. The PEG moieties can be covalently linked to the N-terminus and / or the C-terminus of the Fc-binding peptide. For example, the Fc-binding agent comprises an Fc-binding peptide and a PEG moiety, wherein the PEG moiety can be covalently linked to the C-terminus of the Fc-binding peptide. In another example, the Fc-binding agent comprises an Fc-binding peptide and a PEG moiety, wherein the PEG moiety can be covalently linked to the N-terminus of the Fc-binding peptide. The PEG moieties can comprise, for example, 1 to 30, 1 to 20, or 1 to 10 ethylene glycol units.

[0492] Below, as an example, the structure of an Fc-binding peptide with or without a modification added to the N-terminus and / or C-terminus of the amino acid sequence of SEQ ID NO: 01 is illustrated through a chemical formula.

[0493] For example, the Fc binding agent may have the structure of Chemical Formula 3 below:

[0494] [Chemical Formula 3]

[0495] (M F1 ) 0-1 -(Xaa) 1-3 -C-(Xaa)2-H-Xa 1 -G-Xa 2 -LV-Xa 3 -C-(Xaa) 1-3 -(M F2 ) 0-1 ,

[0496] At this time, M F1 and M F2 are each independently modified,

[0497] Each Xaa is independently any amino acid other than cysteine,

[0498] Xa 1 silver , and here, m is an integer from 1 to 10, and X f is -NH2, -SH, or -C(=O)OH,

[0499] Xa 2 is a glutamic acid residue or an asparagine residue,

[0500] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue.

[0501] In Chemical Formula 3, the cysteine ​​residue adjacent to the N-terminus (i.e., the cysteine ​​residue positioned 2 to 4 from the N-terminus) and the cysteine ​​residue adjacent to the C-terminus (i.e., the cysteine ​​residue positioned 2 to 4 from the C-terminus) may optionally be covalently linked. For example, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus may optionally be linked via a disulfide bond.

[0502] In some embodiments, Xa 1 The residue may be a diaminopropionic acid residue, a diaminobytyric acid residue, an ornithine residue, a lysine residue, a 2,7-diaminoheptanoic acid residue, a 2,8-diaminooctanoic acid residue, a 2,9-diaminononanoic acid residue, a cysteine ​​residue, or a 2-aminosuberic acid residue.

[0503] As shown in chemical formula 3, M F1 may or may not exist.

[0504] As shown in chemical formula 3, M F2 may or may not exist.

[0505] As mentioned above, the modifications can be selected from, for example, addition of a hydrophilic moiety, addition of a hydrophobic moiety, addition of a PEG moiety (e.g., pegylation), addition of an amide group (e.g., amidation of the C-terminus), addition of a carbohydrate group, addition of a hydroxyl group, addition of a phosphate group, addition of a prenyl group (e.g., prenylation), and addition of a farnesyl group (e.g., farnesylation).

[0506] In some embodiments, M F1 may be selected from a PEG moiety, an amide group, a carbohydrate group, a hydroxyl group, a phosphate group, a prenyl group, and a farnesyl group. In certain embodiments, M F1 may be a PEG moiety.

[0507] In some embodiments, M F2 may be selected from a PEG moiety, an amide group, a carbohydrate group, a hydroxyl group, a phosphate group, a prenyl group, and a farnesyl group. In certain embodiments, M F2 may be an amide group.

[0508] At this time, the PEG moiety may include 1 to 30, 1 to 20, or 1 to 10 ethylene glycol units (e.g., -CH2OCH2-, -OCH2CH2-, or -CH2CH2O-).

[0509] For example, a PEG moiety can have the following structure:

[0510] .

[0511] At this time, D PEG is a spacer of the PEG moiety.

[0512] For example, D PEG can be a group whose main chain length is 0 to 6 (i.e., a group whose sum of the number of atoms located in the main chain is 0 to 6).

[0513] For example, D PEG is a combination, or C 1-6 Substituted or unsubstituted alkylene, C 1-6 Substituted or unsubstituted heteroalkylene, C 1-6 Substituted or unsubstituted alkenylene, C 1-6Substituted or unsubstituted heteroalkenylene, substituted or unsubstituted alkynylene, or substituted or unsubstituted heteroalkynylene, wherein the substituted alkylene, substituted heteroalkylene, substituted alkenylene, or substituted heteroalkynylene may include one or more substituents, and may be selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, =O, =S, and -SH. In certain embodiments, the substituent may be selected from -C 1-4 Alkyl, -C(=O)H, -C(=O)CH3, -C(=O)OH, -C(=O)NH2, -NH2, =O, =S, -OH, -NO2, and -SH. Here, heteroalkylene, heteroalkenylene, and heteroalkynylene each independently contain one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0514] In certain embodiments, D PEG is a bond, substituted or unsubstituted C 1-3 Alkylene, or substituted or unsubstituted C 1-3 It may be heteroalkylene, wherein the substituted alkylene or substituted heteroalkylene comprises one or more substituents, wherein the substituents are =O, and wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be independently selected from N, O, and S.

[0515] In the structure of the above PEG moiety, R PEGis a PEG capping group. At this time, the PEG capping group is absent, -CH3, C2 alkyl, C3 alkyl, -NH2, -CH2NH2, -SC(=O)CH3, -SC(=O)CH2CH3, -CH2SC(=O)CH3, -CH2SC(=O)CH2CH3, -OH, -CH2OH, -SH, -CH2SH, -OCH3, -CH2OCH3, -CH2OCH2CH3, -C(=O)CH3, -C(=O)CH2CH3, -CH2C(=O)CH3, -CH2C(=O)CH2CH3, -NHC(=O)CH3, -NHC(=O)CH2CH3, -CH2NHC(=O)CH3, -CH2CH2NHC(=O)CH3, -CH2CH2NHC(=O)CH3, -CH2CH2NHC(=O)CH2CH3, -CH2NHC(=O)CH2CH3, -CH2CH2COOH, glucose or -O-glucose, but is not limited thereto. In certain embodiments, the PEG capping group may be absent, -CH3, -OCH3, -CH2OCH3, -C(=O)CH3, -CH2C(=O)CH3, -NHC(=O)CH3, -CH2NHC(=O)CH3 or -CH2CH2COOH. In some embodiments, the sum of the atomic masses of the atoms belonging to the PEG capping group may be, but is not limited to, 300 daltons or less, 200 daltons or less, 150 daltons or less, 100 daltons or less, or 50 daltons or less.

[0516] In the structure of the above PEG moiety, p may be an integer of 1 to 30, preferably 1 to 10.

[0517] In the structure of the above PEG moiety, [EG] refers to an ethylene glycol unit, and the ethylene glycol unit is as described above in the relevant paragraph.

[0518] The structure of the above-mentioned chemical formula 3 is an example of a case where the Fc-binding substance or Fc-binding peptide includes a modification, and the modifications shown in the structure of the chemical formula 3 (for example, M) are described below. F1 and / or M F2) is not indicated, it will be understood that the Fc-binding agent or Fc-binding peptide may further comprise modifications.

[0519]

[0520] In some embodiments, the Fc binding agent can comprise the amino acid sequence of SEQ ID NO: 02.

[0521] L PDC-(Xaa)2-H-Xa 1 -G-Xa 2 -LV-Xa 3 -CT- D P (SEQ ID NO: 02),

[0522] At this time, Xaa, Xa 1 , Xa 2 , and Xa 3 Each is as described above.

[0523] At this time, the N-terminal L P (L-proline residue) and D P (D-proline residue) can optionally form a D-proline-L-proline template.

[0524]

[0525] In some embodiments, the Fc binding agent can comprise the amino acid sequence of SEQ ID NO: 03.

[0526] CDC-(Xaa)2-H-Xa 1 -G-Xa 2 -LV-Xa 3 -CTC (SEQ ID NO: 03),

[0527] At this time, Xaa, Xa 1 , Xa 2 , and Xa 3 Each is as described above.

[0528] At this time, the cysteine ​​at the N terminal and the cysteine ​​at the C terminal can be optionally covalently linked.

[0529]

[0530] In certain embodiments, the Fc binding agent may comprise the amino acid sequence of SEQ ID NO: 04:

[0531] DC-(Xaa)2-H-Xa 1 -G-Xa 2 -LV-Xa 3 -CT (SEQ ID NO: 04),

[0532] At this time, Xaa, Xa 1 , Xa 2 , and Xa 3 Each is as described above.

[0533] At this time, the cysteine ​​residue adjacent to the N terminus (the cysteine ​​connected to D in SEQ ID NO: 04) and the cysteine ​​residue adjacent to the C terminus (the cysteine ​​connected to T in SEQ ID NO: 04) can optionally be covalently linked (e.g., via a disulfide bond).

[0534] In certain embodiments, Xa 1 It may be a diaminopropionic acid (Dap) residue, a diaminobytyric acid (Dab) residue, an ornithine (Orn) residue, or a lysine (Lys) residue.

[0535]

[0536] In certain embodiments, the Fc binding agent may comprise the amino acid sequence of SEQ ID NO: 05:

[0537] DCAWH-Xa 1 -GELVWCT (SEQ ID NO: 05),

[0538] At this time,

[0539] Xa 1 silver , and here, m is an integer from 1 to 10, and X f is -NH2, -SH, or -C(=O)OH. In some embodiments, m can be an integer from 1 to 4.

[0540] In some embodiments, Xa 1 may be a diaminopropionic acid (Dap) residue, a diaminobytyric acid (Dab) residue, an ornithine (Orn) residue, a lysine (Lys) residue, a 2,7-diaminoheptanoic acid residue, a 2,8-diaminooctanoic acid residue, a 2,9-diaminononanoic acid residue, a cysteine ​​residue, or a 2-aminosuberic acid residue. In certain embodiments, Xa 1 It may be a diaminopropionic acid (Dap) residue, a diaminobytyric acid (Dab) residue, an ornithine (Orn) residue, or a lysine (Lys) residue.

[0541] At this time, the cysteine ​​residue adjacent to the N terminus (the cysteine ​​connected to D in SEQ ID NO: 05) and the cysteine ​​residue adjacent to the C terminus (the cysteine ​​connected to T in SEQ ID NO: 05) can optionally be covalently linked (e.g., via a disulfide bond).

[0542]

[0543] In certain embodiments, the Fc binding agent may comprise any one of the amino acid sequences selected from SEQ ID NO: 06 to SEQ ID NO: 09:

[0544] DCAWH-Dap-GELVWCT (SEQ ID NO: 06);

[0545] DCAWH-Dab-GELVWCT (SEQ ID NO: 07);

[0546] DCAWH-Orn-GELVWCT (SEQ ID NO: 08); and

[0547] DCAWH-Lys-GELVWCT (SEQ ID NO: 09).

[0548] At this time, the cysteine ​​residue adjacent to the N terminus (e.g., a cysteine ​​linked to D) and the cysteine ​​residue adjacent to the C terminus (e.g., a cysteine ​​linked to T) can optionally be covalently linked (e.g., via a disulfide bond).

[0549]

[0550] Design of Fc-binding materials

[0551] As described above, the Fc-binding substance of the present application has binding activity with the Fc region of an antibody. At this time, the Fc-binding substance and the Fc region can be arranged in a specific positional relationship through interactions between amino acid residues. Representative interactions between the Fc-binding peptide of the Fc-binding substance and the Fc region of the antibody include (1) salt linkage with histidine at position 433 of the Fc region, (2) hydrogen bonding with asparagine at position 434, (3) salt linkage with glutamic acid at position 380, and (4) salt linkage with arginine at position 255. These interactions and the specific positional relationships resulting from them can be identified through already known research results. (See DeLano, WL, Ultsch, MH, de, AM, Vos, N., & Wells, JA (2000). Convergent solutions to binding at a protein-protein interface. Science, 287(5456), 1279-1283.)

[0552] In designing the Fc-binding material of the present application, it is important to ensure that the Fc-binding material forms a stable positional relationship with the Fc region of the antibody. This is because the compound comprising the Fc-binding unit of the present application and the labeling process of the antibody using the same (i.e., the process of transferring the target group) were designed based on the positional relationship between the Fc-binding unit and the Fc region discovered through research.

[0553] An example of the design principle of an Fc binding material is the amino acid sequence of SEQ ID NO: 05 (DCAWHXa 1 This is explained through an Fc-binding peptide having GELVWCT.

[0554] The results of simulating the positional relationship between the Fc-binding peptide having the amino acid sequence of SEQ ID NO: 05 and the Fc region based on academic papers, etc., are as shown in Figures 01 to 03. At this time, the 5th histidine residue of the Fc-binding peptide forms a salt linkage with the 380th glutamic acid residue of the Fc region, and it was confirmed that this has a significant effect on the positional relationship between the Fc-binding peptide and the Fc region (see the dotted line in Figure 03). Therefore, it is desirable not to change the histidine residue and its position when designing the Fc-binding peptide. Furthermore, the 8th glutamic acid residue is an electronegative residue and forms a salt linkage with the 255th arginine residue of the Fc region, which has an electronegative charge, and it was confirmed that this has a significant effect on the positional relationship between the Fc-binding peptide and the Fc region (see the dotted line in Figure 03). Therefore, the residue is preferably an acidic amino acid that can correspond to glutamic acid, and can be replaced with asparagine. If the amino acid residues are changed to other amino acid residues or substituted with any functional group, it will affect the intermolecular interaction and affect the positional relationship between the Fc-binding peptide and the Fc region. In addition, glycine at position 7 of SEQ ID NO: 05 is a small amino acid and is necessary to form a folded structure of the Fc-binding peptide. Therefore, it is preferable not to change glycine and its position in the design of the Fc-binding peptide. Figure 04 illustrates the positional relationship between lysine residues of the Fc domain and the Fc-binding peptide. In the positional relationship, Xa 1 It was confirmed that the lysines in the Fc region located most adjacent to each other were lysine 246 and lysine 248 (Figure 04).

[0555]

[0556] Antibodies that have affinity for Fc-binding substances

[0557] As described above, the Fc binding agent has binding affinity to the Fc region of an antibody.

[0558] In this specification, the term "antibody" is used to encompass all antibodies or fragments thereof that have affinity for Fc-binding substances, and even those that do not bind to a specific antigen, such as Fc fragments, can be recognized as antibodies. Furthermore, the term "antibody" can be interpreted to encompass monospecific antibodies, bispecific antibodies, trispecific antibodies, monoclonal antibodies, human antibodies, humanized antibodies, recombinant antibodies, chimeric antibodies, and antibody variants.

[0559] Furthermore, since antibodies contain an Fc region, they may be referred to as Fc proteins.

[0560] In some embodiments, the antibody may comprise an Fc region of IgG. In some embodiments, the Fc region of the antibody may be an Fc region of IgG.

[0561] In some embodiments, the antibody may be an IgG antibody. IgG antibodies include human IgG antibodies, humanized IgG antibodies, and chimeric IgG antibodies.

[0562] IgG is known to be classified into IgG1, IgG2, IgG3, and IgG4.

[0563] In some embodiments, the antibody may be an IgG1 antibody. IgG1 antibodies include human IgG1 antibodies, humanized IgG1 antibodies, and chimeric IgG1 antibodies.

[0564] In some embodiments, the antibody may comprise an Fc region of IgG1. The Fc region of the antibody may be an Fc region of IgG1.

[0565] In some embodiments, the antibody may be an IgG2 antibody. IgG2 antibodies include human IgG2 antibodies, humanized IgG2 antibodies, and chimeric IgG2 antibodies.

[0566] In some embodiments, the antibody may comprise an Fc region of IgG2. The Fc region of the antibody may be an Fc region of IgG2.

[0567] In some embodiments, the antibody may be an IgG3 antibody. IgG3 antibodies include human IgG3 antibodies, humanized IgG3 antibodies, and chimeric IgG3 antibodies.

[0568] In some embodiments, the antibody may comprise an Fc region of IgG3. The Fc region of the antibody may be an Fc region of IgG3.

[0569] In some embodiments, the antibody may be an IgG4 antibody. IgG4 antibodies include human IgG4 antibodies, humanized IgG4 antibodies, and chimeric IgG4 antibodies.

[0570] In some embodiments, the antibody may comprise an Fc region of IgG4. The Fc region of the antibody may be an Fc region of IgG4.

[0571] In some embodiments, the antibody can have an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In particular embodiments, the antibody can have an amino acid sequence of SEQ ID NO: 14 or an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto.

[0572] In some embodiments, the antibody comprises an IgG Fc region (e.g., the Fc region of the antibody is an Fc region of IgG), wherein the IgG Fc region can have an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In particular embodiments, the antibody comprises an Fc region of IgG, wherein the IgG Fc region can have an amino acid sequence of SEQ ID NO: 14 or an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto.

[0573] In some embodiments, the antibody or Fc region of the antibody may comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNH (SEQ ID NO: 11).

[0574] In some embodiments, the antibody or Fc region of the antibody may comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNHY (SEQ ID NO: 12).

[0575] In some embodiments, the antibody or FC region of the antibody may comprise the amino acid sequence GPSVFLFPPKPKDTLM (SEQ ID NO: 13).

[0576] In some embodiments, the antibody has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNH (SEQ ID NO: 11). In some embodiments, the antibody comprises an IgG Fc region, wherein the IgG Fc region has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNH (SEQ ID NO: 11).

[0577] In some embodiments, the antibody has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 15 and SEQ ID NOs: 17 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, and may essentially comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNHY (SEQ ID NO: 12). In some embodiments, the antibody comprises an IgG Fc region, wherein the IgG Fc region has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 15 and SEQ ID NOs: 17 to 18, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, and may essentially comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNHY (SEQ ID NO: 12).

[0578] In some embodiments, the antibody has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises GPSVFLFPPKPKDTLM (SEQ ID NO: 13). In some embodiments, the antibody comprises an IgG Fc region, wherein the IgG Fc region has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises GPSVFLFPPKPKDTLM (SEQ ID NO: 13).

[0579] In some embodiments, the antibody having binding affinity for the Fc-binding peptide of the present application may be an antibody of the IgG isotype. In some embodiments, the antibody having binding affinity for the Fc-binding peptide of the present application may be an antibody of the IgG1 isotype, an antibody of the IgG2 isotype, an antibody of the IgG3 isotype, or an IgG4 isotype. In some embodiments, the antibody having binding affinity for the Fc-binding peptide of the present application may be an antibody of the IgG1 isotype, an antibody of the IgG2 isotype, or an IgG4 isotype.

[0580] As described above, the antibody herein may be an antibody having binding affinity for an Fc-binding entity. For example, even if one or more of the Fab, antigen-binding portion, light chain, and heavy chain variable region are altered, an antibody having binding affinity for an Fc-binding entity (e.g., an antibody comprising an Fc region of IgG1) can still have binding affinity for the Fc-binding entity because it still includes an Fc region to which the Fc-binding entity can bind. Furthermore, even in the case of a protein having only an Fc region to which the Fc-binding entity can bind, it can be recognized as an antibody in the present application because it has binding affinity for the Fc-binding entity. In the present application, a protein that essentially includes an Fc region is referred to as an Fc protein, and in some embodiments, for convenience, an antibody can be recognized as encompassing aspects of an Fc protein.

[0581] Furthermore, the present application provides the amino acid sequence of amino acids 237 to 444 of the Fc region of human IgG1 (SEQ ID NO: 14), the corresponding amino acid sequence of the human IgG2 Fc region (SEQ ID NO: 15), the amino acid sequence of the IgG3 Fc region (SEQ ID NO: 16), the amino acid sequence of the IgG4 Fc region (SEQ ID NO: 17), and the amino acid sequence of the Fc region of trastuzumab, an antibody well known in the art (SEQ ID NO: 18).

[0582] An Fc-binding agent (e.g., an Fc-binding agent comprising an amino acid sequence of any one of SEQ ID NOs: 06 to 09) is known to interact with the amino acid sequence KPKDTLM (SEQ ID NO: 10) of the Fc region (a sequence of amino acid residues 246 to 252 based on the sequence of the Fc region of trastuzumab), and the amino acid sequence MHEALHNHY (SEQ ID NO: 12) of the Fc region (a sequence of amino acid residues 428 to 436 based on the sequence of the Fc region of trastuzumab).

[0583]

[0584] Structure of the Fc binding unit

[0585] A compound comprising an Fc-binding unit comprises an Fc-binding unit. The Fc-binding unit of the compound is derived from an Fc-binding substance. Specifically, the Fc-binding substance or Fc-binding unit comprises Xa 1 Through amino acid residues expressed as (e.g., Xa 1 ), is linked to another part of the compound comprising the Fc binding unit.

[0586] The Fc-binding material from which the Fc-binding unit is derived has been described in detail in the preceding paragraph. Below, the structure of the Fc-binding unit is described in detail.

[0587]

[0588] In some embodiments, the Fc binding unit can have the structure:

[0589] ,

[0590] At this time,

[0591] Each Xaa is independently any amino acid residue,

[0592] Xa 1 'silver , where m is an integer from 1 to 10, and J f is -NH-, -S-, -O-, or -C(=O)-, and *** is Xa 1 ' and an attachment site (or linkage site) to a non-Fc-binding unit among compounds containing an Fc-binding unit, and * represents Xa 1 ' represents an attachment site with an adjacent amino acid residue (e.g., a histidine residue), and ** represents Xa 1 ' represents an attachment site with an adjacent amino acid residue (e.g., a glycine residue),

[0593] Xa 2is a glutamic acid residue or an asparagine residue,

[0594] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue.

[0595] In the above structure, the cysteine ​​residue adjacent to the N-terminus (i.e., the cysteine ​​residue located 2 to 4 from the N-terminus) and the cysteine ​​residue adjacent to the C-terminus (i.e., the cysteine ​​residue located 2 to 4 from the C-terminus) may be optionally covalently linked. For example, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus may be optionally linked via a disulfide bond. For example, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus may optionally have a structure including a carbonyl group. can be shared through .

[0596] In some embodiments, each Xaa can independently be any amino acid other than cysteine.

[0597] In some embodiments, Xa 1 ' may be a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated 2,8-diaminooctanoic acid residue, a conjugated 2,9-diaminononanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue. In certain embodiments, Xa 1' may be a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, or a conjugated lysine (Lys) residue.

[0598] In the structure of the above Fc binding unit, Xa refers to an amino acid residue conjugated to another part of the compound comprising the Fc binding unit. 1 'In relation to, for convenience, the term 'conjugated' may be omitted when describing. For example, a conjugated diaminopropionic acid residue may be referred to as a diaminopropionic acid residue or diaminopropionic acid. For example, a conjugated diaminobutyric acid residue may be referred to as a diaminobutyric acid residue or diaminobutyric acid. For example, a conjugated ornithine residue may be referred to as an ornithine residue or ornithine. For example, a conjugated lysine residue may be referred to as a lysine residue or lysine.

[0599]

[0600] When chemical formula 2 is depicted together with the structure of the above Fc binding unit, chemical formula 2 can be expressed as chemical formula 4 below:

[0601] [Chemical Formula 4]

[0602] .

[0603]

[0604] In some embodiments, Xa 1 ' can have the following structure:

[0605] ,

[0606] Here, m is an integer from 1 to 4,

[0607] *** is Xa 1' and represents an attachment portion with a non-Fc-binding peptide unit among compounds containing an Fc-binding peptide unit,

[0608] * is Xa 1 ' represents an attachment site with an adjacent amino acid residue (e.g., a histidine residue),

[0609] ** is Xa 1 ' represents an attachment site with an adjacent amino acid residue (e.g., a glycine residue).

[0610]

[0611] As described above, the Fc-binding entity from which the Fc-binding unit is derived may comprise modifications. For example, the Fc-binding entity comprises an Fc-binding peptide, wherein one or more modifications may be added to the Fc-binding peptide. As described above, the modifications may be at a level that does not impair the inherent function of the Fc-binding peptide. The Fc-binding unit may be derived from the Fc-binding entity. For example, the Fc-binding unit may further comprise a PEG moiety, wherein the PEG moiety may be linked to the N-terminus and / or the C-terminus of the Fc-binding unit.

[0612] Below, by way of example, the structure of an Fc-binding unit derived from an Fc-binding agent that includes or does not include a modification (e.g., an Fc-binding peptide with or without a modification added to the N-terminus and / or C-terminus of the amino acid sequence) is illustrated.

[0613] For example, an Fc binding unit may have the following structure:

[0614] ,

[0615] At this time, each Xaa, Xa 1 ', Xa 2 , and Xa 3 Each is as described above,

[0616] M F1 and M F2 Each is as described in the paragraphs describing the modifications added to the Fc-binding peptide.

[0617] The structure illustrated above is an example of a structure of an Fc-binding unit derived from an Fc-binding material including a modification or an Fc-binding peptide to which a modification has been added, and the modifications illustrated in the structure (e.g., M) are described below. F1 and / or M F2 ) Although not shown in the structure of the Fc-binding unit, it will be understood that the Fc-binding unit may further comprise modifications.

[0618]

[0619] In some embodiments, the Fc binding unit can have the structure:

[0620] .

[0621] At this time, Xaa, Xa 1 ', Xa 2 , and Xa 3 Each is as described above.

[0622] At this time, the N-terminal L P (L-proline residue) and D P (D-proline residue) can optionally form a D-proline-L-proline template.

[0623]

[0624] In some embodiments, the Fc binding unit can have the structure:

[0625] .

[0626] At this time, Xaa, Xa 1 ', Xa 2 , and Xa 3 Each is as described above.

[0627] At this time, the cysteine ​​at the N terminal and the cysteine ​​at the C terminal can be optionally covalently linked (e.g., via a disulfide bond).

[0628]

[0629] In certain embodiments, the Fc binding unit can have the structure:

[0630] .

[0631] At this time, Xaa, Xa 1 ', Xa 2 , and Xa 3 Each is as described above.

[0632] At this time, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus can optionally be covalently linked (e.g., via a disulfide bond).

[0633]

[0634] In certain embodiments, the Fc binding unit can have the structure:

[0635] .

[0636] At this time,

[0637] Xa 1 'silver , where m is an integer from 1 to 10, and J f is -NH-, -S-, -O-, or -C(=O)-, and *** is Xa 1 ' and an attachment site (or linkage site) to a non-Fc-binding unit among compounds containing an Fc-binding unit, and * represents Xa 1 ' represents an attachment site with an adjacent amino acid residue (e.g., a histidine residue), and ** represents Xa 1 ' may represent an attachment site with an adjacent amino acid residue (e.g., a glycine residue).

[0638] At this time, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus can optionally be covalently linked (e.g., via a disulfide bond).

[0639] In certain embodiments, Xa 1 ' may be a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated 2,8-diaminooctanoic acid residue, a conjugated 2,9-diaminononanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue.

[0640] In certain embodiments, Xa 1 'silver , and here m can be an integer from 1 to 4.

[0641] In certain embodiments, Xa 1 ' may be a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, or a conjugated lysine (Lys) residue.

[0642]

[0643] For example, in a compound comprising an Fc-binding unit, the Fc-binding unit (FcBU) may be described through the Fc-binding substance from which it originates, examples of such descriptions include, but are not limited to:

[0644] The Fc-binding unit is derived from an Fc-binding substance,

[0645] At this time, the Fc binding material contains the amino acid sequence of sequence number 01,

[0646] At this time, the Fc-binding unit and the other parts other than the Fc-binding unit are Xa of the Fc-binding substance. 1 Through (specifically, Xa 1 (via side chains) connected or joined.

[0647] As another example, in a compound comprising an Fc-binding unit, the Fc-binding unit (FcBU) may be described as follows:

[0648] The Fc-binding unit comprises an amino acid sequence of SEQ ID NO: 01, wherein the Fc-binding unit and other parts other than the Fc-binding unit comprise Xa of the amino acid sequence of SEQ ID NO: 01. 1 Through (specifically, Xa 1 (via side chains) connected or joined.

[0649]

[0650] In the preceding paragraphs, the elements of a compound comprising an Fc-binding unit have been described in detail. Below, the reaction between a compound comprising an Fc-binding unit and an antibody is described in detail.

[0651]

[0652] Reaction of a compound containing an Fc binding unit with an antibody

[0653] Delivery location of the target group

[0654] As described above, a compound comprising an Fc binding unit can be used to deliver a target substance to an antibody in a location-specific manner.

[0655] For example, a compound comprising an Fc-binding unit can be used to deliver a bioorthogonal functional group or drug to the target region of an antibody. The region within the antibody to which the desired group is delivered (e.g., the target region) can be located within the Fc region of the antibody. That is, when using a compound comprising the Fc-binding unit of the present application, the desired group can be delivered to the Fc region of the antibody.

[0656] In some embodiments, when utilizing a compound comprising the Fc binding unit of the present application, the desired group can be delivered to one or more selected from lysine 246 (K246) and lysine 248 (K248) of the Fc region of the antibody.

[0657] In some embodiments, when using a compound comprising the Fc binding unit of the present application, a group of interest can be positionally delivered to one or more selected from K246 and K248 of the Fc region of the antibody.

[0658] For example, when a compound comprising an Fc-binding unit of the present application contacts or reacts with an antibody, the percentage of cases in which a target group is delivered to the target region of the antibody may be 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, 99.5% or more, or 100% of all cases (i.e., cases encompassing both cases in which a target group is delivered to the target region and cases in which a target group is delivered to a region other than the target region). For example, when a compound comprising 100 Fc-binding units is used (wherein a compound comprising one Fc-binding unit has one target group), if 100 target groups are delivered to the target region, the percentage of cases in which the target group is delivered to the target region can be described as 100%. For example, when using a compound containing 100 Fc-binding units, if 80 of the target groups are delivered to the target region, and a compound containing 20 Fc-binding units does not react with the antibody (i.e., 20 of the target groups are not delivered to the antibody), the delivery of the target group to the target region can be described as 100%. For example, when using a compound containing 100 Fc-binding units, if 80 of the target groups are delivered to the target region, and 20 of the target groups are delivered to the antibody but to a location other than the target region of the antibody, the delivery of the target group to the target region can be described as 80%. In this case, for example, the delivery of the target group to the target region of the antibody can be measured based on the reaction time (e.g., 3 hours) of the compound containing the Fc-binding unit and the antibody. At this time, the target region may be a region consisting of five or three amino acid residues, including lysine residue 246 (K246) of the Fc region and lysine residue 248 (K248) of the Fc region.For example, if the target region is a region consisting of five amino acid residues, including lysine residue 246 (K246) of the Fc region and lysine residue 248 (K248) of the Fc region, the target region is PK. 246 PK 248 D (SEQ ID NO: 27). For example, if the target region is a region consisting of three amino acid residues, including lysine residue 246 (K246) of the Fc region and lysine residue 248 (K248) of the Fc region, the target region is K 246 PK 248 (Sequence number 28) may be.

[0659] In some embodiments, among the cases in which the target group is delivered to the target region, the percentage of cases in which the target group is delivered to lysine residue 246 (K246) of the Fc region is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 99.5% or more, or may be 100%. In some embodiments, among the cases in which the target group is delivered to lysine residue 248 (K248) of the Fc region, the percentage of cases in which the target group is delivered to lysine residue 248 (K248) of the Fc region is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 99.5% or more, or may be 100%.

[0660] For example, when a compound comprising the Fc binding unit of the present application contacts or reacts with an antibody, the percentage of cases in which the target group is transferred to at least one of K246 and K248 of the Fc region of the antibody may be 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, 99.5% or more, or 100% of all cases (i.e., cases encompassing all cases in which the target group is transferred to at least one of K246 and K248 of the Fc region of the antibody and cases in which the target group is transferred to a location other than K246 and K248).

[0661] For example, when a compound comprising the Fc binding unit of the present application contacts or reacts with an antibody, the percentage of cases in which a target group is transferred to K246 of the Fc region of the antibody may be 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, 99.5% or more, or 100% of all cases (i.e., cases encompassing both cases in which a target group is transferred to K246 of the Fc region of the antibody and cases in which a target group is transferred to a location other than K246).

[0662] For example, when a compound comprising the Fc binding unit of the present application contacts or reacts with an antibody, the percentage of cases in which a target group is transferred to K248 of the Fc region of the antibody may be 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, 99.5% or more, or 100% of all cases (i.e., cases encompassing both cases in which a target group is transferred to K248 of the Fc region of the antibody and cases in which a target group is transferred to a location other than K248).

[0663] In some embodiments, when using a compound comprising the Fc-binding unit of the present application, the target group can be delivered to one or more selected from K246 and K248 of the Fc region of the antibody, whereas the target group may not be delivered to other positions other than one or more selected from K246 and K248. In some embodiments, when using a compound comprising the Fc-binding unit of the present application, delivery of the target group to other positions other than one or more selected from K246 and K248 may be inhibited.

[0664] Compounds comprising an Fc-binding unit are designed to react with free amino groups of lysine residues of antibodies. That is, in some embodiments, when utilizing a compound comprising an Fc-binding unit of the present application, a desired group can be delivered to a lysine in the Fc region of an antibody.

[0665] Furthermore, as described in the design of the Fc-binding agent, when the antibody and the Fc-binding agent interact (e.g., bind), Xa 1 It was explained that the lysines in the Fc region located adjacent to each other are lysine 246 and lysine 248 (the numbering of lysines is set according to Eu numbering).

[0666] For example, the amino acid sequence contained in trastuzumab, indicating lysines 246 and 248, is as follows: GPSVFLFPPK 246 PK 248 DTLM (SEQ ID NO: 13).

[0667] In some literature or antibodies, the lysine residues corresponding to positions 246 and 248 are designated by numbers other than 246 and 248. If the designated residues in some literature are amino acid residues corresponding to positions 246 and 248 of lysine, these positions will be understood as positions 246 and 248 of lysine. For example, some literature refers to the residues corresponding to K246 and K248 of denosumab as K247 and K249, respectively. As another example, some literature refers to the amino acid residues corresponding to K246 and K248 of dupliumab as K251 and K253, respectively (see European Patent Application No. 19818561.3, Publication No. EP 3811978; and European Patent Application No. 18791007.0, Publication No. EP 3617235). Thus, lysine 246 and lysine 248, respectively, may be understood to refer to the lysine residues corresponding to lysine 246 and lysine 248, respectively, in addition to lysine 246 and lysine 248, respectively.

[0668] In some embodiments, when using a compound comprising the Fc-binding unit of the present application, a group of interest can be delivered to lysine 246 (Lys246; K246) and / or lysine 248 (Lys248; K248) of the Fc region of an antibody. In a particular embodiment, when using a compound comprising the Fc-binding unit of the present application, a group of interest can be delivered to lysine 246 (Lys246; K246) of the Fc region of an antibody. In a particular embodiment, when using a compound comprising the Fc-binding unit of the present application, a group of interest can be delivered to lysine 248 (Lys248; K248) of the Fc region of an antibody. By reaction of a compound containing an Fc binding unit with an antibody, the position at which the desired substance is delivered may be lysine 246 (Lys246; K246) and / or lysine 248 (Lys248; K248).

[0669]

[0670] Below, the reaction of a compound comprising the Fc binding unit of the present application with an antibody is described in detail.

[0671]

[0672] Description of the reaction of a compound containing an Fc binding unit with an antibody

[0673] A compound containing an Fc-binding unit is directed to the Fc region of an antibody by the Fc-binding unit. At this time, the effect of directing to a specific region of the antibody by the Fc-binding unit may be referred to as the proximity effect. When explaining based on an Fc-binding substance, the Xa of the Fc-binding substance 1The amino acid residue is adjacent to lysine 246 or lysine 248 of the Fc region of the antibody. The reaction of a compound comprising the Fc-binding unit of the present application with a lysine residue (e.g., lysine 246 or lysine 248) of the antibody can be exemplarily illustrated by the following reaction scheme. Note that the lysine residues of the antibody involved in the reaction are separately illustrated below for illustration purposes.

[0674] [Reaction scheme 1]

[0675] .

[0676] As illustrated in the above Reaction Scheme 1, the reaction of the antibody and the compound comprising the Fc-binding unit (illustrated as the compound of Formula 2 in the above Reaction Scheme) may be, specifically, a reaction of a primary amine group (e.g., an amino group, -NH2) of the antibody with a carbonyl group of the compound comprising the Fc-binding unit. For example, the reaction of the antibody and the compound comprising the Fc-binding unit may be a reaction of a primary amine group of the Fc region of the antibody with a carbonyl group of the compound comprising the Fc-binding unit. For example, the reaction of the antibody and the compound comprising the Fc-binding unit may be a reaction of a primary amine group of lysine residue at position 246 of the Fc region of the antibody with a carbonyl group of the compound comprising the Fc-binding unit. For example, the reaction of the antibody and the compound comprising the Fc-binding unit may be a reaction of a primary amine group of lysine residue at position 248 of the Fc region of the antibody with a carbonyl group of the compound comprising the Fc-binding unit. As illustrated in Reaction Scheme 1, a desired group (e.g., a bioorthogonal functional group, etc.) is transferred to the antibody through the reaction of a compound comprising an antibody and an Fc binding unit.

[0677] For example, the reaction of a compound comprising an antibody and an Fc-binding unit may be referred to as a nucleophilic substitution reaction. In this case, the primary amine group of the antibody may be understood to act as a nucleophile in the nucleophilic substitution reaction. For example, the reaction of a compound comprising an antibody and an Fc-binding unit may be referred to as S N 2 Reaction (S N 2 reaction). For example, the reaction of a compound comprising an antibody and an Fc-binding unit may be referred to as an acyl transfer reaction. As such, the terminology referring to the reaction is not otherwise limited, and the reaction of an antibody and an Fc-binding unit may be described using terms that can be understood by those of ordinary skill in the art.

[0678] The carbonyl group of the compound comprising the Fc binding unit, which is shown to react with the primary amine group of the antibody in Reaction Scheme 1, is separately represented in Chemical Formula 2 as follows:

[0679] [Chemical Formula 2]

[0680] .

[0681] In Chemical Formula 2, the carbonyl group (-C(=O)-) marked with * represents a reaction site with a primary amine group of the antibody. In some embodiments, the carbonyl group marked with * may be referred to as, but is not limited to, an "antibody-reactive group" or "antibody-reactive site" or "reactive carbonyl" of a compound comprising an Fc-binding unit.

[0682] Meanwhile, as illustrated in reaction scheme 1, the group comprising the Fc-binding unit is released from the product by the reaction of the antibody and the compound comprising the Fc-binding unit. Thus, conjugation utilizing a reaction mechanism in which the Fc-binding unit is released through the reaction is referred to as traceless cross-linking, traceless reaction, or traceless conjugation. The advantages of such traceless conjugation and conjugates (e.g., antibody conjugates) prepared via traceless conjugation have been described in detail in previous studies (see, e.g., European Patent Application No. 19818561.3, Publication No. EP 3811978; PCT Patent Application No. PCT / KR2020 / 003282, Publication No. WO2020 / 184944; and Korean Patent Application No. 10-2020-0009162, Publication No. 10-2020-0091826). Furthermore, the document [PCT Patent Application No. PCT / KR2020 / 003282, Publication No. WO2020 / 184944], the entire contents of which are incorporated by reference herein, confirmed that when using a compound comprising an Fc-binding unit disclosed in the document, norbornene, which is one of the examples of the desired group, was transferred to K248 of the Fc region of the antibody.

[0683] As illustrated in Reaction Scheme 1, the groups that are released through the reaction of the antibody and the compound comprising the Fc-binding unit in the compound comprising the Fc-binding unit can be exemplarily illustrated as follows:

[0684] .

[0685] In some embodiments, in the group shown above, -N(R a1 )-O- may be referred to as a leaving group, but is not limited thereto.

[0686] As illustrated in Reaction Scheme 1, through the reaction of a compound comprising an antibody and an Fc-binding unit, the group transferred from the compound comprising an Fc-binding unit to the antibody can be exemplarily illustrated as follows:

[0687] .

[0688]

[0689] Furthermore, to aid understanding, the present application provides a schematic diagram of the reaction between an antibody and a compound comprising an Fc-binding unit, using the compound of Chemical Formula 2-1 as an example of a compound comprising an Fc-binding unit.

[0690] [Reaction scheme 2]

[0691] .

[0692]

[0693] Hereinafter, specific embodiments of compounds comprising the Fc-binding unit of the present application are disclosed. Meanwhile, specific embodiments of compounds comprising the Fc-binding unit of the present application are not limited to the chemical formulas or structures disclosed below, and specific embodiments can be derived or newly created based on the contents of compounds comprising the Fc-binding unit described above.

[0694]

[0695] Specific embodiments of compounds comprising an Fc binding unit

[0696] As described above, the compound comprising the Fc binding unit of the present application may have the structure of Chemical Formula 2:

[0697] [Chemical Formula 2]

[0698] .

[0699]

[0700] Hereinafter, specific examples of compounds comprising the Fc binding unit of the present application are described in detail.

[0701]

[0702] Some embodiments of the present application provide a compound of formula 2-1.

[0703] In some embodiments, Formula 2 can be represented by Formula 2-1 below. In Formula 2, J a When -C(=O)-, chemical formula 2 is expressed as chemical formula 2-1 below.

[0704] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-1:

[0705] [Chemical Formula 2-1]

[0706] ,

[0707] At this time,

[0708] FcBU stands for Fc binding unit,

[0709] GOI is the group of interest,

[0710] L a is linker A,

[0711] D a is spacer A,

[0712] X is C, O, or N (i.e., -X- is -CH2-, -O-, or -NH-),

[0713] R a1 is H or C 1-6 It is alkyl,

[0714] R a2 is H or C 1-6 Alkyl, and

[0715] R a3 is H or C 1-6 It's alkyl.

[0716]

[0717] In Chemical Formula 2-1, FcBU is an Fc-binding unit. The Fc-binding unit (FcBU) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, the Fc-binding unit (FcBU) is as described in the subsections "Overview of Compounds Comprising Fc-binding Units of the Present Application" and "Fc-binding unit (FcBU)" of the section "Compounds Comprising Fc-binding Units".

[0718] In Chemical Formula 2-1, GOI is a group of interest. The group of interest (GOI) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, the group of interest (GOI) is as described in the subsections "Overview of Compounds Comprising Fc-Binding Units of the Present Application" and "Group of Interest (GOI)" of the section "Compounds Comprising Fc-Binding Units".

[0719] In chemical formula 2-1, L a is linker A. Linker A (L a ) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, linker A (L a ) is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Linker A (L"a)" of the section "Compounds Comprising the Fc-Binding Unit".

[0720] In chemical formula 2-1, D a is spacer A. Spacer A (D a ) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, spacer A (D a ) is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Spacer A (spacer A; D"a)" of the section "Compounds Comprising an Fc-Binding Unit".

[0721]

[0722] Some embodiments of the present application provide a compound of formula 2-2.

[0723] In some embodiments, Formula 2 can be represented by Formula 2-2 below. In Formula 2, when the desired group (GOI) is a reactive group (RG), Formula 2 is represented by Formula 2-2 below.

[0724] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-2:

[0725] [Chemical Formula 2-2]

[0726] ,

[0727] At this time,

[0728] FcBU stands for Fc binding unit,

[0729] RG is a reactive group,

[0730] L a is linker A,

[0731] D a is spacer A,

[0732] X is C, O, or N (i.e., -X- is -CH2-, -O-, or -NH-),

[0733] J a is -C(=O)-, -S-, -NH-, or -C(=NH)-,

[0734] R a1 is H or C 1-6 It is alkyl,

[0735] R a2 is H or C 1-6 Alkyl, and

[0736] R a3 is H or C 1-6 It's alkyl.

[0737]

[0738] In Chemical Formula 2-2, FcBU is an Fc binding unit. The Fc binding unit (FcBU) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, the Fc binding unit (FcBU) is as described in the subsections "Overview of Compounds Comprising Fc Binding Units of the Present Application" and "Fc Binding Unit (FcBU)" of the section "Compounds Comprising Fc Binding Units".

[0739] In Chemical Formula 2-2, RG is a reactive group. The reactive group (RG) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, the reactive group (RG) is as described in the subsections "Overview of Compounds Comprising Fc-Binding Units of the Present Application" and "Group of Interest (GOI)" of the section "Compounds Comprising Fc-Binding Units."

[0740] In chemical formula 2-2, L a is linker A. Linker A (L a ) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, linker A (L a ) is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Linker A (L"a)" of the section "Compounds Comprising the Fc-Binding Unit".

[0741] In chemical formula 2-2, D a is spacer A. Spacer A (D a ) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, spacer A (D a) is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Spacer A (spacer A; D"a)" of the section "Compounds Comprising an Fc-Binding Unit".

[0742]

[0743] Some embodiments of the present application provide a compound of formula 2-3.

[0744] In some embodiments, Formula 2 can be represented by Formula 2-3 below. In Formula 2, the desired group (GOI) is a reactive group (RG), wherein the reactive group is In this case, chemical formula 2 is expressed as chemical formula 2-3 below.

[0745] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-3:

[0746] [Chemical Formula 2-3]

[0747] ,

[0748] At this time,

[0749] FcBU stands for Fc binding unit,

[0750] H RG is a reactive moiety,

[0751] D RG is the spacer of the reactive group (spacer RG),

[0752] L a is linker A,

[0753] D a is spacer A,

[0754] X is C, O, or N (i.e., -X- is -CH2-, -O-, or -NH-),

[0755] J ais -C(=O)-, -S-, -NH-, or -C(=NH)-,

[0756] R a1 is H or C 1-6 It is alkyl,

[0757] R a2 is H or C 1-6 Alkyl, and

[0758] R a3 is H or C 1-6 It's alkyl.

[0759]

[0760] In Chemical Formula 2-3, FcBU is an Fc-binding unit. The Fc-binding unit (FcBU) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, the Fc-binding unit (FcBU) is as described in the subsections "Overview of Compounds Comprising Fc-binding Units of the Present Application" and "Fc-binding unit (FcBU)" of the section "Compounds Comprising Fc-binding Units".

[0761] In chemical formula 2-3, H RG is a reactive moiety. Reactive moiety (H RG ) are described in detail in the previous paragraphs, and are as described in the previous paragraphs. For example, the reactive moiety is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Group of Interest (GOI)" of the section "Compounds Comprising an Fc-Binding Unit".

[0762] In chemical formula 2-3, D RG is a spacer of a reactive group. The spacer of a reactive group (D RG) are described in detail in the previous paragraphs, and are as described in the previous paragraphs. For example, the spacer of the reactive group is as described in the subsections "Overview of compounds comprising Fc-binding units of the present application" and "Group of interest (GOI)" of the section "Compounds comprising Fc-binding units".

[0763] In chemical formula 2-3, L a is linker A. Linker A (L a ) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, linker A (L a ) is as described in detail in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Linker A (L"a)" of the section "Compounds Comprising the Fc-Binding Unit".

[0764] In chemical formula 2-3, D a is spacer A. Spacer A (D a ) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, spacer A (D a ) is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Spacer A (spacer A; D"a)" of the section "Compounds Comprising an Fc-Binding Unit".

[0765]

[0766] Some embodiments of the present application provide a compound of formula 2-4.

[0767] In some embodiments, Formula 2 can be represented by Formula 2-4 below. In Formula 2, J a is -C(=O)-, and spacer A (D a ) This C 1-10 is unsubstituted alkylene, and R a3 When H, chemical formula 2 is expressed as chemical formula 2-4 below.

[0768] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-4:

[0769] [Chemical Formula 2-4]

[0770] ,

[0771] At this time,

[0772] aa is an integer from 1 to 10,

[0773] FcBU stands for Fc binding unit,

[0774] GOI is the group of interest,

[0775] L a is linker A,

[0776] X is C or O (i.e., -X- is -CH2- or -O-),

[0777] R a1 is H or C 1-6 Alkyl, and

[0778] R a2 is H or C 1-6 It's alkyl.

[0779]

[0780] In Chemical Formula 2-4, FcBU is an Fc-binding unit. The Fc-binding unit (FcBU) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, the Fc-binding unit (FcBU) is as described in the subsections "Overview of Compounds Comprising Fc-binding Units of the Present Application" and "Fc-binding unit (FcBU)" of the section "Compounds Comprising Fc-binding Units".

[0781] In Chemical Formula 2-4, GOI is a group of interest. The group of interest (GOI) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, the group of interest (GOI) is as described in the subsections "Overview of Compounds Comprising Fc-Binding Units of the Present Application" and "Group of Interest (GOI)" of the section "Compounds Comprising Fc-Binding Units".

[0782] In chemical formula 2-4, L a is linker A. Linker A(L a ) is described in detail in the previous paragraphs, and is as described in the previous paragraphs. For example, linker A(L a ) is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Linker A (L"a)" of the section "Compounds Comprising the Fc-Binding Unit".

[0783]

[0784] Some embodiments of the present application provide compounds of formula 2-5.

[0785] In some embodiments, Formula 2 can be represented by Formula 2-5 below.

[0786] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-5:

[0787] [Chemical Formula 2-5]

[0788] ,

[0789] At this time,

[0790] aa is an integer from 1 to 10,

[0791] FcBU stands for Fc binding unit,

[0792] RG is a reactive group,

[0793] L a is linker A,

[0794] X is C or O (i.e., -X- is -CH2- or -O-),

[0795] R a1 is H or C 1-6 Alkyl, and

[0796] R a2 is H or C 1-6 It's alkyl.

[0797]

[0798] In Chemical Formula 2-5, FcBU is an Fc binding unit. FcBU is as described in the previous paragraphs. For example, FcBU is as described in the subsections "Overview of Compounds Comprising Fc Binding Units of the Present Application" and "Fc Binding Unit (FcBU)" of the section "Compounds Comprising Fc Binding Units".

[0799] In certain embodiments, FcBU may have the structure:

[0800] ,

[0801] At this time,

[0802] Each Xaa is independently any amino acid residue other than a cysteine ​​residue,

[0803] Xa 1 'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue,

[0804] Xa 2 is a glutamic acid residue or an asparagine residue,

[0805] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue,

[0806] In the above structure, the cysteine ​​residue adjacent to the N terminus (i.e., the cysteine ​​residue located 2 to 4 from the N terminus) and the cysteine ​​residue adjacent to the C terminus (i.e., the cysteine ​​residue located 2 to 4 from the C terminus) may optionally be covalently linked.

[0807] For example, a cysteine ​​residue adjacent to the N-terminus and a cysteine ​​residue adjacent to the C-terminus can optionally be linked via a disulfide bond.

[0808] In certain embodiments, the Fc binding unit can have the structure:

[0809] ,

[0810] At this time,

[0811] Each Xaa is independently any amino acid other than cysteine,

[0812] Xa 1 'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue,

[0813] Xa 2 is a glutamic acid residue or an asparagine residue,

[0814] Xa 3is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue,

[0815] At this time, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus are optionally covalently linked (e.g., via a disulfide bond).

[0816] In certain embodiments, the Fc binding unit can have the structure:

[0817] ,

[0818] At this time,

[0819] Xa 1 'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue.

[0820]

[0821] In Chemical Formula 2-5, RG is a reactive group. RG is as described in the preceding paragraphs. For example, RG is as described in the subsections "Overview of Compounds Comprising Fc-Binding Units of the Present Application" and "Group of Interest (GOI)" of the section "Compounds Comprising Fc-Binding Units."

[0822] In certain embodiments, RG may comprise a reactive moiety.

[0823] In certain embodiments, RG comprises a reactive moiety, wherein the sum of the atomic masses of all atoms constituting RG may be 3000 Daltons or less, 2500 Daltons or less, 2000 Daltons or less, 1500 Daltons or less, 1000 Daltons or less, 900 Daltons or less, 800 Daltons or less, 700 Daltons or less, 600 Daltons or less, 500 Daltons or less, 400 Daltons or less, 300 Daltons or less, 200 Daltons or less, or 100 Daltons or less.

[0824] In certain embodiments, RG may have the following structure:

[0825] ,

[0826] At this time, D RG is the spacer of the reactive group (spacer R), and H RG is a reactive moiety.

[0827] In certain embodiments, D RG is a bond, or substituted or unsubstituted C 1-6 Alkylene, substituted or unsubstituted C 1-6 It may be heteroalkylene. In this case, the substituted alkylene or substituted heteroalkylene may include one or more substituents, and the substituents are each independently -C 1-4 Alkyl and =O may be selected from, wherein heteroalkylene comprises one or more heteroatoms, wherein the heteroatoms may each be independently selected from O, N, and S. In a particular embodiment, D RG is a bond, or substituted or unsubstituted C 1-3 Alkylene, substituted or unsubstituted C 1-3 It may be heteroalkylene. In this case, the substituted alkylene or substituted heteroalkylene may include one or more substituents, and the substituents are each independently -C 1-4Alkyl and =O may be selected from. Here, heteroalkylene contains one or more heteroatoms, and here, the heteroatoms may be independently selected from O, N, and S.

[0828]

[0829] In chemical formula 2-5, L a is linker A. Linker A is as described in the previous paragraphs. For example, L a is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Linker A (L"a)" of the section "Compounds Comprising the Fc-Binding Unit".

[0830] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be independently selected from O, N, and S.

[0831] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be heteroalkylene, wherein the heteroalkylene contains one or more heteroatoms, and wherein each heteroatom may be O.

[0832] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 20 ethylene glycol units.

[0833] In certain embodiments, linker A is a bond, unsubstituted C 1-50Alkylene, or unsubstituted C 1-50 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 16 ethylene glycol units.

[0834] In certain embodiments, linker A is a bond, unsubstituted C 1-30 Alkylene, or unsubstituted C 1-30 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 10 ethylene glycol units.

[0835] In certain embodiments, linker A is a bond, unsubstituted C 1-25 Alkylene, or unsubstituted C 1-25 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 8 ethylene glycol units.

[0836] In certain embodiments, linker A is a bond, unsubstituted C 1-15 Alkylene, or unsubstituted C 1-15 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 5 ethylene glycol units.

[0837] In certain embodiments, linker A is a bond, or unsubstituted C 1-6 It may be alkylene.

[0838] In certain embodiments, linker A is bonded or unsubstituted C 1-3 It may be alkylene.

[0839]

[0840] Some embodiments of the present application provide a compound of formula 2-6.

[0841] In some embodiments, Formula 2 can be represented by Formula 2-6 below.

[0842] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-6:

[0843] [Chemical Formula 2-6]

[0844] ,

[0845] At this time,

[0846] aa is an integer from 1 to 10,

[0847] FcBU stands for Fc binding unit,

[0848] H RG is a reactive moiety,

[0849] L a is linker A,

[0850] X is C or O (i.e., -X- is -CH2- or -O-),

[0851] R a1 is H or C 1-6 Alkyl, and

[0852] R a2 is H or C 1-6 It's alkyl.

[0853]

[0854] In Chemical Formula 2-6, FcBU is an Fc binding unit. The Fc binding unit is as described in the previous paragraphs. For example, FcBU is as described in the subsections "Overview of Compounds Comprising Fc Binding Units of the Present Application" and "Fc Binding Unit (FcBU)" of the section "Compounds Comprising Fc Binding Units".

[0855] In certain embodiments, FcBU may have the structure:

[0856] ,

[0857] At this time,

[0858] Each Xaa is independently any amino acid residue other than a cysteine ​​residue,

[0859] Xa 1 'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue,

[0860] Xa 2 is a glutamic acid residue or an asparagine residue,

[0861] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue,

[0862] In the above structure, the cysteine ​​residue adjacent to the N terminus (i.e., the cysteine ​​residue located 2 to 4 from the N terminus) and the cysteine ​​residue adjacent to the C terminus (i.e., the cysteine ​​residue located 2 to 4 from the C terminus) may optionally be covalently linked.

[0863] For example, a cysteine ​​residue adjacent to the N-terminus and a cysteine ​​residue adjacent to the C-terminus can optionally be linked via a disulfide bond.

[0864] In certain embodiments, the Fc binding unit can have the structure:

[0865] ,

[0866] At this time,

[0867] Each Xaa is independently any amino acid other than cysteine,

[0868] Xa 1 'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue,

[0869] Xa 2 is a glutamic acid residue or an asparagine residue,

[0870] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue.

[0871] At this time, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus can optionally be covalently linked (e.g., via a disulfide bond).

[0872] In certain embodiments, the Fc binding unit can have the structure:

[0873] ,

[0874] At this time,

[0875] Xa 1'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue.

[0876]

[0877] In chemical formula 2-6, H RG is a reactive moiety. The reactive moiety is as described in the previous paragraphs. For example, H RG is as described in the subsections "Overview of compounds comprising Fc-binding units of the present application" and "group of interest (GOI)" of the section "Compounds comprising Fc-binding units".

[0878] In certain embodiments, H RG may be a bioorthogonal functional group.

[0879] In certain embodiments, H RG may have any of the following structures:

[0880]

[0881] ,

[0882] At this time, hn is an integer from 1 to 3,

[0883] R H are each independently H or -R, =O, =S, -NO2, -CR3, -NR2, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6Alkyl, C 3-10 Cycloalkyl, C 3-10 Selected from heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, =O, =S, and -SH.

[0884] In certain embodiments, H RG may be selected from an azide group, a terminal alkyne group, a terminal alkene group, a cyclic alkyne (e.g., cyclooctyne) group, a tetrazine group, a norbornene group, a cycloalkene (e.g., cyclooctene) group, an oxime group, and an isocyanide group. At this time, the cyclooctyne may be any one selected from OCT cyclooctyne, BCN (Bicyclononyne), DBCO (Dibenzocyclooctyne), DIBAC (aza-dibenzocyclooctynes), DIBO (dibenzocyclooctynol), DIFO (difluorinated cyclooctynes), BARAC (biarylazacyclooctynone), DIMAC (dimethoxyazacyclooctyne), and DIFBO (difluorobenzocyclooctyne), but is not otherwise limited. At this time, cyclooctene can be any one selected from, for example, a cis-cyclooctene group and a trans-cyclooctene group.

[0885] In certain embodiments, H RG may be an azide group or a norbornene group.

[0886] In certain embodiments, H RG may be a click chemical functional group.

[0887] In certain embodiments, H RG can be selected from among diels-alder diene, diels-alder dienophile, IEDDA diene, and IEDDA dienophile.

[0888]

[0889] In chemical formula 2-6, L a is linker A. Linker A is as described in the previous paragraphs. For example, L a is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Linker A (L"a)" of the section "Compounds Comprising the Fc-Binding Unit".

[0890] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be independently selected from O, N, and S.

[0891] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be heteroalkylene, wherein the heteroalkylene contains one or more heteroatoms, and wherein each heteroatom may be O.

[0892] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 20 ethylene glycol units.

[0893] In certain embodiments, linker A is a bond, unsubstituted C 1-50 Alkylene, or unsubstituted C 1-50It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 16 ethylene glycol units.

[0894] In certain embodiments, linker A is a bond, unsubstituted C 1-30 Alkylene, or unsubstituted C 1-30 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 10 ethylene glycol units.

[0895] In certain embodiments, linker A is a bond, unsubstituted C 1-25 Alkylene, or unsubstituted C 1-25 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 8 ethylene glycol units.

[0896] In certain embodiments, linker A is a bond, unsubstituted C 1-15 Alkylene, or unsubstituted C 1-15 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 5 ethylene glycol units.

[0897] In certain embodiments, linker A is a bond, or unsubstituted C 1-6 It may be alkylene.

[0898] In certain embodiments, linker A is bonded or unsubstituted C 1-3 It may be alkylene.

[0899] In certain embodiments, R a1 Silver C 1-3may be alkyl. In certain embodiments, R a2 is H or C 1-3 It could be an alkyl.

[0900]

[0901] Some embodiments of the present application provide compounds of formula 2-7.

[0902] In some embodiments, Formula 2 can be represented by Formula 2-7 below.

[0903] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-7:

[0904] [Chemical Formula 2-7]

[0905] ,

[0906] At this time,

[0907] aa is an integer from 1 to 10,

[0908] FcBU stands for Fc binding unit,

[0909] H RG is a reactive moiety,

[0910] L a is linker A,

[0911] X is either C or O.

[0912]

[0913] In Chemical Formula 2-7, FcBU is an Fc binding unit. The Fc binding unit is as described in the previous paragraphs. For example, FcBU is as described in the subsections "Overview of Compounds Comprising Fc Binding Units of the Present Application" and "Fc Binding Unit (FcBU)" of the section "Compounds Comprising Fc Binding Units".

[0914] In certain embodiments, FcBU may have the structure:

[0915] ,

[0916] At this time,

[0917] Each Xaa is independently any amino acid residue other than a cysteine ​​residue,

[0918] Xa 1 'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue,

[0919] Xa 2 is a glutamic acid residue or an asparagine residue,

[0920] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue,

[0921] In the above structure, the cysteine ​​residue adjacent to the N terminus (i.e., the cysteine ​​residue located 2 to 4 from the N terminus) and the cysteine ​​residue adjacent to the C terminus (i.e., the cysteine ​​residue located 2 to 4 from the C terminus) may optionally be covalently linked.

[0922] For example, a cysteine ​​residue adjacent to the N-terminus and a cysteine ​​residue adjacent to the C-terminus can optionally be linked via a disulfide bond.

[0923] In certain embodiments, the Fc binding unit can have the structure:

[0924] ,

[0925] At this time,

[0926] Each Xaa is independently any amino acid other than cysteine,

[0927] Xa 1 'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue,

[0928] Xa 2 is a glutamic acid residue or an asparagine residue,

[0929] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue,

[0930] At this time, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus are optionally covalently linked (e.g., via a disulfide bond).

[0931] In certain embodiments, the Fc binding unit can have the structure:

[0932] ,

[0933] At this time,

[0934] Xa 1'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue.

[0935]

[0936] In chemical formula 2-7, H RG is a reactive moiety. The reactive moiety is as described in the previous paragraphs. For example, H RG is as described in the subsections "Overview of compounds comprising Fc-binding units of the present application" and "group of interest (GOI)" of the section "Compounds comprising Fc-binding units".

[0937] In certain embodiments, H RG may be a bioorthogonal functional group.

[0938] In certain embodiments, H RG may have any of the following structures:

[0939]

[0940] ,

[0941] At this time, hn is an integer from 1 to 3,

[0942] R H are each independently H or -R, =O, =S, -NO2, -CR3, -NR2, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6Alkyl, C 3-10 Cycloalkyl, C 3-10 Selected from heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, =O, =S, and -SH.

[0943] In certain embodiments, H RG may be selected from an azide group, a terminal alkyne group, a terminal alkene group, a cyclic alkyne (e.g., cyclooctyne) group, a tetrazine group, a norbornene group, a cycloalkene (e.g., cyclooctene) group, an oxime group, and an isocyanide group. At this time, the cyclooctyne may be any one selected from OCT cyclooctyne, BCN (Bicyclononyne), DBCO (Dibenzocyclooctyne), DIBAC (aza-dibenzocyclooctynes), DIBO (dibenzocyclooctynol), DIFO (difluorinated cyclooctynes), BARAC (biarylazacyclooctynone), DIMAC (dimethoxyazacyclooctyne), and DIFBO (difluorobenzocyclooctyne), but is not otherwise limited. At this time, cyclooctene can be any one selected from, for example, a cis-cyclooctene group and a trans-cyclooctene group.

[0944] In certain embodiments, H RG may be an azide group or a norbornene group.

[0945] In certain embodiments, H RG may be a click chemical functional group.

[0946] In certain embodiments, H RG can be selected from among diels-alder diene, diels-alder dienophile, IEDDA diene, and IEDDA dienophile.

[0947]

[0948] In chemical formula 2-7, L a is linker A. Linker A is as described in the previous paragraphs. For example, L a is as described in the subsections "Overview of Compounds Comprising the Fc-Binding Unit of the Present Application" and "Linker A (L"a)" of the section "Compounds Comprising the Fc-Binding Unit".

[0949] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be independently selected from O, N, and S.

[0950] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be heteroalkylene, wherein the heteroalkylene contains one or more heteroatoms, and wherein each heteroatom may be O.

[0951] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 20 ethylene glycol units.

[0952] In certain embodiments, linker A is a bond, unsubstituted C 1-50 Alkylene, or unsubstituted C 1-50It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 16 ethylene glycol units.

[0953] In certain embodiments, linker A is a bond, unsubstituted C 1-30 Alkylene, or unsubstituted C 1-30 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 10 ethylene glycol units.

[0954] In certain embodiments, linker A is a bond, unsubstituted C 1-25 Alkylene, or unsubstituted C 1-25 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 8 ethylene glycol units.

[0955] In certain embodiments, linker A is a bond, unsubstituted C 1-15 Alkylene, or unsubstituted C 1-15 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 5 ethylene glycol units.

[0956] In certain embodiments, linker A is a bond, or unsubstituted C 1-6 It may be alkylene.

[0957] In certain embodiments, linker A is bonded or unsubstituted C 1-3 It may be alkylene.

[0958]

[0959] Hereinafter, specific examples of compounds comprising the Fc-binding unit of the present application are additionally illustrated. The description of each element of the Fc-binding unit has been described in detail in the preceding paragraphs, and the description of each element is as described above.

[0960]

[0961] Some embodiments of the present application provide a compound of formula 2-8.

[0962] In some embodiments, Formula 2 can be represented by Formula 2-8 below.

[0963] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-8:

[0964] [Chemical Formula 2-8]

[0965] ,

[0966] At this time,

[0967] FcBU stands for Fc binding unit,

[0968] H RG is a reactive moiety,

[0969] L a is linker A,

[0970] X is either C or O.

[0971]

[0972] In certain embodiments, FcBU may have the structure:

[0973] ,

[0974] At this time,

[0975] Each Xaa is independently any amino acid residue other than a cysteine ​​residue,

[0976] Xa 1'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue,

[0977] Xa 2 is a glutamic acid residue or an asparagine residue,

[0978] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue,

[0979] In the above structure, the cysteine ​​residue adjacent to the N terminus (i.e., the cysteine ​​residue located 2 to 4 from the N terminus) and the cysteine ​​residue adjacent to the C terminus (i.e., the cysteine ​​residue located 2 to 4 from the C terminus) may optionally be covalently linked.

[0980] For example, a cysteine ​​residue adjacent to the N-terminus and a cysteine ​​residue adjacent to the C-terminus can optionally be linked via a disulfide bond.

[0981] In certain embodiments, the Fc binding unit can have the structure:

[0982] ,

[0983] At this time,

[0984] Each Xaa is independently any amino acid other than cysteine,

[0985] Xa 1'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue,

[0986] Xa 2 is a glutamic acid residue or an asparagine residue,

[0987] Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue,

[0988] At this time, the cysteine ​​residue adjacent to the N-terminus and the cysteine ​​residue adjacent to the C-terminus are optionally covalently linked (e.g., via a disulfide bond).

[0989] In certain embodiments, the Fc binding unit can have the structure:

[0990] ,

[0991] At this time,

[0992] Xa 1 'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, a conjugated lysine (Lys) residue, a conjugated 2,7-diaminoheptanoic acid residue, a conjugated cysteine ​​residue, or a conjugated 2-aminosuberic acid residue.

[0993]

[0994] In certain embodiments, HRG may be a bioorthogonal functional group.

[0995] In certain embodiments, H RG may have any of the following structures:

[0996]

[0997] ,

[0998] At this time, hn is an integer from 1 to 3,

[0999] R H are each independently H or -R, =O, =S, -NO2, -CR3, -NR2, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Selected from heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, =O, =S, and -SH.

[1000] In certain embodiments, H RGmay be selected from an azide group, a terminal alkyne group, a terminal alkene group, a cyclic alkyne (e.g., cyclooctyne) group, a tetrazine group, a norbornene group, a cycloalkene (e.g., cyclooctene) group, an oxime group, and an isocyanide group. At this time, the cyclooctyne may be any one selected from OCT cyclooctyne, BCN (Bicyclononyne), DBCO (Dibenzocyclooctyne), DIBAC (aza-dibenzocyclooctynes), DIBO (dibenzocyclooctynol), DIFO (difluorinated cyclooctynes), BARAC (biarylazacyclooctynone), DIMAC (dimethoxyazacyclooctyne), and DIFBO (difluorobenzocyclooctyne), but is not otherwise limited. At this time, cyclooctene can be any one selected from, for example, a cis-cyclooctene group and a trans-cyclooctene group.

[1001] In certain embodiments, H RG may be an azide group or a norbornene group.

[1002] In certain embodiments, H RG may be a click chemical functional group.

[1003] In certain embodiments, H RG can be selected from among diels-alder diene, diels-alder dienophile, IEDDA diene, and IEDDA dienophile.

[1004]

[1005] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60It may be heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom can be independently selected from O, N, and S.

[1006] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be heteroalkylene, wherein the heteroalkylene contains one or more heteroatoms, and wherein each heteroatom may be O.

[1007] In certain embodiments, linker A is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 20 ethylene glycol units.

[1008] In certain embodiments, linker A is a bond, unsubstituted C 1-50 Alkylene, or unsubstituted C 1-50 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 16 ethylene glycol units.

[1009] In certain embodiments, linker A is a bond, unsubstituted C 1-30 Alkylene, or unsubstituted C 1-30 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 10 ethylene glycol units.

[1010] In certain embodiments, linker A is a bond, unsubstituted C 1-25 Alkylene, or unsubstituted C1-25 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 8 ethylene glycol units.

[1011] In certain embodiments, linker A is a bond, unsubstituted C 1-15 Alkylene, or unsubstituted C 1-15 It may be a heteroalkylene, wherein the heteroalkylene comprises one or more heteroatoms, wherein each heteroatom may be O, and the heteroalkylene may comprise 0 to 5 ethylene glycol units.

[1012] In certain embodiments, linker A is a bond, or unsubstituted C 1-6 It may be alkylene.

[1013] In certain embodiments, linker A is bonded or unsubstituted C 1-3 It may be alkylene.

[1014]

[1015] Hereinafter, specific examples of compounds comprising the Fc-binding unit of the present application are additionally illustrated, and specific examples of compounds comprising the Fc-binding unit are not limited to the structures provided below. The description of each element of the Fc-binding unit has been described in detail in the preceding paragraphs, and the description of each element is as described above.

[1016]

[1017] Some embodiments of the present application provide a compound of formula 2-9.

[1018] In some embodiments, Formula 2 can be represented by Formula 2-9 below.

[1019] In some embodiments, a compound comprising an Fc binding unit of the present application may have the structure of Formula 2-9:

[1020] [Chemical Formula 2-9]

[1021] .

[1022]

[1023] Some embodiments of the present application provide a compound of formula 2-10.

[1024] In some embodiments, Formula 2 can be represented by Formula 2-10 below.

[1025] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-10:

[1026] [Chemical Formula 2-10]

[1027] .

[1028]

[1029] Some embodiments of the present application provide a compound of formula 2-11.

[1030] In some embodiments, Formula 2 can be represented by Formula 2-11 below.

[1031] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-11:

[1032] [Chemical Formula 2-11]

[1033] .

[1034]

[1035] Some embodiments of the present application provide a compound of formula 2-12.

[1036] In some embodiments, Formula 2 can be represented by Formula 2-12 below.

[1037] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-12:

[1038] [Chemical Formula 2-12]

[1039] .

[1040]

[1041] Some embodiments of the present application provide a compound of formula 2-13.

[1042] In some embodiments, Formula 2 can be represented by Formula 2-13 below.

[1043] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-13:

[1044] [Chemical Formula 2-13]

[1045] .

[1046]

[1047] Some embodiments of the present application provide a compound of formula 2-14.

[1048] In some embodiments, Formula 2 can be represented by Formula 2-14 below.

[1049] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-14:

[1050] [Chemical Formula 2-14]

[1051] .

[1052]

[1053] Some embodiments of the present application provide a compound of formula 2-15.

[1054] In some embodiments, Formula 2 can be represented by Formula 2-15 below.

[1055] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-15:

[1056] [Chemical Formula 2-15]

[1057] .

[1058]

[1059] Some embodiments of the present application provide a compound of formula 2-16.

[1060] In some embodiments, Formula 2 can be represented by Formula 2-16 below.

[1061] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-16:

[1062] [Chemical Formula 2-16]

[1063] ,

[1064] At this time,

[1065] sf is an integer from 0 to 8,

[1066] sg is an integer from 0 to 15, and

[1067] sh is an integer from 0 to 8.

[1068]

[1069] Some embodiments of the present application provide a compound of formula 2-17.

[1070] In some embodiments, Formula 2 can be represented by Formula 2-17 below.

[1071] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-17:

[1072] [Chemical Formula 2-17]

[1073] ,

[1074] At this time,

[1075] sf is an integer from 0 to 8,

[1076] sg is an integer from 0 to 15, and

[1077] sh is an integer from 0 to 8.

[1078]

[1079] Some embodiments of the present application provide a compound of formula 2-18.

[1080] In some embodiments, Formula 2 can be represented by Formula 2-18 below.

[1081] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-18:

[1082] [Chemical Formula 2-18]

[1083] ,

[1084] At this time,

[1085] sa is an integer from 1 to 6, and

[1086] se is an integer from 0 to 15.

[1087]

[1088] Some embodiments of the present application provide a compound of formula 2-19.

[1089] In some embodiments, Formula 2 can be represented by Formula 2-19 below.

[1090] In some embodiments, a compound comprising the Fc binding unit of the present application may have the structure of Formula 2-19:

[1091] [Chemical Formula 2-19]

[1092] ,

[1093] At this time,

[1094] sa is an integer from 1 to 6,

[1095] sb is an integer from 0 to 3,

[1096] sc is an integer from 1 to 15, and

[1097] sd is an integer from 0 to 3.

[1098]

[1099] Below, specific structures that compounds containing an Fc-binding unit may have are illustrated. The chemical formulas below are to be understood as specific examples that compounds containing the Fc-binding unit of the present application may take, and the scope of compounds containing the Fc-binding unit provided by the present application is not to be construed as being limited to the following examples.

[1100] In some embodiments, the compound comprising an Fc binding unit can have any one of the structures of the following formulae:

[1101]

[1102]

[1103]

[1104] At this time,

[1105] sa is an integer from 1 to 6,

[1106] sb is an integer from 0 to 3,

[1107] sc is an integer from 1 to 15,

[1108] sd is an integer from 0 to 3,

[1109] se is an integer from 0 to 15,

[1110] FcBU is a structure Have,

[1111] At this time, Xa 1 'is a conjugated diaminopropionic acid (Dap) residue, a conjugated diaminobytyric acid (Dab) residue, a conjugated ornithine (Orn) residue, or a conjugated lysine (Lys) residue.

[1112]

[1113] Preparation of a compound comprising an Fc binding unit

[1114] The compound comprising the Fc-binding unit described above can be prepared by reacting an Fc-binding material and a compound for preparing a compound comprising the Fc-binding unit (e.g., which may be referred to as a precursor compound of the compound comprising the Fc-binding unit).

[1115] Fc binding agents have been described in detail in the previous paragraphs.

[1116] Below, compounds for preparing compounds containing an Fc binding unit are described in detail.

[1117]

[1118] A compound for preparing a compound comprising an Fc-binding unit may be referred to as a precursor compound of a compound comprising an Fc-binding unit.

[1119] Below, precursor compounds of compounds containing an Fc binding unit are described, using compounds that can be used to prepare compounds of Chemical Formula 2-1 as examples.

[1120] A precursor compound of a compound comprising an Fc binding unit may have, for example, a structure of formula 5:

[1121] [Chemical Formula 5]

[1122] ,

[1123] At this time, R pre is an -OH, N-Hydroxysuccinimide (NHS) group, or pentafluorophenol group.

[1124] For example, R pre may have any of the following structures:

[1125] .

[1126] For example, Xa of the amino acid sequence included in the Fc binding substance 1 Amino acid residue and R of the compound of chemical formula 5 preA compound including an Fc binding unit (e.g., a compound of formula 2-1) can be prepared through the reaction of a carbonyl group adjacent to the Fc binding unit, but is not limited thereto.

[1127]

[1128] Furthermore, some embodiments of the present application provide methods for preparing a compound comprising an Fc binding unit.

[1129] In some embodiments, a method of preparing a compound comprising an Fc binding unit may comprise:

[1130] Contacting or reacting a precursor compound of a compound comprising an Fc-binding substance and an Fc-binding unit (e.g., a compound of formula 5).

[1131] Here, the contact between the precursor of the compound including the Fc-binding unit and the Fc-binding material can be accomplished through various methods. For example, the contact can be achieved by mixing a composition including the precursor of the compound including the Fc-binding unit and a composition including the Fc-binding material. In another example, the contact can be achieved by adding the precursor of the compound including the Fc-binding unit and the Fc-binding material to a pre-prepared solution, but is not limited thereto.

[1132] Furthermore, the method for preparing a compound comprising an Fc-binding unit may further comprise a step of obtaining a compound comprising an Fc-binding unit. Furthermore, the method for preparing a compound comprising an Fc-binding unit may further comprise a step of incubating a composition or solution comprising a precursor of a compound comprising an Fc-binding unit and an Fc-binding substance.

[1133] In some embodiments, the contact or reaction of the precursor of the compound comprising the Fc-binding unit and the Fc-binding agent may be performed under appropriate conditions. For example, the contact or reaction may be performed at a pH of 4 to 12. For example, the contact or reaction may be performed at a temperature of 10°C to 50°C. For example, the contact or reaction may be performed for 10 minutes to 3 days.

[1134]

[1135] In addition to the precursor compound of the above-described chemical formula 5, various compounds can be used to prepare a compound comprising an Fc-binding unit, and the embodiments of the precursor compound of the compound comprising an Fc-binding unit and the embodiments of the method for preparing the compound comprising an Fc-binding unit are not limited to the examples described above.

[1136]

[1137] Composition or kit comprising a compound comprising an Fc binding unit

[1138] Some embodiments of the present application provide a composition comprising a compound comprising an Fc binding unit.

[1139] In some embodiments, a composition comprising a compound comprising an Fc binding unit can be used to prepare an antibody conjugate comprising the desired group described below.

[1140] In some embodiments, a composition comprising a compound comprising an Fc binding unit can be used to deliver a desired group to an antibody.

[1141] The compound comprising the Fc-binding unit is as described above. In some embodiments, the compound comprising the Fc-binding unit can have a structure of any one of Formula 2 and Formulas 2-1 to 2-27.

[1142] In some embodiments, a composition comprising a compound comprising an Fc-binding unit may further comprise additional components in addition to the compound comprising the Fc-binding unit. For example, additional components included in the composition may include, but are not limited to, pharmaceutically acceptable salts, excipients, diluents, stabilizers, and pH adjusters.

[1143] Some embodiments of the present application provide a kit comprising a compound comprising an Fc binding unit.

[1144] In some embodiments, a kit comprising a compound comprising an Fc binding unit can be used to prepare an antibody comprising the desired group described below.

[1145] In some embodiments, a kit comprising a compound comprising an Fc binding unit can be used to deliver a desired group to an antibody.

[1146] The compound comprising the Fc-binding unit is as described above. In some embodiments, the compound comprising the Fc-binding unit can have a structure of any one of Formula 2 and Formulas 2-1 to 2-27.

[1147] In some embodiments, a kit comprising a compound comprising an Fc-binding unit may further comprise additional components in addition to the compound comprising the Fc-binding unit. For example, additional components included in the kit may include, but are not limited to, pharmaceutically acceptable salts, excipients, diluents, stabilizers, and pH adjusters.

[1148]

[1149] Compounds comprising the Fc-binding unit of the present application described above may exhibit high reaction efficiency in reaction with antibodies. For example, compounds comprising the Fc-binding unit may exhibit enhanced delivery efficiency of a target group in reaction with antibodies. Compounds comprising the Fc-binding unit according to some embodiments may exhibit high stability. For example, high stability may have a positive effect on the storage, distribution, and / or reaction with antibodies of the compound.

[1150] Hereinafter, a method for preparing an antibody conjugate using a compound comprising an Fc-binding unit is described in detail. The product produced by the reaction of an antibody with a compound comprising an Fc-binding unit may be referred to as, but is not limited to, an antibody comprising a target group, a conjugate comprising a target group (e.g., an antibody conjugate), or a modified antibody.

[1151]

[1152] Length design of partial structures of compounds containing Fc binding units

[1153] Compounds comprising the Fc-binding unit of the present application can be used to site-specifically deliver a target group to an antibody. For example, the Fc-binding unit of the present application can be used to deliver a target group to K246 and / or K248 of an antibody.

[1154] The following content is provided solely for the purpose of explaining the length design of the partial structure of the compound including the Fc binding unit of the present application, and the scope of the present application should not be limited by the following content.

[1155] As described above, the reaction between a compound containing an Fc-binding unit and an antibody is induced by the proximity effect of the Fc-binding unit derived from the Fc-binding substance, and the interaction between the Fc-binding substance and the antibody can be partially confirmed through simulation. For example, the Fc-binding substance and the Fc region of the antibody can be arranged in a specific positional relationship, and this positional relationship can be confirmed through simulation.

[1156] Meanwhile, a person skilled in the art will appreciate that the reaction between the *-marked reactive carbonyl of a compound comprising an Fc-binding unit and the primary amine group can occur more successfully when the reaction site (reactive carbonyl marked with *) of the compound comprising an Fc-binding unit with the antibody is adjacent to the primary amine group of the target lysine (K246 and / or K248) of the antibody.

[1157] Below, the amino acid sequence of sequence number 05 (DCAWHXa 1 The positional relationship with the antibody is explained using an Fc-binding substance having GELVWCT as an example.

[1158] Figures 05 and 06 illustrate the positional relationship between the primary amine groups of lysines 246 and 248 of the Fc region of an antibody and the Fc-binding substance when the antibody and the Fc-binding substance interact. Specifically, Figures 05 and 06 illustrate the Xa of the Fc-binding substance. 1 The minimum and maximum distances to the beta carbon of Xa1' of the Fc-binding unit of the compound including the Fc-binding unit and the reactive carbonyl carbon indicated by * are shown. Through FIG. 05 and FIG. 06, (1) when the distance between the beta carbon of Xa1' of the Fc-binding unit and the reactive carbonyl carbon indicated by * (hereinafter collectively referred to as distance A) is within the range of distances shown in FIG. 05 and FIG. 06 or is similar to the distances shown, it can be expected that the target substance can be more successfully delivered to K246 and / or K248.

[1159] For illustration, in the structure of chemical formula 2, Xa 1 The structure from the beta carbon of ' to the carbon of the reactive carbonyl is shown below:

[1160] ,

[1161] At this time, ** is Xa 1 ' indicates the beta carbon, * indicates the carbon of the reactive carbonyl, and m1 is an integer from 0 to 9.

[1162] In the above structure, when the sum of the number of atoms located in the main chain (hereinafter, the length of the main chain) relative to the distance A is 8 or more and 20 or less, it is expected that the length formed by the above structure will fall within the range of distances shown in FIGS. 05 and 06 or be similar to the distances shown, so that the target group can be more successfully transferred to K246 and / or K248.

[1163] For example, if m1 is 0, D a If J is C1 alkylene, the number of atoms located in the main chain in the above structure is counted as 8. That is, in this case, the length of the main chain is 8. According to the above, J f and J a - Each atom is located in the main chain, and therefore, when counting the length of the main chain, J f is counted as 1, and J a is counted as 1. For example, if m1 is 2, D a- When m1 is C3 alkylene, the length of the main chain of the above structure is 12. For example, if m1 is 2, and D a- When it is C3 heteroalkenylene, the length of the main chain of the above structure is 12.

[1164] D aThe number of atoms located in the main chain is conveniently referred to as k. In some embodiments, the sum of m1 and k may be 1 or more and 13 or less. For example, m1 may be 0, and k may be an integer of 1 or more and 13 or less. For example, m1 may be 2, and k may be an integer of 1 or more and 11 or less. For example, m1 may be 3, and k may be an integer of 1 or more and 10 or less. For example, m1 may be 4, and k may be an integer of 1 or more and 9 or less. For example, m1 may be 4, and k may be an integer of 1 or more and 8 or less. In certain embodiments, the sum of m1 and k may be 1 or more and 12 or less. In certain embodiments, the sum of m1 and k may be 1 or more and 11 or less. In certain embodiments, the sum of m1 and k may be 1 or more and 10 or less.

[1165]

[1166]

[1167] Preparation of antibody conjugates using compounds containing Fc binding units

[1168] Reaction of a compound comprising an Fc binding unit with an antibody and an antibody conjugate prepared thereby

[1169] A compound comprising an Fc-binding unit can contact or react with an antibody to transfer a target group to the antibody. Specifically, a compound comprising an Fc-binding unit can contact or react with an antibody to site-specifically transfer a target group to the antibody. Here, the site to which the target group is transferred (e.g., a target site or a labeling site) may be K246 and / or K248 of the Fc region of the antibody. That is, an antibody conjugate comprising (e.g., site-specifically comprising) a target group can be prepared by contacting, mixing, or reacting a compound comprising an Fc-binding unit with an antibody. In this case, in the prepared antibody conjugate, the target group may be linked to at least one of K246 and K248. Specifically, since antibodies have two heavy chains and can each contain lysine 246 and lysine 248 (e.g., trastuzumab has a total of four labeling positions), antibody conjugates containing one to four desired groups can be prepared by reacting, contacting, or mixing an antibody with a compound comprising an Fc-binding unit.

[1170] For example, an antibody conjugate comprising one target group can be prepared, wherein the antibody conjugate comprising one target group can be referred to as a Group of interest-to-Antibody ratio (GAR) 1 antibody conjugate.

[1171] For example, an antibody conjugate comprising two desired groups can be prepared, wherein the antibody conjugate comprising two desired groups can be referred to as a GAR2 antibody conjugate.

[1172] For example, an antibody conjugate comprising three desired groups can be prepared, wherein the antibody conjugate comprising three desired groups can be referred to as a GAR3 antibody conjugate.

[1173] For example, an antibody conjugate comprising four target groups can be prepared, wherein the antibody conjugate comprising four target groups can be referred to as a GAR4 antibody conjugate.

[1174] The term "antibody conjugate comprising a target group" encompasses all aspects of antibody conjugates comprising one to four target groups as described above. Preferably, the antibody conjugate comprising a target group may be an antibody conjugate comprising two target groups.

[1175]

[1176] FIG. 07 illustrates a reaction of a compound comprising the Fc-binding unit of the present application with an antibody, and an antibody conjugate comprising a target group produced by the reaction. As illustrated in FIG. 07, the reaction of the antibody with a compound comprising the Fc-binding unit can deliver the target group to a target site within the antibody (e.g., either K246 or K248 of the Fc region of the antibody).

[1177]

[1178] Below, antibody conjugates comprising one to four desired groups are described in more detail.

[1179] Before describing antibody conjugates comprising one to four target groups, we will give names to target sites that may be present in antibodies. Antibodies are generally known to have two heavy chains and two light chains. One of the two heavy chains may be referred to as the first heavy chain, and the other may be referred to as the second heavy chain. K246 and K248 present in one of the two heavy chains (the first heavy chain) may be referred to as the first K246 and the first K248, respectively. K246 and K248 present in the other of the two heavy chains (the second heavy chain) may be referred to as the second K246 and the second K248, respectively.

[1180] In some embodiments, an antibody conjugate comprising a target group may comprise one target group. In some embodiments, one target group may be linked to any one of the first K246, the first K248, the second K246, and the second K248. In a particular embodiment, one target group may be linked to any one of the first K246 and the second K246. In a particular embodiment, one target group may be linked to any one of the first K248 and the second K248.

[1181] In some embodiments, an antibody conjugate comprising a group of interest may comprise two groups of interest (e.g., a first group of interest and a second group of interest). In this case, the two groups of interest may be linked via two lysines selected from K246 of the first heavy chain (first K246), K248 of the first heavy chain (first K248), K246 of the second heavy chain (second K246), and K248 of the second heavy chain (second K248). In some embodiments, the first group of interest of the two groups of interest may be linked to one heavy chain of the antibody (the first heavy chain) and the second group of interest may be linked to the other heavy chain of the antibody (the second heavy chain). In some embodiments, the first group of interest may be linked to either K246 or K248 of the first heavy chain and the second group of interest may be linked to either K246 or K248 of the second heavy chain. In a particular embodiment, both groups of interest may be linked to K246. In this case, one of the two groups of interest may be linked to K246 of the first heavy chain, and the other may be linked to K246 of the second heavy chain. In a particular embodiment, both groups of interest may be linked to K248. In this case, one of the two groups of interest may be linked to K248 of the first heavy chain, and the other may be linked to K248 of the second heavy chain. In a particular embodiment, the two groups of interest may be linked to K246 and K248, respectively. In this case, one of the two groups of interest (e.g., the first group of interest) may be linked to K246 of the first heavy chain, and the other (e.g., the second group of interest) may be linked to K248 of the second heavy chain.By way of example, through FIGS. 08 to 10, an antibody comprising a target group in which two target groups are linked to K246 (FIG. 08); an antibody conjugate comprising a target group in which two target groups are linked to K248 (FIG. 09); and an antibody conjugate comprising two target groups, wherein one of the two target groups (the first target group) is linked to K246 and the other target group (the second target group) is linked to K248 (FIG. 10).

[1182] In some embodiments, an antibody conjugate comprising a target group may comprise three target groups. In this case, the three target groups may be linked via three lysines selected from the first K246, the first K248, the second K246, and the second K248, respectively.

[1183] In some embodiments, an antibody conjugate comprising a target group may comprise four target groups. In this case, the four target groups may be linked to the first K246, the first K248, the second K246, and the second K248, respectively.

[1184]

[1185] As described above, the preparation of a conjugate using a compound comprising an Fc-binding unit (e.g., an antibody conjugate comprising a group of interest) essentially involves the use of the following two elements:

[1186] (1) a compound comprising an Fc binding unit; and

[1187] (2) Antibodies.

[1188]

[1189] Below, the components used in the preparation of a conjugate using a compound comprising an Fc binding unit are described in detail.

[1190]

[1191] Element 1 used in the manufacture of a conjugate - a compound comprising an Fc binding unit

[1192] As described above, compounds comprising an Fc-binding unit of the present application are used in the preparation of conjugates (e.g., antibody conjugates comprising a group of interest). Compounds comprising an Fc-binding unit of the present application are described in detail in the section "Compounds Comprising an Fc-binding unit" of the present application, and compounds comprising an Fc-binding unit used in the preparation of conjugates are as described in that section.

[1193]

[1194] Element 2 used in conjugate manufacturing - Antibody

[1195] As mentioned above, antibodies are used in the preparation of conjugates (e.g., antibody conjugates comprising a group of interest). The Fc-binding agent or Fc-binding unit has binding affinity for the Fc region of an antibody.

[1196] In some embodiments, the antibody may comprise an Fc region of IgG. In some embodiments, the Fc region of the antibody may be an Fc region of IgG.

[1197] In some embodiments, the antibody may be an IgG antibody. IgG antibodies include human IgG antibodies, humanized IgG antibodies, and chimeric IgG antibodies.

[1198] IgG is known to be classified into IgG1, IgG2, IgG3, and IgG4.

[1199] In some embodiments, the antibody may be an IgG1 antibody. IgG1 antibodies include human IgG1 antibodies, humanized IgG1 antibodies, and chimeric IgG1 antibodies.

[1200] In some embodiments, the antibody may comprise an Fc region of IgG1. The Fc region of the antibody may be an Fc region of IgG1.

[1201] In some embodiments, the antibody may be an IgG2 antibody. IgG2 antibodies include human IgG2 antibodies, humanized IgG2 antibodies, and chimeric IgG2 antibodies.

[1202] In some embodiments, the antibody may comprise an Fc region of IgG2. The Fc region of the antibody may be an Fc region of IgG2.

[1203] In some embodiments, the antibody may be an IgG3 antibody. IgG3 antibodies include human IgG3 antibodies, humanized IgG3 antibodies, and chimeric IgG3 antibodies.

[1204] In some embodiments, the antibody may comprise an Fc region of IgG3. The Fc region of the antibody may be an Fc region of IgG3.

[1205] In some embodiments, the antibody may be an IgG4 antibody. IgG4 antibodies include human IgG4 antibodies, humanized IgG4 antibodies, and chimeric IgG4 antibodies.

[1206] In some embodiments, the antibody may comprise an Fc region of IgG4. The Fc region of the antibody may be an Fc region of IgG4.

[1207] In some embodiments, the antibody can have an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In particular embodiments, the antibody can have an amino acid sequence of SEQ ID NO: 14 or an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto.

[1208] In some embodiments, the antibody comprises an IgG Fc region (e.g., the Fc region of the antibody is an Fc region of IgG), wherein the IgG Fc region can have an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In particular embodiments, the antibody comprises an Fc region of IgG, wherein the IgG Fc region can have an amino acid sequence of SEQ ID NO: 14 or an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto.

[1209] In some embodiments, the antibody or Fc region of the antibody may comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNH (SEQ ID NO: 11).

[1210] In some embodiments, the antibody or Fc region of the antibody may comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNHY (SEQ ID NO: 12).

[1211] In some embodiments, the antibody or FC region of the antibody may comprise the amino acid sequence GPSVFLFPPKPKDTLM (SEQ ID NO: 13).

[1212] In some embodiments, the antibody has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNH (SEQ ID NO: 11). In some embodiments, the antibody comprises an IgG Fc region, wherein the IgG Fc region has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNH (SEQ ID NO: 11).

[1213] In some embodiments, the antibody has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 15 and SEQ ID NOs: 17 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, and may essentially comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNHY (SEQ ID NO: 12). In some embodiments, the antibody comprises an IgG Fc region, wherein the IgG Fc region has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 15 and SEQ ID NOs: 17 to 18, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, and may essentially comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and the amino acid sequence of MHEALHNHY (SEQ ID NO: 12).

[1214] In some embodiments, the antibody has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises GPSVFLFPPKPKDTLM (SEQ ID NO: 13). In some embodiments, the antibody comprises an IgG Fc region, wherein the IgG Fc region has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises GPSVFLFPPKPKDTLM (SEQ ID NO: 13).

[1215] In some embodiments, the antibody having binding affinity for the Fc binding agent or peptide of the present application may be an antibody of the IgG isotype. In some embodiments, the antibody having binding affinity for the Fc binding agent or peptide of the present application may be an antibody of the IgG1 isotype, an antibody of the IgG2 isotype, an antibody of the IgG3 isotype, or an IgG4 isotype. In some embodiments, the antibody having binding affinity for the Fc binding agent or peptide of the present application may be an antibody of the IgG1 isotype, an antibody of the IgG2 isotype, or an IgG4 isotype.

[1216] The inventors of the present application have confirmed that a desired group (e.g., a bioorthogonal functional group) is transferred to an antibody by a reaction between a compound comprising the Fc-binding unit of the present application and the antibody for various types of antibodies. More specifically, the inventors of the present application have confirmed that when a compound comprising the Fc-binding unit of the present application (compound 9 of the embodiment) is used, a desired group (e.g., an azide group) is transferred to each of trastuzumab (an IgG1 type antibody), an IgG1 type anti-CLDN18.2 antibody, an IgG1 type anti-CD154 antibody, Denosumab (an IgG2 type antibody), and Dupliumab.

[1217] In some embodiments, the antibody is selected from the group consisting of adalimumab (Humira), rituximab (Rituxan), trastuzumab (Herceptin), bevacizumab (Avastin), infliximab (Remicade), pembrolizumab (Keytruda), nivolumab (Opdivo), eculizumab (Soliris), alemtuzumab (Lemtrada, Campath), daratumumab (Darzalex), ipilimumab (Yervoy), golimumab (Simponi), tocilizumab (Actemra), ranibizumab (Lucentis), Secukinumab (Cosentyx), Ixekizumab (Taltz), Dupliumab (Dupixent), Denosumab, Ustekinumab (Stelara), Palivizumab (Synagis), Durvalumab (Imfinzi), Atezolizumab (Tecentriq), Omalizumab (Xolair), Vedolizumab (Entyvio), Abciximab (ReoPro), Basiliximab (Simulect), Alefacept (Amevive), Daclizumab (Zinbryta), and Elotuzumab (Empliciti).

[1218] In some embodiments, the antibody is EpCAM, CD2, CD3, CD4, CD5, CD6, CD11, CD19, CD20, CD22, CD26, CD30, CD33, CD37, CD38, CD40, CD44, CD56, CD79, CD105, CD138, EphA receptors, EphB receptors, EGFR, EGFRvIII, HER2, HER3, mesothelin, cripto, alphavbeta3, alphavbeta5, Nectin-4, TROP2, PD1, PD-L1, BCMA, B3H7, FOLR-a, Tissue factor, CLDN 1 (claudin 1), CLDN 3 (claudin 3), CLDN 4 (claudin 4), CLDN 6 (claudin 6), CLDN 18.2 (claudin 18.2) and alpha v It may be an antibody that binds to any one of the beta 6 integrins.

[1219] In some embodiments, the antibody may be an anti-CLDN18.2 antibody (see literature [Korean Patent Application No. 10-2021-7023724]).

[1220] In some embodiments, the heavy chain of the anti-CLDN18.2 antibody can comprise a CDRH1 having an amino acid sequence of SEQ ID NO: 19 (TYGVH) or having at least 90% sequence identity thereto, a CDRH2 having an amino acid sequence of SEQ ID NO: 20 (VIWAGGSTNYNSALMS) or having at least 90% sequence identity thereto, and a CDRH3 having an amino acid sequence of SEQ ID NO: 21 (AAYYGNGLDY) or having at least 90% sequence identity thereto. In certain embodiments, the anti-CLDN18.2 antibody can have two heavy chains comprising a CDRH1 having the amino acid sequence of SEQ ID NO: 19, a CDRH2 having the amino acid sequence of SEQ ID NO: 20, and a CDRH3 having the amino acid sequence of SEQ ID NO: 21.

[1221] In some embodiments, the light chain of the anti-CLDN18.2 antibody can comprise a CDRL1 having an amino acid sequence of SEQ ID NO: 22 (KSSQTLLNSGNQKNYLT) or having at least 90% sequence identity thereto, a CDRL2 having an amino acid sequence of SEQ ID NO: 23 (WASTGES) or having at least 90% sequence identity thereto, and a CDRL3 having an amino acid sequence of SEQ ID NO: 24 (QNAYFYPFT) or having at least 90% sequence identity thereto. In particular embodiments, the anti-CLDN18.2 antibody can have two light chains comprising a CDRL1 having the amino acid sequence of SEQ ID NO: 22, a CDRL2 having the amino acid sequence of SEQ ID NO: 23, and a CDRL3 having the amino acid sequence of SEQ ID NO: 24.

[1222] In some embodiments, the heavy chain of the anti-CLDN18.2 antibody may comprise a CDRH1 having an amino acid sequence of SEQ ID NO: 19 or at least 90% sequence identity thereto, a CDRH2 having an amino acid sequence of SEQ ID NO: 20 or at least 90% sequence identity thereto, and a CDRH3 having an amino acid sequence of SEQ ID NO: 21 or at least 90% sequence identity thereto, and the light chain of the anti-CLDN18.2 antibody may comprise a CDRL1 having an amino acid sequence of SEQ ID NO: 22 or at least 90% sequence identity thereto, a CDRL2 having an amino acid sequence of SEQ ID NO: 23 or at least 90% sequence identity thereto, and a CDRL3 having an amino acid sequence of SEQ ID NO: 24 or at least 90% sequence identity thereto. In certain embodiments, the anti-CLDN18.2 antibody may have two heavy chains comprising CDRH1 having the amino acid sequence of SEQ ID NO: 19, CDRH2 having the amino acid sequence of SEQ ID NO: 20, and CDRH3 having the amino acid sequence of SEQ ID NO: 21, and two light chains comprising CDRL1 having the amino acid sequence of SEQ ID NO: 22, CDRL2 having the amino acid sequence of SEQ ID NO: 23, and CDRL3 having the amino acid sequence of SEQ ID NO: 24.

[1223] In some embodiments, the anti-CLDN18.2 antibody may be an antibody comprising a heavy chain having an amino acid sequence of SEQ ID NO: 25 or having at least 90% sequence identity thereto, and a light chain having an amino acid sequence of SEQ ID NO: 26 or having at least 90% sequence identity thereto. In a particular embodiment, the anti-CLDN18.2 antibody may be an antibody comprising a heavy chain having an amino acid sequence of SEQ ID NO: 25 and a light chain having an amino acid sequence of SEQ ID NO: 26. Specifically, the anti-CLDN18.2 antibody may have two heavy chains having the amino acid sequence of SEQ ID NO: 25 and two light chains having the amino acid sequence of SEQ ID NO: 26.

[1224]

[1225] The binding affinity of an Fc binding agent to an antibody or the Fc region of an antibody can be expressed as a dissociation constant (Kd). In some embodiments, the Kd (dissociation constant) of an Fc binding agent to an antibody is 10 μM, 1 μM (1x10 -6 M), 900nM, 800nM, 700nM, 600nM, 500nM, 450nM, 400nM, 350nM, 300nM, 250nM, 200nM, 180nM, 160nM, 140nM, 120nM, 100nM, 90nM, 80nM, 70nM, 60nM, 50nM, 40nM, 30nM, 20nM, 10nM, 5nM, 2nM, 1nM (1x10 -9 M), 0.5 nM, 0.1 nM, 0.05 nM, or 0.01 nM or less, or within a range set by any two values ​​selected from the foregoing values. In some embodiments, the Kd (dissociation constant) for the Fc domain of an antibody of the Fc binding agent is 10 μM, 1 μM (1x10 -6 M), 900nM, 800nM, 700nM, 600nM, 500nM, 450nM, 400nM, 350nM, 300nM, 250nM, 200nM, 180nM, 160nM, 140nM, 120nM, 100nM, 90nM, 80nM, 70nM, 60nM, 50nM, 40nM, 30nM, 20nM, 10nM, 5nM, 2nM, 1nM (1x10 -9 M), 0.5 nM, 0.1 nM, 0.05 nM, or 0.01 nM or less, or within a range set by any two values ​​selected from the aforementioned values, but is not limited thereto.

[1226]

[1227] Method for preparing antibody conjugate using compound containing Fc binding unit

[1228] Some embodiments of the present application provide methods for preparing antibody conjugates comprising a group of interest. Methods for preparing antibody conjugates comprising a group of interest may be referred to as methods for preparing antibody conjugates that site-specifically comprise a group of interest, methods for transferring a group of interest to an antibody, methods for site-specifically transferring a group of interest to an antibody, and methods for site-specifically transferring a reactive group (e.g., a bioorthogonal functional group) to an antibody. As such, the methods may be freely referred to as such, consistent with the purpose to be achieved by reacting a compound comprising an Fc-binding unit with an antibody.

[1229]

[1230] Some embodiments of the present application provide a method of preparing an antibody conjugate comprising a desired group comprising:

[1231] Contacting a compound and an antibody comprising an Fc binding unit.

[1232] Here, the term "contact" may be replaced with terms such as "reaction" or "mixing".

[1233] Here, the compound comprising an Fc-binding unit is a compound comprising an Fc-binding unit provided by the present application. For example, the compound comprising an Fc-binding unit may be a compound of formula 2. For example, the compound comprising an Fc-binding unit may be any one of compounds of formula 2 and formula 2-1 to formula 2-27 (i.e., the compound comprising an Fc-binding unit may be a compound having a structure of any one of formulas 2 and formula 2-1 to formula 2-27). The compound comprising an Fc-binding unit of the present application and the elements included therein are described in detail in the section “Compound comprising an Fc-binding unit”, and the compound comprising an Fc-binding unit and the elements included therein are as described in the preceding paragraphs.

[1234] Here, the contact between the compound comprising the Fc-binding unit and the antibody can be accomplished through various methods. For example, the contact can be achieved by mixing a composition comprising the compound comprising the Fc-binding unit and a composition comprising the antibody. In another example, the contact can be achieved by adding the compound comprising the Fc-binding unit and the antibody to a pre-prepared solution, but is not limited thereto.

[1235] In some embodiments, an antibody conjugate comprising a desired group can be prepared by contacting a compound comprising an Fc-binding unit and an antibody. In some embodiments, in the antibody conjugate comprising a desired group, the desired group can be linked to at least one selected from K246 and K248 of the Fc region of the antibody. In some embodiments, in the antibody conjugate comprising a desired group, the desired group can be linked to K246 of the Fc region of the antibody. In some embodiments, in the antibody conjugate comprising a desired group, the desired group can be linked to K248 of the Fc region of the antibody. In some embodiments, an antibody conjugate comprising a desired group can comprise one to four desired groups. In some embodiments, an antibody conjugate comprising a desired group can comprise one desired group. In certain embodiments, an antibody conjugate comprising a desired group can comprise two desired groups.

[1236] In some embodiments, the target group can be site-specifically transferred to the antibody (e.g., to a target region of the antibody) by contacting a compound comprising an Fc-binding unit with an antibody. For example, the target region can consist of 1 to 20, 1 to 10, 1 to 5, or 1 to 3 consecutive amino acid residues, including K246 and K248 of the Fc region. In some embodiments, the target group can be transferred to any one or more lysine residues selected from K246 and K248 of the Fc region of the antibody by contacting a compound comprising an Fc-binding unit with the antibody. In some embodiments, the target group can be transferred to K246 of the Fc region of the antibody by contacting a compound comprising an Fc-binding unit with the antibody. In some embodiments, the target group can be transferred to K248 of the Fc region of the antibody by contacting a compound comprising an Fc-binding unit with the antibody.

[1237] In some embodiments, the contacting or reaction of a compound comprising an Fc binding unit and an antibody can be performed in a solution or composition.

[1238] In some embodiments, the contacting or reaction of the compound comprising an Fc-binding unit and the antibody can be carried out at a pH selected from any one of pH 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, and 12, or a pH range set by two values ​​selected from the foregoing. In some embodiments, the contacting or reaction of the compound comprising an Fc-binding unit and the antibody can be carried out at a pH of from 5 to 10, from 6 to 10, from 7 to 10, from 5 to 9, from 5 to 8, from 6 to 9, from 6 to 8, from 7 to 9, or from 7 to 8. In particular embodiments, the reaction of the compound comprising an Fc-binding unit and the antibody can be carried out at a pH of from 6.5 to 8.5 or from 7 to 8. In certain embodiments, the reaction of a compound comprising an Fc binding unit and an antibody can be performed at about pH 7.4.

[1239] In some embodiments, the contacting or reaction of the compound comprising an Fc-binding unit and the antibody can be performed at 10° C. to 50° C., 10° C. to 45° C., 15° C. to 45° C., 15° C. to 40° C., 20° C. to 45° C., 20° C. to 40° C., 20° C. to 35° C., 20° C. to 30° C., 25° C. to 45° C., 25° C. to 40° C., 25° C. to 35° C., or 20° C. to 30° C. In some embodiments, the reaction of the compound comprising an Fc-binding unit and the antibody can be performed at room temperature.

[1240] In some embodiments, the method for preparing an antibody conjugate comprising a group of interest may further comprise incubating a solution (or composition) comprising a compound comprising an Fc-binding unit and the antibody conjugate. During the incubation, optionally, but not limited to, stirring or vortexing may be performed. The incubation may be performed for a period of time of about, but not limited to, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 42 hours, 48 ​​hours, 60 hours, or 72 hours or more.

[1241] In some embodiments, the method for preparing an antibody conjugate comprising a target group may further comprise a step of obtaining an antibody conjugate comprising the target group. In this case, obtaining an antibody conjugate comprising the target group may include, but is not limited to, a step of purifying an antibody conjugate comprising the target group.

[1242] By way of example, a method for preparing an antibody conjugate comprising a desired group may include the following (see FIG. 11):

[1243] (a) mixing a compound comprising an Fc-binding unit and an antibody to prepare a composition comprising a compound comprising an Fc-binding unit and an antibody;

[1244] (b) incubating a composition comprising a compound comprising the Fc binding unit and an antibody; and

[1245] (c) Obtaining an antibody conjugate comprising the desired group from the incubated composition.

[1246]

[1247] Advantages of using a compound comprising the Fc binding unit of the present application

[1248] As described above, the compound comprising the Fc-binding unit of the present application can exhibit enhanced reaction efficiency in reaction with an antibody. For example, the reaction of the compound comprising the Fc-binding unit of the present application with an antibody can have an enhanced reaction rate. Accordingly, the production yield of the conjugate prepared by the reaction of the compound comprising the Fc-binding unit with an antibody can be increased, or the time required for preparing the conjugate can be reduced. Hereinafter, the advantages of using the compound comprising the Fc-binding unit of the present application, i.e., the advantages of the method for preparing an antibody conjugate comprising a desired group provided by the present application, will be described.

[1249]

[1250] As described above, when using a compound containing a conventional Fc binding unit (a compound of Chemical Formula 1-1), there was a problem that an antibody conjugate containing a desired group was not produced or took a long time to produce because the reaction efficiency of the compound of Chemical Formula 1-1 with an antibody was not good. According to an experiment conducted by the inventors of the present application, even though the antibody and the compound of Chemical Formula 1-1 were reacted for 3 hours, only 5% of the antibody conjugate was obtained, and even the obtained antibody conjugate was confirmed to be a GAR1 antibody conjugate (see Example 02).

[1251] On the other hand, it is confirmed that the production yield of the antibody conjugate is improved when a compound comprising the Fc binding unit of the present application is used.

[1252]

[1253] In some embodiments, the reaction efficiency can be calculated based on the amount of antibody conjugate comprising a target group produced through the reaction for a predetermined time. The reaction efficiency can be calculated, for example, from the yield of antibody conjugates produced through the reaction for a predetermined time. For example, if an antibody conjugate comprising 60 target groups is produced from 100 antibodies initially added through a 1-hour reaction, the yield of the antibody conjugate comprising the target group is 60%. For example, if an antibody conjugate comprising 80 target groups is produced from 100 antibodies initially added through a 3-hour reaction, the yield of the antibody conjugate comprising the target group is 80%. The antibody conjugate comprising the target group here may include all of the GAR1, GAR2, GAR3, and GAR4 antibody conjugates. The antibody conjugate comprising the target group here may refer to the GAR2 antibody conjugate. For example, if 60 GAR1 antibody conjugates and 20 GAR2 antibody conjugates are produced from 100 antibodies initially added through a 3-hour reaction, the yield of GAR2 antibody conjugates is 20%. In some embodiments, the reaction efficiency can be calculated based on the amount of antibody-payload conjugates described below. Here, the antibody-payload conjugates can include all of a PAR (payload to antibody ratio) 1 antibody conjugate in which one payload (i.e., one cargo moiety) is linked to an antibody unit, a PAR2 antibody conjugate in which two payloads are linked to an antibody unit, a PAR3 antibody conjugate in which three payloads are linked to an antibody unit, and a PAR4 antibody conjugate in which four payloads are linked to an antibody unit. For example, if 60 PAR2 antibody conjugates are produced from 100 antibodies initially added through a 3-hour reaction, the yield is 60%.The antibody conjugate that serves as the reference for the yield can be appropriately selected. For example, the antibody conjugate that serves as the reference for the yield can be the GAR1 and GAR2 antibody conjugates. For example, the antibody conjugate that serves as the reference for the yield can be the PAR1 and PAR2 antibody conjugates. For example, the antibody conjugate that serves as the reference for the yield can be the GAR2 antibody conjugate. For example, the antibody conjugate that serves as the reference for the yield can be the PAR2 antibody conjugate.

[1254] In some embodiments, the yield of the antibody conjugate comprising the desired group obtained through the reaction for a predetermined time period can be greater than or equal to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95%. Here, the predetermined time period can be, for example, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 42 hours, 48 ​​hours, 60 hours, or 72 hours or more.

[1255]

[1256] Composition or kit for the preparation of an antibody conjugate comprising a desired group

[1257] Some embodiments of the present application provide a composition or kit for preparing an antibody conjugate comprising a desired group. The composition or kit for preparing an antibody conjugate comprising a desired group can be used to prepare an antibody conjugate comprising the desired group. The composition or kit for preparing an antibody conjugate comprising a desired group comprises a compound comprising an Fc-binding unit and an antibody. The compound comprising an Fc-binding unit and the antibody are each as described above.

[1258] Some embodiments of the present application provide a composition for the preparation of an antibody conjugate comprising a desired group.

[1259] In some embodiments, a composition for preparing an antibody conjugate comprising a desired group may further comprise additional components in addition to the compound comprising the Fc binding unit and the antibody. For example, additional components included in the composition may include, but are not limited to, carriers, excipients, diluents, stabilizers, and pH adjusters.

[1260] Some embodiments of the present application provide a kit for preparing an antibody conjugate comprising a desired group.

[1261] In some embodiments, a kit for preparing an antibody conjugate comprising a desired group may further comprise additional components in addition to the compound and antibody comprising the Fc-binding unit. For example, additional components included in the kit may include, but are not limited to, carriers, excipients, diluents, stabilizers, and pH adjusters.

[1262]

[1263] Hereinafter, antibody conjugates comprising a desired group prepared by reaction or contact of a compound comprising an Fc binding unit and an antibody are described in detail.

[1264]

[1265]

[1266] Antibody conjugate containing the desired group

[1267] Overview of antibody conjugates containing the target group

[1268] As described above, an antibody conjugate comprising a desired group can be prepared by reacting a compound comprising an Fc binding unit with an antibody.

[1269]

[1270] The compound of the following chemical formula 6 may be referred to as an antibody conjugate containing the desired group.

[1271] Some embodiments of the present application provide a compound having the structure of Formula 6:

[1272] [Chemical Formula 6]

[1273] .

[1274] In chemical formula 6, Ab is an antibody unit.

[1275] In chemical formula 6, L b is linker B.

[1276] In Chemical Formula 6, GOI is a group of interest. The group of interest is described in detail in the previous paragraphs, and is as described in the previous paragraphs.

[1277] In chemical formula 6, n is an integer from 1 to 4.

[1278] At this time, in Chemical Formula 6, the desired group may be linked to at least one of lysine residues 246 (K246) and lysine residues 248 (K248) of the Fc region of the antibody unit.

[1279]

[1280] Below, each element of the compound of chemical formula 6 is described in detail.

[1281]

[1282] antibody unit

[1283] An antibody unit is derived from an antibody and may be referred to as a conjugated antibody. Furthermore, the structure of an antibody unit is identical to the antibody from which it is derived, except for the conjugated portion, and thus, it may be referred to as an antibody, and the descriptions provided in the paragraphs describing antibodies may be applied as is. Figure 12 compares the structures of an antibody and an antibody unit by specifying the conjugated portion. As illustrated in Figure 12, an antibody unit derived from an antibody and an antibody from which an antibody unit is derived are structurally nearly identical.

[1284] In some embodiments, the antibody (or antibody unit) may comprise an Fc region of IgG. In some embodiments, the Fc region of the antibody may be an Fc region of IgG.

[1285] In some embodiments, the antibody may be an IgG antibody. IgG antibodies include human IgG antibodies, humanized IgG antibodies, and chimeric IgG antibodies.

[1286] IgG is known to be classified into IgG1, IgG2, IgG3, and IgG4.

[1287] In some embodiments, the antibody may be an IgG1 antibody. IgG1 antibodies include human IgG1 antibodies, humanized IgG1 antibodies, and chimeric IgG1 antibodies.

[1288] In some embodiments, the antibody may comprise an Fc region of IgG1. The Fc region of the antibody may be an Fc region of IgG1.

[1289] In some embodiments, the antibody may be an IgG2 antibody. IgG2 antibodies include human IgG2 antibodies, humanized IgG2 antibodies, and chimeric IgG2 antibodies.

[1290] In some embodiments, the antibody may comprise an Fc region of IgG2. The Fc region of the antibody may be an Fc region of IgG2.

[1291] In some embodiments, the antibody may be an IgG3 antibody. IgG3 antibodies include human IgG3 antibodies, humanized IgG3 antibodies, and chimeric IgG3 antibodies.

[1292] In some embodiments, the antibody may comprise an Fc region of IgG3. The Fc region of the antibody may be an Fc region of IgG3.

[1293] In some embodiments, the antibody may be an IgG4 antibody. IgG4 antibodies include human IgG4 antibodies, humanized IgG4 antibodies, and chimeric IgG4 antibodies.

[1294] In some embodiments, the antibody may comprise an Fc region of IgG4. The Fc region of the antibody may be an Fc region of IgG4.

[1295] In some embodiments, the antibody can have an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In particular embodiments, the antibody can have an amino acid sequence of SEQ ID NO: 14 or an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto.

[1296] In some embodiments, the antibody comprises an IgG Fc region, wherein the IgG Fc region can have an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In particular embodiments, the antibody comprises an Fc region of an IgG, wherein the IgG Fc region can have an amino acid sequence selected from SEQ ID NOs: 14 or an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto.

[1297] In some embodiments, the antibody or Fc region of the antibody may comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNH (SEQ ID NO: 11).

[1298] In some embodiments, the antibody or Fc region of the antibody may comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNHY (SEQ ID NO: 12).

[1299] In some embodiments, the antibody or FC region of the antibody may comprise the amino acid sequence GPSVFLFPPKPKDTLM (SEQ ID NO: 13).

[1300] In some embodiments, the antibody has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNH (SEQ ID NO: 11). In some embodiments, the antibody comprises an IgG Fc region, wherein the IgG Fc region has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNH (SEQ ID NO: 11).

[1301] In some embodiments, the antibody has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 15 and SEQ ID NOs: 17 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, and may comprise the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNHY (SEQ ID NO: 12). In some embodiments, the antibody comprises an IgG Fc region (e.g., the Fc region of the antibody is an Fc region of IgG), wherein the IgG Fc region has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 15 and SEQ ID NOs: 17 to 18, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, and wherein the antibody comprises the amino acid sequence of KPKDTLM (SEQ ID NO: 10) and MHEALHNHY (SEQ ID NO: 12).

[1302] In some embodiments, the antibody has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises GPSVFLFPPKPKDTLM (SEQ ID NO: 13). In some embodiments, the antibody comprises an IgG Fc region, wherein the IgG Fc region has an amino acid sequence selected from any one of SEQ ID NOs: 14 to 18 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, wherein the antibody essentially comprises GPSVFLFPPKPKDTLM (SEQ ID NO: 13).

[1303] In some embodiments, the antibody having binding affinity for the Fc binding agent or peptide of the present application may be an antibody of the IgG isotype. In some embodiments, the antibody having binding affinity for the Fc binding agent or peptide of the present application may be an antibody of the IgG1 isotype, an antibody of the IgG2 isotype, an antibody of the IgG3 isotype, or an IgG4 isotype. In some embodiments, the antibody having binding affinity for the Fc binding agent or peptide of the present application may be an antibody of the IgG1 isotype, an antibody of the IgG2 isotype, or an IgG4 isotype.

[1304] In some embodiments, the antibody is selected from the group consisting of adalimumab (Humira), rituximab (Rituxan), trastuzumab (Herceptin), bevacizumab (Avastin), infliximab (Remicade), pembrolizumab (Keytruda), nivolumab (Opdivo), eculizumab (Soliris), alemtuzumab (Lemtrada, Campath), daratumumab (Darzalex), ipilimumab (Yervoy), golimumab (Simponi), tocilizumab (Actemra), ranibizumab (Lucentis), Secukinumab (Cosentyx), Ixekizumab (Taltz), Dupliumab (Dupixent), Denosumab, Ustekinumab (Stelara), Palivizumab (Synagis), Durvalumab (Imfinzi), Atezolizumab (Tecentriq), Omalizumab (Xolair), Vedolizumab (Entyvio), Abciximab (ReoPro), Basiliximab (Simulect), Alefacept (Amevive), Daclizumab (Zinbryta), and Elotuzumab (Empliciti).

[1305] In some embodiments, the antibody is EpCAM, CD2, CD3, CD4, CD5, CD6, CD11, CD19, CD20, CD22, CD26, CD30, CD33, CD37, CD38, CD40, CD44, CD56, CD79, CD105, CD138, EphA receptors, EphB receptors, EGFR, EGFRvIII, HER2, HER3, mesothelin, cripto, alphavbeta3, alphavbeta5, Nectin-4, TROP2, PD1, PD-L1, BCMA, B3H7, FOLR-a, Tissue factor, CLDN 1 (claudin 1), CLDN 3 (claudin 3), CLDN 4 (claudin 4), CLDN 6 (claudin 6), CLDN 18.2 (claudin 18.2) and alpha v It may be an antibody that binds to any one of the beta 6 integrins.

[1306] In some embodiments, the antibody may be an anti-CLDN18.2 antibody (see literature [Korean Patent Application No. 10-2021-7023724]).

[1307] In some embodiments, the heavy chain of the anti-CLDN18.2 antibody can comprise a CDRH1 having an amino acid sequence of SEQ ID NO: 19 (TYGVH) or having at least 90% sequence identity thereto, a CDRH2 having an amino acid sequence of SEQ ID NO: 20 (VIWAGGSTNYNSALMS) or having at least 90% sequence identity thereto, and a CDRH3 having an amino acid sequence of SEQ ID NO: 21 (AAYYGNGLDY) or having at least 90% sequence identity thereto. In certain embodiments, the anti-CLDN18.2 antibody can have two heavy chains comprising a CDRH1 having the amino acid sequence of SEQ ID NO: 19, a CDRH2 having the amino acid sequence of SEQ ID NO: 20, and a CDRH3 having the amino acid sequence of SEQ ID NO: 21.

[1308] In some embodiments, the light chain of the anti-CLDN18.2 antibody can comprise a CDRL1 having an amino acid sequence of SEQ ID NO: 22 (KSSQTLLNSGNQKNYLT) or having at least 90% sequence identity thereto, a CDRL2 having an amino acid sequence of SEQ ID NO: 23 (WASTGES) or having at least 90% sequence identity thereto, and a CDRL3 having an amino acid sequence of SEQ ID NO: 24 (QNAYFYPFT) or having at least 90% sequence identity thereto. In particular embodiments, the anti-CLDN18.2 antibody can have two light chains comprising a CDRL1 having the amino acid sequence of SEQ ID NO: 22, a CDRL2 having the amino acid sequence of SEQ ID NO: 23, and a CDRL3 having the amino acid sequence of SEQ ID NO: 24.

[1309] In some embodiments, the heavy chain of the anti-CLDN18.2 antibody may comprise a CDRH1 having an amino acid sequence of SEQ ID NO: 19 or at least 90% sequence identity thereto, a CDRH2 having an amino acid sequence of SEQ ID NO: 20 or at least 90% sequence identity thereto, and a CDRH3 having an amino acid sequence of SEQ ID NO: 21 or at least 90% sequence identity thereto, and the light chain of the anti-CLDN18.2 antibody may comprise a CDRL1 having an amino acid sequence of SEQ ID NO: 22 or at least 90% sequence identity thereto, a CDRL2 having an amino acid sequence of SEQ ID NO: 23 or at least 90% sequence identity thereto, and a CDRL3 having an amino acid sequence of SEQ ID NO: 24 or at least 90% sequence identity thereto. In certain embodiments, the anti-CLDN18.2 antibody may have two heavy chains comprising CDRH1 having the amino acid sequence of SEQ ID NO: 19, CDRH2 having the amino acid sequence of SEQ ID NO: 20, and CDRH3 having the amino acid sequence of SEQ ID NO: 21, and two light chains comprising CDRL1 having the amino acid sequence of SEQ ID NO: 22, CDRL2 having the amino acid sequence of SEQ ID NO: 23, and CDRL3 having the amino acid sequence of SEQ ID NO: 24.

[1310] In some embodiments, the anti-CLDN18.2 antibody may be an antibody comprising a heavy chain having an amino acid sequence of SEQ ID NO: 25 or having at least 90% sequence identity thereto, and a light chain having an amino acid sequence of SEQ ID NO: 26 or having at least 90% sequence identity thereto. In a particular embodiment, the anti-CLDN18.2 antibody may be an antibody comprising a heavy chain having an amino acid sequence of SEQ ID NO: 25 and a light chain having an amino acid sequence of SEQ ID NO: 26. Specifically, the anti-CLDN18.2 antibody may have two heavy chains having the amino acid sequence of SEQ ID NO: 25 and two light chains having the amino acid sequence of SEQ ID NO: 26.

[1311]

[1312] Linker of antibody conjugate containing the desired group (Linker B; L'b)

[1313] The reaction of a compound comprising an Fc binding unit with an antibody has been described in detail in the previous paragraphs, whereby a moiety comprising a group of interest is site-specifically transferred to the antibody. The linker of the antibody conjugate comprising the group of interest (linker B; L b ) has a structure identical to that of some compounds comprising an Fc binding unit.

[1314]

[1315] Linker B can have the following structure:

[1316] .

[1317] In the above structure, R a2 , R a3 , X, and L a Each of these has been described in detail in the preceding paragraphs, and each of these is as described in the preceding paragraphs.

[1318]

[1319] In certain embodiments, linker B can be represented by the following structure:

[1320] .

[1321] In the above structure, R a2 , X, and L a Each of these has been described in detail in the preceding paragraphs, and each of these is as described in the preceding paragraphs.

[1322]

[1323] In certain embodiments, linker B can be represented by the following structure:

[1324] .

[1325] In the above structure, X, and L a Each of these has been described in detail in the preceding paragraphs, and each of these is as described in the preceding paragraphs.

[1326]

[1327] Location where the target group is connected

[1328] In Chemical Formula 6, n is an integer from 1 to 4. In some embodiments, n may be an integer from 1 to 2. In a particular embodiment, n may be 2.

[1329] The desired group may be linked (via linker B) to one or more of lysine residues 246 (K246) and lysine residues 248 (K248) of the Fc region of the antibody unit. More specifically, the antibody unit may comprise two heavy chains (a first heavy chain and a second heavy chain), wherein "-L b -GOI" can be linked to any one or more of K246 of the first heavy chain, K248 of the first heavy chain, K246 of the second heavy chain, and K248 of the second heavy chain.

[1330] For example, if one target group is linked to K246 of the first heavy chain of the antibody unit (via linker B), n is 1. For example, if one target group is linked to K248 of the first heavy chain of the antibody unit, n is 1.

[1331] For example, if each of the two target groups is linked to K246 of the first heavy chain and K246 of the second heavy chain of the antibody unit, n is 2. For example, if each of the two target groups is linked to K248 of the first heavy chain and K248 of the second heavy chain of the antibody unit, n is 2. For example, if each of the two target groups is linked to K246 of the first heavy chain and K248 of the second heavy chain of the antibody unit, n is 2.

[1332] For example, if each of the three desired groups is linked to K246 of the first heavy chain, K248 of the first heavy chain, and K248 of the second heavy chain of the antibody unit, then n is 3. For example, if each of the three desired groups is linked to K246 of the first heavy chain, K246 of the second heavy chain, and K248 of the second heavy chain of the antibody unit, then n is 3.

[1333] For example, if each of the four target groups is linked to K246 of the first heavy chain, K248 of the first heavy chain, K246 of the second heavy chain, and K248 of the second heavy chain of the antibody unit, then n is 4.

[1334]

[1335] Specific examples of antibody conjugates comprising a target group - antibody conjugates comprising a reactive group

[1336] The desired group may include a reactive group or a functional group. For example, if the desired group includes a reactive group, an antibody conjugate including the desired group may be referred to as an antibody conjugate including a reactive group.

[1337]

[1338] For example, the compound of chemical formula 6 may be the compound of chemical formula 6-1.

[1339] Some embodiments of the present application provide an antibody conjugate comprising a desired group (i.e., an antibody conjugate comprising a reactive group herein) having the structure of the following chemical formula 6-1:

[1340] [Chemical Formula 6-1]

[1341] .

[1342] At this time, L b is linker B, and linker B is as described above.

[1343] At this time, RG is a reactive group, and the reactive group is as described above.

[1344]

[1345] Use of antibody conjugates containing reactive groups

[1346] Overview of the Use of Antibody Conjugates Containing Reactive Groups

[1347] An antibody-payload conjugate (e.g., an antibody-drug conjugate) can be prepared using an antibody conjugate containing a reactive group. The antibody-payload conjugate can be prepared by contacting, reacting, or mixing an antibody containing a reactive group with a payload.

[1348]

[1349] An antibody conjugate comprising a reactive group refers to a group capable of reacting with another group. For example, the reactive group may be or include a reactive moiety, wherein the reactive moiety is a moiety that is reactive with another group. For example, the reactive moiety may be a bioorthogonal functional group (e.g., azide or norbornene).

[1350] The payload may comprise a reactive group capable of reacting with a reactive group of the antibody conjugate.

[1351] The reactive group of the antibody conjugate comprising the reactive group may be referred to as the first reactive group, and the reactive group of the payload may be referred to as the second reactive group.

[1352] When an antibody conjugate containing a reactive group and a payload are contacted, reacted, or mixed, an antibody-payload conjugate can be prepared by reaction of the first reactive group and the second reactive group.

[1353]

[1354] Below, the payload is described in detail.

[1355]

[1356] payload

[1357] Payload Overview

[1358] The payload may comprise a second reactive group and an active moiety. The payload may comprise one or more active moieties, wherein each of the one or more active moieties is independently selected. The active moieties are described in detail in the preceding paragraphs and are as described in the preceding paragraphs. For example, the active moiety may be, but is not limited to, a drug, an imaging moiety, a radioactive moiety, a protein having a specific function, a peptide having a specific function, an affinity agent (e.g., biotin, streptavidin, and aptamers), a stabilizer, a vitamin, a nucleic acid (e.g., DNA or RNA), or a PEG moiety. In some embodiments, the active moiety may be a drug moiety, an imaging moiety, a radioactive moiety, or an affinity agent.

[1359] The second reactive group may be a group capable of reacting with the first reactive group. For example, if the first reactive group is or comprises an azide, the second reactive group may be or comprises a DBCO capable of reacting with an azide. As another example, if the first reactive group is or comprises a norbornene, the second reactive group may be or comprises a tetrazine.

[1360] In some embodiments, the sum of the atomic masses of all atoms comprising the payload may be less than or equal to 10,000 Daltons, 9,000 Daltons, 8,000 Daltons, 7,000 Daltons, 6,000 Daltons, 5,000 Daltons, 4,500 Daltons, 4,000 Daltons, 3,500 Daltons, 3,000 Daltons, 2,500 Daltons, 2,000 Daltons, 1,500 Daltons, 1,000 Daltons, or 500 Daltons. It will be apparent to one of ordinary skill in the art that the sum of the atomic masses of all atoms comprising the payload will be greater than or equal to 20 Daltons.

[1361]

[1362] For example, the payload may have the structure of Chemical Formula 7 below:

[1363] [Chemical Formula 7]

[1364] ,

[1365] At this time, CM is a cargo moiety (CM),

[1366] RG 2 is the second reactive group.

[1367] The cargo moiety is characterized by comprising one or more active moieties.

[1368]

[1369] Below, the aspects of the second reactive group are described.

[1370]

[1371] Second reactive group

[1372] For a description of the second reactive group, reference may be made to the description relating to the reactive group described in the section “Compounds Comprising an Fc Binding Unit”.

[1373] The second reactive group comprises a reactive moiety. Here, the reactive moiety of the second reactive group is referred to as a second reactive moiety.

[1374] In some embodiments, the second reactive group may further comprise a reactive moiety (e.g., a second reactive moiety) as described in the paragraphs describing the reactive group, in addition to a reactive group spacer (e.g., a second reactive group spacer). In this case, the reactive moiety is linked to a moiety other than the reactive group via the reactive group spacer. For example, the second reactive group may have the following structure:

[1375] ,

[1376] At this time D RG2is a spacer of the second reactive group, and H RG2 is a second reactive moiety.

[1377]

[1378] In some embodiments, the second reactive moiety may be a bioorthogonal functional group (e.g., a second bioorthogonal functional group). The description of the bioorthogonal functional group is the same as the description of the bioorthogonal functional group in the section "Compounds Comprising an Fc Binding Unit" of the present application.

[1379] In some embodiments, the reactive moiety may be a click chemistry functional group (e.g., a second click chemistry functional group). The description of the click chemistry functional group is the same as the description of the click chemistry functional group in the section "Compounds Comprising an Fc-Binding Unit" of the present application.

[1380] In some embodiments, the reactive moiety can be selected from an azide group, a terminal alkyne group, a cyclic alkyne (e.g., cyclooctyne) group, a tetrazine group, a norbornene group, a cycloalkene (e.g., cyclooctene) group, a tetrazole group, an oxime group, and an isocyanide group, a halogen group, an aldehyde group, a nitrone group, a hydroxylamine group, a nitrile group, a hydrazine group, a ketone group, a bronic acid group, a cyanobenzothiazole group, an allyl...

Claims

1. A compound comprising an Fc binding unit having a structure of Chemical Formula 2-2: [Chemical Formula 2-2] , At this time, D a is a spacer A, wherein the spacer A is a bond, substituted or unsubstituted C 1-20 Alkylene, substituted or unsubstituted C 1-20 Heteroalkylene, substituted or unsubstituted C 2-20 Alkenylene, substituted or unsubstituted C 2-20 Heteroalkenylene, substituted or unsubstituted C 2-20 Alkynylene, or substituted or unsubstituted C 2-20 Heteroalkynylene, wherein substituted indicates that one or more hydrogen atoms in the group described by the term substituted is replaced by one or more substituents, wherein the substituents are each independently selected from -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH, wherein the substituent is not H, and wherein the heteroalkylene, heteroalkenylene, heteroalkynylene, heterocycloalkyl, or heteroaryl comprises one or more heteroatoms, wherein each heteroatom is independently selected from N, O, and S, L a is a linker A, wherein the linker A is a bond, substituted or unsubstituted C 1-100 Alkylene, substituted or unsubstituted C 1-100 Heteroalkylene, substituted or unsubstituted C 2-100 Alkenylene, substituted or unsubstituted C 2-100 Heteroalkenylene, substituted or unsubstituted C 2-100 Alkynylene, or substituted or unsubstituted C 2-100 Heteroalkynylene, wherein substituted means that one or more hydrogen atoms in the group described by the term substituted are substituted with one or more substituents, wherein the substituents are each independently selected from -R, =O, =S, -NO2, -CR3, -NR2, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH, wherein the substituent is not H, and wherein the heteroalkylene, heteroalkenylene, heteroalkynylene, heterocycloalkyl, or heteroaryl comprises one or more heteroatoms, wherein each heteroatom is independently selected from N, O, and S, X is -CH2-, -O-, or -NH-, R a1 is H or C 1-6 It is alkyl, R a2 is H or C 1-6 It is alkyl, R a3 is H or C 1-6 It is alkyl, J a is -C(=O)-, -S-, -NH-, or -C(=NH)-, RG is a reactive group, wherein the reactive group comprises a reactive moiety, FcBU is an Fc binding unit, wherein the Fc binding unit has a structure Have, At this time, Each Xaa is independently any amino acid residue, Xa 2 is a glutamic acid residue or an asparagine residue, Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue, The cysteine ​​residue adjacent to the N terminus and the cysteine ​​residue adjacent to the C terminus are optionally covalently linked, Xa 1 'silver and, Here, m is an integer greater than or equal to 1 and less than or equal to 5, J f is -NH-, -S-, or -C(=O)-, * and ** are each Xa 1 ' represents the attachment point with the adjacent amino acid residue, *** is Xa 1 ', and represents an attachment site with a non-Fc-binding unit portion of a compound including an Fc-binding unit.

2. In paragraph 1, A compound comprising an Fc binding unit, wherein X is -O- or -CH2-.

3. In paragraph 1, A compound comprising an Fc binding unit, wherein X is -O-.

4. In paragraph 1, R a1 Silver C 1-3 A compound comprising an alkylene, Fc binding unit.

5. In paragraph 1, R a1 A compound comprising a methyl, Fc-binding unit.

6. In paragraph 1, R a2 and R a3 are each independently H and C 1-3 A compound comprising an Fc binding unit, wherein the Fc binding unit is one selected from the group consisting of alkyl.

7. In paragraph 1, R a2 and R a3 A compound comprising an Fc binding unit, all of which are H.

8. In paragraph 1, J a A compound comprising an Fc binding unit, which is -C(=O)-.

9. In paragraph 1, D a is unsubstituted C 1-10 alkylene, unsubstituted C 1-10 Heteroalkylene, unsubstituted C 2-10 Alkenylene, or unsubstituted C 2-10 A compound comprising an Fc-binding unit, which is a heteroalkenylene.

10. In paragraph 1, D a is unsubstituted C 1-10 A compound comprising an alkylene, Fc binding unit.

11. In paragraph 1, A compound comprising an Fc-binding unit, wherein the Fc-binding unit has the following structure: .

12. In paragraph 1, J f A compound comprising an Fc binding unit, which is -NH-.

13. In paragraph 1, A compound comprising an Fc binding unit, wherein m is an integer greater than or equal to 1 and less than or equal to 4.

14. In paragraph 1, m is a compound comprising a 3-membered Fc binding unit.

15. In paragraph 1, L a is a bond, unsubstituted C 1-60 alkylene, unsubstituted C 1-60 Heteroalkylene, unsubstituted C 2-60 Alkenylene, or unsubstituted C 2-60 A compound comprising an Fc-binding unit, which is a heteroalkenylene.

16. In paragraph 1, L a is a bond, unsubstituted C 1-60 Alkylene, or unsubstituted C 1-60 A compound comprising an Fc binding unit, wherein the heteroalkylene is a heteroalkylene, wherein the unsubstituted heteroalkylene comprises 0 to 20 ethylene glycol units.

17. In paragraph 1, L a is a bond, unsubstituted C 1-30 Alkylene, or unsubstituted C 1-30 A compound comprising an Fc binding unit, wherein the heteroalkylene is a heteroalkylene, wherein the unsubstituted heteroalkylene comprises 0 to 10 ethylene glycol units.

18. In paragraph 1, L a is a bond, unsubstituted C 1-24 Alkylene, or unsubstituted C 1-24 A compound comprising an Fc binding unit, wherein the heteroalkylene is a heteroalkylene, wherein the unsubstituted heteroalkylene comprises 0 to 8 ethylene glycol units.

19. In paragraph 1, L a Is And, At this time, sf is an integer from 0 to 8, sg is an integer from 0 to 15, sh is an integer from 0 to 8, A compound comprising an Fc binding unit.

20. In paragraph 19, sf is an integer from 0 to 3, sg is an integer from 0 to 10, sh is an integer from 0 to 3, A compound comprising an Fc binding unit.

21. In paragraph 1, RG is a compound comprising an Fc binding unit represented by the following structure: , At this time, D RG is the spacer of the reactive group (spacer RG), At this time, the spacer of the above reactive group is bonded, substituted or unsubstituted C 1-6 Alkylene, substituted or unsubstituted C 1-6 Heteroalkylene, substituted or unsubstituted C 2-6 Alkenylene, or substituted or unsubstituted C 2-6 Heteroalkenylene, wherein substituted means that one or more hydrogen atoms in the group modified by the term substituted are replaced by one or more substituents, wherein the substituents are each independently -C 1-4 alkyl, -C(=O)H, -C(=O)CH3, -C(=O)OH, -C(=O)NH2, -NH2, =NH, =O, =S, -OH, -NO2, and -SH, wherein the heteroalkylene or heteroalkenylene contains one or more heteroatoms, wherein the heteroatoms are each independently selected from O, N, and S, H RG is a reactive moiety.

22. In paragraph 1, A compound comprising an Fc binding unit, wherein the reactive moiety is a bioorthogonal functional group.

23. In paragraph 1, The reactive moiety is a compound comprising an Fc binding unit represented by any one of the following structures: , At this time, hn is an integer from 1 to 3, R H are each independently H or -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Selected from heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH.

24. In paragraph 1, The reactive moiety is selected from an azide group, a terminal alkyne group, a terminal alkene group, a cyclooctyne group, a tetrazine group, a norbornene group, a cyclooctene group, an oxime group, and an isocyanide group, wherein the cyclooctyne group is any one selected from OCT cyclooctyne, BCN (Bicyclononyne), DBCO (Dibenzocyclooctyne), DIBAC (aza-dibenzocyclooctynes), DIBO (dibenzocyclooctynol), DIFO (difluorinated cyclooctynes), BARAC (biarylazacyclooctynone), DIMAC (dimethoxyazacyclooctyne), and DIFBO (difluorobenzocyclooctyne), wherein the cyclooctene group is any one selected from a cis-cyclooctene group and a trans-cyclooctene group.

25. A compound comprising an Fc binding unit having a structure of the following chemical formula 2-15: [Chemical Formula 2-15] , At this time, aa is an integer greater than or equal to 1 and less than or equal to 10, X is -O- or -CH2-, L a is a linker A, wherein the linker A is a bond, unsubstituted C 1-30 alkylene, or unsubstituted C containing 0 to 10 ethylene glycol units 1-30 It is heteroalkylene, is a reactive group, D RG is a spacer of a reactive group (spacer RG), wherein the spacer of the reactive group is a bond, substituted or unsubstituted C 1-6 Alkylene, substituted or unsubstituted C 1-6 Heteroalkylene, substituted or unsubstituted C 2-6 Alkenylene, or substituted or unsubstituted C 2-6 Heteroalkenylene, wherein substituted means that one or more hydrogen atoms in the group modified by the term substituted are replaced by one or more substituents, wherein the substituents are each independently -C 1-4 alkyl, -C(=O)H, -C(=O)CH3, -C(=O)OH, -C(=O)NH2, -NH2, =NH, =O, =S, -OH, -NO2, and -SH, wherein the heteroalkylene or heteroalkenylene contains one or more heteroatoms, wherein the heteroatoms are each independently selected from O, N, and S, H RG is a reactive moiety, FcBU is an Fc binding unit, wherein the Fc binding unit has a structure Have, At this time, Each Xaa is independently any amino acid, Xa 2 is a glutamic acid residue or an asparagine residue, Xa 3 is a tryptophan residue, a naphthylalanine residue, or a phenylalanine residue, The cysteine ​​residue adjacent to the N terminus and the cysteine ​​residue adjacent to the C terminus are optionally covalently linked, Xa 1 'silver and, Here, m is an integer greater than or equal to 1 and less than or equal to 4, * and ** are each Xa 1 ' represents the attachment point with the adjacent amino acid residue, *** is Xa 1 ', and represents an attachment site with a non-Fc-binding unit portion of a compound including an Fc-binding unit.

26. In paragraph 25, A compound comprising an Fc binding unit, wherein aa is an integer greater than or equal to 1 and less than or equal to 6.

27. In paragraph 25, aa is a compound containing a 3-membered Fc binding unit.

28. In paragraph 25, A compound comprising an Fc-binding unit, wherein the Fc-binding unit has the following structure: .

29. In paragraph 25, m is a compound comprising a 3-membered Fc binding unit.

30. In paragraph 25, A compound comprising an Fc binding unit, wherein the reactive moiety is a bioorthogonal functional group.

31. In paragraph 25, The reactive moiety is a compound comprising an Fc binding unit represented by any one of the following structures: , At this time, hn is an integer from 1 to 3, R H are each independently H or -R, =O, =S, -NO2, -CR3, -NR2, =NR, -OR, -SR, -C(=O)R, -C(=O)CR3, -C(=O)OR, and -C(=O)NR2, wherein R is each independently H, halogen, C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Selected from heterocycloalkyl, aryl, heteroaryl, -OH, -NH2, -COOH, =O, =S, and -SH.

32. In paragraph 25, The reactive moiety is selected from an azide group, a terminal alkyne group, a terminal alkene group, a cyclooctyne group, a tetrazine group, a norbornene group, a cyclooctene group, an oxime group, and an isocyanide group, wherein the cyclooctyne group is any one selected from OCT cyclooctyne, BCN (Bicyclononyne), DBCO (Dibenzocyclooctyne), DIBAC (aza-dibenzocyclooctynes), DIBO (dibenzocyclooctynol), DIFO (difluorinated cyclooctynes), BARAC (biarylazacyclooctynone), DIMAC (dimethoxyazacyclooctyne), and DIFBO (difluorobenzocyclooctyne), wherein the cyclooctene group is any one selected from a cis-cyclooctene group and a trans-cyclooctene group.

33. A method for preparing an antibody conjugate comprising a reactive group comprising: Contacting a compound comprising an Fc binding unit of any one of claims 1 to 32 with an antibody.

34. In paragraph 33, A method for preparing an antibody conjugate comprising a reactive group, wherein the antibody is an IgG antibody.

35. In paragraph 33, A method for preparing an antibody conjugate comprising a reactive group, wherein the antibody is an IgG antibody, and the IgG antibody is a human IgG antibody, a humanized IgG antibody, or a chimeric IgG antibody.

36. In paragraph 33, A method for producing an antibody conjugate comprising a reactive group, wherein the Fc region of the antibody comprises an amino acid sequence of SEQ ID NO: 14 or an amino acid sequence having at least 80% sequence identity thereto, wherein the Fc region of the antibody comprises an amino acid sequence of KPKDTLM (SEQ ID NO: 10) and an amino acid sequence of MHEALHNHY (SEQ ID NO: 12).

37. In paragraph 33, By contacting an antibody with a compound containing an Fc binding unit, the reactive group is transferred to the target region of the antibody, A method for producing an antibody conjugate comprising a reactive group, wherein the target region is composed of five consecutive amino acid residues and includes K246 and K248 of the Fc region of the antibody.

38. In paragraph 33, A method for preparing an antibody conjugate comprising a reactive group, wherein the reactive group is transferred to at least one of K246 and K248 of the Fc region of the antibody by contacting the antibody with a compound comprising an Fc binding unit.

39. In paragraph 33, A method for preparing an antibody conjugate comprising a reactive group, the method further comprising obtaining an antibody conjugate comprising a reactive group.

40. In paragraph 33, By contacting an antibody with a compound comprising an Fc binding unit, an antibody conjugate comprising a reactive group is prepared, At this time, the antibody conjugate containing the reactive group contains 1 to 4 reactive groups. A method for preparing an antibody conjugate comprising a reactive group.

41. In paragraph 33, By contacting an antibody with a compound comprising an Fc binding unit, an antibody conjugate comprising a reactive group is prepared, At this time, the antibody conjugate containing the above reactive group contains two reactive groups, In the antibody conjugate containing the reactive group at this time, one of the two reactive groups (the first reactive group) is linked to one of K246 and K248 of one of the two heavy chains of the antibody (the first heavy chain), In the antibody conjugate containing the reactive group at this time, the other reactive group (the second reactive group) of the two reactive groups is linked to either K246 or K248 of the other heavy chain (the second heavy chain) of the two heavy chains of the antibody. A method for preparing an antibody conjugate comprising a reactive group.

42. In paragraph 33, By contacting an antibody with a compound comprising an Fc binding unit, an antibody conjugate comprising a reactive group is prepared, At this time, the antibody conjugate containing the above reactive group contains two reactive groups, At this time, each of the two reactive groups is linked to K246 of one heavy chain of the antibody and K246 of the other heavy chain, or to K248 of one heavy chain of the antibody and K248 of the other heavy chain, respectively. A method for preparing an antibody conjugate comprising a reactive group.

43. In paragraph 33, A method for preparing an antibody conjugate comprising a reactive group, wherein contacting of a compound comprising an Fc binding unit with an antibody is achieved by a method comprising: Mixing a composition comprising a compound comprising an Fc binding unit and a composition comprising an antibody.

44. In paragraph 43, A method for preparing an antibody conjugate comprising a reactive group, wherein mixing a composition comprising a compound comprising an Fc binding unit and a composition comprising an antibody is performed under conditions of pH 6 to pH 8.5.

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