Peptide transmembrane delivery systems and their applications

Abstract

The present invention relates to a peptide conjugate, a pharmaceutical composition containing the same, and a method for using the same, for example, for the treatment and / or prevention of a disease in a subject.

Description

[Technical Field]

[0001] Reference to electronic sequence listings The contents of the electronic sequence listing (APSN-P-013-PCT.xml; size: 6,714 bytes; and creation date: March 27, 2024) are incorporated herein by reference in their entirety.

[0002] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 454,786, filed on 27 March 2023, entitled “Peptide Transmembrane Delivery System and Use thereof,” and to U.S. Provisional Patent Application No. 63 / 566,959, filed on 19 March 2024, entitled “Peptide Transmembrane Delivery System and Use thereof.” The contents thereof are incorporated herein by reference in their entirety. [Background technology]

[0003] Many peptides exhibit remarkable biological activity and potentially have the potential to be used as therapeutically active material components in pharmaceutical preparations. Current pharmaceutical research focuses on peptides that facilitate the targeting of intracellular protein-protein interactions, which is virtually impossible with small molecule-based drugs.

[0004] Peptides are more stable in biological culture media compared to other biomolecules such as oligonucleotides; however, most of them lack the ability to cross cell membranes.

[0005] Currently, the primary approach to improving cell permeability involves conjugating therapeutic peptide sequences with cell-permeable peptides (CPPs) that lack therapeutic activity but act as carriers by helping the therapeutic peptides cross the cell membrane. Many CPPs exhibit their potent permeability only at high micromolar concentrations (>10 μM). For the delivery of medium or large-sized drugs, these concentrations are unacceptable in a clinical setting. Therefore, there is a need to enhance permeability efficiency to achieve lower, meaningful clinical concentrations.

[0006] To achieve this objective, the main obstacle to peptide delivery in disease treatment is their extremely low cellular permeability. Therefore, safe and efficient means for intracellular delivery of peptides have been needed for many years, but remain unresolved. [Overview of the project]

[0007] In one aspect of the invention, a compound comprising any pharmaceutically acceptable salt, any hydrate, or any solvate of Formula 1: [ka] A conjugate is provided which includes a polymer covalently bonded to the portion represented by the formula, a and b each independently represent integers from 1 to 10; L represents a linker containing at least one -N(R5)- group, with linkers having a single CC bond length from 1 to 30; R represents one or more substituents, each independently representing halo, -NO2, -CN, -OH, -CONH2, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, oxo, -NHCOR, -NHCSR, -NHCNR, -NC(=O)OR, -NC(=O)NR', ​​-NC(=S)OR', -NC(=S)N R', -SO2R', -SOR', -SR', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', C1-C6 haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(C1-C6 alkyl), hydroxy(C1-C6 alkoxy), alkoxy(C1-C6 alkyl), alkoxy(C1-C6 alkoxy), C1-C6 alkyl-NR '2, C1-C6 alkyl-SR', -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -CO2R', -OCOR, -OCOR', -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​or a combination thereof; each R' independently represents hydrogen, or optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally placed Selected from the group including substituted heteroaryls, optionally substituted aryls, or combinations thereof; R5 is H, or a linear or branched optionally substituted C1-C5 alkyl (optionally containing one or more heteroatoms); the wavy bond represents an attachment site to the polymer; the polymer includes peptides, or polymers containing a thio group, an amino group, a 1,3-nitrone, an azide, a diene, a tetrazine, a maleimide, an active ester, an α,β-unsaturated keto acid derivative, or any combination thereof.

[0008] In one embodiment, part is Equation 2: [ka] is represented by, wherein, c is an integer independently selected from 0 to 10; L1 is a linker selected from a bond, or -N-C(=O)-, -C(=O)N-, -S-S-, -S-C(=O), Y-(C 0-10 )alkyl-X-(C 0-10 )alkyl-Y, and click reaction products, or any combination thereof; each X and Y is either absent or independently selected from heteroatoms, oligomers, click reaction products, -CONR’-, -CNNR’-, -CSNR’-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR’, -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-; the oligomer contains 2 to 15 repeating units; and, W represents (i) an amide, (ii) a click reaction product, or both.

[0009] In one embodiment, the repeating unit is selected from: alkylene oxide, natural or non-natural amino acid derivatives, α-hydroxycarboxylic acid residues, and amino groups.

[0010] In one embodiment, L1 is -N-C(=O), -C(=O)N, or Y-(C 0-10 )alkyl-X-(C 0-10 )alkyl-Y.

[0011] In one embodiment, the moiety is of formula 3:

Chemical formula

[0012] In one embodiment, the click reaction product comprises a succinimide-thioether, an azidoalkyne addition cyclization product, a direct and / or inverse electron demand Diels Alder reaction product, a dibenzylcyclooctyne 1,3-nitrone addition cyclization product, or any combination thereof.

[0013] In one embodiment, the polymer is a polynucleotide substituted by one or more of the following: a thio group, an amino group, a 1,3-nitrone, an azide, a diene, a tetrazine, a maleimide, an active ester, an α,β-unsaturated keto acid derivative, or any combination thereof.

[0014] In one embodiment, the moiety is attached to (i) the N-terminus of a peptide, or the C-terminus of a peptide, (ii) the C-terminal amino acid of a peptide, (iii) at least one of the following: an amino group and a thio group of an amino acid residue of a peptide, or any combination of (i)-(iii).

[0015] In one embodiment, the moiety is of formula 4:

Chemical formula

[0016] In one embodiment, Y1 is -N-C(=O)- and Y2 is -N-C(=O)- or -C(O=)O-.

[0017] In one embodiment, R5 is methyl.

[0018] In one embodiment, R is halogen.

[0019] In one embodiment, the conjugate is [ka] Select from the following options.

[0020] In one embodiment, the conjugate is derived from any of the compounds listed in Table A or B (i.e., the conjugate is obtained by conjugation with a polymer of a precursor from Table A or B).

[0021] In another embodiment, a compound comprising any pharmaceutically acceptable salt, any hydrate, or any solvate thereof is provided; the compound is given by formula 5: [ka] It is expressed by, in the formula, a and b each independently represent integers from 1 to 10; M represents a linker containing at least one -N(R5)- group; the linker has a single CC bond length from 1 to 30; Z is a moiety having reactivity to a functional group selected from thio groups, amino groups, 1,3-nitrones, azides, dienes, tetrazines, succinimides, α,β-unsaturated carbonyls, α,β-unsaturated keto acid derivatives, and active esters, or any combination thereof; R represents one or more substituents, each independently representing halo, -NO2, -CN -OH, -CONH2, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, -NHCOR, -NHCSR, -NHCNR, -NC(=O)OR, -NC(=O)NR', ​​-NC(=S)OR', -NC(=S)NR', ​​-SO2R', -SOR', -SR', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', C1-C6 haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1-C6 alkyl), -N(C 1-C6alkyl)2, C1-C6alkoxy, C1-C6haloalkoxy, Hydroxy(C1-C6alkyl), Hydroxy(C1-C6alkoxy), Alkoxy(C1-C6alkyl), Alkoxy(C1-C6alkoxy), C1-C6alkyl-NR'2, C1-C6alkyl-SR', -CONH(C1-C6alkyl), -CON(C1-C6alkyl)2, -CO2H, -CO2R', -OCOR, -OCOR', -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)O R' is selected from R', -OC(=S)NR', ​​or a combination thereof; each R' independently represents hydrogen, or is selected from the group including optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or a combination thereof; R5 is H, or a linear or branched optionally substituted C1-C5 alkyl (optionally containing one or more heteroatoms).

[0022] In one embodiment, the compound is of formula 6: [ka] It is expressed by, in the formula, c is an integer selected from 1 to 10; L1 is a bond, or -NC(=O)-, -C(=O)N-, -SS-, -SC(=O), Y-(C 0-10 )alkyl-X-(C 0-10 ) A linker selected from alkyl-Y (including any combination thereof); Each X and Y is either absent or independently selected from heteroatoms, oligomers, click reaction products, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, -OC(=S)N-; Oligomers contain 2 to 15 repeating units.

[0023] In one embodiment, L1 is -NC(=O), -C(=O)N, or Y-(C0-10)alkyl-X-(C0-10)alkyl-Y.

[0024] In one embodiment, the compound is of formula 7: [ka] It is represented by [this].

[0025] In one embodiment, the compound is of formula 8: [ka] This is expressed as follows, where n is an integer independently chosen from 1 to 10; Each Y1 and Y2 is independently selected from -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-.

[0026] In one embodiment, Y1 is -NC(=O)- and Y2 is -NC(=O)- or -C(O=)O-.

[0027] In one embodiment, Z is selected from active esters, α-β unsaturated keto compounds, and maleimides.

[0028] In one embodiment, R is a halo and R5 is a methyl group.

[0029] In one embodiment, the compound is [ka] Selected from.

[0030] In one embodiment, Z is reactive to a polymer containing a functional group; the compound is a precursor of the conjugate of the invention.

[0031] In another embodiment, a pharmaceutical composition comprising the conjugate of the invention and a pharmaceutically acceptable carrier is provided.

[0032] In one embodiment, the pharmaceutical composition comprises a therapeutically effective amount of conjugate.

[0033] In one embodiment, the pharmaceutical composition is intended for use in the treatment or prevention of a disease in a subject.

[0034] In one embodiment, the disease is cancer.

[0035] In another embodiment, a kit is provided comprising the compound of the invention and (i) a peptide; and (i) a polymer selected from polymers comprising a thio group, an amino group, a 1,3-nitrone, an azide, a diene, a tetrazine, or any combination thereof.

[0036] In one embodiment, the molar ratio between the polymer and the compound in the kit is approximately 1:1.

[0037] In one embodiment, the kit further includes instructions for reacting a polymer and a compound to obtain the conjugate of the invention.

[0038] In one embodiment, the kit further includes a liquid carrier.

[0039] In another embodiment, a method is provided for treating a disease or condition in a subject, comprising administering a therapeutically effective amount of the pharmaceutical composition of the invention to a subject to treat the disease.

[0040] In one embodiment, the disease is a cell proliferation-related disorder.

[0041] In another embodiment, a method is provided for internalizing a peptide into the cells of a subject, comprising contacting cells with the pharmaceutical composition of the invention, thereby internalizing the peptide into the cells.

[0042] In one embodiment, the cells are cancer cells.

[0043] The subject matter considered to be the invention is specifically pointed out and explicitly claimed in the final part of the specification. However, the invention, with respect to both its structure and method of operation, along with its purpose, features and advantages, can be best understood by referring to the detailed description below, when read together with the accompanying drawings. [Brief explanation of the drawing]

[0044] [Figure 1] This graph bar shows the B16-melanoma cell viability (normalized to the relevant control) after exposure to an exemplary conjugate of the invention, which includes the JNK-peptide as an exemplary therapeutic sequence. The amino acid sequences of the tested conjugate and control are presented in Table 1. [Figure 2]This graph bar shows the HeLa cell viability after exposure to an exemplary conjugate of the invention. #1, 2, and 3 represent CPP, control 2, and control 1, respectively. The amino acid sequences of the tested conjugates and controls are presented in Table 1. [Figure 3] This graph bar shows the B16-melanoma cell viability (normalized to the relevant control) after exposure to an exemplary conjugate of the invention, which includes the JNK-peptide as an exemplary therapeutic sequence. The amino acid sequences of the tested conjugate and control are presented in Table 3. [Modes for carrying out the invention]

[0045] The detailed description below includes many specific details to provide a complete understanding of the invention. However, those skilled in the art will understand that the invention can be carried out without these specific details. In other cases, well-known methods, procedures, and components are not described in detail so as not to obscure the invention.

[0046] In one aspect of the invention, formula A': [ka] A conjugate is provided which contains a polymer covalently bonded to the portion represented by the formula, where a and b each independently represent integers between 1 and 10; A represents a sterol; R Tis an optionally substituted alkyl or optionally substituted heteroalkyl; L represents a linker containing at least one -N(R5)- group, the linker having a bond length of 1 to 30 single CCs; each R independently represents a substituent or H; the wavy bond represents an attachment site to the polymer; the polymer is a polyamino acid (e.g., cyclic or linear peptides, proteins, or any copolymer thereof), polyethyleneimine (PEI), or a polymer comprising amino acids, further comprising thio groups, amino groups, 1,3-nitrones, azides, dienes, tetrazines, maleimides, active esters, α,β-unsaturated keto acid derivatives, or any combination thereof, or comprising the same;Each R represents hydrogen or one or more substituents, each independently being halo, -NO2, -CN, -OR', -OH, -CONH2, HCONH-, oxo, carbonyl, amino, imino, thioxo, phosphate (i.e., -OPO(OR')2), phosphonate, phosphine, phosphine, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, -NHCOR', -NHCSR' -NHCNR', -NC(=O)OR', -NC(=O)NR', ​​-NC(=S)OR', -NC(=S)NR', ​​-SO2R', -SOR', -SR', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', C1-C6 haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C 6-haloalkoxy, hydroxy(C1-C6 alkyl), hydroxy(C1-C6 alkoxy), alkoxy(C1-C6 alkyl), alkoxy(C1-C6 alkoxy), C1-C6 alkyl-NR'2, C1-C6 alkyl-SR', -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -COR', -CO2R', -OCOR', -OCOR', -OC(=O)OR', -OC(= Selected from O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​amino(C1-C6 alkyl), C1-C6 mercaptoalkyl, -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -CO2R, -OCOR, -OC(=O)OR, -OC(=O)NR, -OC(=S)OR, -OC(=S)NR, alkyl-aryl, alkyl-heteroaryl, or combinations thereof;Each R’ independently represents hydrogen or is selected from the group consisting of optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or combinations thereof. In some embodiments, the sterol is selected from phytosterol, zymosterol, ergosterol, and steroids. In some embodiments, the polymer is attached to a plurality of moieties (e.g., 2, 3, 4, or 5 moieties).;

[0047] In another aspect of the invention, formula B’:

Chemical formula

[0048] In some embodiments, the moiety of the invention is formula A’’:

Chemical formula

Chemical formula

[0049] In some embodiments, part of the invention is formula A'1: [ka] It is represented by the formula, where each X independently represents H or one or more alkyl groups (e.g., one or more methyl groups). In some embodiments, the sterol is cholesterol.

[0050] In some embodiments, part of the invention is formula 1A: [ka] By, or formula 1: [ka] It is represented by [this].

[0051] In some embodiments, a portion of the invention is represented by Formula 1 or any of the formulas disclosed herein, and includes any pharmaceutically acceptable salt, any hydrate, or any solvate thereof. In some embodiments, the polymer of the invention includes any stereoisomer, any salt, or any hydrate thereof.

[0052] In some embodiments, R is H, or represents one or more substituents, each independently being halo, -NO2, -CN, -OR', -OH, -CONH2, HCONH-, oxo, carbonyl, amino, imino, thioxo, phosphate (i.e., -OPO(OR')2), phosphonate, phosphine, phosphite, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, -NHCOR', -NHCSR', -NHCNR', -NC(=O)OR', -NC(= O)NR', ​​-NC(=S)OR', -NC(=S)NR', ​​-SO2R', -SOR', -SR', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', C1-C6 haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(C1-C6 alkyl), hydroxy(C1-C6 alkoxy), alkoxy(C1-C6 alkyl), Alkoxy(C1-C6 alkoxy), C1-C6 alkyl-NR'2, C1-C6 alkyl-SR', -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -COR', -CO2R', -OCOR', -OCOR', -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​Amino(C1-C6 alkyl), C1-C6 mercaptoalkyl, -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H -CO2R, -OCOR, -OC(=O)OR, -OC(=O)NR, -OC(=S)OR, -OC(=S)NR, alkyl-aryl, alkyl-heteroaryl, or combinations thereof; each R' independently represents hydrogen, or is selected from the group including optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or combinations thereof. In some embodiments, the conjugate comprises multiple parts of the invention (e.g., two, three, or four parts) covalently bonded to a single polymer molecule.

[0053] The term "polymer comprising amino acids, and further comprising thio groups, amino groups, 1,3-nitrones, azides, dienes, tetrazines, maleimides, active esters, and / or α,β-unsaturated keto acid derivatives" refers to the polymer in its non-conjugated form (i.e., the original form before being conjugated to the portion of the invention), as disclosed herein. Those skilled in the art will recognize that in the conjugated form (i.e., within the conjugate of the invention), the thio groups, amino groups, α,β-unsaturated keto acid derivatives, etc., of the polymer form covalent bonds with the portion. Thus, the above groups of the polymer in the conjugated form undergo chemical modification, for example: the amino group reacts with the active ester to form an amide bond, and the α,β-unsaturated keto acid derivative undergoes Michael addition to form compounds of formula A or formula B, as disclosed below.

[0054] In some embodiments, the non-conjugated polymer comprises a functional group selected from thio groups, amino groups, 1,3-nitrones, azides, dienes, tetrazines, maleimides, active esters, and / or α,β-unsaturated keto acid derivatives, or any combination thereof. In some embodiments, the conjugated polymer is bonded to L via a functional group (i.e., a chemically modified functional group as disclosed above). In some embodiments, the wavy bond of the conjugate of the invention represents the bond between the functional group of the polymer (i.e., a chemically modified functional group) and the L of the portion.

[0055] In some embodiments, the α,β-unsaturated keto acid derivative is given by formula A: [ka] , Therefore, equation B: [ka] Represented by or by both formulas A and B, where X is NH, S or O; R'' represents hydrogen, or optionally substituted C1-C 10 Alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C3-C 10 R'' is selected from the group including heterocyclyls, optionally substituted heteroaryls, optionally substituted aryls, or combinations thereof; and dashed bonds represent attachment sites to the polymer (e.g., peptides), wavy bonds represent attachment sites to the linker L of the part of the invention (or vice versa), or any combination thereof. In some embodiments, R'' is a substituted aryl, an aryl. In some embodiments, R'' is benzyl.

[0056] In some embodiments, the conjugate of the invention comprises a polymer conjugated to at least one part, or multiple parts selected from two parts, 1 to 4, or 1 to 3 parts of the invention (as defined by Formula 1, or any subsequent formula disclosed herein), including any range in between. In some embodiments, the polymer is a peptide, and the multiple parts are conjugated to the peptide at the same location within the peptide sequence, or at different locations (e.g., via any of the N-terminus, C-terminus, and / or side-chain residues within the peptide sequence).

[0057] In some embodiments, the polymer is covalently bonded to the linker via bonds selected from amide bonds, ester bonds, SS bonds, and click reaction products (including any combination thereof). In some embodiments, the linker is optionally substituted C1-C10 alkyl, heteroatom (e.g., O, N, NH, or S), -C(O)NH-, -C(O)O-, -C(O)-, -C(O)S-, -C(NH)NH-, -C(NH)O-, -C(NH)S-, C1-C10 aminoalkyl, C1-C10 alkoxy, C1-C10 mercaptoalkyl, -SS-, -SC(=O), Y-(C0-10)alkyl-X-(C0-10)alkyl-Y , or click reaction products (including any combination thereof); each X and Y is absent or independently selected from heteroatoms, oligomers, click reaction products, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-. In some embodiments, the oligomer contains 2 to 15 repeating units. In some embodiments, the repeating units are selected from alkylene oxides, natural or unnatural amino acid residues (i.e., amino acids lacking a hydrogen atom in the amino group and a hydroxyl moiety in the carboxyl group; exemplary amino acid residues are glycine, alanine, and / or β-alanine residues), α-hydroxycarboxylic acid residues (i.e., α-hydroxycarboxylic acids lacking a hydrogen atom in the α-hydroxy group and a hydroxyl moiety in the carboxyl group; e.g., lactate residues and / or glycolic acid residues) (including any copolymer) and any combination thereof. In some embodiments, the linker lacks disulfide bonds.

[0058] In some embodiments, the repeating units of the oligomer include amines. In some embodiments, the repeating units of the oligomer include amines selected from the following (including any copolymers and any combination thereof): [ka] (wherein each n is an independent integer between 1 and 10); or [ka] (In the formula, Ch is H, or an attachment site to an ethyleneamine or polyethyleneimine chain).

[0059] In some embodiments, the linker has single CC bond lengths of 1-30, 1-20, 1-10, 1-5, 5-10, 5-15, and 5-25 (including any range in between).

[0060] In some embodiments, the linker is characterized by an average molecular weight of 30-10,000 Da, 30-5,000 Da, 30-1,000 Da, 100-1,500 Da, 300-1,000 Da, 300-5,000 Da, 300-2,000, 300-3,000 Da, or 300-10,000 Da (including any range in between). In some embodiments, the linker is characterized by an average molecular weight of 300-3,000 Da, with CC bond lengths of 2-30, 2-10, 2-20 (including any range in between).

[0061] In some embodiments, the oligomer comprises repeating units of 2-15, 2-5, 2-10, 2-7, 5-15, and 3-8 (including any range in between), where the repeating units are as described above. In some embodiments, the oligomer comprises or is essentially composed of repeating units having the same chemical composition. In some embodiments, the oligomer comprises or is essentially composed of chemically different repeating units (e.g., in the form of copolymers, random copolymers, block copolymers, etc.).

[0062] In some embodiments, the oligomers are characterized by an average molecular weight of 30-1000Da, 30-100Da, 30-200Da, 30-300Da, 30-400Da, 30-500Da, 30-600Da, 30-700Da, 30-800Da, 30-900Da, 100-300Da, 50-350Da, 100-500Da, 150-500Da, or 100-900Da (including any range in between).

[0063] In some embodiments, the oligomer is or comprises a plurality of ethylene oxide repeating units, i.e., polyethylene glycol (PEG).

[0064] In some embodiments, the term "click reaction" indicates a highly efficient and selective reaction with a reaction yield of nearly 100% and minimal byproduct formation. A click reaction forms a covalent bond (conjugation) between two reactive groups attached to the same or different molecules. The reactive groups exhibit high reactivity towards each other and very low reactivity towards other functional groups, resulting in a highly specific / selective reaction. In some embodiments, click reactions enable specific conjugation of two different molecules (e.g., two polymers or one polymer and a small molecule).

[0065] In some embodiments, click reactions are well known in the art and include, among others, Michael addition of maleimide to thiol (resulting in the formation of succinimide-thioether); Michael addition of α,β-unsaturated keto acid derivatives to thiol; azido-alkyne cycloaddition; Diels-Alder reactions (e.g., direct and / or inverse electron-required Diels-Alder); dibenzylcyclooctin 1,3-nitrone (or azide) cycloaddition; alkenetetrazole photoclick reactions, and the like. In some embodiments, the αβ-unsaturated keto acid derivatives are represented by formulas A and B, as disclosed above.

[0066] In some embodiments, the click reaction product includes succinimide-thioether, azidoalkyne cycloaddition products, direct and / or inverse electron-required Diels-Alder reaction products, dibenzylcyclooctin 1,3-nitrone cycloaddition products, or compounds of formula A and / or formula B.

[0067] In some embodiments, succinimide-thioether is [ka] In the formula, (i) the wavy bonds represent attachment points of the part of the invention to linker L, and the dashed bonds represent attachment points to the polymer (e.g., peptide); or (ii) the wavy bonds represent attachment points to the polymer (e.g., peptide), and the dashed bonds represent attachment points of the part of the invention to linker L.

[0068] In some embodiments, the click reaction product includes a portion formed via the click reaction, which is as described above. In some embodiments, the click reaction product includes a product formed by any of the following: Michael addition of maleimide to thiol (succinimide-thioether formed); azidoalkyne cycloaddition; Diels-Alder reaction (e.g., direct and / or inverse electron-required Diels-Alder); dibenzylcyclooctin 1,3-nitrone (or azide) cycloaddition; alkenetetrazole photoclick reaction, or any combination thereof.

[0069] In some embodiments, part is Equation 2: [ka] This is expressed by the formula, where W, R, R5, a and b are as disclosed above; m is an integer independently selected from 1 to 5; each X1 is independently O, S, NR5, CRR or CH2; R T1c may be an optionally substituted linear or branched alkyl, or a linear or branched haloalkyl; c is an integer independently selected from 0 to 10; L1 is a bond, or -NC(=O)-, -C(=O)N-, -SS-, -SC(=O), Y-(C 0-10 )alkyl-X-(C 0-10 The linker is selected from alkyl-Y, cycloalkyl, heterocyclyl, and click reaction products, or any combination thereof.

[0070] In some embodiments, part is formula 2': [ka] This is expressed by the formula, where R, R5, a, b, and m are as disclosed above; c is an integer independently selected from 0 to 10; and L1 is a combination of -NC(=O)-, -C(=O)N-, -SS-, -SC(=O), and Y-(C 0-10 )alkyl-X-(C 0-10 ) is a linker selected from alkyl-Y (including any combination thereof); W is absent or represents (i) an amide, (ii) a click reaction product, or both; wavy bonds represent attachment sites to the polymer; the polymer is selected from polymers containing peptides and thio groups, amino groups, 1,3-nitrones, azides, dienes, tetrazines, or any combination thereof.

[0071] In some embodiments, R represents at least two substituents, each independently selected from F, Cl, Br, and I. In some embodiments, R represents two, three, or four substituents, each independently selected from F, Cl, Br, and I. In some embodiments, R represents two, three, or four fluoro substituents. In some embodiments, at least one R is F.

[0072] In some embodiments, R5 is H or a linear or branched C1-C5 alkyl group. In some embodiments, the C1-C5 alkyl group contains 1, 2, 3, 4, or 5 carbon atoms. In some embodiments, R5 is methyl.

[0073] In some embodiments, part is formula 2A: [ka] Represented by the formula, where R, R5, L1, W, a, b, and c are as disclosed above; d and e are each an integer independently selected from 1 to 10. In some embodiments, W is an amide, succinimide-thioether, azidoalkyne cycloaddition product, direct and / or inverse electron-required Diels-Alder reaction product, dibenzylcyclooctin 1,3-nitrone cycloaddition product, or a compound of formula A and / or formula B, or comprises thereof.

[0074] In some embodiments, part is formula 2B: [ka] It is represented by [this].

[0075] In some embodiments, part is formula 2C: [ka] By and / or, formula 2C1: [ka] It is expressed as follows, where R' is as disclosed above. In some embodiments, R' is benzyl.

[0076] In some embodiments, part is formula 2D: [ka] It is represented by [this].

[0077] In some embodiments, L1 is -NC(=O), -C(=O)N, or Y-(C 0-10 )alkyl-X-(C 0-10 ) It is alkyl-Y.

[0078] In some embodiments, X is [ka] The formula is as disclosed above, where R' is an integer independently selected from 1 to 15; and the wavy bond represents an attachment point of the polymer (e.g., a peptide) or part of the invention to the linker. In some embodiments, R' is hydrogen.

[0079] In some embodiments, n is 1 to 15, 1 to 3, 1 to 5, 1 to 7, 1 to 10, 2 to 15 (including any range in between).

[0080] In some embodiments, part is Equation 3: [ka] (wherein R, R5, Wa, b, c are as disclosed above; d and e are each independent integers between 1 and 10, and optionally at least one X1 is -O-) by; or formula 3': [ka] (wherein R, R5, Wa, b, c, d, and e are as disclosed above) is expressed by:

[0081] In some embodiments, part is formula 3A: [ka] This is expressed as follows, where R, R5, W, b, c, d, and e are as disclosed above.

[0082] In some embodiments, part is Equation 4: [ka] Represented by the formula, where each Y1 and Y2 is either absent or independently selected from cycloalkyl, heterocyclyl, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-.

[0083] In some embodiments, part is formula 4': [ka] Represented by the formula, where R, R5, W, a, b, c, d, e, and n are as disclosed above; and each Y1 and Y2 is independently selected from -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, -OC(=S)N-, cycloalkyl, heterocyclyl, where R' is as disclosed above. In some embodiments, Y1 is -C(=O)NH-. In some embodiments, Y1 and Y2 are -C(=O)NH-. In some embodiments, Y2 is -C(=O)O. In some embodiments, Y2 is absent. In some embodiments, the part is represented by formula 4 or 4', where b is 1 and a is 3.

[0084] In some embodiments, part is formula 9: [ka] Represented by the formula, where each X2 is independently N or CR; each R is independently hydrogen or one or more substituents, each independently halo, -NO2, -CN, -OR', -OH, -CONH2, HCONH-, oxo, carbonyl, amino, imino, thioxo, phosphate (i.e., -OPO(OR')2), phosphonate, phosphine, phosphine, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, -NHCOR', -NHCSR', -NHCNR' -NC(=O)OR', -NC(=O)NR', ​​-NC(=S)OR', -NC(=S)NR', ​​-SO2R', -SOR', -SR', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', C1-C6 haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(C1-C6 alkyl), hydroxy(C1-C6 alkoxy), alkoxy(C1- C6 alkyl), alkoxy (C1-C6 alkoxy), C1-C6 alkyl-NR'2, C1-C6 alkyl-SR', -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -COR', -CO2R', -OCOR', -OCOR', -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​amino(C1-C6 alkyl), C1-C6 mercaptoalkyl, -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -CO2R, -OCOR, -OC(=O)OR, -OC(=O)NR, -OC(=S)OR, -OC(=S)NR, alkyl-aryl, alkyl-heteroaryl, or combinations thereof; each R' independently represents hydrogen, or selected from the group including optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or combinations thereof; as well as L and R T This is as described above.

[0085] In some embodiments, part is formula 10: [ka] It is expressed by the formula, where X1, c, L1, m, R, b, L and R T1 This is as described above. In some embodiments, the part is represented by formula 10, where L1 is a bond; at least one X1 is CRR, each R independently is hydrogen, or one or more substituents, each independently being -NO2, -CN, -OR', -OH, -CONH2, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, -NC(=O)OR', -NC(=O )NR', ​​-NC(=S)OR', -NC(=S)NR', ​​-SO2R', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -CO2H, -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​or a combination thereof. In some embodiments, the part is represented by formula 10, where L1 is a bond; X1 is a CRR (e.g., methylene).

[0086] In some embodiments, part of formula 10A: [ka] It is expressed by the formula, where c, m, b, X1, L1, R and R T1 This is as described above; and X' represents O-Et.

[0087] In some embodiments, the part is represented by one of the above formulas, where R T1 teeth [ka] That is the case.

[0088] In some embodiments, part is formula 10B: [ka] This is expressed by the formula, where c, m, b, X1, X', L1, and R are as described above; and R T1 It is -C(CF3)3.

[0089] In some embodiments, a portion of the invention is derived from one of the precursors in Table A (i.e., chemically modified by conjugation with a polymer).

[0090] In some embodiments, the polymers disclosed herein include an amino group bonded thereto. In some embodiments, the polymer includes a side chain bonded to the polymer backbone, the side chain being any one of the following: a thio group, an amino group, a 1,3-nitro group, an azide group, a diene group, a tetrazine group, or any combination thereof. In some embodiments, the amino group is a side chain bonded to the polymer backbone. In some embodiments, the amino group is a primary amine. The term “primary amine” refers to an amine whose amino group is directly bonded to only one carbon. In some embodiments, the conjugate of the invention is a reaction product of an amino group or thio group of the polymer and a precursor Z, as disclosed below.

[0091] In some embodiments, the polymer is or contains polyethyleneimine. In some embodiments, the polymer contains at least one amino acid.

[0092] In some embodiments, the polymer is characterized by an average molecular weight of 1000-5000 Da, 1500-4500 Da, 2000-4000 Da, 2500-3500 Da, 1000-2000 Da, 1000-3000 Da, 1000-4000 Da, or 3000-5000 Da (including any range in between).

[0093] In some embodiments, the polymer is a peptide (also used herein as “polyamino acid”). In some embodiments, the polymer is a random polyamino acid. In some embodiments, the peptide comprises a therapeutically effective amino acid sequence, or further comprises a cell-permeable peptide essentially composed thereof and conjugated to a therapeutically effective amino acid sequence.

[0094] In this specification, the terms “polyamino acid,” “peptide,” “polypeptide,” and “protein” are used interchangeably and refer to polymers of amino acid residues. In some embodiments, the polymers have an amino acid sequence. In some embodiments, all polymer species of the composition / conjugate / precursor of the invention have the same amino acid sequence.

[0095] The terms “peptide,” “polypeptide,” and “protein” as used herein encompass natural peptides, peptide derivatives, e.g., β-peptides, peptide mimetic drugs (typically including non-peptide bonds or other synthetic modifications), and peptide analogs peptoids and semipeptoids or any combination thereof. In another embodiment, the terms “peptide,” “polypeptide,” and “protein” apply to amino acid polymers in which at least one amino acid residue is an artificial chemical analog of a corresponding naturally occurring amino acid.

[0096] The term “derivative” or “chemical derivative” includes any chemical derivative of a polypeptide having one or more residues that have been chemically derivatized by a reaction on a side chain or on any functional group within the peptide. Examples of such derivatized molecules include peptides having one or more protecting groups (e.g., side chain protecting groups and / or N-terminal protecting groups), and / or peptides in which a free amino group is derivatized to form an amine hydrochloride, a p-toluenesulfonyl group, a carbobenzoxy group, a t-butyloxycarbonyl group, an acetyl group, or a formyl group. Free carboxyl groups can be derivatized to form their amides, salts, methyl and ethyl esters or other types of esters or hydrazides. Free hydroxyl groups can be derivatized to form O-acyl or O-alkyl derivatives. Free thiol groups can be derivatized to form SS bonds, thioesters, or S-alkyl derivatives. The imidazole nitrogen of histidine can be derivatized to form N-im-benzylhistidine. These peptides are also included as chemical derivatives, which include one or more naturally occurring amino acid derivatives of 20 standard amino acid residues. For example: 4-hydroxyproline can replace proline; 5-hydroxylysine can replace lysine; 3-methylhistidine can replace histidine; homoserine can replace serine; and Dab, Daa, and / or ornithine(O) can replace lysine.

[0097] In addition, peptide derivatives may differ from the natural sequence of the peptide of the invention by chemical modification, such as terminal-NH2 acylation, acetylation, or thioglycolic acid amidation, and amidation of terminal and / or side-chain carboxyl groups by, for example, ammonia, methylamine, etc. The peptide can be linear, cyclic, or branched, and may have any higher-order structure, which can be achieved using methods known in the art.

[0098] The term "amino acid" as used herein means an organic compound containing both a basic amino group and an acidic carboxyl group. This term includes naturally occurring amino acids, protected amino acids (e.g., those containing one or more protecting groups in the carboxyl, amine, and / or side chains of the amino acid), unusual, unnaturally occurring amino acids (e.g., D-amino acids), and amino acids that are known to exist biologically in free or bound forms but are not typically present in proteins. This term also includes modified and unusual amino acids, for example, those disclosed in Roberts and Vellaccio (1983) The Peptides. 5:342-429.Modified, abnormal, or non-naturally occurring amino acids include, but are not limited to, D-amino acids, hydroxylysine, 4-hydroxyproline, N-Cbz-protected aminovaleric acid (Nva), ornithine (O), aminooctanoic acid (Aoc), 2,4-diaminobutyric acid (Abu), homoarginine, norleucine (Nle), N-methylaminobutyric acid (MeB), 2-naphthylalanine (2Np), aminoheptanoic acid (Ahp), phenylglycine, β-phenylproline, tert-leucine, 4-aminocyclohexylalanine (Cha), N-methyl-norleucine, 3,4-dehydroproline, N,N-dimethylaminoglycine, N-methylaminoglycine, 4-aminopipetdine-4-carboxylic acid, 6-aminocaproic acid, tran s-4-(aminomethyl)-cyclohexanecarboxylic acid, 2-,3-, and 4-(aminomethyl)-benzoic acid, 1-aminocyclopentanecarboxylic acid, 1-aminocyclopropanecarboxylic acid, cyanopropionic acid, 2-benzyl-5-aminopentanoic acid, norvaline (Nva), 4-O-methylthreonine (TMe), 5-O-methylhomoserine (hSM), tert-butylalanine (tBu), cyclopentylalanine (Cpa), 2-aminoisobutyric acid (Aib), N-methylglycine (MeG), N-methylalanine (MeA), N-methylphenylalanine (MeF), 2-thienylalanine (2Th), 3-thienylalanine (3Th), O-methyltyrosine (YMe), 3-benzothienylalanine (Bzt), and D-alanine (DAl).

[0099] In some embodiments, the peptide sequence is or includes a D-amino acid sequence. In some embodiments, at least 70%, at least 80%, at least 90%, and at least 95% of the amino acids in the peptide sequence are in the D-configuration. In some embodiments, the amino acids in the peptide sequence are in the D-configuration. In some embodiments, the peptide is linked to one or more parts of the invention. In some embodiments, the parts of the invention are linked covalently via: (i) the N-terminus of the peptide; (ii) the C-terminus of the peptide and / or (iii) a side chain in the peptide sequence (e.g., an amino or thio group of an amino acid residue). In some embodiments, the N-terminus of the peptide is linked to W. In some embodiments, the C-terminus of the peptide is linked to W. In some embodiments, the side chains of amino acids in the peptide sequence are linked to W.

[0100] In some embodiments, the peptide sequence is a therapeutic sequence. Hereinafter, the term “therapeutic sequence” refers to a polyamino acid sequence configured to induce a therapeutic effect in a subject (e.g., to treat, prevent, or reduce the symptoms of a disease). Furthermore, the term “therapeutic sequence” encompasses any polyamino acid sequence that can modify the activity, functionality, survival, fitness, appearance, structure, development, behavior, or any combination thereof of a cell. In some embodiments, the therapeutic sequence can bind to an intracellular target and control (upward or downward) the activity of the intracellular target. In some embodiments, the intracellular target is selected from intracellular proteins, enzymes, receptors, or any combination thereof. In some embodiments, the therapeutic sequence is endogenous or exogenous. Hereinafter, the term “endogenous” refers to the fact that the compound is produced in or naturally within the cell to which it is contacted.

[0101] In some embodiments, the therapeutic sequence is an apoptosis-inducing factor. The term “apoptosis-inducing factor” encompasses any polyamino acid sequence that can induce or promote programmed cell death.

[0102] In some embodiments, the conjugate further comprises (i) a cell-permeable peptide (CPP) covalently bonded to the portion of the invention via a linker disclosed herein, or (ii) a cell-permeable peptide covalently bonded to the peptide (i.e., not necessarily bonded to the portion of the invention). In some embodiments, the CPP is covalently bonded to the portion via a reactive group of the linker, e.g., OH, carboxy, amino, thio, sulfo, etc. In some embodiments, the portion is represented by formula 2, where X1 is a CRR, and one of the Rs is a reactive group bonded to or reactive with respect to the CPP (e.g., to an amino, carboxy, or thio group of the CPP).

[0103] In some embodiments, the CPP is covalently bonded to the polymer (peptide) and / or to the linker of the invention. In some embodiments, the CPP is bonded to the linker of the invention and to the peptide as shown below: [ka] In the formula, the wavy bond represents attachment to a part of the invention, where W is as described herein, and “peptide” is a peptide sequence (also referred to herein as “cargo”). In some embodiments, the CPP is bonded to the N-terminus of the peptide. In some embodiments, the CPP is bonded to the C-terminus of the peptide. In some embodiments, the CPP is bonded to the side chain of an amino acid in the peptide sequence, or to the backbone of the peptide.

[0104] In some embodiments, the conjugate of the invention comprises a cargo covalently bonded to a carrier, the carrier being a part of the invention or comprising it. In some embodiments, the cargo comprises a peptide or a polymer disclosed herein that is delivered into the cells of a subject. In some embodiments, the cargo is a peptide or polymer disclosed herein or comprising it, and the cargo is a therapeutic cargo. The term “therapeutic cargo” refers to any peptide having a therapeutically effective amino acid sequence configured to induce a therapeutic effect in a subject (e.g., to treat, prevent, or reduce the symptoms of a disease). Furthermore, the term “therapeutic cargo” encompasses any polyamino acid sequence or polymer sequence (e.g., oligonucleotide sequence) that can modify the activity, functionality, survival, fitness, appearance, structure, development, behavior, or any combination thereof of cells. In some embodiments, the therapeutic cargo can bind to an intracellular target and control (upward or downward) the activity of the intracellular target. In some embodiments, the conjugate further comprises a CPP that is (i) covalently bonded to the part of the invention via a linker disclosed herein, or (ii) covalently bonded to the polymer or peptide of the invention (i.e., not necessarily bonded to the part of the invention). In some embodiments, the carrier comprises the part of the invention and the CPP. As shown in the Examples section below, the inventors were surprised to observe that a conjugate containing a combination of the CPP and the part of the invention as a carrier has better cellular internalization than a similar conjugate containing the part of the invention as a carrier alone (without the CPP).

[0105] The term "cell-permeable peptide (CPP)" refers to a peptide covalently bound to a cargo molecule that can enhance or induce the permeability of the cargo molecule (i.e., the peptide) into the cell. CPPs can have amino acid residue lengths of 4–30, 4–50, 8–20, 8–25, or 8–30 (including any range in between). In some embodiments, CPPs are linear or cyclic peptides. CPPs typically have multiple positive charges at neutral pH and / or contain specific sequences configured to penetrate the cell membrane (usually by endocytosis) and move the cargo into the cell.

[0106] The term "CPP" encompasses positively charged CPPs, negatively charged CPPs, and amphiphilic CPPs.

[0107] Positively charged CPPs contain multiple (at least three) positively charged amino acid residues (i.e., Lys, Arg, and / or His). Positively charged CPPs are configured, among other things, to electrostatically bind to negatively charged cell membranes under physiological conditions (e.g., pH values ​​of approximately 6.5–7.5). Non-limiting examples of positively charged CPPs include peptides based on multiple lysines and / or arginines, e.g., R4-9, K4-9, Xentry, and TAT.

[0108] Amphiphilic CPPs typically contain both positively charged amino acid residues and hydrophobic amino acid residues (e.g., Trp, Phe, Ile, Tyr, Phe, and Met). The positively charged and hydrophobic amino acid residues may be separated into two distinct blocks, or they may be mixed within the CPP sequence.

[0109] Non-restrictive examples of amphipathic CPPs include penetratin, transportan, pVEC, MAP, Pep-1, MPG, and VT5.

[0110] Non-limiting examples of negatively charged CPPs (i.e., negatively charged at a neutral pH of approximately 6.5–7.5 or 6–8) include (i) sweet arrow peptide-SAP(E) having a high proline content and configured to adopt a helical secondary structure, and (ii) azurin-derived p28.

[0111] In some embodiments, the portion of the invention (defined by Formula 1, or any subsequent formula disclosed herein) is a carrier configured to enhance or induce cellular internalization of a polymer, the polymer being as described herein. In some embodiments, the portion of the invention is a peptide carrier. In some embodiments, the carrier comprises the portion of the invention and a CPP. In some embodiments, the CPP is covalently bonded to the portion (e.g., via L). In some embodiments, the CPP and the portion are bonded to the polymer. In some embodiments, the CPP and the portion are independently bonded to the polymer at different locations. In some embodiments, the carrier comprises a CPP and a portion covalently bonded to each other (e.g., via L), and the carrier is covalently bonded to the polymer via the CPP (e.g., at the N-terminus of the CPP, at the C-terminus of the CPP, or via the side chains of amino acid residues located within the sequence of the CPP).

[0112] In some embodiments, the polymer of the invention is a peptide. In some embodiments, the polymer of the invention is a therapeutic peptide sequence, or comprises a therapeutic peptide sequence. In some embodiments, the peptide is an amino acid sequence selected from the sequences disclosed in Table 1, or comprises a therapeutic peptide sequence. Sequence ID No. 1 shows an exemplary non-limiting therapeutic peptide sequence (JNK) used in Example 7. Sequence ID No. 2 shows a JNK conjugated to an exemplary CPP (Tat, Sequence ID No. 4). [Table 1]

[0113] precursor In another embodiment, a compound is provided comprising any pharmaceutically acceptable salt, any hydrate, or any solvate thereof; the compound has the formula: [ka] This is expressed by the formula, where A represents a sterol; R TR is optionally substituted alkyl or optionally substituted heteroalkyl; each R is independently hydrogen or one or more substituents, each independently halo, -NO2, -CN, -OR', -OH, -CONH2, HCONH-, oxo, carbonyl, amino, imino, thioxo, phosphate (i.e., -OPO(OR')2), phosphonate, phosphine, phosphine, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, -NHCOR', -NHCSR', -NHCNR', -NC (=O)OR', -NC(=O)NR', ​​-NC(=S)OR', -NC(=S)NR', ​​-SO2R', -SOR', -SR', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', C1-C6 haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(C1-C6 alkyl), hydroxy(C1-C6 alkoxy), alkoxy(C1-C6 alkyl), alkoxy(C 1-C6 alkoxy), C1-C6 alkyl-NR'2, C1-C6 alkyl-SR', -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -COR', -CO2R', -OCOR', -OCOR', -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​amino(C1-C6 alkyl), C1-C6 mercaptoalkyl, -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -CO2R, -OCOR, -OC(=O)OR, -OC( -O)NR, -OC(=S)OR, -OC(=S)NR, alkyl-aryl, alkyl-heteroaryl, or combinations thereof; each R' independently represents hydrogen, or selected from the group including optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or combinations thereof; M represents a linker containing at least one -N(R5)- group; the linker has a single CC bond length of 1 to 30;a, b, and R5 are as disclosed above; and Z is reactive to functional groups selected from thio groups, amino groups, 1,3-nitrones, azides, dienes, succinimides, α-β unsaturated carbonyls, α-β unsaturated keto acid derivatives, carbonyls, carboxylates, and active esters. In some embodiments, Z is reactive to small molecules or to polymers containing functional groups. In some embodiments, Z is reactive to the functional groups of the polymer of the invention.

[0114] In some embodiments, the compound is represented by the following formula: [ka] In the formula, b represents an integer between 1 and 10; A represents a sterol; B represents at least one of optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; R T is an optionally substituted alkyl or optionally substituted heteroalkyl; and M and Z are as disclosed herein.

[0115] In some embodiments, the compound is formulated with the formula: [ka] By, or by formula: [ka] This is expressed as follows, where R, B, b, Rt, M, and Z are as disclosed above.

[0116] In some embodiments, the compounds disclosed herein are precursors to the conjugates of the invention. In some embodiments, the compounds disclosed herein are configured to react with the polymers disclosed herein to obtain the conjugates of the invention.

[0117] In some embodiments, the precursor is formula 5A: [ka] By, or formula 5: [ka] It is represented by [this].

[0118] In some embodiments, the precursor is formula 11: [ka] This is expressed as follows, where each X² is independently N or CR; and R, Z, M, and Rt are as described herein.

[0119] In some embodiments, the precursor is formula 12: [ka] Represented by the formula, where m is an integer independently selected from 1 to 5; each X1 is independently O, S, NR5, CRR, or CH2; RT1 includes optionally substituted linear or branched alkyl; c is an integer independently selected from 0 to 10; L1 is a bond, or -NC(=O)-, -C(=O)N-, -SS-, -SC(=O), Y-(C0-10)alkyl-X-(C0-10)alkyl-Y, cycloalkyl, heterocyclyl, and click reaction products, or The linker is selected from any combination of these; each X and Y is either absent or independently selected from heteroatoms, oligomers, click reaction products, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-; the oligomers contain 2 to 15 repeating units.

[0120] In some embodiments, Z is or comprises active esters (e.g., thio-esters, pentofluorophenyl esters, N-hydroxysuccinimide esters, hydroxybenzotriazole esters, etc.); α,β-unsaturated carbonyls; α,β-unsaturated keto acid derivatives; maleimides; 1,3-nitrones; thiols; amines; azides; alkynes; dienes; alkenes; tetrazoles; or any combination thereof. In some embodiments, Z is or comprises acyl halides, chloroformates, anhydrides, aldehydes, epoxides, isocyanates, isothiocyanates, maleimides, carbonates (e.g., nitrophenyl carbonate), sulfonyl chlorides, iodoacetamides, acyl azides, imide esters, vinyl sulfones, ortho-pyridyl disulfides; or any combination thereof.

[0121] In some embodiments, Z is an active ester. The term "active ester" refers to an ester that has enhanced reactivity (faster kinetics) to nucleophilic attack compared to ordinary alkyl esters. Active esters react with nucleophiles at room temperature to obtain nearly quantitative amide bond formation. Active esters have an alcohol component that induces a greater electron-withdrawing effect compared to ordinary alkyl esters. Electron withdrawal enhances the electrophilicity of the carbonyl carbon, thereby promoting the formation of a tetrahedral intermediate with the nucleophile. Typically, the alcohol component of active esters is a better leaving group than the alcohol of ordinary alkyl esters. In this specification, the term "active ester" refers to a storage-stable compound.

[0122] The term "heteroalkyl" refers to an alkyl group (i.e., a linear, branched, or cyclic alkyl chain) that contains one or more heteroatoms (e.g., O, S, N, and / or NH) within the alkyl chain's backbone.

[0123] In some embodiments, Z is selected from active esters (e.g., NHS esters), maleimides, and α,β unsaturated keto acids (e.g., benzoyl acrylic anhydride). In some embodiments, the α,β unsaturated keto acid or its derivatives is represented by the following formula: [ka] In the formula, R'' represents hydrogen, or optionally a substituted C1-C 10 Alkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C3-C 10 Selected from the group including heterocyclyls, optionally substituted heteroaryls, optionally substituted aryls, or combinations thereof; and, wavy bonds represent attachment points to M.

[0124] In some embodiments, R'' is an aryl or substituted aryl. In some embodiments, R'' is benzyl.

[0125] In some embodiments, the compound is formulated with formula 6: [ka] This is expressed as follows, where R, R5, L1, a, b, c, and Z are as disclosed above.

[0126] In some embodiments, the compound is formula 6A: [ka] This is expressed as follows, where R, R5, L1, Z, a, b, c, d, and e are as disclosed above.

[0127] In some embodiments, L1 is -NC(=O), -C(=O)N, or Y-(C 0-10 )alkyl-X-(C 0-10 ) It is alkyl-Y.

[0128] In some embodiments, X is [ka] The formula is as disclosed above, where R' is an integer independently selected from 1 to 15; and the wavy bond represents an attachment point of the polymer (e.g., a peptide) or part of the invention to the linker. In some embodiments, R' is hydrogen.

[0129] In some embodiments, the compound is formulated with formula 7: [ka] This is expressed as follows, where R, R5, Z, a, b, c, d, and e are as disclosed above.

[0130] In some embodiments, the compound is of formula 7A: [ka] This is expressed as follows, where R, R5, Z, b, c, d, and e are as disclosed above.

[0131] In some embodiments, the compound is formula 10: [ka] This is expressed as follows, where R, R5, Z, Y1, Y2a, b, c, d, e, and n are as disclosed above.

[0132] In some embodiments, the compound (precursor) is one of the compounds listed in Table A or Table B.

[0133] [Table 2] JPEG2026513268000060.jpg177159

[0134] [Table 3]

[0135] The MNM type (as assigned in Table 2) is highlighted in bold.

[0136] In another embodiment, a kit is provided comprising the compound of the invention (i.e., a precursor), and a polymer selected from (i) polyamino acids; and (i) polymers comprising a thio group, an amino group, a 1,3-nitrone, an azide, a diene, a tetrazine, or any combination thereof.

[0137] In some embodiments, the polymer is present in a composition (e.g., a solid composition or a liquid composition, e.g., a solution, dispersion, or suspension). In some embodiments, the composition comprises a plurality of nanoparticles (lipid nanoparticles, liposomes, micelles, vesicles, nanocapsules, etc.), each of which comprises the polymer.

[0138] In some embodiments, the molar ratio between the polymer and the precursor in the kit is approximately 1:1, approximately 1.5:1, approximately 5:1, and approximately 5:1 to 1:1, approximately 4:1 to 1:1, approximately 3:1 to 1:1, approximately 2:1 to 1:1, and 1.2:1 to 1:1 (including any range in between).

[0139] In some embodiments, the kit further includes instructions for reacting the polymer and precursor under preferred conditions to obtain the conjugate of the invention. In some embodiments, preferred conditions include conditions suitable for reacting the polymer and the precursor of the invention, such as reaction time (e.g., 0.1 to 10 hours (including any range therein)) and temperature (e.g., 5 to 90°C (including any range therein)).

[0140] In some embodiments, the kit further comprises a solvent. In some embodiments, the solvent is suitable for dissolving the polymer and precursor. In some embodiments, the polymer and precursor have a solubility in the solvent of at least 0.5 g / L, at least 10 g / L, or 0.5 to 100 g / L (including any range in between). In some embodiments, the solvent is selected from aqueous buffer, water, or organic solvents, but is not limited.

[0141] Pharmaceutical composition In another embodiment, a pharmaceutical composition comprising a conjugate of the invention and a pharmaceutically acceptable carrier is provided. In some embodiments, the pharmaceutical composition comprises one or more conjugates of the invention and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of one or more conjugates of the invention.

[0142] In some embodiments, the pharmaceutical composition essentially comprises the conjugate of the invention and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition essentially comprises the conjugate of the invention as a therapeutically active material component. In some embodiments, the pharmaceutical composition lacks additional therapeutically active material components.

[0143] In this specification, the term “essentially composed of” means that a composition, method, or structure may include additional material components, steps, and / or parts, provided that the additional material components, steps, and / or parts do not substantially alter the basic and novel properties of the claimed composition, method, or structure.

[0144] In some embodiments, the pharmaceutical composition is intended for use in the treatment or prevention of a disease or disorder in a subject requiring such treatment. In some embodiments, the pharmaceutical composition is intended for use in the treatment of cancer.

[0145] In some embodiments, pharmaceutically acceptable carriers are also called excipients or adjuvants. Hereinafter, the terms “carrier,” “excipient,” or “adjuvant” refer to any component of a pharmaceutical composition that is not an activator. Hereinafter, the term “pharmaceutically acceptable carrier” refers to a non-toxic, inert solid, semi-solid liquid filler, diluent, mounting material, any type of formulation aid, or simply a sterile aqueous medium, such as physiological saline. Some examples of materials that can function as pharmaceutically acceptable carriers are listed below: sugars, e.g., lactose, glucose and sucrose; starches, e.g., corn starch and potato starch; cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; tragacanth powder; malt, gelatin, talc; excipients, e.g., cocoa butter and suppository wax; oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, e.g., propylene glycol; polyols, e.g., glycerin, sorbitol, mannitol and polyethylene glycol; esters, e.g., ethyl oleate and ethyl laurate, agar; buffers, e.g., magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline, Ringer's solution; ethyl alcohol and phosphate buffer solution; and other non-toxic and suitable substances used in pharmaceutical formulations. Some non-limiting examples of substances that can function as carriers in this specification include: sugars, starches, cellulose and its derivatives, tragacanth powder, malt, gelatin, talc, stearic acid, magnesium stearate, calcium sulfate, vegetable oils, polyols, alginic acid, pyrogen-free water, isotonic saline, phosphate buffer solution, cocoa butter (suppository base), emulsifiers, and other non-toxic, pharmaceutically compatible substances used in other pharmaceutical formulations. Wetting agents and lubricants, such as sodium lauryl sulfate, as well as colorants, flavoring agents, excipients, stabilizers, antioxidants, and preservatives may also be present. Any non-toxic, inert, and effective carrier may be used to formulate the compositions intended herein.Suitable pharmaceutically acceptable carriers, excipients, and diluents in this regard are well known to those skilled in the art, and are described, for example, in: The Merck Index, 13th edition, Budavari et al., Eds., Merck & Co., Inc., Rahway, NJ (2001); the CTFA (Cosmetic, Toiletry, and Fragrance Association) International Cosmetic Ingredient Dictionary and Handbook, 10th edition (2004); and the “Inactive Ingredient Guide,” U.S. Food and Drug Administration (FDA) Center for Drug Evaluation and Research (CDER) Administration, all of which are incorporated by reference as a whole. Examples of pharmaceutically acceptable excipients, carriers, and diluents useful in this composition include distilled water, physiological saline, Hartmann solution, Ringer's solution, dextrose solution, Hanks' solution, and DMSO. These additional inactive components, as well as effective formulations and administration procedures, are well known in the art and are described in standard textbooks, e.g., Goodman and Gillman's: The Pharmacological Bases of Therapeutics, 8th edition, Gilman et al. Eds. Pergamon Press (1990); Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Co., Easton, Pa. (1990); and Remington: The Science and Practice of Pharmacy, 21st edition, Lippincott Williams & Wilkins, Philadelphia, Pa. (2005), each of which is incorporated herein by reference in whole. The compositions described herein may also be contained in artificially created structures, e.g., liposomes, ISCOMS, sustained-release particles, and other vehicles that increase the half-life of peptides or polypeptides in serum.Liposomes for use with the peptides described herein are formed from standard vesicle-forming lipids, which generally include neutral and negatively charged phospholipids and sterols, such as cholesterol. The selection of lipids is generally determined by considerations such as liposome size and stability in blood. Various methods are available for liposome preparation, as outlined, for example, by Coligan, JE et al., Current Protocols in Protein Science, 1999, John Wiley & Sons, Inc., New York, and also see U.S. Patents 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

[0146] In some embodiments, the carrier may, in total, constitute about 0.1% to about 99.99% by weight of the pharmaceutical composition presented herein. In some embodiments, the carrier may, in total, constitute 0.1 to 99.99% by weight, 0.1 to 50% by weight, 0.2 to 30% by weight, 0.1 to 70% by weight, 30 to 99.99% by weight, 50 to 99.99% by weight (including any range in between) of the pharmaceutical composition presented herein.

[0147] In some embodiments, the pharmaceutical compositions disclosed herein include a conjugate of the therapeutically effective dose of the invention. The term "therapeutically effective dose" refers to the dose required in a subject to achieve the desired therapeutic or prophylactic outcome, and the amount effective for that period of time. The exact dosage form and regimen will be determined by the physician in accordance with the patient's condition.

[0148] In this specification, the term “pharmaceutically acceptable salt” refers to any non-toxic salt of the conjugate of the present invention that, when administered to a subject, e.g., a human, can directly or indirectly provide the compound of the present invention or its therapeutically active metabolites or residues. For example, the term “pharmaceutically acceptable” can mean that it is approved by a federal or state government regulatory authority or is listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeias for use in animals, more specifically in humans.

[0149] pharmaceutically acceptable salts are well known in the art. For example, SMBerge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19. pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases.

[0150] Non-limiting examples of pharmaceutically acceptable salts include, but are not limited to, alkali metal salts, alkaline earth metal salts, acetates, aspartates, benzenesulfons, benzoates, bicarbonates, carbonates, halides (e.g., bromides, chlorides, iodides, fluorides), hydrogen tartrates, citrates, salicylates, stearates, succinates, sulfates, tartrates, decanoates, edetates, fumarates, glucons, and lactates, or any combination thereof.

[0151] Examples of pharmaceutically acceptable, non-toxic acid addition salts include salts of amino groups formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by other methods used in the art, such as ion exchange.

[0152] Other pharmaceutically acceptable salts include: adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphosulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptone, glycerophosphate, glycolate, gluconate, glycolate, hemisulfate, heptaneate, hexanoate, hydrochloride, hydrobromide, hydroiodide, and 2-hydroxy - Ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc.

[0153] In some embodiments, the conjugate of the invention is present in a pharmaceutical composition having pharmaceutical-grade purity, i.e., characterized by a chemical purity of at least about 90%, at least about 95%, greater than 95%, or greater than 99%, 90-99.999%, 90-95%, 90-97%, 95-99%, (including any range in between), and the pharmaceutical composition is intended for use in the treatment of a disease or disorder in a subject requiring such treatment.

[0154] method In one embodiment, a method is provided for treating a disease or condition in a subject, comprising administering a therapeutically effective amount of the pharmaceutical composition of the invention to a subject, thereby treating the disease or condition.

[0155] In another embodiment, a method is provided for internalizing a polymer of the invention (e.g., a polynucleic acid or peptide disclosed herein) into cells, comprising contacting cells with a pharmaceutical composition of the invention or a conjugate of the invention. In some embodiments, the cells are isolated cells. In some embodiments, the cells are cultured cells. In some embodiments, the cells are tissue cells. In some embodiments, the cells are mammalian cells (e.g., human cells). In some embodiments, the cells comprise any cells of the subject. In some embodiments, the cells are attached to, dispersed in, or suspended on a support. In some embodiments, the cells are in contact with a cell culture medium.

[0156] In some embodiments, contact involves adding an effective amount of conjugate or pharmaceutical composition to cells, such that the effective amount induces the internalization of a therapeutic amount of polymer into the cytosol of the cells. In some embodiments, contact involves adding the conjugate or pharmaceutical composition to a cell medium that comes into contact with the cells. In some embodiments, contact involves administering an effective amount of conjugate or pharmaceutical composition to a subject (topically or systemically). Cellular internalization can be determined or quantified spectroscopically, for example, by labeling the conjugate with a fluorescent probe and measuring the intracellular fluorescence signal (e.g., by an ELISA reader, FACS, etc.).

[0157] In some embodiments, contact involves administering the pharmaceutical composition of the invention to a subject. In some embodiments, contact involves administering a therapeutically effective amount of the pharmaceutical composition to a subject.

[0158] In some embodiments, administration includes systemic administration. In some embodiments, administration includes local administration. In some embodiments, administration includes intravenous administration.

[0159] In some embodiments, the method is for reducing at least one symptom associated with a disease or condition in a subject. In some embodiments, the subject suffers from a cell proliferation disease. In some embodiments, the disease is a cell proliferation disease. In some embodiments, the cell proliferation disease is cancer.

[0160] In some embodiments, the disease is selected from the following: retinal excitotoxicity, neuropathic pain, intraocular inflammation, conditions associated with cerebral ischemia / intracerebral hemorrhage, hearing loss, and sudden sensorineural hearing loss caused by any one of the following: cochlear ischemia, semicircular canal damage, trauma during cochlear implant surgery, acute otitis interna, and aminoglycoside ototoxicity (including any combination thereof).

[0161] Definitions As used herein, the term "alkyl" also encompasses saturated or unsaturated hydrocarbons, and thus further encompasses alkenyl and alkynyl. The term "alkyl" encompasses straight-chain, branched, or cyclic hydrocarbon chains.

[0162] As used herein, the term "alkenyl" refers to an unsaturated alkyl as defined herein having at least two carbon atoms, from 2 to 30 carbon atoms, and at least one carbon-carbon double bond. The alkenyl may be substituted or unsubstituted by one or more substituents as described above.

[0163] As used herein, the term "alkynyl" refers to an unsaturated alkyl having at least two carbon atoms, from 2 to 30 carbon atoms, and at least one carbon-carbon triple bond as defined herein. The alkynyl may be substituted or unsubstituted by one or more substituents as described above.

[0164] As used herein, the term "C1-C6 alkyl" including any C1-C6 alkyl-related compound refers to any straight-chain or branched alkyl chain containing 1 to 6, 1 to 2, 2 to 3, 3 to 4, 4 to 5, 5 to 6 carbon atoms (including any range therebetween). In some embodiments, C1-C6 alkyl includes any one of methyl, ethyl, propyl, butyl, pentyl, iso-pentyl, hexyl, and tert-butyl, or any combination thereof. In some embodiments, the C1-C6 alkyl described herein further includes an unsaturated bond, and the unsaturated bond is located at the 1st, 2nd, 3rd, 4th, 5th, or 6th position of the C1-C6 alkyl.

[0165] The term "C1-C6 haloalkyl" refers to a C1-C6 alkyl as described herein substituted by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 halide atoms, and the halide is selected from F, Br, Cl, and I, or a combination thereof.

[0166] As used herein, the terms "halogen", "halo" and "halide" are referred to in the same sense herein, representing an atom of a halogen, which is fluorine, chlorine, bromine, or iodine, and is also referred to as fluoride, chloride, bromide, and iodide herein.

[0167] The term "(C3-C10) cycloalkyl" refers to an optionally substituted C3, C4, C5, C6, C7, C8, C9 or C10 ring. In some embodiments, the (C3-C10) ring includes an optionally substituted cyclopropane, cyclobutene, cyclopentane, cyclohexane, or cycloheptane.

[0168] In some embodiments, the terms “hydroxy(C1-C6 alkyl)” and “C1-C6 alkoxy” are used herein to mean the same thing and refer to the C1-C6 alkyls described herein that are substituted with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 hydroxyl groups, the hydroxyl groups being located at the 1, 2, 3, 4, 5, or 6 positions of the C1-C6 alkyl (including any combination thereof).

[0169] The term "aryl" refers to an all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group having a fully conjugated π-electron system. The aryl group may be substituted or unsubstituted, as shown herein.

[0170] The term "alkoxy" refers to both O-alkyl and -O-cycloalkyl groups as defined herein.

[0171] The term "aryloxy" represents -O-aryl as defined herein.

[0172] Each of the alkyl, cycloalkyl, and aryl groups in the general formulas herein may be substituted with one or more substituents, each substituent independently of the substituted group and its position within the molecule, for example, a halide, alkyl, alkoxy, cycloalkyl, nitro, amino, hydroxyl, thiol, thioalkoxy, carboxy, amide, aryl, and aryloxy. Additional substituents are also conceivable.

[0173] The term "hydroxyl" or "hydroxy" refers to the -OH group.

[0174] The terms "mercapto" or "thiol" represent the -SH group.

[0175] The term "thioalkoxy" refers to both -S-alkyl groups and -S-cycloalkyl groups, as defined herein.

[0176] The term "thioaryloxy" refers to both -S-aryl and -S-heteroaryl groups, as defined herein.

[0177] The term "amino" represents the -NR'R'' group, where R' and R'' are as described herein.

[0178] The term "C3-C10 heterocyclyl" refers to an optionally substituted C3, C4, C5, C6, C7, C8, C9, or C10 heterocyclic aromatic and / or aliphatic or unsaturated ring, representing a monocyclic or fused ring group having one or more atoms such as nitrogen, oxygen, and sulfur within the ring(s). The ring may also have one or more double bonds. However, the ring does not have a fully conjugated π-electron system. Representative examples include piperidine, piperazine, tetrahydrofuran, tetrahydropyran, and morpholino.

[0179] The terms "carboxy" or "carboxylate" represent the -C(O)OR' group, where R' is hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl (bonded via a ring carbon), or heterocyclyl (bonded via a ring carbon), as defined herein.

[0180] The term "carbonyl" represents the -C(O)R' group, where R' is defined as described above.

[0181] The above terms also include their thio derivatives (thiocarboxy and thiocarbonyl).

[0182] The term "thiocarbonyl" represents the -C(S)R' group, where R' is defined as described above.

[0183] The "thiocarboxy" group represents a -C(S)OR' group, where R' is as defined herein.

[0184] The "sulfinyl" group represents a -S(O)R' group, where R' is as defined herein.

[0185] The "sulfonyl" or "sulfonate" group represents a -S(O)2R' group, where R' is as defined herein.

[0186] The "carbamyl" or "carbamate" group represents an -OC(O)NR'R'' group, where R' is as defined herein, and R'' is as defined for R'.

[0187] The "nitro" group denotes a -NO2 group.

[0188] As used herein, the term "amide" encompasses C-amides and N-amides.

[0189] As used herein, the term "C-amide" represents a -C(O)NR'R'' terminal group or a -C(O)NR'-linking group, where R' and R'' are as defined herein.

[0190] As used herein, the term "N-amide" represents a -NR''C(O)R' terminal group or a -NR'C(O)-linking group, where R' and R'' are as defined herein.

[0191] As used herein, the term "carboxylic acid derivative" encompasses carboxy, amide, carbonyl, anhydride, carbonate ester, and carbamate.

[0192] The "cyano" or "nitrile" group denotes a -CN group.

[0193] The terms "azo" or "diazo" represent an -N=NR' terminal group or -N=N- linking group, as defined above, where R' is as defined above.

[0194] The term "guanidine" represents the -R'NC(N)NR"R"' terminal group or the -R'NC(N)NR"- linking group, as defined herein, where R', R" and R'" are as defined herein.

[0195] In this specification, the term "azide" refers to the -N3 group.

[0196] The term "sulfonamide" refers to the -S(O)2NR'R'' group, where R' and R'' are as defined herein.

[0197] The terms "phosphonyl" or "phosphonate" represent the -OP(O)-(OR')2 group, where R' is as defined above.

[0198] The term "phosphenyl" represents the -PR'R'' group, where R' and R'' are as defined above.

[0199] The term "alkylaryl" refers to an alkyl group as defined herein, substituted with an aryl group as described herein. An exemplary alkylaryl is benzyl.

[0200] The term "heteroaryl" refers to a monocyclic (e.g., C5-C6 heteroaryl ring) or fused (i.e., a ring sharing adjacent pairs of atoms) group having one or more atoms in the ring(s), such as nitrogen, oxygen, and sulfur, and also having a fully conjugated π-electron system. In some embodiments, the terms "heteroaryl" and "C5-C6 heteroaryl" are used interchangeably herein. Examples of heteroaryl groups, but not limited to, include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, and purine. Heteroaryl groups may be substituted or unsubstituted with one or more substituents, as described above. Representative examples include thiadiazole, pyridine, pyrrole, oxazole, indole, and purine.

[0201] In this specification, the terms “treatment” or “to treat” a disease, disorder, or condition encompass alleviation of at least one symptom, reduction of its severity, or inhibition of its progression. Treatment does not necessarily mean that the disease, disorder, or condition is completely cured. For a treatment to be effective, a useful composition as used herein only needs to reduce the severity of the disease, disorder, or condition, reduce the severity of its associated symptoms, or improve the quality of life of the patient or subject.

[0202] In this specification, the term “prevention” of a disease, disorder, or condition encompasses delaying, preventing, suppressing, or inhibiting the onset of the disease, disorder, or condition. As used in accordance with the subject matter described herein, the term “prevention” relates to a preventive process in which a subject is exposed to the active material components described herein before the induction or onset of a disease / disorder process. This is possible when an individual has a genetic lineage that shows a predisposition to the development of the disease / disorder to be prevented. For example, this is also true for an individual whose ancestors show a predisposition to a certain type of inflammatory disorder.

[0203] The term "suppression" is used to describe a state where a disease / disorder process has already begun, but the obvious symptoms of the condition are not yet recognized. Thus, an individual's cells may have the disease / disorder, but the external signs of the disease / disorder are not yet clinically recognized. In either case, the term "prevention" can be applied to encompass both prevention and suppression.

[0204] Conversely, the term "treatment" refers to the clinical application of an active agent to combat an existing condition in which the clinical symptoms are already recognized in the patient.

[0205] In the discussion, unless otherwise noted, adjectives such as “substantially” and “about” modifying the state or relational characteristics of one or more features of an embodiment of the invention are understood to mean that the state or characteristic is defined within the permissible limits for the behavior of the embodiment for the intended application. Unless otherwise noted, the word “or” in the specification and claims is considered to be an inclusive, not exclusive, “or” indicating at least one or any combination of the items it combines with.

[0206] The term “a, an” used above or anywhere else in this specification should be understood to mean “one or more” of the enumerated components. It will be obvious to those skilled in the art that the use of the singular form includes the plural form unless otherwise specified. Thus, the terms “a, an” and “at least one” are used interchangeably in this application.

[0207] To better understand this instruction, and without in any way limiting its scope, all numbers representing quantities, percentages, or proportions, and other numerical values ​​used in the specification and claims, should be understood in all cases to be modified by the term "approximately," unless otherwise noted. Therefore, unless otherwise indicated, the numerical parameters specified in the following specification and attached claims are approximations that may vary depending on the desired properties to be obtained. At a minimum, each numerical parameter should be interpreted by applying the usual rounding technique, taking into account at least the reported number of significant figures.

[0208] In the description and claims of this application, each of the verbs “comprise,” “include,” and “have,” and their conjugations, are used to indicate that the one or more objects of the verb are not necessarily a complete list of the components, elements, or parts of the one or more objects of the verb.

[0209] Other terms used herein are to be defined by their well-known meanings in the art.

[0210] Unless otherwise stated and evident from the context, the term “or” is understood to be inclusive in this specification.

[0211] Throughout this specification and claims, the word “comprise” or variations thereof, such as “comprises” or “comprising,” indicates the inclusion of any enumerated integer or set of integers, but not the exclusion of any other integer or set of integers.

[0212] In this specification, the term "consists essentially of" or variations thereof, as used throughout the specification and claims, such as "consist essentially of" or "consisting essentially of," indicate the inclusion of any enumerated integer or set of integers, and the optional inclusion of any enumerated integer or set of integers that does not substantially alter the basic or novel characteristics of the identified method, structure, or composition.

[0213] In this specification, terms such as “comprises,” “comprising,” “containing,” and “having” may mean “includes,” “including,” etc.; similarly, “consisting essentially of” or “consists essentially” have the meanings described in U.S. patent law, and the term is unrestricted, allowing for entities beyond those enumerated, as long as the basic or novel characteristics of the enumerated are not altered by entities beyond those enumerated, but prior art embodiments are excluded. In one embodiment, the terms “comprises,” “comprising,” and “having” are interchangeable with “consisting.”

[0214] While the invention has been described in conjunction with its specific embodiments, it is clear that many substitutions, modifications, and alterations will be apparent to those skilled in the art. Therefore, it is intended to encompass all such substitutions, modifications, and alterations that fall within the spirit and broad scope of the appended claims.

[0215] All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent as each individual publication, patent, and patent application is specifically and individually indicated as being incorporated herein by reference. In addition, any citation or reference of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. Section headings, to the extent that they are used, should not necessarily be construed as restrictive. [Examples]

[0216] In general, the nomenclature used herein and the laboratory procedures used in the present invention include biochemical, cell biological, and molecular biological techniques. Such techniques are fully described in the literature.

[0217] A non-limiting, exemplary synthesis procedure for the conjugate of the invention is provided below.

[0218] Example 1 Synthesis of Apo-Si-K-1011-2 Step 1: Synthesis of 1011-5 [ka]

[0219] Step 2: Synthesis of 170A-6a [ka]

[0220] Step 3: Synthesis of Apo-Si-K-1011-2 [ka]

[0221] Exemplary precursors that have been synthesized and successfully incorporated into exemplary conjugates are presented in Tables A and B above.

[0222] Example 2 Synthesis of Apo-Si-K-1012 and Apo-Si-K-1021 [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0223] The precursor of the invention is referred to herein as "MNM".

[0224] Example 3 N-terminal MNM-JNKI Step 1: Synthesis of Compound 2 [ka]

[0225] Step 2: Peptide synthesis Peptide synthesis was carried out using standard Fmoc chemistry on a solid support using Dde-side-chain protected Fmoc-lysine. After completing SPPS synthesis including N-terminal Fmoc deprotection, the Dde protecting group was cleaved with hydrazine and subsequently treated with Fmoc-Osu to obtain peptides with Fmoc-protected lysine side chains. After cleavage from the resin, the obtained peptides were purified by HPLC. Peptide sequence: NH2-ggdqsrpvqpflnlttprk(Fmoc)pr-C(=O)NH2(Sequence ID 1)

[0226] Step 3: Conjugate Synthesis [ka]

[0227] DIEA (2.98 μL, 17.1 μmol) was added to a solution of compound 2 (11.4 mg, 11.4 μmol) and peptide (11.4 μmol) in DMF (100 μL). The reaction mixture was stirred at 25°C for 0.5 hours. LC-MS showed that compound 2 was completely consumed and a single main peak with the desired mass was detected.

[0228] Next, 200 μL of TEA was added and the mixture was stirred for 16 hours. The reaction was monitored by LC-MS. The reaction mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by prep-HPLC (TFA conditions) to obtain N-terminal MNM-JNKI as a white solid.

[0229] Example 4 Lys-MNM-JNKI Step 1: Peptide synthesis Peptide synthesis was performed using standard Fmoc chemistry on a solid support using Boc-side-chain protected Fmoc-lysine. After completing SPPS synthesis (without N-terminal Fmoc deprotection), the peptide was cleaved from the resin and purified by HPLC to obtain a peptide with an Fmoc-protected N-terminus. Peptide sequence: Fmoc-NH-ggdqsrpvqpflnlttprkpr-C(=O)NH2(Sequence ID 1)

[0230] Step 2: Conjugate Synthesis [ka]

[0231] Step 2 was carried out as disclosed above.

[0232] Example 5 MNM-CPP-JNKI Step 1: Conjugate Synthesis Peptide synthesis was carried out on a solid support using standard Fmoc chemistry, with Dde side-chain protected lysine, which was Fmoc-protected on the solid support as disclosed above. Peptide sequence: H2N-ggdqsrpvqpflnlttprkprpprrrqrrk(Fmoc)k(Fmoc)rg-C(=O)NH2(Sequence ID 2)

[0233] Step 2: Conjugate Synthesis [ka]

[0234] Step 2 was carried out as disclosed above.

[0235] Example 6 C-terminal MNM-JNKI Step 1: Conjugate Synthesis The peptide was synthesized on a solid support using standard Fmoc chemistry (Rink amide MBHA Resin (0.1 mmol, 0.3 g, sub: 0.33 mmol / g)). Peptide sequence: dqsrpvqpflnlttprkprppcc (SEQ ID NO: 3)

[0236] Step 2: Conjugate Synthesis [ka]

[0237] All peptide sequences mentioned herein were synthesized using D amino acids.

[0238] Example 7 Survival rate test Four exemplary conjugates of the invention were tested in viability studies. The conjugates contained a JNK inhibitor (D-JNKI-1), and their effect on cell viability was investigated on B16 cells (mouse melanoma) and Hela-GFP cells (human cervical cancer) based on an XTT-based assay (Biological Industries; #20-300-1000).

[0239] B16 (8,000 cells / well) and Hela (5,000 cells / well) cells were seeded in 96-well flat-bottom plates and grown overnight at 37°C / 5% CO2 in cell medium supplemented with 10% serum. The medium was then replaced with low-serum medium (Opti-MEM, Gibco; #31985070), and the cells were treated with the peptides mentioned in Tables 2 and 3, respectively (Table 3 shows all sequences used in the experiments presented in Figures 1-3). All peptide stock solutions were prepared in DMSO unless otherwise specified (stock solution concentration 1 mM). The day after treatment, XTT reagent was added to each well according to the manufacturer's instructions, and the colorimetric analysis results were measured using a colorimetric reader (Infinite M200 PRO; Tecan). Absorbance values ​​were normalized to individual controls (vehicles), and the results are summarized in Figures 1, 3, and 2 for B16 and Hela cells, respectively.

[0240] Figure 1 summarizes the results of viability tests on melanoma B16 cells. Control experiments showed that CPP and D-JNKI-1 had a slight effect on cell viability up to 10 μM (up to 10%), and that when CPP and JNKI were conjugated (CPP-JNKI), viability decreased by a further 20%. Exposure of cells to 25 μM CPP-JNKI reduced viability to 25%. Investigation of the four inventive conjugates—N-terminal MNM-JNKI, C-terminal MNM-JNKI, Lys-MNM-JNKI, and MNM-CPP-JNKI—shows that the attachment site within the peptide affects the reactivity of the inhibitor. When the conjugate is attached to the C-terminus of the peptide, the viability becomes equal to that of CPP-JNKI. On the other hand, when JNKI was attached to the N-terminus, viability rates of 30% and 2% were observed for 5 μM and 10 μM, respectively, indicating that the conjugate enhances JNKI activity. For Lys-MNM-JNKI, the viability decreased by more than half at 10 μM. The best results were achieved when the conjugate was attached to CPP (MNM-CPP-JNKI), with viability rates of 50% and 0% observed for 1 and 5 μM, respectively. IC 50 The values ​​are summarized in Table 2 for each conjugate.

[0241] The inventors observed that conjugates containing a combination of CPP and the inventive component (Molecular Nano-Motor; MNM) as carriers were significantly superior in terms of cellular delivery of cargo (e.g., peptides) (more than 3 times higher activity, e.g., for #Pe21) and induced higher cellular mortality than a similar conjugate containing the same MNM but without CPP (#Pe20). The inventive conjugates tested (including the #No. assigned in Figure 3), as well as the controls used in the experiments, are summarized in Table 3. The structures of the MNMs used in the experiments are presented in Tables A and B. Graphs summarizing the in vitro results of the experiments are presented in Figures 1-3.

[0242] [Table 4]

[0243] [Table 5]

[0244] The same trend was observed in Hela cells (Figure 2). Of particular note is MNM1026: [ka] It was completely inactive in vitro.

[0245] While the invention has been described in conjunction with its specific embodiments, it is clear that many substitutions, modifications, and alterations will be apparent to those skilled in the art. Therefore, it is intended to encompass all such substitutions, modifications, and alterations that fall within the spirit and broad scope of the appended claims.

[0246] All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent as each individual publication, patent, and patent application is specifically and individually indicated as being incorporated herein by reference. In addition, any citation or reference of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. Section headings, to the extent that they are used, should not necessarily be construed as restrictive.

Claims

1. Formula A', comprising any pharmaceutically acceptable salt, any hydrate, or any solvate thereof: 【Chemistry 1】 A conjugate comprising a polymer covalently bonded to the portion represented by the formula, where, a and b each independently represent integers from 1 to 10; A represents a sterol; R Ｔ is an optionally substituted alkyl or optionally substituted heteroalkyl; L represents a linker containing at least one -N(R5)- group, wherein the linker has a single C-C bond length of 1 to 30; R is hydrogen or each is independently halo, -NO₂, -CN, -OR', -OH, -CONH₂, HCONH-, oxo, carbonyl, amino, imino, thioxo, phosphate (i.e., -OPO(OR')₂), phosphonate, phosphine, phosphite, -CONR'₂, -CNNR'₂, -CSNR'₂, -CONH-OH, -CONH-NH₂, -NHCOR', -NHCSR', -NHCNR', -NC(=O)OR', -NC(=O)NR', -NC(=S)OR', -NC(=S)NR', -SO₂R', -SOR', -SR', -SO₂OR', -SO₂N(R')₂, -NHNR'₂, -NNR', C₁-C₆ haloalkyl, optionally substituted C₁-C₆ alkyl, -NH₂, -NR'R', -NH(C₁-C₆ alkyl), -N(C₁-C₆ alkyl)₂, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, hydroxy(C₁-C₆ alkyl), hydroxy(C₁-C₆ alkoxy), alkoxy(C₁-C₆ alkyl), alkoxy(C₁-C₆ alkoxy), C₁-C₆ alkyl-NR'₂, C₁-C₆ alkyl-SR', -CONH(C₁-C₆ alkyl), -CON(C₁-C₆ alkyl)₂, -CO₂H, -COR', -CO₂R', -O COR', -O COR', -OC(=O)OR', -OC(=O)NR', -OC(=S)OR', -OC(=S)NR', amino(C １ -C ６ alkyl), C １ -C ６ mercaptoalkyl, -CONH(C １ -C ６ alkyl), -CON(C １ -C ６ alkyl) ２ , -CO ２ H, -CO ２ R represents one or more substituents selected from -OCOR, -OC(=O)OR, -OC(=O)NR, -OC(=S)OR, -OC(=S)NR, alkyl-aryl, alkyl-heteroaryl, or combinations thereof; each R' independently represents hydrogen, or selected from the group including optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or combinations thereof; The wavy bonds represent attachment points to the polymer; the polymer contains polyamino acids. Conjugate.

2. The aforementioned part is Equation 1A: 【Chemistry 2】 By, or formula 2: 【Transformation 3】 It is expressed by, in the formula, m is an integer independently selected from 1 to 5; each X １ These are independently O, S, NR ５ , CRR or CH ２ And; R Ｔ１ This includes optionally substituted linear or branched alkyl groups; R5 is either H, or a linear or branched optionally substituted C1-C5 alkyl group which may optionally contain one or more heteroatoms; c is an integer independently selected from 0 to 10; L1 is a bond, or -N-C(=O)-, -C(=O)N-, -S-S-, -S-C(=O), Y-(C ０－１０ ) Alkyl-X-(C ０－１０ ) A linker selected from alkyl-Y, cycloalkyl, heterocyclyl and click reaction products, or any combination thereof; Each X and Y is either absent or independently selected from heteroatoms, oligomers, click reaction products, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-; The oligomer comprises 2 to 15 repeating units; and W represents (i) amide, (ii) click reaction product, or both. The conjugate according to claim 1.

3. The repeating unit is selected from the following conjugates according to claim 2: alkylene oxide, natural or unnatural amino acid derivative, α-hydroxycarboxylic acid residue, and amino group.

4. L1 is -N-C(=O), -C(=O)N, or Y-(C ０－１０ ) Alkyl-X-(C ０－１０ ) alkyl-Y; at least one X1 is CRR, each R independently is hydrogen or one or more substituents, each independently being -NO2, -CN, -OR', -OH, -CONH2, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, -NC(=O)OR', -NC(=O)NR', ​​-NC(=S)OR', -NC(=S)NR A conjugate according to any one of claims 1 to 3, selected from ', -SO2R', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -CO2H, -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​or a combination thereof.

5. The aforementioned part is Equation 3: 【Chemistry 4】 The conjugate according to any one of claims 2 to 4, which is expressed as follows, where d and e are each independently integers between 1 and 10, and at least one X1 is -O-.

6. The click reaction product comprises any one of the following conjugates according to any one of claims 2 to 5: succinimide-thioether; azidoalkyne cycloaddition product; Diels-Alder reaction product; reverse electron-required Diels-Alder reaction product; dibenzylcyclooctin 1,3-nitrone cycloaddition product; or any combination thereof.

7. The conjugate according to any one of claims 1 to 6, wherein the polymer is a peptide, the peptide comprises a therapeutically effective amino acid sequence, and further comprises a cell-permeable peptide bound to the therapeutically effective amino acid sequence.

8. The conjugate according to claim 7, wherein the portion is bonded to (i) the N-terminus or C-terminus of the peptide, (ii) the C-terminal amino acid of the peptide, (iii) at least one of the amino group and thio group of an amino acid residue of the peptide, or any combination of (i)-(iii).

9. The aforementioned part is Equation 4: 【Transformation 5】 The conjugate according to any one of claims 1 to 4, wherein n is an integer independently selected from 1 to 10, and each Y1 and Y2 is absent or independently selected from cycloalkyl, heterocyclyl, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-.

10. The conjugate according to claim 9, wherein Y1 is -N-C(=O)- and Y2 is -N-C(=O)- or -C(O=)O-.

11. The conjugate according to any one of claims 1 to 10, wherein R5 is methyl.

12. The conjugate according to any one of claims 1 to 11, wherein R is a halogen.

13. The aforementioned conjugate is 【Transformation 6】 A conjugate according to any one of claims 1 to 12, selected from the above.

14. A compound comprising any pharmaceutically acceptable salt, any hydrate, or any solvate thereof; the compound is of formula C: 【Transformation 7】 By, or formula 5A: 【Transformation 8】 It is expressed by, in the formula, a and b each independently represent integers from 1 to 10; A represents a sterol; R Ｔ is an optionally substituted alkyl or optionally substituted heteroalkyl; M represents a linker containing at least one -N(R5)- group; the linker has a single C-C bond length of 1 to 30; Z is a moiety that is reactive to a functional group selected from a thio group, an amino group, a 1,3-nitrone, an azide, a diene, a tetrazine, a succinimide, an α-β unsaturated carbonyl, an α-β unsaturated keto acid derivative, a carbonyl and an active ester, or any combination thereof; R is either hydrogen, or each independently a halo, -NO2, -CN, -OR', -OH, -CONH2, HCONH-, oxo, carbonyl, amino, imino, thioxo, phosphate (i.e., -OPO(OR')2), phosphonate, phosphine, phosphine, -CONR'2, -CNNR'2, -CSNR'2, -C ONH-OH, -CONH-NH2, -NHCOR', -NHCSR', -NHCNR', -NC(=O)OR', -NC(=O)NR', ​​-NC(=S)OR', -NC(=S)NR', ​​-SO2R', -SOR', -SR', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', C1-C6 haloalkyl, Substitutable C1-C6 alkyl, -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(C1-C6 alkyl), hydroxy(C1-C6 alkoxy), alkoxy(C1-C6 alkyl), alkoxy(C1-C6 alkoxy), C1-C6 alkyl-NR'2, C1-C6 alkyl-SR', -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -COR', -CO2R', -OCOR', -OCOR', -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​amino(C １ -C ６ Alkyl), C １ -C ６ Mercaptoalkyl, -CONH(C １ -C ６ Alkyl), -CON(C １ -C ６ Alkyl) ２ , -CO ２ H, -CO ２ Represents one or more substituents selected from R, -OCOR, -OC(=O)OR, -OC(=O)NR, -OC(=S)OR, -OC(=S)NR, alkyl-aryl, alkyl-heteroaryl, or combinations thereof; Each R' independently represents hydrogen, or is selected from the group including optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or combinations thereof; R5 is either H or a linear or branched optionally substituted C1-C5 alkyl group which may optionally contain one or more heteroatoms. compound.

15. The aforementioned compound has formula 6: 【Chemistry 9】 It is expressed by, in the formula, m is an integer independently selected from 1 to 5; each X １ These are independently O, S, NR ５ , CRR or CH ２ And; R Ｔ１ This includes optionally substituted linear or branched alkyl groups; c is an integer selected from 1 to 10; L1 is a bond, or -N-C(=O)-, -C(=O)N-, -S-C(=O), Y-(C ０－１０ ) Alkyl-X-(C ０－１０ ) A linker selected from alkyl-Y, cycloalkyl, heterocyclyl, and click reaction products (including any combination thereof); Each X and Y is either absent or independently selected from heteroatoms, oligomers, click reaction products, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, -OC(=S)N-; The oligomer contains 2 to 15 repeating units. The compound according to claim 14.

16. L1 is -N-C(=O), -C(=O)N, or Y-(C0-10)alkyl-X-(C0-10)alkyl-Y; and at least one X1 is CRR, where each R is independently hydrogen or one or more substituents, each independently being -NO2, -CN, -OR', -OH, -CONH2, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, -NC(=O)OR', The compound according to claim 15, selected from -NC(=O)NR', ​​-NC(=S)OR', -NC(=S)NR', ​​-SO2R', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -CO2H, -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​or a combination thereof.

17. The aforementioned compound is given by formula 7: 【Chemistry 10】 The compound according to any one of claims 14 to 16, as represented by [the above].

18. The aforementioned compound is formula 8: 【Chemistry 11】 It is expressed by, in the formula, n is an integer independently selected from 1 to 10; The compound according to any one of claims 14 to 16, wherein each Y1 and Y2 is independently selected from cycloalkyl, heterocyclyl, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-.

19. The compound according to claim 18, wherein Y1 is -N-C(=O)- and Y2 is -N-C(=O)- or -C(O=)O-.

20. Z is a compound according to any one of claims 14 to 19, selected from carboxylates, active esters, carbonates, isocyanates, α-β unsaturated keto compounds, thioacetyls, and maleimides.

21. The compound according to any one of claims 14 to 20, wherein R is a halo and R5 is methyl.

22. The compound according to any one of claims 14 to 21, wherein the compound comprises any one of the compounds in Table A.

23. The compound according to any one of claims 14 to 22, wherein Z is reactive to a polymer containing the functional group, and the compound is a precursor of the conjugate according to any one of claims 1 to 13.

24. A pharmaceutical composition comprising a conjugate according to any one of claims 1 to 13 and a pharmaceutically acceptable carrier.

25. The pharmaceutical composition according to claim 24, comprising a therapeutically effective amount of the conjugate.

26. A pharmaceutical composition according to claim 24 or 25 for use in the treatment or prevention of a disease in a subject.

27. The pharmaceutical composition according to claim 26, wherein the disease is cancer.

28. A kit comprising a compound according to any one of claims 14 to 23, and (i) a polyamino acid; and (ii) a polymer selected from polymers comprising an amino acid and further comprising a thio group, an amino group, a 1,3-nitrone, an azide, a diene, a tetrazine, or any combination thereof.

29. The kit according to claim 28, wherein the molar ratio between the polymer and the compound in the kit is approximately 1:

1.

30. The kit according to claim 28 or 29, further comprising instructions for reacting the polymer and the compound to obtain the conjugate according to any one of claims 1 to 13.

31. The kit according to any one of claims 28 to 30, further comprising a liquid carrier.

32. A method for treating a disease or condition in a subject, comprising administering a therapeutically effective amount of the pharmaceutical composition according to claim 24 or 25 to the subject, thereby treating the disease.

33. The method according to claim 32, wherein the disease is a cell proliferation-related disease.

34. A method for internalizing polyamino acids into cells, comprising contacting the cells with the pharmaceutical composition described in claim 24 or 25, thereby internalizing the peptide into the cells.

35. The method according to claim 34, wherein the cells are cancer cells.

36. Formula B', comprising any pharmaceutically acceptable salt, any hydrate, or any solvate: 【Chemistry 12】 A conjugate comprising a polymer covalently bonded to the portion represented by the formula, where, b represents an integer between 1 and 10; A represents a sterol; B represents an optionally substituted C3-C10 heterocycline; R Ｔ is an optionally substituted alkyl or optionally substituted heteroalkyl; L represents a linker with a single C-C bond length of 1 to 30; The wavy bonds represent attachment points to the polymer; and, The polymer contains polyamino acids. Conjugate.

37. The aforementioned part is Equation 9: 【Chemistry 13】 It is expressed by, in the formula, each X ２ These are independently N or CR; Each R is independently hydrogen, or each is independently halo, -NO2, -CN, -OR', -OH, -CONH2, HCONH-, oxo, carbonyl, amino, imino, thioxo, phosphate (i.e., -OPO(OR')2), phosphonate, phosphine, phosphine, -CONR'2, -CNNR'2, -CSNR' 2, -CONH-OH, -CONH-NH2, -NHCOR', -NHCSR', -NHCNR', -NC(=O)OR', -NC(=O)NR', ​​-NC(=S)O R', -NC(=S)NR', ​​-SO2R', -SOR', -SR', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', C1-C6 haloalkyl , optionally substituted C1-C6 alkyl, -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(C1-C6 alkyl), hydroxy(C1-C6 alkoxy), alkoxy(C1-C6 alkyl), alkoxy(C1-C6 alkoxy), C1-C6 alkyl-NR'2, C1-C6 alkyl-SR', -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -COR', -CO2R', -OCOR', -OCOR', -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​amino(C １ -C ６ Alkyl), C １ -C ６ Mercaptoalkyl, -CONH(C １ -C ６ Alkyl), -CON(C １ -C ６ Alkyl) ２ , -CO ２ H, -CO ２ The conjugate according to claim 1, wherein R represents one or more substituents selected from -OCOR, -OC(=O)OR, -OC(=O)NR, -OC(=S)OR, -OC(=S)NR, alkyl-aryl, alkyl-heteroaryl, or combinations thereof; each R' independently represents hydrogen or is selected from the group comprising optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or combinations thereof.

38. The aforementioned part is Equation 10: 【Chemistry 14】 It is expressed by, in the formula, m is an integer independently selected from 1 to 5; each X １ These are independently O, S, NR ５ , CRR or CH ２ And; R Ｔ１ This includes optionally substituted linear or branched alkyl groups; c is an integer independently selected from 0 to 10; L1 is either a bond or -N-C(=O)-, -C(=O)N-, -S-C(=O), Y-(C ０－１０ ) Alkyl-X-(C ０－１０ ) A linker selected from alkyl-Y, cycloalkyl, heterocyclyl and click reaction products, or any combination thereof; Each X and Y is either absent or independently selected from heteroatoms, oligomers, click reaction products, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-; the oligomers contain 2 to 15 repeating units; and W represents (i) amide, (ii) click reaction product, or both. The conjugate according to claim 37.

39. L1 is a bond; at least one X1 is a CRR, where each R is independently a hydrogen or one or more substituents, each independently -NO2, -CN, -OR', -OH, -CONH2, -CONR'2, -CNNR'2, -CSNR'2, -CONH-OH, -CONH-NH2, -NC(=O)OR', -NC(=O)NR', ​​-NC(=S)OR', -NC(=S) The conjugate according to claim 38, selected from NR', -SO2R', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -CO2H, -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​or a combination thereof.

40. A compound comprising any pharmaceutically acceptable salt, any hydrate, or any solvate thereof, wherein the compound is of formula D: 【Chemistry 15】 Represented by (including any pharmaceutically acceptable salt, any hydrate, or any solvate thereof), where, b represents an integer between 1 and 10; A represents a sterol; B represents an optionally substituted C3-C10 heterocycline; R Ｔ is an optionally substituted alkyl or optionally substituted heteroalkyl; M represents a linker with a single C-C bond length of 1 to 30; Z is a moiety that is reactive to a functional group selected from a thio group, an amino group, a 1,3-nitrone, an azide, a diene, a tetrazine, a succinimide, an α-β unsaturated carbonyl, an α-β unsaturated keto acid derivative, a carbonyl and an active ester, or any combination thereof. compound.

41. Formula 11: 【Chemistry 16】 It is expressed by, in the formula, each X ２ These are independently N or CR; Each R independently represents hydrogen or one or more substituents, each independently representing halo, -NO2, -CN, -OR', -OH, -CONH2, HCONH-, oxo, carbonyl, amino, imino, thioxo, phosphate (i.e., -OPO(OR')2), phosphonate, phosphine, phosphine, -CONR'2, -CNNR' 2, -CSNR'2, -CONH-OH, -CONH-NH2, -NHCOR', -NHCSR', -NHCNR', -NC(=O)OR', -NC(=O)NR', ​​- NC(=S)OR', -NC(=S)NR', ​​-SO2R', -SOR', -SR', -SO2OR', -SO2N(R')2, -NHNR'2, -NNR', C1-C6 Haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NR'R', -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(C1-C6 alkyl), hydroxy(C1-C6 alkoxy), alkoxy(C1-C6 alkyl), alkoxy(C1-C6 alkoxy), C1-C6 alkyl-NR'2, C1-C6 alkyl-SR', -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -COR', -CO2R', -OCOR', -OCOR', -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​amino(C １ -C ６ Alkyl), C １ -C ６ Mercaptoalkyl, -CONH(C １ -C ６ Alkyl), -CON(C １ -C ６ Alkyl) ２ , -CO ２ H, -CO ２ R' is selected from -OCOR, -OC(=O)OR, -OC(=O)NR, -OC(=S)OR, -OC(=S)NR, alkyl-aryl, alkyl-heteroaryl, or combinations thereof; each R' independently represents hydrogen, or is selected from the group including optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or combinations thereof. The compound according to claim 40.

42. Formula 12: 【Chemistry 17】 It is expressed by, in the formula, m is an integer independently selected from 1 to 5; each X １ These are independently O, S, NR ５ , CRR or CH ２ And; R Ｔ１ This includes optionally substituted linear or branched alkyl groups; c is an integer independently selected from 0 to 10; L1 is either a bond or -N-C(=O)-, -C(=O)N-, -S-C(=O), Y-(C ０－１０ ) Alkyl-X-(C ０－１０ ) A linker selected from alkyl-Y, cycloalkyl, heterocyclyl and click reaction products, or any combination thereof; Each X and Y is either absent or independently selected from heteroatoms, oligomers, click reaction products, -CONR'-, -CNNR'-, -CSNR'-, -NC(=O)O-, -NC(=S)O-, -NC(=S)N-, -SO2-, -SO-, -SR', -C(=O)-, -OC(=O)-, -OC(=O)O-, -OC(=S)O-, and -OC(=S)N-; The oligomer contains 2 to 15 repeating units. The compound according to claim 41.

43. The compound according to any one of claims 40 to 42, wherein the compound comprises any one of the compounds in Table B.

Images