Targeted linear conjugates containing polyethyleneimine and polyethylene glycol and polyplexes containing same

Abstract

The present invention relates to polyplexes comprising linear conjugates of LPEI and PEG. The LPEI and PEG fragments of the linear conjugates are preferably linked via a [3+2] cycloaddition between an azide and an alkene or alkyne to produce a 1,2,3 triazole or a 4,5-dihydro-1H-[1,2,3]triazole. The linear conjugates are preferably further conjugated to a targeting fragment to enable selective interaction with specific cell types. The conjugates can form polyplexes with therapeutic agents, such as nucleic acids, to deliver the therapeutic agents to cells.

Description

[Background technology]

[0001] Cancer remains a leading cause of death worldwide. For most solid tumors after surgical removal, chemotherapy is an important treatment option for managing remaining cancer cells. The main reason for chemotherapy failure is inefficient targeting and uptake of chemotherapeutic agents by tumors (Vasir & Labhasetwar Technology in Cancer Research & Treatment 4(4), 363-374(2005)). Insufficient accessibility to tumors requires higher doses, and due to the nature of chemotherapeutic agents, this leads to nonspecific uptake and toxicity of healthy cells. Targeted drug delivery strategies, in which therapeutic agents reversibly bind to targeting ligands and are selectively delivered to cells for treatment, are currently applied to many chemotherapeutic agents in clinical use. This strategy shows promise for maximizing the safety and efficacy of a given chemotherapeutic agent, as their selective delivery to target cells avoids nonspecific uptake and associated toxicity in healthy cells (Srinivasarao & Low, Chem. Rev., 117, 12133-12164, (2017)) and may result in a higher maximum tolerated dose.

[0002] Cationic polymers are known to form supramolecular polyplexes with negatively charged nucleic acids in solution. For example, linear polyethyleneimine (LPEI) is protonated at physiological pH and therefore has a net positive charge. When LPEI is incubated with nucleic acids, which have a net negative charge at physiological pH, the LPEI and nucleic acids can form polyplexes held together by electrostatic interactions. These supramolecular polyplexes can be taken up by cells in vivo, where they can deliver nucleic acid sequences into cells. Therefore, supramolecular polyplexes containing cationic polymers and nucleic acids can be used as vectors for therapy.

[0003] Despite their promise, technical challenges remain regarding the formation of homogeneous, well-characterized cationic polymers. Polyplexes containing only LPEI are prone to aggregation and interactions with serum proteins, potentially limiting their potential as nucleic acid delivery agents. To overcome these challenges, the polymer LPEI can be conjugated or copolymerized with polyethylene glycol (PEG). PEG fragments can help shield LPEI from the surrounding matrix, improving the biocompatibility and blood circulation of the resulting polyplexes.

[0004] However, coupling of PEG to LPEI generally occurs via the formation of a covalent bond between an electrophilic PEG fragment and a secondary amine embedded within the LPEI backbone fragment, resulting in branched, heterogeneous conjugates with random inclusion of PEG fragments, characterized based on the average PEG inclusion density. In such conjugates, the PEG fragments, whether single or multiple, are generally orthogonally attached to the LPEI fragment without site specificity. Such random synthesis and inaccurate characterization of LPEI-PEG conjugates can make it difficult to establish a clear structure-activity relationship (SAR) between the conjugate structure and the activity of the resulting supramolecular polyplex. Therefore, there is a need for homogeneous LPEI-PEG conjugates with well-defined chemical structures.

[0005] Overexpression of EGFR has been observed in advanced stages of melanoma and correlates with disease progression and resistance to vemurafenib (a BRAF inhibitor) (Kovacs, E., et al., (2015). Annu. Rev. Biochem. 84, 739-764; Gross, A., et al., (2014). Target. Oncol. 10, 77-84).

[0006] EGFR is overexpressed in over 90% of head and neck tumors (Kalyankrishna S and Grandis, JR. J Clin Oncol 2006;24:2666-72), and this overexpression is associated with decreased overall survival (Byeon HK, et al., Exp Mol Med. 2019 Jan 16;51(1):1-14). Summary of the Invention

[0007] The present invention provides targeted conjugates comprising specifically defined, discrete molecular weight PEG fragments linked by discrete linkages formed by defined chemoselective reactions, instead of the random and uncontrolled attachment of electrophilic PEG fragments to multiple nucleophiles on an LPEI backbone fragment. Thus, the present invention provides more homogeneous targeting conjugates with defined chemical structures. The discrete, specifically defined building blocks and linkages not only ensure a consistent and predictable ratio of all components of the conjugates of the present invention, including a consistent and predictable ratio of LPEI to PEG fragments, but also ensure a defined, linear conjugate instead of a randomly branched conjugate. Thus, the LPEI fragment is linked in a linear, end-to-end manner to a single, specifically defined, discrete PEG fragment with a defined, discrete molecular weight, which is further linked to a targeting fragment. The chemoselective attachment of the LPEI fragment to the specifically defined, discrete PEG fragment can be carried out using any suitable chemical precursor capable of forming a chemoselective bond. In a preferred embodiment, the chemoselective conjugation of the LPEI fragment with a specifically defined, discrete PEG fragment occurs via a [3+2] cycloaddition between an azide and an alkyne or alkene to yield a 1,2,3-triazole or a 4,5-dihydro-1H-[1,2,3]triazole.

[0008] In preferred conjugates of the present invention, the PEG fragment is further selectively linked to a targeting fragment to target specific cell types and promote uptake of the compositions, conjugates, and / or polyplexes of the present invention in those specific cell types. Thus, preferred embodiments include one or more, typically and preferably one, targeting fragment, such as hEGF, a HER2 ligand, DUPA, or folate, specifically linked to the LPEI-PEG diconjugate to form an LPEI-PEG-targeting fragment triconjugate, capable of targeting a corresponding receptor, such as hEGFR, HER2, PSMA, or folate, on a specific cell type, typically a cancer cell type. In the case of polyplexes of the present invention, such triconjugates are combined with polyanions, such as nucleic acids, preferably RNA, and more preferably dsRNA, such as polyinosinic:polycytidylic acid (poly(IC)), or mRNA or pDNA. Polyanions, such as poly(IC), can serve as cytotoxic and / or immunostimulatory payloads that are delivered to and taken up by cells.

[0009] Even more surprisingly, the inventors have advantageously found that the resulting preferred conjugates and polyplexes according to the present invention, which have significantly reduced heterogeneity and therefore a significantly reduced number of potentially biologically active conjugates and polyplexes due to the defined chemoselective attachment of LPEI fragments to specifically defined, discrete PEG fragments, not only form polyplexes of appropriate size, but also maintain or even increase their overall biological activity, such as potency and selectivity for reducing survival and inducing cell death of targeted cancer cells. Furthermore, the compositions and polyplexes of the present invention, which comprise nucleic acids encoding peptides or proteins of interest, particularly pharmaceutically active peptides or proteins such as cytokines, interferons, or toxins, not only selectively deliver the pharmaceutically active nucleic acids encoding the pharmaceutically active peptides or proteins to target cells, particularly cancer cells, but also result in high expression and efficient protein translation and secretion of the encoded pharmaceutically active proteins.

[0010] Thus, in one aspect, the present invention provides a composition comprising a conjugate, the conjugate comprising: a linear polyethyleneimine fragment containing an alpha end and an omega end; a polyethylene glycol fragment comprising a first end terminus and a second end terminus, the polyethylene glycol fragment comprising, and preferably consisting of, a discrete number m of repeating -(O-CH-CH)- units, wherein the discrete number m of repeating -(O-CH-CH)- units is any discrete number between 25 and 100, preferably between 25 and 60; The alpha terminus of the polyethyleneimine fragment is the initiating residue; The omega end of the polyethyleneimine fragment is linked to the first end of the polyethylene glycol fragment by a divalent covalent linking group -ZX 1 -ZX 1 - is not a single bond and -Z- is not an amide; The second end of the polyethylene glycol fragment is capable of binding to a targeting fragment, and preferably the second end of the polyethylene glycol fragment is a bivalent covalent linking moiety X 2 and more preferably, the targeting fragment is capable of binding to a cell.

[0011] In another aspect, the invention provides a composition comprising a conjugate, wherein the conjugate is of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, ZX 1 - is not a single bond and Z is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell.

[0012] In a further aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A is one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0013] Although the HN-N=N fragment of the bicyclic ring in Formula I is typically depicted herein using one single bond and one double bond for simplicity, those skilled in the art will recognize that Formula I and related conjugate structures depicted herein can alternatively be depicted as shown below. Such depictions of Formula I are used interchangeably herein. [ka] During the ceremony, fragment [ka] is the fragment R 1 (NR 2 CH2CH2) n Two different regioisomeric bonds, i.e., [ka] and [ka] where the wavy line represents the chemical bond to ring A. Thus, Formula I as depicted herein represents two regioisomeric embodiments, namely, fragment R 1 (NR 2 CH2CH2) nis attached to the top nitrogen atom of the structure or to the bottom nitrogen atom of the structure, but not to the middle nitrogen atom. One of skill in the art will understand that the same applies to other formulas herein, including Formula IA, Formula IB, Formula IC, Formula ID, Formula IE, Formula IH, Formula IJ, Formula IK, etc.

[0014] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A is one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0015] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A is one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23-, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell, and preferably the composition consists of the conjugate.

[0016] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2-CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A is one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q-, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell.

[0017] In a further aspect, the present invention provides methods for synthesizing compositions comprising conjugates of Formula I, comprising reacting an azide-containing LPEI fragment with an alkene- or alkyne-containing PEG fragment at a pH below about 5, preferably about 4 or less. In some preferred embodiments, the LPEI fragment comprises an azide at the omega terminus and the PEG fragment comprises an alkene or alkyne at its first terminus.

[0018] In a further aspect, the present invention provides a polyplex comprising a composition described herein and a polyanion, preferably wherein the polyanion is a nucleic acid, more preferably wherein the nucleic acid is RNA, and again more preferably wherein the polyanion is polyinosinic:polycytidylic acid (poly(IC)).

[0019] In a further aspect, the invention provides a polyplex comprising a composition described herein and a nucleic acid. In a further aspect, the invention provides a polyplex comprising a composition described herein and a nucleic acid, wherein the nucleic acid is RNA. In a further aspect, the invention provides a polyplex comprising a composition described herein and polyinosinic:polycytidylic acid (poly(IC)).

[0020] In another aspect, the present invention provides a polyplex comprising a triconjugate as described herein, preferably such a conjugate of Formula I* or Formula I, and a polyanion, e.g., a nucleic acid, preferably polyinosinic:polycytidylic acid (poly(IC)).

[0021] In one aspect, the present invention provides pharmaceutical compositions comprising a trichouate described herein, preferably such a conjugate of Formula I* or Formula I, and / or a polyplex, and pharmaceutically acceptable salts thereof.

[0022] In one aspect, the invention provides a polyplex as described herein, or a pharmaceutical composition comprising a polyplex as described herein, for use in the treatment of a disease or disorder, preferably cancer.

[0023] In one aspect, the invention provides the use of a polyplex described herein in the manufacture of a medicament for treating a disease or disorder, such as cancer.

[0024] In another aspect, the present invention provides a method of treating a disease or disorder, such as cancer, in a subject in need thereof, comprising administering to the subject an effective amount of a polyplex described herein.

[0025] In one aspect, the present invention provides a composition comprising a conjugate for use in treating head and neck cancer, the conjugate comprising: a linear polyethyleneimine (LPEI) fragment comprising an alpha end and an omega end; a polyethylene glycol (PEG) fragment, preferably a linear polyethylene glycol (PEG) fragment, comprising a first end and a second end; the omega end of the LPEI fragment is linked to the first end of the PEG fragment by a covalent linking moiety; the covalent linking moiety is not an amide; preferably, the alpha end of the LPEI fragment is bonded to a methyl group or a hydrogen atom, more preferably, the alpha end of the LPEI fragment is bonded to a hydrogen atom; and preferably, the second end of the PEG fragment is bonded to a targeting fragment.

[0026] In one aspect, the present invention provides a composition comprising a conjugate for use in treating head and neck cancer, the conjugate comprising a linear polyethyleneimine fragment comprising an alpha end and an omega end; a polyethylene glycol fragment comprising a first end and a second end; the alpha end of the polyethyleneimine fragment is an initiating residue; the omega end of the polyethyleneimine fragment is linked to the first end of the polyethylene glycol fragment by a covalent linking moiety; the covalent linking moiety is not a single bond or an amide; preferably, the second end of the polyethylene glycol fragment is capable of reacting, and preferably the second end is capable of binding to a targeting fragment.

[0027] In one aspect, the present invention provides a composition comprising a conjugate for use in treating head and neck cancer, the conjugate comprising: a linear polyethyleneimine fragment containing an alpha end and an omega end; a polyethylene glycol fragment comprising a first end and a second end; The alpha end of the polyethyleneimine fragment is the initiating residue; the omega end of the polyethyleneimine fragment is the first terminal end of the polyethylene glycol fragment, which is linked to a covalent linking group -ZX 1-, -Z- is not a single bond, -Z- is not an amide; -X 1 - is a divalent covalent linking moiety; The second end of the polyethylene glycol fragment is capable of binding to a targeting fragment, and preferably the polyethylene glycol fragment binds to the targeting fragment. In a preferred embodiment of this aspect, the composition consists of the conjugate.

[0028] In another aspect, the present invention provides a composition comprising a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of head and neck cancer, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is any integer from 1 to 200, and preferably, m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor; and Preferably, the composition consists of the conjugate.

[0029] In another aspect, the present invention provides a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of head and neck cancer, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is any integer from 1 to 200, and preferably, m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0030] In another aspect, the present invention provides a composition comprising a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is any integer from 1 to 200; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0031] In another aspect, the present invention provides a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is any integer from 1 to 200; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0032] In another aspect, the present invention provides a composition comprising a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is any integer from 1 to 200, and preferably, m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell, and preferably the composition consists of the conjugate.

[0033] In another aspect, the present invention provides a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is any integer from 1 to 200, and preferably, m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell.

[0034] In a further aspect, the present invention provides a polyplex comprising a composition described herein and a polyanion, preferably wherein the polyanion is a nucleic acid, more preferably wherein the nucleic acid is RNA, and again more preferably wherein the polyanion is polyinosinic:polycytidylic acid (poly(IC)), for use in treating head and neck cancer.

[0035] In a further aspect, the present invention provides a polyplex comprising a composition described herein and a nucleic acid for use in treating head and neck cancer. In a further aspect, the present invention provides a polyplex comprising a composition described herein and a nucleic acid, wherein the nucleic acid is RNA, for use in treating head and neck cancer. In a further aspect, the present invention provides a polyplex comprising a composition described herein and polyinosinic:polycytidylic acid (poly(IC)) for use in treating head and neck cancer.

[0036] In another aspect, the present invention provides a polyplex comprising a triconjugate as described herein, preferably such a conjugate of Formula I* or Formula I, and a polyanion, e.g., a nucleic acid, preferably polyinosinic:polycytidylic acid (poly(IC)), for use in treating head and neck cancer.

[0037] In one aspect, the present invention provides a pharmaceutical composition comprising a tri-conjugate described herein, preferably such a conjugate of Formula I* or Formula I, and / or a polyplex, and pharmaceutically acceptable salts thereof, for use in the treatment of head and neck cancer.

[0038] In one aspect, the invention provides a polyplex as described herein, or a pharmaceutical composition comprising a polyplex as described herein, for use in the treatment of head and neck cancer.

[0039] In one aspect, the invention provides use of a polyplex described herein in the manufacture of a medicament for treating head and neck cancer.

[0040] In another aspect, the present invention provides a method of treating head and neck cancer in a subject in need thereof, comprising administering to the subject an effective amount of a polyplex described herein.

[0041] In another aspect, the present invention provides a composition comprising a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of head and neck cancer, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor; and Preferably, the composition consists of the conjugate.

[0042] In another aspect, the present invention provides a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of head and neck cancer, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0043] In another aspect, the present invention provides a composition comprising a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with RA2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0044] In another aspect, the present invention provides a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two RA1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0045] In another aspect, the present invention provides a composition comprising a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22-, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell, and preferably the composition consists of the conjugate.

[0046] In another aspect, the present invention provides a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2)n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q-, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell.

[0047] In one aspect, the present invention provides a composition comprising a conjugate for use in treating melanoma, the conjugate comprising: a linear polyethyleneimine (LPEI) fragment comprising an alpha end and an omega end; a polyethylene glycol (PEG) fragment, preferably a linear polyethylene glycol (PEG) fragment, comprising a first end and a second end; the omega end of the LPEI fragment is linked to the first end of the PEG fragment by a covalent linking moiety; the covalent linking moiety is not an amide; preferably, the alpha end of the LPEI fragment is bonded to a methyl group or a hydrogen atom, more preferably, the alpha end of the LPEI fragment is bonded to a hydrogen atom; and preferably, the second end of the PEG fragment is bonded to a targeting fragment.

[0048] In one aspect, the present invention provides a composition comprising a conjugate for use in treating melanoma, the conjugate comprising a linear polyethyleneimine fragment comprising an alpha end and an omega end; a polyethylene glycol fragment comprising a first end and a second end; the alpha end of the polyethyleneimine fragment is an initiating residue; the omega end of the polyethyleneimine fragment is linked to the first end of the polyethylene glycol fragment by a covalent linking moiety; the covalent linking moiety is not a single bond or an amide; preferably, the second end of the polyethylene glycol fragment is capable of reacting, and preferably the second end is capable of binding to a targeting fragment.

[0049] In one aspect, the present invention provides a composition comprising a conjugate for use in the treatment of melanoma, the conjugate comprising: a linear polyethyleneimine fragment containing an alpha end and an omega end; a polyethylene glycol fragment comprising a first end and a second end; The alpha end of the polyethyleneimine fragment is the initiating residue; the omega end of the polyethyleneimine fragment is the first terminal end of the polyethylene glycol fragment, which is linked to a covalent linking group -ZX 1 -, -Z- is not a single bond, -Z- is not an amide; -X 1 - is a divalent covalent linking moiety; The second end of the polyethylene glycol fragment is capable of binding to a targeting fragment, and preferably the polyethylene glycol fragment binds to the targeting fragment. In a preferred embodiment of this aspect, the composition consists of the conjugate.

[0050] In another aspect, the present invention provides a composition comprising a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of melanoma, R 1 -(NR 2 -CH2-CH2) n -ZX 1-(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is any integer from 1 to 200, and preferably, m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor; and Preferably, the composition consists of the conjugate.

[0051] In another aspect, the present invention provides a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of melanoma, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is any integer from 1 to 200, and preferably, m is any integer from 1 to 100; R 1is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0052] In another aspect, the present invention provides a composition comprising a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is any integer from 1 to 200; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more RA1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0053] In another aspect, the present invention provides a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is any integer from 1 to 200; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0054] In another aspect, the present invention provides a composition comprising a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is any integer from 1 to 200, and preferably, m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell, and preferably the composition consists of the conjugate.

[0055] In another aspect, the present invention provides a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is any integer from 1 to 200, and preferably, m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell.

[0056] In a further aspect, the present invention provides a polyplex comprising a composition described herein and a polyanion, preferably wherein the polyanion is a nucleic acid, more preferably wherein the nucleic acid is RNA, and again more preferably wherein the polyanion is polyinosinic:polycytidylic acid (poly(IC)), for use in treating melanoma.

[0057] In a further aspect, the present invention provides a polyplex comprising a composition described herein and a nucleic acid for use in treating melanoma. In a further aspect, the present invention provides a polyplex comprising a composition described herein and a nucleic acid, wherein the nucleic acid is RNA, for use in treating melanoma. In a further aspect, the present invention provides a polyplex comprising a composition described herein and polyinosinic:polycytidylic acid (poly(IC)) for use in treating melanoma.

[0058] In another aspect, the present invention provides a polyplex comprising a triconjugate as described herein, preferably such a conjugate of Formula I* or Formula I, and a polyanion, e.g., a nucleic acid, preferably polyinosinic:polycytidylic acid (poly(IC)), for use in treating melanoma.

[0059] In one aspect, the present invention provides a pharmaceutical composition comprising a tri-conjugate described herein, preferably said conjugate of Formula I* or Formula I, and / or a polyplex, and pharmaceutically acceptable salts thereof, for use in the treatment of melanoma.

[0060] In one aspect, the invention provides a polyplex described herein, or a pharmaceutical composition comprising a polyplex described herein, for use in the treatment of melanoma.

[0061] In one aspect, the invention provides the use of a polyplex described herein in the manufacture of a medicament for treating melanoma.

[0062] In another aspect, the present invention provides a method of treating melanoma in a subject in need thereof, comprising administering to the subject an effective amount of a polyplex described herein.

[0063] In another aspect, the present invention provides a composition comprising a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of melanoma, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor; and Preferably, the composition consists of the conjugate.

[0064] In another aspect, the present invention provides a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of melanoma, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0065] In another aspect, the present invention provides a composition comprising a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0066] In another aspect, the present invention provides a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0067] In another aspect, the present invention provides a composition comprising a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell, and preferably the composition consists of the conjugate.

[0068] In another aspect, the present invention provides a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell.

[0069] In another aspect, the present invention provides a composition comprising a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor; and Preferably, the composition consists of the conjugate.

[0070] In another aspect, the present invention provides a conjugate of formula I*, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0071] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0072] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0073] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell, and preferably the composition consists of the conjugate.

[0074] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell.

[0075] In another aspect, the present invention provides a composition comprising a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR; and Preferably, the composition consists of the conjugate.

[0076] In another aspect, the present invention provides a conjugate of formula I*, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond and -Z- is not -NHC(O)-; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0077] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2is a divalent covalent linking moiety; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0078] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0079] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR, and preferably the composition consists of the conjugate.

[0080] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0081] In another aspect, the present invention provides a composition comprising a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF), and Preferably, the composition consists of the conjugate.

[0082] In another aspect, the present invention provides a conjugate of formula I*, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: R 1 -(NR 2-CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond and -Z- is not -NHC(O)-; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0083] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0084] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0085] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; RA1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF), and preferably the composition consists of the conjugate.

[0086] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n-The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0087] In another aspect, the present invention provides a composition comprising a conjugate of formula IA, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0088] In another aspect, the present invention provides a conjugate of formula IA, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0089] In another aspect, the present invention provides a composition comprising a conjugate of formula IA, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF), and preferably the composition consists of the conjugate.

[0090] In another aspect, the present invention provides a conjugate of formula IA, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0091] In another aspect, the present invention provides a composition comprising a conjugate of formula IA-3 or IA-4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0092] In another aspect, the present invention provides a conjugate of formula IA-3 or IA-4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0093] In another aspect, the present invention provides a composition comprising a conjugate of formula IA-3 or IA-4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF), and preferably the composition consists of the conjugate.

[0094] In another aspect, the present invention provides a conjugate of formula IA-3 or IA-4, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl;14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0095] In another aspect, the present invention provides a composition comprising a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of head and neck cancer, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR; and Preferably, the composition consists of the conjugate.

[0096] In another aspect, the present invention provides a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of head and neck cancer, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond and -Z- is not -NHC(O)-; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0097] In another aspect, the present invention provides a composition comprising a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0098] In another aspect, the present invention provides a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0099] In another aspect, the present invention provides a composition comprising a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more RA1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23-, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR, and preferably the composition consists of the conjugate.

[0100] In another aspect, the present invention provides a conjugate of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0101] In another aspect, the present invention provides a composition comprising a conjugate of formula IA or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0102] In another aspect, the present invention provides a conjugate of formula IA or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0103] In another aspect, the present invention provides a composition comprising a conjugate of formula IA or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 )q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF), and preferably the composition consists of the conjugate.

[0104] In another aspect, the present invention provides a conjugate of formula IA or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23-, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0105] In another aspect, the present invention provides a composition comprising a conjugate of formula IA-3 or IA-4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0106] In another aspect, the present invention provides a conjugate of formula IA-3 or IA-4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0107] In another aspect, the present invention provides a composition comprising a conjugate of formula IA-3 or IA-4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer, [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23-, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF), and preferably the composition consists of the conjugate.

[0108] In another aspect, the present invention provides a conjugate of formula IA-3 or IA-4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of head and neck cancer: [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1is -H or -CH3; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0109] In another aspect, the present invention provides a composition comprising a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of melanoma, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond, -Z- is not -NHC(O)-; L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR; and Preferably, the composition consists of the conjugate.

[0110] In another aspect, the present invention provides a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof for use in the treatment of melanoma, R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L(formula I*); During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90% of which is H; X 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety, -ZX 1 - is not a single bond and -Z- is not -NHC(O)-; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0111] In another aspect, the present invention provides a composition comprising a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0112] In another aspect, the present invention provides a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0113] In another aspect, the present invention provides a composition comprising a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR, and preferably the composition consists of the conjugate.

[0114] In another aspect, the present invention provides a conjugate of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90% of which is H; Ring A may have one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, wherein the targeting fragment is capable of binding to epidermal growth factor receptor (EGFR), preferably the targeting fragment is capable of binding to a cell expressing EGFR, more preferably the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR.

[0115] In another aspect, the present invention provides a composition comprising a conjugate of formula IA or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0116] In another aspect, the present invention provides a conjugate of formula IA or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0117] In another aspect, the present invention provides a composition comprising a conjugate of formula IA or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF), and preferably the composition consists of the conjugate.

[0118] In another aspect, the present invention provides a conjugate of formula IA or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; R A1 are independently selected from one or more of C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0119] In another aspect, the present invention provides a composition comprising a conjugate of formula IA-3 or IA-4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0120] In another aspect, the present invention provides a conjugate of formula IA-3 or IA-4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1is -H or -CH3; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0121] In another aspect, the present invention provides a composition comprising a conjugate of formula IA-3 or IA-4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma, [ka] Formula IA-3 [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF), and preferably the composition consists of the conjugate.

[0122] In another aspect, the present invention provides a conjugate of formula IA-3 or IA-4 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof for use in the treatment of melanoma: [ka] [ka] During the ceremony, n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60, and more preferably, the discrete number m of repeating -(O-CH-CH)- units is 36; R 1 is the starting residue, preferably R 1 is -H or -CH3; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C1-C6 alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23-, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, —H, —COH, C-C alkyl, or oxo; and L is a targeting fragment, the targeting fragment is epidermal growth factor (EGF), preferably the targeting fragment is human EGF (hEGF).

[0123] The compositions and polyplexes of the invention comprising the linear, non-random LPEI-PEG diconjugates, and thus triconjugates, described herein not only ensure a consistent and predictable ratio of LPEI to PEG fragments, but also typically and preferably ensure a structurally defined, linear conjugate of LPEI fragments to PEG fragments. Thus, they offer greater batch-to-batch consistency, ease of manufacturing, and more predictable SAR compared to the branched LPEI-PEG diconjugates currently prepared using the random, uncontrolled synthetic strategies described above.

[0124] Even more advantageously and surprisingly, when the linear, non-random conjugates of the invention described herein are combined with polyanions and nucleic acids (e.g., poly(IC)) to form polyplexes and administered to cells, the polyplexes surprisingly not only maintain but even increase their antitumor activity as polyplexes made using random, branched conjugates. Thus, despite the significant reduction in variability and number of conjugate structures used and therefore in the number of possible (biological) activities, including targeting and presentation of their targeting fragments to the surface of target cells and subsequent uptake, there is no loss in the efficacy of the linear LPEI-l-PEG:nucleic acid polyplexes described herein. In contrast, the conjugates and compositions of the invention can even increase their overall biological activity.

[0125] Further features and advantages of the present technology will be apparent to those skilled in the art upon reading the following detailed description of the invention, and further aspects and embodiments of the invention will become apparent as the description continues. [Brief explanation of the drawings]

[0126] [Figure 1] DLS backscattering plots of triplicate Me-LPEI-l-[N3:BCN]-PEG36-DUPA:poly(IC) polyplexes measuring size distribution and zeta potential in 20 mM HEPES, 5% glucose (pH 7.2), 0.1875 mg / mL, 1.0 mL volume, and an N / P ratio of 4. The z-average diameter was 130 nm and the polydispersity index (PDI) was 0.134. The zeta potential was 26.6 mV. [Figure 2]DLS backscattering plots of triplicate Me-LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC) polyplexes measuring size distribution and zeta potential in 20 mM HEPES, 5% glucose (pH 7.2), 0.1875 mg / mL, 1.0 mL volume, and an N / P ratio of 4. The z-average diameter was 140 nm and the polydispersity index (PDI) was 0.132. The zeta potential was 28.2 mV. [Figure 3]

[0023] Figure 1 depicts differential PSMA expression as determined in vitro by flow cytometry for a panel of human prostate cancer cell lines (LNCaP, VCaP, PC-3, DU145). Staggered histograms of fluorescence intensity are shown, with mean fluorescence intensity (MFI) indicated. [Figure 4A] Flow cytometry analysis of MHC I expression on the cell surface of a prostate cancer cell line with high PSMA expression (LNCaP) as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(Glu) polyplexes at 0.0125 and 0.125 μg / mL payload, or without treatment (untreated control). Isotype and unstained controls show background fluorescence. Staggered histograms of fluorescence intensity are shown, and mean fluorescence intensity (MFI) is indicated. [Figure 4B] Flow cytometry analysis of MHC I expression on the cell surface of a prostate cancer cell line with low PSMA expression (DU145) as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(Glu) polyplexes at 0.0125 and 0.125 μg / mL payload, or without treatment (untreated control). Isotype and unstained controls show background fluorescence. Staggered histograms of fluorescence intensity are shown, and mean fluorescence intensity (MFI) is indicated. [Figure 5A]1 is a plot of cell viability in LNCaP cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG24-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG24-DUPA:poly(Glu). The x-axis shows the logarithm of the concentration of poly(IC) or poly(Glu) delivered. [Figure 5B] 1 is a plot of cell viability in PC-3 cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG24-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG24-DUPA:poly(Glu). The x-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 5C] 1 is a plot of cell viability in DU145 cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG24-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG24-DUPA:poly(Glu). The x-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 5D] 1 is a plot of cell survival in LNCaP cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG24-folate:poly(IC) and LPEI-l-[N3:DBCO]-PEG24-folate:poly(Glu). The x-axis shows the logarithm of the concentration of poly(IC) or poly(Glu) delivered. [Figure 5E] 1 is a plot of cell viability in DU145 cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG24-folate:poly(IC) and LPEI-l-[N3:DBCO]-PEG24-folate:poly(Glu). The x-axis shows the logarithm of the concentration of poly(IC) or poly(Glu) delivered. [Figure 6A] 1 is a plot of cell survival in LNCaP cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(Glu). The x-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 6B] 1 is a plot of cell viability in PC-3 cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(Glu). The x-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 6C] 1 is a plot of cell viability in DU145 cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(Glu). The x-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 7] 1 is a plot of cell viability in LNCaP cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC), LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(Glu), Me-LPEI[N3:DBCO]PEG36-[MAL-S]-DUPA:poly(IC), and Me-LPEI[N3:DBCO]PEG36-[MAL-S]-DUPA:poly(Glu). The X-axis shows the logarithm of the concentration of poly(IC) or poly(Glu) delivered. [Figure 8] 1 is a plot of cell viability in LNCaP cells as a function of treatment with LPEI-1-[N3:BCN]-PEG-[MAL-S]-DUPA:poly(IC), LPEI-1-[N3:BCN]-PEG-[MAL-S]-DUPA:poly(Glu), Me-LPEI[N3:BCN]PEG-[MAL-S]-DUPA:poly(IC), and Me-LPEI[N3:BCN]PEG-[MAL-S]-DUPA:poly(Glu). The X-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 9]1 is a plot of cell survival in DU145 prostate cancer cells with low PSMA expression as a function of treatment with LPEI-1-[N3:DBCO]-PEG36-DUPA:poly(IC), LPEI-1-[N3:DBCO]-PEG36-DUPA:poly(Glu), Me-LPEI[N3:DBCO]PEG36-[MAL-S]-DUPA:poly(IC), and Me-LPEI[N3:DBCO]PEG36-[MAL-S]-DUPA:poly(Glu). The X-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 10] 1 is a plot of cell survival in DU145 prostate cancer cells with low PSMA expression as a function of treatment with LPEI-1-[N3:BCN]-PEG36-DUPA:poly(IC), LPEI-1-[N3:BCN]-PEG36-DUPA:poly(Glu), Me-LPEI[N3:BCN]PEG36-[MAL-S]-DUPA:poly(IC), and Me-LPEI[N3:BCN]PEG36-[MAL-S]-DUPA:poly(Glu). The X-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 11] 1 is a plot of cell survival in LNCaP cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC); LPEI-l-[N3:DBCO]-PEG36-[(NH2)MAL-S]-DUPA:poly(IC); LPEI-l-[N3:BCN]-PEG36-DUPA:poly(IC); LPEI-l-[N3:SCO]-PEG36-[MAL-S]-DUPA:poly(IC); LPEI-l-[N3:DBCO]-PEG36-[CONH]-DUPA:poly(IC); and LPEI-l-[N3:DBCO]-PEG36-[S-MAL]-DUPA:poly(IC) polyplexes. The X-axis represents the logarithm of the concentration of delivered poly(IC). [Figure 12]1 is a plot of cell survival in VCaP prostate cancer cells with moderate PSMA cell surface expression as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(Glu). The X-axis indicates the concentration of delivered poly(IC) or poly(Glu). [Figure 13] 1 is a plot of cell viability in DU145 cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC); LPEI-l-[N3:DBCO]-PEG36-[(NH2)MAL-S]-DUPA:poly(IC); LPEI-l-[N3:BCN]-PEG36-DUPA:poly(IC); LPEI-l-[N3:SCO]-PEG36-[MAL-S]-DUPA:poly(IC); LPEI-l-[N3:DBCO]-PEG36-[CONH]-DUPA:poly(IC); and LPEI-l-[N3:DBCO]-PEG36-[S-MAL]-DUPA:poly(IC) polyplexes. The X-axis indicates the concentration of delivered poly(IC). [Figure 14A] 1 is a plot of IP-10 secretion as a function of LPEI-1-[N3:DBCO]-PEG24-DUPA:poly(IC) concentration in LNCaP and PC-3 cells. [Figure 14B] 1 is a plot of IP-10 secretion as a function of LPEI-1-[N3:DBCO]-PEG36-DUPA:poly(IC) concentration in LNCaP and PC-3 cells. [Figure 14C] 1 is a plot of IP-10 secretion as a function of LPEI-1-[N3:DBCO]-PEG36-DUPA:poly(IC) concentration in LNCaP and DU145 cells. [Figure 15A] 1 is a plot of RANTES secretion as a function of LPEI-1-[N3:DBCO]-PEG24-DUPA:poly(IC) concentration in LNCaP and PC-3 cells. [Figure 15B]1 is a plot of RANTES secretion as a function of LPEI-1-[N3:DBCO]-PEG36-DUPA:poly(IC) concentration in LNCaP and PC-3 cells. [Figure 15C] 1 is a plot of RANTES secretion as a function of LPEI-1-[N3:DBCO]-PEG36-DUPA:poly(IC) concentration in LNCaP and DU145 cells. [Figure 16A] 1 is a plot of IFNβ secretion as a function of LPEI-1-[N3:DBCO]-PEG24-DUPA:poly(IC) concentration in LNCaP and PC-3 cells. [Figure 16B] 1 is a plot of IFNβ secretion as a function of LPEI-1-[N3:DBCO]-PEG36-DUPA:poly(IC) concentration in LNCaP and PC-3 cells. [Figure 16C] 1 is a plot of IFNβ secretion as a function of LPEI-1-[N3:DBCO]-PEG36-DUPA:poly(IC) concentration in LNCaP and DU145 cells. [Figure 17] Western blot imaging analysis showing the qualitative levels of caspase-3, cleaved caspase-3, PARP, cleaved PARP, RIG-1;MDA5, and ISG15 as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(Glu) polyplexes at 0, 0.0625, and 0.625 μg / mL. GAPDH served as a protein loading control. [Figure 18]Immunoblot analysis of prostate cancer cells with high PSMA (LNCaP) and low PSMA expression (DU145) as a function of treatment with LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC) and LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(Glu) polyplexes at 0.02 and 0.2 μg / mL payload (poly(IC) and poly(Glu)), respectively, for 5 and 24 hours. The analysis shows qualitative levels of IκB, phospho-IκB, IRF3, phospho-IRF3, NFκB, phospho-NFκB, and PD-L1. GAPDH served as a protein loading control. [Figure 19] SEM image of polyplex particles containing compounds 31 and 31b and poly(IC), i.e., LPEI-l-[N3:DBCO]-PEG36-DUPA:poly(IC), formed at an N / P ratio of 4 and a concentration of 0.1875 mg / mL in 20 mM HEPES buffer, 5% glucose (HBG), pH 7.2. [Figure 20] Luminescence normalized to viability is shown in human prostate cell lines differing in cell surface expression of PSMA: PSMA-high-expressing LNCaP cells and PSMA-low-expressing DU145 cells, after transfection with PSMA-targeted polyplexes containing mRNA encoding luciferase. The X-axis indicates the concentration of mRNA in the polyplex (0.25, 0.5, and 1.0 μg / mL). The Y-axis indicates luminescence normalized to viability in arbitrary units (AU). Selective transfection of PSMA-overexpressing cells with Luc mRNA and selective expression of luciferase were demonstrated. [Figure 21] Figure 1 shows the levels of secreted human IL-2 from two cell lines with different PSMA expression: PSMA-high expressing LNCaP cells and PSMA-low expressing DU145 cells, after transfection with PSMA-targeted polyplexes containing hIL-2 mRNA. Selective expression of human IL-2 from PSMA-overexpressing cells is demonstrated. [Figure 22]Figure 1 shows the levels of secreted human IFN-β from two cell lines with different PSMA expression: PSMA-high expressing LNCaP cells and PSMA-low expressing DU145 cells, after transfection with PSMA-targeted polyplexes containing hIFN-β mRNA. Selective expression of human IFN-β from PSMA-high expressing cells is demonstrated. [Figure 23] This figure shows the inhibition of protein biosynthesis by DT-A protein in two cell lines with different PSMA expression: high-PSMA-expressing LNCaP cells and low-PSMA-expressing DU145 cells, after transfection with the PSMA-targeting polyplex LPEI-l-[N3:DBCO]PEG36-DUPA containing mRNA DT-A. Western blot analysis using anti-puromycin antibody as a probe was used to detect the inhibition of protein biosynthesis. GAPDH was used as a loading control. Selective inhibition of protein biosynthesis in PSMA-overexpressing cells is demonstrated. [Figure 24] Figure 1 shows luminescence from human prostate cell lines with different cell surface expression of PSMA: high-PSMA-expressing LNCaP cells and low-PSMA-expressing DU145 cells. Cells were treated with PSMA-targeted polyplexes containing plasmid DNA encoding luciferase. The X-axis shows the concentration of pGreenFire-CMV in the polyplex (0.25, 0.5, and 1.0 μg / mL). The Y-axis shows luminescence in arbitrary units (AU). The mean and standard deviation from triplicate samples are shown. Selective expression of luciferase after transfection of PSMA-overexpressing cells with plasmid DNA encoding luciferase (pGreenFire-CMV) is demonstrated. [Figure 25]Figure 1 shows the levels of secreted human IL-2 normalized to cell viability in cell lines with different PSMA expression: high-expressing LNCaP and C4-2 cells, and low-expressing DU145 cells, after transfection with PSMA-targeted polyplexes containing a plasmid encoding the IL-2 protein. The X-axis shows the concentration of hIL-2 plasmid DNA (0.25, 0.5, and 1.0 μg / mL) in the polyplex. The Y-axis shows the concentration of secreted IL-2 normalized to cell viability in arbitrary units (AU). Selective expression / secretion of human IL-2 after transfection of PSMA-overexpressing cells with plasmid DNA encoding hIL-2 is demonstrated. [Figure 26A] 1 is a plot of cell survival in MCF7 cells as a function of treatment with LPEI-1-[N3:DBCO]-PEG36-hEGF:poly(IC) and LPEI-1-[N3:DBCO]-PEG36-hEGF:poly(Glu). The X-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 26B] 1 is a plot of cell survival in A431 cells as a function of treatment with LPEI-1-[N3:DBCO]-PEG36-hEGF:poly(IC) and LPEI-1-[N3:DBCO]-PEG36-hEGF:poly(Glu). The X-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 27A] 1 is a plot of cell survival in MCF7 cells as a function of treatment with LPEI-1-[N3:DBCO]-PEG24-hEGF:poly(IC) and LPEI-1-[N3:DBCO]-PEG24-hEGF:poly(Glu). The X-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 27B] 1 is a plot of cell survival in A431 cells as a function of treatment with LPEI-l-[N3:DBCO]-PEG24-hEGF:poly(IC) and LPEI-l-[N3:DBCO]-PEG24-hEGF:poly(Glu). The x-axis shows the logarithm of the concentration of poly(IC) or poly(Glu) delivered. [Figure 28A]1 is a plot of cell survival in MCF7 cells as a function of treatment with the nontargeted polyplexes LPEI-l-[N3:DBCO]-PEG23-OMe:poly(IC) and LPEI-l-[N3:DBCO]-PEG23-OMe:poly(Glu). The x-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 28B] 1 is a plot of cell survival in A431 cells as a function of treatment with the nontargeted polyplexes LPEI-l-PEG23-OMe:poly(IC) and LPEI-l-PEG23-OMe:poly(Glu). The x-axis shows the logarithm of the concentration of delivered poly(IC) or poly(Glu). [Figure 29A] Plot of luminescence (AU) in Renca parental and Renca EGFR M1 H cells treated with LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] compared to the control delivery vehicle, Messenger MAX. After 24 hours of treatment, luminescence was measured at N / P ratios of 4, 6, and 12, and final concentrations of LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] and Lipofectamine Messenger MAX ranging from 0.125 to 1.0 μg / mL. [Figure 29B] Plot of luminescence (AU) in Renca parental and Renca EGFR M1 H cells treated with LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] compared to the control delivery vehicle, jetPEI. After 24 h of treatment, luminescence was measured at N / P ratios of 4, 6, and 12, and at final concentrations of LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] and jetPEI ranging from 0.125 to 1.0 μg / mL. [Figure 29C]This figure shows a plot of the luminescence ratio (AU) between Renca parental cells and Renca EGFR M1 H cells treated with LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] using Messenger MAX as a comparative delivery vehicle. After 24 hours of treatment, luminescence was measured at N / P ratios of 4, 6, and 12, and at final concentrations of LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] and Lipofectamine Messenger MAX ranging from 0.125 to 1.0 μg / mL. The ratio was calculated by dividing the luminescence signal from Renca EGFR M1 H cells by the luminescence signal from Renca parental cells. [Figure 29D] This figure shows a plot of the luminescence ratio (AU) between Renca parental cells and Renca EGFR M1 H cells treated with LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] using jetPEI as a comparative delivery vehicle. After 24 hours of treatment, luminescence was measured at N / P ratios of 4, 6, and 12 and final concentrations of LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] and jetPEI ranging from 0.125 to 1.0 μg / mL. The ratio was calculated by dividing the mean luminescence signal from Renca EGFR M1 H cells by the mean luminescence signal from Renca parental cells. [Figure 29E] Figure 1 shows a plot of percent survival in Renca parental and Renca EGFR M1 H cells treated with LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] compared to the control delivery vehicle, Messenger MAX. After 24 hours of treatment, percent survival was measured at N / P ratios of 4, 6, and 12, and at final concentrations of LPEI-l-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] and Messenger MAX ranging from 0.125 to 1.0 μg / mL. [Figure 30A] Relative luminescence (AU) in Renca parental and Renca EGFR M1 H cells treated with LPEI-1-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] after 6 hours of treatment at an N / P ratio of 4 is shown. [Figure 30B] Relative luminescence (AU) in Renca parental and Renca EGFR M1 H cells treated with LPEI-1-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] after 6 hours of treatment at an N / P ratio of 6 is shown. [Figure 30C] Relative luminescence (AU) in Renca parental and Renca EGFR M1 H cells treated with LPEI-1-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] after 22 hours of treatment at an N / P ratio of 4 is shown. [Figure 30D] Relative luminescence (AU) in Renca parental and Renca EGFR M1 H cells treated with LPEI-1-[N3:DBCO]PEG36-hEGF:[Fluc mRNA] after 22 hours of treatment at an N / P ratio of 6 is shown. [Figure 31] Figure 1 shows luminescence from cancer cells with different cell surface expression of human folate receptor (FR) (MCF7: low folate receptor expression; SKOV3: high folate receptor expression) after treatment with FR-targeted polyplexes containing mRNA encoding Renilla luciferase (R-Luc). The X-axis shows the concentration of mRNA in the polyplex (0.125, 0.25, 0.5, and 1.0 μg / mL). The Y-axis shows luminescence in arbitrary units (RLU). The standard deviation from quadruplicate samples is shown. Selective expression of Renilla Luc in folate receptor-overexpressing cells is demonstrated. [Figure 32] Figure 1 shows the levels of secreted human IL-2 normalized to survival from two cell lines with different human EGFR (hEGFR) expression: hEGFR-high-expressing RencaEGFR M1 H cells and human EGFR-negative Renca (parent) cells, after transfection with EGFR-targeted polyplexes containing hIL-2 mRNA. The X-axis shows the concentration of mRNA in the polyplex (0.125, 0.25, 0.5, and 1.0 μg / mL). The Y-axis shows the levels of secreted human IL-2 normalized to survival in arbitrary units (AU). Selective expression and secretion of human IL-2 from EGFR-high-expressing cells is demonstrated. [Figure 33]1 shows the levels of secreted human IFNγ (hIFNγ) from RencaEGFR M1 H (high expression of human EGFR) and Renca (parental, negative, no expression of human EGFR) cell lines after transfection with EGFR-targeted polyplexes containing hIFNγ mRNA. Selective transfection of EGFR-overexpressing cells with hIFNγ mRNA and selective expression and secretion of hIFNγ protein are demonstrated. [Figure 34] Figure 1 shows the levels of human EPO secreted by cancer cells with differential expression of human folate receptor (FR) (SKOV3: high FR expression; MCF7: low FR expression) after treatment with FR-targeted polyplexes containing mRNA encoding human EPO. The X-axis shows the concentration of mRNA in the polyplex (0.125, 0.25, 0.5, and 1.0 μg / mL). The Y-axis shows the concentration of hEPO released into the medium (mIU / mL). Standard deviations from quadruplicate samples are shown. Selective expression of hEPO in folate receptor-overexpressing cells is demonstrated. [Figure 35A] Figure 35 shows cell surface expression of human EGFR in various cell lines: RencaEGFR M1 H, WI-38, and MCF-7 cells. The data shown in Figures 35A and 35B are from two separate experiments using different flow cytometers. [Figure 35B] Figure 35 shows cell surface expression of human EGFR in various cell lines: WI-38, U87MG, and MCF-7 cells. The data shown in Figures 35A and 35B are from two separate experiments using different flow cytometers. [Figure 35C] Figure 1 shows luminescence levels normalized to cell viability from high EGFR-expressing RencaEGFR M1 H cells and low EGFR-expressing MCF7 cells after transfection with EGFR-targeted polyplexes containing LPEI-l-[N3:DBCO]PEG36-hEGF and a luciferase-encoding plasmid formulated at an N / P ratio of 6. Selective expression and activity of luciferase in EGFR-overexpressing cells is demonstrated. [Figure 35D]The levels of luminescence normalized to cell viability are shown for additional cell lines: rapidly proliferating cancerous U87MG cells expressing moderate levels of EGFR; slowly proliferating noncancerous WI38 cells expressing moderate levels of EGFR; and slowly proliferating noncancerous HUVEC cells expressing minimal or no EGFR. These cells were transfected with EGFR-targeted polyplexes containing LPEI-1-[N3:DBCO]PEG36-hEGF and a luciferase-encoding plasmid (N / P ratio of 6) in the same experiment as the cells shown in Figure 35C. Selective expression of luciferase in rapidly proliferating cancerous cells expressing moderate levels of EGFR is demonstrated. [Figure 36A] Figure 1 shows luminescence levels in two cell lines with different human EGFR expression: high EGFR-expressing RencaEGFR M1 H cells and human EGFR-negative Renca (parental) cells, after transfection with a linear EGFR-targeted polyplex of the invention containing LPEI-l-[N3:DBCO]PEG36-hEGF and a plasmid (pGreenFire1-CMV) encoding luciferase produced at an N / P ratio of 3. Selective expression of luciferase in EGFR-overexpressing cells is demonstrated. [Figure 36B] Figure 1 shows luminescence levels in two cell lines with different human EGFR expression: high EGFR-expressing RencaEGFR M1 H cells and human EGFR-negative Renca (parental) cells, after transfection with a linear EGFR-targeted polyplex of the invention containing LPEI-l-[N3:DBCO]PEG36-hEGF and a plasmid (pGreenFire1-CMV) encoding luciferase produced at an N / P ratio of 4. Selective expression of luciferase in EGFR-overexpressing cells is demonstrated. [Figure 37A]Figure 1 shows the levels of secreted human IL-2 (hIL-2) from two cell lines with different human EGFR expression: high EGFR-expressing RencaEGFR M1 H cells and human EGFR-negative parental Renca cells, after transfection with EGFR-targeted polyplexes containing plasmids encoding LPEI-l-[N3:DBCO]PEG36-hEGF and hIL-2. Selective expression of hIL-2 from EGFR-overexpressing cells is demonstrated. [Figure 37B] Figure 1 shows the levels of secreted human IL2 after transfection of low numbers of high EFGR-expressing RencaEGFR M1 H cells (600 cells) with EGFR-targeted polyplexes containing plasmids encoding LPEI-l-[N3:DBCO]PEG36-hEGF and hIL2 at the indicated concentrations of plasmid (0.125 and 0.25 μg / ml). Polyplexes were formulated at an N / P ratio of 6, and IL2 secretion was detected after 2, 3, and 4 days. [Figure 38] Figure 1 shows the levels of secreted human IFNβ from RencaEGFR M1 H cancer cells, which have high human EGFR expression, after transfection with two different EGFR-targeting polyplexes containing pCMV-hIFNβ at two different N / P ratios (N / P 3 and N / P 4). The X-axis indicates the concentration of pCMV-hIFNβ plasmid DNA (0.25, 0.5, 1.0, and 2.0 μg / mL) in the polyplexes. The Y-axis indicates the concentration of secreted IFNβ protein (pg / mL), shown as the mean with standard deviation from triplicate samples. Secretion of human IFNβ from EGFR-overexpressing cancer cells was demonstrated for the delivery vectors tested, with a significant advantage of the linear triconjugate vector LPEI-l-[N3:DBCO]-PEG36-hEGF compared to each random delivery vector. [Figure 39] 1 is a plot of fluorescence intensity measured by flow cytometry analysis of EGFR cell surface expression on B16F10 parental cells and B16F10 cells stably transfected with human EGFR (B16F10-hEGFR). [Figure 40]Figure 1 is a plot of B16F10-hEGFR subcutaneous tumor growth in mice as a function of time (days) after intravenous administration of 1.25 mg / kg LPEI-l-[N3:DBCO]-PEG36-hEGF:poly(IC) three times per week compared to treatment with buffer control (5% glucose). Mean tumor volume and standard error of the mean are shown. [Figure 41] Western blot analysis using anti-EGF receptor antibody (D38B1) XP (CST #4267). Protein lysates were prepared for each cell line, electrophoresed, and subjected to immunoblot analysis. Actin (Millipore #MAB1501) indicates equal loading of total protein. [Figure 42] 1 is a plot of HSC-3 cell survival after 72 hours of treatment with LPEI-1-[N3:DBCO]-PEG36-hEGF:poly(IC). The X-axis indicates the concentration of poly(IC) delivered. [Figure 43] Figure 1 shows a plot of head and neck subcutaneous tumor growth as a function of time (days) after intravenous administration of LPEI-l-[N3:DBCO]-PEG36-hEGF:poly(IC) at 1.25 mg / kg three times per week compared to untreated mice. Mice were treated up to day 25, and the mean tumor volume and standard error of the mean are shown. [Figure 44] 1 is a plot of IFN-γ secretion by unstimulated PBMCs or PBMCs pre-stimulated with TransAct™ after incubation with medium from cancer cells treated for 24 hours with vehicle control, or LPEI-l-[N3:DBCO]-PEG36-hEGF:poly(IC) or LPEI-l-[N3:DBCO]-PEG36-hEGF:poly(Glu) control polyplexes at the indicated concentrations. DETAILED DESCRIPTION OF THE INVENTION

[0127] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The embodiments, preferred embodiments, and highly preferred embodiments described and disclosed herein should apply to all aspects and other embodiments, preferred embodiments, and highly preferred embodiments, whether or not specifically mentioned again.

[0128] The present invention provides linear conjugates of LPEI and PEG that can form polyplexes with polyanions and nucleic acids, such as poly(IC), as outlined herein and below. The conjugates preferably comprise an LPEI fragment, a PEG fragment, and a targeting fragment. In preferred embodiments, the LPEI fragment and the PEG fragment are coupled in a discrete end-to-end manner. In some preferred embodiments, the LPEI fragment and the PEG fragment are coupled via covalent attachment of an azide to an alkene or alkyne to form a 1,2,3-triazole or a 4,5-dihydro-1H-[1,2,3]triazole.

[0129] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0130] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0131] The term "and / or" is used in this disclosure to mean either "and" or "or," unless otherwise indicated.

[0132] As used herein, the term "about" shall mean ±10%. For example, about 50% shall mean 45% to 55%. Preferably, as used herein, the term "about" shall mean ±5%. For example, about 50% shall mean 47.5% to 52.5%.

[0133] As used herein, the phrase "number X to number Y" includes number X and number Y. For example, the phrase "0.01 μmol to 50 μmol" refers to 0.01 μmol, 50 μmol, and values ​​therebetween. The same applies to the phrase "about number X to about number Y."

[0134] The term "optionally substituted" is understood to mean that a given chemical moiety (e.g., an alkyl group) can (but is not required to) be bonded to other substituents (e.g., heteroatoms). For example, an optionally substituted alkyl group can be a fully saturated alkyl chain (i.e., pure hydrocarbon). Alternatively, the same optionally substituted alkyl group can have substituents other than hydrogen. For example, it can be bonded at any point along the chain to a halogen atom, an alkoxy group, or any other substituent described herein. Thus, the term "optionally substituted" means that a given chemical moiety has the potential to contain other functional groups, but does not necessarily have additional functional groups.

[0135] The term "optionally replaced" is understood to refer to a situation in which a carbon atom of a methylene group (i.e., -CH-) may, but need not, be replaced by a heteroatom (e.g., -NH-, -O-). For example, a C alkylene (i.e., propylene) group in which one of the methylene groups is "optionally replaced" can have the structure -CH-O-CH- or -O-CH-CH-. Those skilled in the art will understand that a methylene group cannot be replaced if such a replacement would result in an unstable chemical moiety. For example, those skilled in the art will understand that four methylene groups cannot be replaced by oxygen atoms simultaneously. Thus, in some preferred embodiments, when one methylene group of an alkylene fragment is replaced with a heteroatom, one or both of the adjacent carbon atoms are not replaced with a heteroatom.

[0136] The term "aryl" refers to cyclic aromatic hydrocarbon groups having one to two aromatic rings, including monocyclic or bicyclic groups such as phenyl, biphenyl, or naphthyl. 10 An aryl group contains 6 to 10 carbon atoms. When containing two aromatic rings (such as bicyclic), the aromatic rings of the aryl group may be joined at a single point (e.g., biphenyl) or fused (e.g., naphthyl). The aryl group is optionally substituted at any point of attachment with one or more substituents, e.g., 1 to 5 substituents, which are themselves optionally substituted. Furthermore, when containing two fused rings, the aryl group defined herein can have an unsaturated or partially saturated ring fused to a fully saturated ring. Exemplary ring systems of these aryl groups include indanyl, indenyl, tetrahydronaphthalenyl, and tetrahydrobenzoannulenyl. In some preferred embodiments, the aryl group is a phenyl group.

[0137] Unless otherwise specifically defined, "heteroaryl" means a monovalent monocyclic or polycyclic aromatic ring of 5 to 24 ring atoms containing one or more ring heteroatoms selected from N, S, P, or O, with the remaining ring atoms being C. A 5-10 membered heteroaryl group contains 5 to 10 atoms. Heteroaryl, as defined herein, also refers to a bicyclic heteroaromatic group in which the heteroatoms are selected from N, S, P, or O. Aromatic groups are optionally substituted independently with one or more substituents described herein. Examples include, but are not limited to, furyl, thienyl, pyrrolyl, pyridyl, pyrazolyl, pyrimidinyl, imidazolyl, isoxazolyl, oxazolyl, oxadiazolyl, pyrazinyl, indolyl, thiophen-2-yl, quinolyl, benzopyranyl, isothiazolyl, thiazolyl, thiadiazole, indazole, benzimidazolyl, thieno[3,2-b]thiophene, triazolyl, triazinyl, imidazolyl, and the like. Dazo[1,2-b]pyrazolyl, furo[2,3-c]pyridinyl, imidazo[1,2-a]pyridinyl, indazolyl, pyrrolo[2,3-c]pyridinyl, pyrrolo[3,2-c]pyridinyl, pyrazolo[3,4-c]pyridinyl, thieno[3,2-c]pyridinyl, thieno[2,3-c]pyridinyl, thieno[2,3-b]pyridinyl, benzothiazolyl, indolyl, indolinyl, indolinonyl, dihydrobenzothio Phenyl, dihydrobenzofuranyl, benzofuran, chromanyl, thiochromanyl, tetrahydroquinolinyl, dihydrobenzothiazine, dihydrobenzoxanyl, quinolinyl, isoquinolinyl, 1,6-naphthyridinyl, benzo[de]isoquinolinyl, pyrido[4,3-b][1,6]naphthyridinyl, thieno[2,3-b]pyrazinyl, quinazolinyl, tetrazolo[1,5-a]pyridinyl, [1,2,4]thiazolinyl Zolo[4,3-a]pyridinyl, isoindolyl, pyrrolo[2,3-b]pyridinyl, pyrrolo[3,4-b]pyridinyl, pyrrolo[3,2-b]pyridinyl, imidazo[5,4-b]pyridinyl, pyrrolo[1,2-a]pyrimidinyl, tetrahydropyrrolo[1,2-a]pyrimidinyl, 3,4-dihydro-2H-1λ2-pyrrolo[2,1-b]pyrimidine, dibenzo[b,d]thiophene, pyridin-2-one, furo[3,2-c]pyridinyl, furo[2,3-c]pyridinyl, 1H-pyrido[3,4-b][1,4]thiazinyl, benzoxazolyl, benzisoxazolyl, furo[2,3-b]pyridinyl, benzothiophenyl, 1,5-naphthyridinyl, furo[3,2-b]pyridinyl, [1,2,4]triazolo[1,5-a]pyridinyl, benzo[1,2,3]triazolyl, imidazo[1,2-a]pyrimidinyl, [1,2,4]triazolo[4,3-b]pyridazinyl, benzo[c] [1,2,5]thiadiazolyl, benzo[c][1,2,5]oxadiazole, 1,3-dihydro-2H-benzo[d]imidazol-2-one, 3,4-dihydro-2H-pyrazolo[1,5-b][1,2]oxazinyl, 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridinyl, thiazolo[5,4-d]thiazolyl, imidazo[2,1-b][1,3,4]thiadiazolyl, thieno[2,3-b]pyrrolyl, 3H-indolyl, and derivatives thereof. Furthermore, when containing two fused rings, heteroaryl groups as defined herein can have an unsaturated or partially saturated ring fused to a fully saturated ring. Exemplary ring systems of these heteroaryl groups include indolinyl, indolinonyl, dihydrobenzothiophenyl, dihydrobenzofuran, chromanyl, thiochromanyl, tetrahydroquinolinyl, dihydrobenzothiazine, 3,4-dihydro-1H-isoquinolinyl, 2,3-dihydrobenzofuran, indolinyl, indolyl, and dihydrobenzoxanyl.

[0138] The term "alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon. A C1-C6 alkyl group contains 1 to 6 carbon atoms. Examples of C1-C6 alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, and neopentyl.

[0139] The term "alkylene" refers to a straight-chain or branched, saturated divalent hydrocarbon fragment. A C0-C6 alkyl group contains from 0 to 6 carbon atoms. Examples of C0-C6 alkylene groups include, but are not limited to, methylene, ethylene, propylene, butylene, pentylene, isopropylene, isobutylene, sec-butylene, tert-butylene, isopentylene, and neopentylene.

[0140] The term "C1-C6-alkoxy" as used herein refers to a substituted hydroxyl of the formula (-OR'), where R' is an optionally substituted C1-C6 alkyl as defined herein, and the oxygen moiety is directly attached to the parent molecule; thus, the term "C1-C6 alkoxy" as used herein refers to a straight-chain or branched C1-C6 alkoxy, which can be, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, straight-chain or branched pentoxy, straight-chain or branched hexyloxy. Preferred alkoxy is C1-C4 alkoxy, and C1-C3 alkoxy.

[0141] The term "cycloalkyl" means a monocyclic or polycyclic saturated carbocyclic ring containing 3 to 18 carbon atoms. A C3-C8 cycloalkyl contains 3 to 8 carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptanyl, cyclooctanyl, norboranyl, norborenyl, bicyclo[2.2.2]octanyl, or bicyclo[2.2.2]octenyl. A C3-C8 cycloalkyl is a cycloalkyl group containing 3 to 8 carbon atoms.

[0142] The term "cycloalkenyl" means a monocyclic non-aromatic unsaturated carbocyclic ring containing 5 to 18 carbon atoms. Examples of cycloalkenyl groups include, but are not limited to, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, and norborenyl. A C5-C8 cycloalkenyl is a cycloalkenyl group containing 5 to 8 carbon atoms.

[0143] The terms "heterocyclyl" or "heterocycloalkyl" or "heterocycle" refer to monocyclic or polycyclic 3- to 24-membered rings containing carbon and heteroatoms derived from oxygen, nitrogen, or sulfur, and lacking delocalized π-electrons (aromaticity) shared between ring carbons or heteroatoms. 3- to 10-membered heterocycloalkyl groups contain 3-10 atoms. Heterocyclyl rings include, but are not limited to, oxetanyl, azetazinyl, tetrahydrofuranyl, pyrrolidinyl, oxazolinyl, oxazolidinyl, thiazolinyl, thiazolidinyl, pyranyl, thiopyranyl, tetrahydropyranyl, dioxalinyl, piperidinyl, morpholinyl, thiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholinyl S-dioxide, piperazinyl, azepinyl, oxepinyl, diazepinyl, tropanyl, and homotropanyl.

[0144] The term "heterocycloalkenyl" refers to a monocyclic or polycyclic 3- to 24-membered ring containing carbon and heteroatoms derived from oxygen, nitrogen, or sulfur, in which there are no delocalized π-electrons (aromaticity) shared between ring carbons or heteroatoms, but at least one unsaturated element is present within the ring. 3- to 10-membered heterocycloalkenyl groups contain 3 to 10 atoms.

[0145] As used herein, the term "halo" or "halogen" means fluoro (F), chloro (Cl), bromo (Br), or iodo (I).

[0146] The term "carbonyl" refers to a functional group consisting of a carbon atom double-bonded to an oxygen atom. It may be abbreviated herein as "oxo," C(O), or C=O.

[0147] The term "overexpression" refers to increased expression of a gene or protein in a cell or on the cell surface compared to basal or normal expression. In a preferred embodiment, the targeting fragment is capable of binding to a cell that overexpresses a cell surface receptor. In one embodiment, cells that overexpress a cell surface receptor refer to an elevated level of the cell surface receptor expressed in the cells of a particular tissue compared to the level of the cell surface receptor measured in normal healthy cells of the same tissue type under similar conditions. In one embodiment, cells that overexpress a cell surface receptor refer to an increased level of the cell surface receptor in a cell compared to the level in the same cell or a closely related non-malignant cell under normal physiological conditions.

[0148] The term "polyanion" as used herein refers to a polymer, preferably a biopolymer, having two or more negatively charged sites. Typically and preferably, the term "polyanion" as used herein refers to a polymer, preferably a biopolymer, composed of repeating units that contain residues that can carry a negative charge. In a further embodiment, the polyanion is a polymer, preferably a biopolymer, composed of repeating units that contain negatively charged residues. In another preferred embodiment, the polyanion is a nucleic acid, more preferably DNA, RNA, polyglutamic acid, or hyaluronic acid.

[0149] As used herein, the term "nucleic acid" includes deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA), or a combination thereof. In preferred embodiments, the term "nucleic acid" refers to deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA), and as used herein, refers to genomes, viruses, and recombinantly prepared and chemically synthesized molecules. Nucleic acids can be single- or double-stranded and in the form of linear or covalently closed circular molecules, can include chemical derivatization of nucleic acids on the nucleotide bases, sugars, or phosphates, and can contain non-natural nucleotides and nucleotide analogs.

[0150] As used herein, the term "dispersity" (abbreviated as D) refers to the distribution of molar masses in a given polymer sample, such as in polymer fragments, as used herein for the conjugates and polyplexes of the invention. This is defined herein as D = (M w / M n ), where D is the dispersity; M w is the weight average molecular weight of the polymer sample or polymer fragment; M n is the number average molecular weight of the polymer sample or polymer fragment.

[0151] The term "polydispersity index" (abbreviated as PDI) as used herein refers to the polydispersity index in dynamic light scattering measurements of polyplex nanoparticles, such as the polyplexes of the present invention. This index is a number calculated from a simple two-parameter fit to correlation data (cumulant analysis). The polydispersity index is dimensionless and is scaled so that values ​​below 0.05 are rarely found except in highly monodisperse standards. Values ​​above 0.7 indicate a sample with a very broad size distribution and are likely unsuitable for dynamic light scattering (DLS) techniques. Various size distribution algorithms work with data that fall between these two extremes. The zeta-average diameter (z-average diameter) and polydispersity index of the polyplexes of the present invention are determined by dynamic light scattering (DLS) based on the assumption that the polyplexes are isotropic and spherical. The calculation of these parameters is defined and determined in accordance with the ISO standard document ISO 22412:2017.

[0152] The term "amino acid residue" refers to a divalent residue derived from an organic compound containing the functional groups amine (-NH) and carboxylic acid (-COOH), typically and preferably with a side chain specific to each amino acid. In a preferred embodiment of the present invention, the amino acid residue is a divalent residue derived from an organic compound containing the functional groups amine (-NH) and carboxylic acid (-COOH), the divalency being achieved by the amine and the carboxylic acid functional groups, and thus by the -NH- and -CO- moieties. In an alternative preferred embodiment of the present invention, the amino acid residue is a divalent residue derived from an organic compound containing the functional groups amine (-NH) and carboxylic acid (-COOH), the divalency being achieved by the amine or the carboxylic acid functional group and by further functional groups present in the amino acid residue. As a preferred example and embodiment, an amino acid residue according to the present invention derived from cysteine ​​comprises the divalent structure S-(CH)-CH(COOH)-NH-, the divalency being achieved by the amino functionality and the contained thiol functionality. The term "amino acid residue" as used herein typically and preferably includes amino acid residues derived from natural or non-natural amino acids. Furthermore, the term "amino acid residue" as used herein typically and preferably also includes amino acid residues derived from chemically synthesized non-natural amino acids, including alpha- (α-), beta- (β-), gamma- (γ-), or delta- (δ-) amino acids, as well as mixtures thereof in any ratio. Furthermore, the term "amino acid residue" as used herein typically and preferably also includes amino acid residues derived from alpha amino acids, including any isomeric forms thereof, particularly their D- and L-stereoisomers (alternatively addressed by the (R) and (S) nomenclature), as well as mixtures thereof in any ratio, preferably a 1:1 racemic ratio. The terms "D-stereoisomer," "L-stereoisomer," "D-amino acid," or "L-amino acid" refer to the chiral alpha carbon of an amino acid. Thus, in a preferred embodiment, the amino acid residue is a divalent group of the structure -NH-CHR-C(O)-, where R is the amino acid side chain.Two or more consecutive amino acid residues preferably form a peptide (i.e., amide) bond at both the amine and carboxylic acid moieties of each amino acid residue. Di-, tri-, or polypeptide amino acid residues are typically (AA). a When described herein as a nucleotide, the sequence provided is presented from left to right in the NC direction. Thus, for example, Trp-Trp-Gly (AA) a shall refer to the amino acid residues corresponding to the N-terminus of the tripeptide, where Trp has an -NH- valence, and Gly has an -CO- valence, corresponding to the C-terminus of the tripeptide.

[0153] As used herein, the terms "peptide," "polypeptide," and "protein" refer to a substance comprising about two or more consecutive amino acid residues linked together via peptide bonds. The terms "peptide," "polypeptide," and "protein" are used interchangeably herein to refer to polymers of amino acid residues of any length. In one embodiment, the term "protein" refers to large peptides, particularly peptides having at least about 151 amino acids; in one embodiment, the term "peptide" refers to a substance containing about 2 or more, about 3 or more, about 8 or more, or about 20 or more, and up to about 50, about 100, or about 150.

[0154] As used herein, the term "epitope" refers to an antigenic determinant in a molecule such as an antigen. An epitope of a protein preferably comprises a continuous or discontinuous portion of the protein and is preferably 5 to 100, preferably 5 to 50, more preferably 8 to 30, and most preferably 10 to 25 amino acids in length; for example, an epitope may be preferably 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length.

[0155] The term "antibody" refers to any immunoglobulin, whether natural or wholly or partially synthetically produced, as well as derivatives and characteristic portions thereof. Antibodies can be monoclonal or polyclonal. Antibodies can be members of any immunoglobulin class, including any of the human classes: IgG, IgM, IgA, IgD, and IgE. As used herein, an antibody fragment (i.e., a characteristic portion of an antibody) refers to any derivative of an antibody that is less than full-length. Generally, an antibody fragment retains at least a significant portion of the full-length antibody's specific binding ability. Examples of antibody fragments include, but are not limited to, single-chain and double-chain fragments, Fab, Fab', F(ab')2, scFv, Fv, dsFv diabody, and Fd fragments. Antibody fragments can be produced by any means. For example, antibody fragments can be enzymatically or chemically produced by fragmentation of an intact antibody and / or recombinantly produced from a gene encoding a partial antibody sequence. Alternatively, or additionally, antibody fragments can be wholly or partially synthetically produced. Antibody fragments optionally include single-chain antibody fragments. Alternatively or additionally, antibody fragments may comprise multiple chains linked together, for example, by disulfide bonds. Antibody fragments may optionally include multimolecular complexes. Functional antibody fragments typically comprise at least about 50 amino acids, more typically at least about 200 amino acids. In some embodiments, antibodies may include chimeric (e.g., "humanized") antibodies and single-chain (recombinant) antibodies. In some embodiments, antibodies may have reduced effector function and / or bispecific molecules. In some embodiments, antibodies may include fragments produced by a Fab expression library. Single-chain Fvs (scFvs) are recombinant antibody fragments consisting only of a variable light chain (VL) and a variable heavy chain (VH) covalently linked to each other by a polypeptide linker. Either the VL or VH may comprise an NH2-terminal domain. The polypeptide linker may be of variable length and composition, so long as the two variable domains are bridged without significant steric hindrance. Typically, the linker contains a stretch of primarily glycine and serine residues, with some glutamic acid or lysine residues interspersed for solubility.Diabodies are dimeric scFvs. Diabodies typically have shorter peptide linkers than most scFvs and often preferentially associate as dimers. Fv fragments are antibody fragments consisting of one VH and one VL domain held together by noncovalent interactions. As used herein, the term "dsFv" refers to an Fv with an engineered intermolecular disulfide bond to stabilize the VH-VL pair. F(ab')2 fragments are antibody fragments essentially equivalent to those obtained from immunoglobulins by digestion with the enzyme pepsin at pH 4.0-4.5. Fragments can be recombinantly produced. Fab' fragments are antibody fragments essentially equivalent to those obtained by reduction of the disulfide bridge(s) linking the two heavy chain pieces in the F(ab')2 fragment. Fab' fragments can be recombinantly produced. 1. Fab fragments are antibody fragments essentially equivalent to those obtained by digesting immunoglobulins with an enzyme (e.g., papain). Fab fragments can be recombinantly produced. The heavy chain segment of the Fab fragment is the Fd piece.

[0156] As used herein, the term "alpha end of a linear polyethyleneimine fragment" (α-end of an LPEI fragment) refers to the terminal end of an LPEI fragment at which initiation of polymerization occurs using an electrophilic initiator as further described below for the term "initiating residue."

[0157] As used herein, the term "omega end of a linear polyethyleneimine fragment" (ω end of an LPEI fragment) refers to the terminal end of an LPEI fragment at which polymerization termination occurs using a nucleophile, such as an azide, thiol, or other nucleophile, as described herein.

[0158] The term "organic residue" refers to any suitable organic group that can be bonded to a nitrogen atom embedded within an LPEI fragment. In a preferred embodiment, the organic residue is bonded to the nitrogen atom via a carbonyl group to form an amide bond. Without wishing to be bound by theory, the organic residue is incorporated into the nitrogen atom of a poly(2-oxazoline) during ring-opening polymerization of the 2-oxazoline (e.g., Glassner et al., (2018), Poly(2-oxazoline)s: A comprehensive overview of polymer structures and their physical properties. Polym. Int, 67:32-45. https: / / doi.org / 10.1002 / pi.5457). Typically and preferably, the organic residue is cleaved from the poly(2-oxazoline) (i.e., the amide is typically cleaved) to yield the -(NH-CH-CH)- moiety embedded within the LPEI and LPEI fragment, and thus the conjugate of the present invention. However, if the cleavage reaction is not complete, a fraction of the organic residue is not cleaved. Thus, in a preferred embodiment of the present invention, R of the conjugates of the present invention, including those of formula I* and I 1 -(NR 2 -CH2-CH2) n -R in part 2 at least 80%, preferably 90%, of the R of the conjugates of the invention, including those of formula I* or I, are H; 1 -(NR 2 -CH2-CH2) n -R in part 2 Preferably at least 91%, more preferably 92%, more preferably 93%, more preferably 94%, more preferably 95%, more preferably 96%, more preferably 97%, more preferably 98%, and most preferably 99% of is H.

[0159] The term "initial residue" refers to the LPEI fragment and R 1 -(NR 2 -CH2-CH2) n- refers to a residue present in the - moiety, which is derived from any initiator, typically and preferably any electrophilic initiator, capable of initiating the polymerization of poly(2-oxazoline) from 2-oxazoline. As described in Glassner et al., (2018), Poly(2-oxazoline)s: A comprehensive overview of polymer structures and their physical properties. Polym. Int, 67:32-45. https: / / doi.org / 10.1002 / pi.5457, "Different initiator systems can be used, including toluenesulfonic acid (TsOH), or alkylsulfonates such as methyl p-toluenesulfonate (MeOT), which are most frequently found in the literature, p-nitrobenzenesulfonate (nosylate) and trifluoromethanesulfonate (triflate), alkyl, benzyl and acetyl halides, oxazolinium salts and Lewis acids." Thus, in a preferred embodiment, R 1 is -H or -CH3, but those skilled in the art will recognize that R 1 is any other suitable residue, e.g., C where n is greater than 1. n Alkyl groups, typically C 1-6 It will be understood that these groups may also include, but are not limited to, alkyl groups, benzyl groups, or acetyl groups.

[0160] In one aspect, the invention provides a composition comprising a conjugate, the conjugate comprising a linear polyethyleneimine fragment comprising an alpha end and an omega end; a polyethylene glycol fragment comprising a first end and a second end; the alpha end of the polyethyleneimine fragment is an initiating residue; and the omega end of the polyethyleneimine fragment is a covalent linking group, -ZX 1 -ZX 1 - is not a single bond, -Z- is not an amide; -X 1is a divalent covalent linking moiety; the second end of the polyethylene glycol fragment is capable of binding to a targeting fragment, and preferably the polyethylene glycol fragment is bound to a targeting fragment. In a preferred embodiment of this aspect, the composition consists of the conjugate. In a preferred embodiment, the linear polyethyleneimine fragment has the formula R 1 -(NR 2 -CH2-CH2) n -, and n is an integer of 1 to 1500. In a more preferred embodiment, the R 1 -(NR 2 -CH2-CH2) n The - moiety is a dispersed polymer moiety having a repeating unit n of about 115 to about 1150 and a dispersity of about 5 or less, preferably a repeating unit n of about 280 to about 700 and a dispersity of about 3 or less, more preferably a repeating unit n of about 350 to about 630 and a dispersity of about 2 or less, and preferably R 1 is -H or -CH3.

[0161] In another aspect, the present invention provides a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof: R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L (Formula I*); wherein n is any integer from 1 to 1500; m is any integer from 1 to 200, preferably m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90%, of X are H; 1 and X 2 are independently a divalent covalent linking moiety; -ZX 1- is a bivalent covalent linking moiety where -Z- is not -NHC(O)-; L is a targeting fragment preferably capable of binding to a cell, and preferably the composition consists of the conjugate.

[0162] In another aspect, the present invention provides a conjugate of formula I*, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof: R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L (Formula I*); wherein n is any integer from 1 to 1500; m is any integer from 1 to 200, preferably m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90%, of X are H; 1 and X 2 are independently a divalent covalent linking moiety; -ZX 1 - is a bivalent covalent linking moiety where -Z- is not -NHC(O)-; L is a targeting fragment preferably capable of binding to a cell.

[0163] In another aspect, the present invention provides a conjugate of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof: R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L (Formula I*); wherein n is any integer from 1 to 1500; m is any integer from 1 to 200, preferably m is any integer from 1 to 100; R1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90%, of X are H; 1 and X 2 are independently divalent covalent linking moieties; Z is a divalent covalent linking moiety; ZX 1 - is not a single bond; -Z- is not -NHC(O)-; L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor, and preferably the composition consists of the conjugate.

[0164] In another aspect, the present invention provides a conjugate of formula I*, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof: R 1 -(NR 2 -CH2-CH2) n -ZX 1 -(O-CH2-CH2) m -X 2 -L (Formula I*); wherein n is any integer from 1 to 1500; m is any integer from 1 to 200, preferably m is any integer from 1 to 100; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably 90%, of X are H; 1 and X 2 are independently a divalent covalent linking moiety; Z is a divalent covalent linking moiety; -ZX 1- is not a single bond, -Z- is not -NHC(O)-; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0165] Thus, in one aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A is one or more R A1 is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with; R A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with; R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor.

[0166] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single bond or a double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% are H; ring A may contain one or more R A1R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y 1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C-C alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23-, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, -H, -CO2H, C1-C6 alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell, and preferably the composition consists of the conjugate.

[0167] As described herein, the depiction of Formula I above includes the fragment R 1 (NR 2 CH2CH2) n Two different regioisomeric bonds, i.e., [ka] and [ka] represents The wavy line represents a chemical bond to ring A. Thus, Formula I as depicted herein includes two regioisomeric embodiments, namely, fragment R 1 (NR 2 CH2CH2) n is attached to the top nitrogen atom of the structure or to a bottom nitrogen atom of the structure, but not to the middle nitrogen atom. Formula I, depicted above, is used interchangeably herein with an equivalent depiction of Formula I that includes the following fragment: HNN=N, i.e., [ka]

[0168] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single bond or a double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 at least 80%, preferably 90%, of are H; ring A may contain one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is the formula -(Y1 ) p -, where p is an integer from 1 to 20, and Y 1 Each occurrence of represents a chemical bond, -CR 11 R 12 -, -C(O)-, -O-, -S-, -NR 13 -, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl or heteroaryl being selected from one or more R 13 and each divalent heterocycle is optionally substituted with one or more R 14 wherein R 11 , R 12 and R 13 is independently, at each occurrence, H or C1-C6 alkyl; 14 is independently, at each occurrence, H, C-C alkyl, or oxo; X 2 is the formula -(Y 2 ) q -, where q is an integer from 1 to 50, and Y 2 Each occurrence of represents a chemical bond, -CR 21 R 22 -, NR 23 -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R 23 and each divalent heterocyclic moiety is optionally substituted with one or more R 24 wherein R 21 , R 22 and R 23 are each independently, at each occurrence, —H, —COH, or C-C alkyl, and each C-C alkyl may be selected from one or more of —OH, oxo, C-C 10 optionally substituted with aryl or 5- to 8-membered heteroaryl; 24 is independently, at each occurrence, -H, -COH, C-C alkyl, or oxo; and L is preferably a targeting fragment capable of binding to a cell. In a preferred embodiment, the R 1is —H. In a preferred embodiment, the R 1 is -CH3.

[0169] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with RA2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor. 1 is —H. In a preferred embodiment, the R 1 is -CH3.

[0170] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating -(O-CH-CH)- units, and the discrete number m of repeating -(O-CH-CH)- units is any discrete number from 25 to 100, preferably from 25 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor. 1 is —H. In a preferred embodiment, the R 1 is -CH3.

[0171] In some embodiments, the covalent linking moiety Z comprises a triazole.

[0172] In some embodiments, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% of the LPEI in the composition are linked to the PEG fragments by a single covalent linking moiety, preferably the covalent linking moiety produces a linear end-to-end linkage between the LPEI fragments and the PEG fragments. In some embodiments, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% of the LPEI fragments in the composition are included in the conjugate, preferably as determined by UV spectroscopy or mass spectrometry. In some embodiments, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% of the LPEI in the composition are included in the conjugate, preferably as determined by UV spectroscopy or mass spectrometry. In some embodiments, the composition consists essentially of the conjugate. In some embodiments, the composition consists of the conjugate.

[0173] In some embodiments, at least 60% of the LPEI fragments in the composition are linked to a single PEG fragment by a single covalent linking moiety Z, preferably, the covalent linking moiety Z creates a linear end-to-end linkage between the LPEI fragment and the PEG fragment. In some embodiments, at least 60% of the LPEI fragments in the composition are linked to a PEG fragment by a single triazole linker, preferably as determined by UV spectroscopy or mass spectrometry. In some embodiments, at least 70% of the LPEI fragments in the composition are linked to a PEG fragment by a single covalent linking moiety Z, preferably, the covalent linking moiety Z creates a linear end-to-end linkage between the LPEI fragment and the PEG fragment. In some embodiments, at least 70% of the LPEI fragments in the composition are included in the conjugate, preferably, as determined by UV spectroscopy or mass spectrometry. In some embodiments, at least 80% of the LPEI fragments in the composition are linked to a PEG fragment by a single covalent linking moiety Z, preferably, the covalent linking moiety Z creates a linear end-to-end linkage between the LPEI fragment and the PEG fragment. In some embodiments, at least 80% of the LPEI fragments in the composition are comprised in the conjugate, preferably as determined by UV spectroscopy or mass spectroscopy. In some embodiments, at least 90% of the LPEI fragments in the composition are linked to the PEG fragment by a single covalent linking moiety Z, preferably the covalent linking moiety Z creates a linear end-to-end linkage between the LPEI fragment and the PEG fragment. In some embodiments, at least 90% of the LPEI fragments in the composition are comprised in the conjugate, preferably as determined by UV spectroscopy or mass spectroscopy. In some embodiments, at least 95% of the LPEI fragments in the composition are linked to the PEG fragment by a single covalent linking moiety Z, preferably the covalent linking moiety Z creates a linear end-to-end linkage between the LPEI fragment and the PEG fragment. In some embodiments, at least 95% of the LPEI fragments in the composition are comprised in the conjugate, preferably as determined by UV spectroscopy or mass spectroscopy.In some embodiments, at least 99% of the LPEI fragments in the composition are linked to the PEG fragment by a single covalent linking moiety Z, preferably the covalent linking moiety Z creates a linear end-to-end linkage between the LPEI fragment and the PEG fragment. In some embodiments, at least 99% of the LPEI fragments in the composition are included in the conjugate, preferably as determined by UV spectroscopy or mass spectrometry. In some embodiments, the composition consists essentially of the conjugate. In some embodiments, the composition consists of the conjugate. In some embodiments, the LPEI fragment does not include substitutions beyond its first and second termini.

[0174] In some embodiments, Formula I* has the structure: R 1 -(NH-CH2-CH2) n -NHC(O)-(CH2-CH2-O) m -X 2 In some embodiments, formula I* does not include the structure R 1 -(NR 2 -CH2-CH2) n -NHC(O)-X 1 -(O-CH2-CH2) m -X 2 In some embodiments, the composition does not include the structure R 1 -(NH-CH2-CH2) n -NHC(O)-X 1 -(O-CH2-CH2) m -X 2 In some embodiments, the composition does not include a conjugate of structure R 1 -(NR 2 -CH2-CH2) n -NHC(O)-(CH2-CH2-O) m -X 2 -L conjugates are not included.

[0175] In some embodiments, R 1 is -H.

[0176] In some embodiments, R in the composition 2 At least 80% of the R groups in the composition are —H. 2 At least 85%, preferably 90%, preferably 95%, more preferably 99% of R are -H. 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 At least 85%, preferably 90% of R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 At least 90% of R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 At least 90% of R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 In another preferred embodiment, at least 91%, preferably at least 92%, more preferably at least 93% of R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 At least 94%, preferably at least 95%, more preferably 96% of R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in the part 2 At least 95%, preferably at least 97%, even more preferably at least 98%, and even more preferably 99% of the total is H.

[0177] In some embodiments, ring A is an 8-membered cycloalkenyl, a 5-membered heterocycloalkyl, or a 7-8-membered heterocycloalkenyl, and each cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl may have one or more R A1 is optionally replaced by

[0178] In some embodiments, ring A is cyclooctene, succinimide, or a 7-8 membered heterocycloalkenyl, wherein the heterocycloalkyl or heterocycloalkenyl does not contain heteroatoms other than N, O, and S, and each cyclooctene, heterocycloalkyl, or heterocycloalkenyl may contain one or more R A1 is optionally replaced by

[0179] In some embodiments, ring A is cyclooctene, succinimide, or a 7-8 membered heterocycloalkenyl, wherein the heterocycloalkyl or heterocycloalkenyl contains one or more heteroatoms, preferably one or two heteroatoms selected from N, O, and S, and each cyclooctene, heterocycloalkyl, or heterocycloalkenyl contains one or more R A1 is optionally replaced by

[0180] In some embodiments, ring A is a cyclooctene, a succinimide, or an 8-membered heterocycloalkene, wherein the heterocycloalkene contains exactly one heteroatom selected from N, O, and S, and each cyclooctene or heterocycloalkene contains one or more R A1 is optionally replaced by

[0181] In some embodiments, R A1 is -H, oxo, or fluorine, or two R A1 combine to form one or more fused phenyl rings, preferably one or two fused phenyl rings, each phenyl ring optionally substituted with one or more -OSO3H or -SO3H.

[0182] In some embodiments, ring A is a cyclooctene, a succinimide, or an 8-membered heterocycloalkene, wherein the heterocycloalkene contains exactly one heteroatom selected from N, O, and S, and each cyclooctene or heterocycloalkene contains one or more R A1 optionally substituted with R A1 is oxo or fluorine, or two R A1 combine to form one or more fused phenyl rings, preferably one or two fused phenyl rings.

[0183] In some embodiments, ring A is cyclooctene, succinimide, or an 8-membered heterocycloalkene, wherein the heterocycloalkene contains exactly one heteroatom selected from N, and each cyclooctene or heterocycloalkene contains one or two R A1 is optionally replaced by

[0184] In some embodiments, R A1 is -H, oxo, or fluorine, or two R A1 combine to form one or more fused phenyl rings, preferably one or two fused phenyl rings, each phenyl ring containing one or more R A2 is optionally replaced by

[0185] In some embodiments, ring A is cyclooctene, succinimide, or an 8-membered heterocycloalkene, wherein the heterocycloalkene contains exactly one heteroatom selected from N, and each cyclooctene or heterocycloalkene contains one or two R A1 optionally substituted with R A1 is -H, oxo, or fluorine, or two R A1 combine to form one or more fused phenyl rings, preferably one or two fused phenyl rings, each phenyl ring optionally substituted with one or more -OSO3H or SO3H.

[0186] In some preferred embodiments, ring A is cyclooctene, succinimide, or an 8-membered heterocycloalkene, the heterocycloalkene containing exactly one heteroatom selected from N, and each cyclooctene or heterocycloalkene containing one or two R A1 optionally substituted with R A1 is -H or two R A1 combine to form one or more fused phenyl rings, preferably one or two fused phenyl rings, each phenyl ring optionally substituted with one or more -OSO3H or SO3H.

[0187] Preparation of linear conjugates The conjugates of the present invention can be prepared by several methods well known to those skilled in the art of polymer synthesis. For example, the compounds of the present invention can be synthesized using the methods described below, along with synthetic methods known in the field of polymer chemistry, or variations thereof recognized by those skilled in the art. Methods include, but are not limited to, the methods described below. The conjugates of the present invention can be synthesized according to the steps outlined in general schemes 1, 2, 3, 4, 5, 6, 7 and 8, or can be prepared using alternative sequences for assembling intermediates without departing from the present invention. The conjugates of the present invention can also be synthesized using slight variations of the steps outlined below. For example, while scheme 3 shows the use of a tetrafluorophenyl ester as an electrophilic functional group for coupling with hEGF, those skilled in the art will recognize other suitable electrophilic functional groups that can be used for the same purpose.

[0188] In some preferred embodiments, the LPEI fragment and the PEG fragment are coupled via a [3+2] cycloaddition between an azide and an alkene or alkyne to form a 1,2,3 triazole or a 4,5-dihydro-1H-[1,2,3]triazole. In some preferred embodiments, the LPEI fragment contains an azide functional group and the PEG fragment contains an alkene or alkyne functional group.

[0189] LPEI fragment The conjugates of the present invention can comprise LPEI fragments, PEG fragments. Linear polyethyleneimine (LPEI) has the chemical formula -[NH-CH-CH]-. Thus, linear polyethyleneimine (LPEI) is -[NH-CH-CH] n - repeating units n. LPEIs can be synthesized according to many methods known in the art, including, inter alia, polymerization of 2-oxazolines followed by hydrolysis of the pendant amide bonds (see, e.g., Brissault et al., Bioconjugate Chem., 2003, 14, 581-587). As noted above, polymerization of poly(2-oxazoline) from 2-oxazolines (i.e., suitable precursors for LPEIs) can be initiated with any suitable initiator. In some embodiments, the initiator leaves an initiator residue at the alpha-terminus of the poly(2-oxazoline). In preferred embodiments, the initiator residue (i.e., Formula I* or R of Formula I) is a 2-oxazoline. 1 ) is a hydrogen atom or a C1-C6 alkyl, preferably a hydrogen or a C1-C4 alkyl, more preferably a hydrogen or methyl group; most preferably a hydrogen atom). In a preferred embodiment, the initial residue R of formula I 1 is a hydrogen atom or a C1-C6 alkyl, preferably a hydrogen or a C1-C4 alkyl, more preferably a hydrogen or a methyl group; most preferably a hydrogen atom. In a preferred embodiment, the initial residue (i.e., R 1 ) is -H or -CH3, most preferably -H. In a preferred embodiment, the initial residue R of formula I* 1 is —H. In a preferred embodiment, the initial residue R of formula I 1 is -H. In a preferred embodiment, the initial residue R of formula I* 1 In a preferred embodiment, the initial residue R of formula I is 1 is —CH3. However, one skilled in the art will understand that the initiating residue can be the residue remaining from any suitable initiator capable of initiating the polymerization of 2-oxazoline to poly(2-oxazoline).

[0190] In some embodiments, an LPEI fragment can be attached to a PEG fragment via a [3+2] cycloaddition between an azide and an alkene or alkyne to form a 1,2,3 triazole or a 4,5-dihydro-1H-[1,2,3]triazole, and the LPEI fragment contains an azide (-N3) functional group at the omega-terminus of the chain. In some preferred embodiments, the LPEI fragment is not further substituted except for a single substitution at the alpha-terminus. For example, in some preferred embodiments, the LPEI fragment contains the repeating formula -[NH-CH2-CH2]- and is substituted at the omega-terminus with an azide group that can be coupled to an alkyne or alkene substituent on the PEG fragment. In some preferred embodiments, the alpha-terminus of the LPEI fragment can be substituted with a hydrogen atom or a C1-C6 alkyl, preferably hydrogen or C1-C4 alkyl, more preferably hydrogen or methyl; most preferably a hydrogen atom.

[0191] For example, in some preferred embodiments, the LPEI fragment may be substituted at the alpha terminus with a hydrogen atom or a C1-C6 alkyl, preferably a hydrogen atom or a C1-C4 alkyl, more preferably a hydrogen atom or a methyl group, and at the omega terminus with an azide group; in some preferred embodiments, there are no additional substitutions on the LPEI fragment. For example, the conjugates of the present invention can be prepared from an LPEI fragment of the following formula: [ka] In the formula, R 1 may be any suitable initial residue, preferably hydrogen or C1-C6 alkyl, preferably hydrogen or C1-C4 alkyl, more preferably hydrogen or methyl, most preferably hydrogen.

[0192] In some embodiments, the LPEI fragment can be terminated with a thiol group, and thus in some embodiments, the omega-terminus of the LPEI fragment comprises, and preferably is, a thiol group, which can be coupled to a reactive alkene group on a PEG fragment via a thiol-ene reaction. Thus, in some embodiments, the conjugates of the present invention can be prepared from an LPEI fragment of the formula: [ka] In the formula, R 1 may be any suitable starting residue, preferably hydrogen or methyl, preferably hydrogen.

[0193] In some embodiments, the LPEI fragment can be terminated with an alkene group, and thus in some embodiments, the omega-terminus of the LPEI fragment comprises, and preferably is, an alkene group, which can be coupled to a reactive thiol group on a PEG fragment via a thiol-ene reaction. Thus, in some embodiments, the conjugates of the present invention can be prepared from an LPEI fragment of the formula: [ka] In the formula, R 1 may be any suitable starting residue, preferably hydrogen or methyl, preferably hydrogen.

[0194] LPEI fragments can include a range of lengths (i.e., repeat units, as represented above by the variable "n"). For example, LPEI fragments can include 1 to 1000 repeat units (i.e., -NH-CH-CH-). In some embodiments, LPEI fragments can be present as dispersed polymer moieties and do not include a discrete number of -NH-CH-CH- repeat units. For example, LPEI fragments can be present as dispersed polymer moieties having a molecular weight of about 5 to 50 KDa, preferably about 5 KDa or less, preferably about 4 KDa or less, preferably about 3 KDa or less, preferably about 2 KDa or less, and preferably about 1.5 KDa or less. In some embodiments, LPEI fragments can be present as dispersed polymer moieties having a molecular weight of about 10 to 40 KDa and a dispersity of about 4 or less, preferably about 3 or less, preferably about 2 or less, and preferably about 1.5 or less. In some embodiments, LPEI fragments can be present as dispersed polymer moieties having a molecular weight of about 12-30 KDa and a dispersity of about 3 or less, preferably about 2 or less, and preferably about 1.5 or less. In some embodiments, LPEI fragments can be present as dispersed polymer moieties having a molecular weight of about 15-27 KDa and a dispersity of about 2 or less, preferably about 1.5 or less. In some embodiments, LPEI fragments can be present as dispersed polymer moieties having a molecular weight of about 17-25 KDa and a dispersity of about 1.2 or less.

[0195] For example, the LPEI fragments can be present as dispersed polymer portions containing about 115-1150 repeating units, preferably with a dispersity of about 5 or less, preferably about 4 or less, preferably about 3 or less, preferably about 2 or less, and preferably about 1.5 or less. In some embodiments, the LPEI fragments can be present as dispersed polymer portions containing about 230-930 repeating units with a dispersity of about 4 or less, preferably about 3 or less, preferably about 2 or less, and preferably about 1.5 or less. In some embodiments, the LPEI fragments can be present as dispersed polymer portions containing about 280-700 repeating units with a dispersity of about 3 or less, preferably about 2 or less, and preferably about 1.5 or less. In some embodiments, the LPEI fragments can be present as dispersed polymer portions containing about 350-630 repeating units with a dispersity of about 2 or less, preferably about 1.5 or less. In some embodiments, the LPEI fragments can be present as dispersed polymer portions containing about 400-580 repeating units with a dispersity of about 1.2 or less.

[0196] In some embodiments, the R 1 -(NR 2 -CH2-CH2) n The R - moiety is a dispersion polymer moiety having a repeating unit n of 115 to 1150 and a dispersity of about 5 or less, and preferably 1 -(NR 2 -CH2-CH2) n The R - moiety is a dispersed polymer moiety having repeating units n of 280 to 700 and a dispersity of about 3 or less, and more preferably 1 -(NR 2 -CH2-CH2) n The R - moiety is a dispersion polymer moiety having repeat units n of 350 to 630 and a dispersity of about 2 or less, and again more preferably 1 -(NR 2 -CH2-CH2) n The - portion is a disperse polymer portion having repeat units n of 400 to 580 and a dispersity of about 1.2 or less.

[0197] In a preferred embodiment, the polyethyleneimine fragment is a disperse polymer moiety having about 115 to about 1150 repeating units and a dispersity of about 5 or less, preferably about 230 to about 930 repeating units and a dispersity of about 4 or less; more preferably about 280 to about 700 repeating units and a dispersity of about 3 or less; again more preferably about 350 to about 630 repeating units and a dispersity of about 2 or less; even more preferably about 400 to about 580 repeating units and a dispersity of about 1.2 or less.

[0198] In a preferred embodiment, the polyethyleneimine fragment is a dispersion polymer moiety having about 115 to about 1150 repeating units and a dispersity of about 5 or less, preferably about 4 or less, preferably about 3 or less, preferably about 2 or less, and preferably about 1.5 or less. In a preferred embodiment, the polyethyleneimine fragment is a dispersion polymer moiety having about 230 to about 930 repeating units and a dispersity of about 4 or less, preferably about 3 or less, preferably about 2 or less, and preferably about 1.5 or less. In a preferred embodiment, the polyethyleneimine fragment is a dispersion polymer moiety having about 280 to about 700 repeating units and a dispersity of about 3 or less, preferably about 2 or less, and preferably about 1.5 or less. In a preferred embodiment, the polyethyleneimine fragment is a dispersion polymer moiety having about 350 to about 630 repeating units and a dispersity of about 2 or less, preferably about 1.5 or less. In a preferred embodiment, the polyethyleneimine fragment is a dispersion polymer moiety having about 400 to about 580 repeating units and a dispersity of about 1.2 or less.

[0199] As discussed above, those skilled in the art will understand that, in some embodiments, LPEI fragments can include organic residues (i.e., pendant amide groups) linked by nitrogen atoms embedded within the LPEI chain. Those skilled in the art will understand that such organic residues (i.e., amide groups) can be formed during the ring-opening polymerization of 2-oxazolines to form poly(2-oxazolines). Without wishing to be bound by theory, LPEIs can be formed from poly(2-oxazolines) by cleavage of amide groups (e.g., using an acid such as HCl). However, in some cases, not all amide bonds can be cleaved under these conditions. Thus, in some embodiments, about 5% or less of the nitrogen atoms in LPEI fragments can be bonded to organic residues to form amides. In some embodiments, about 4% or less, about 3% or less, about 2% or less, about 1% or less, about 0.5% or less, about 0.4% or less, about 0.3% or less, about 0.2% or less, or about 0.1% or less of the nitrogen atoms in LPEI fragments can be bonded to organic residues to form amides. Those skilled in the art will understand that the molecular weight of an LPEI fragment includes the percentage of the LPEI fragment that is attached to an organic residue as an amide. Furthermore, those skilled in the art will understand that while the chemical structures depicted herein show repeating -NH-CH-CH- fragments, trace amounts of residual organic residues, such as pendant amide groups (e.g., as defined above), may still be present in the resulting triconjugates or polyplexes of the present disclosure. The term "triconjugate," as sometimes used herein, is intended to refer to the conjugates of the present invention. The prefix "tri-" is due to the three components included in the conjugates of the present invention: the LPEI fragment, the PEG fragment, and the targeting fragment.

[0200] PEG fragment Polyethylene glycol (PEG) has the chemical formula -[O-CH-CH]-. Therefore, polyethylene glycol (PEG) is -[O-CH-CH] mIn some preferred embodiments, the PEG fragment can be coupled to the LPEI fragment via a [3+2] cycloaddition between an azide and an alkene or alkyne to form a 1,2,3 triazole or a 4,5-dihydro-1H-[1,2,3]triazole, and each reactive precursor molecule comprising the PEG fragment further comprises an alkene or alkyne functional group. For example, in some preferred embodiments, the reactive precursor molecule comprising the PEG fragment comprises the repeating formula -[O-CH-CH]-, an alkene or alkyne group (e.g., a linking moiety "X" as discussed herein) that can be coupled to the azide group of the corresponding reactive precursor molecule comprising the LPEI fragment. 1 ") at the first end (i.e., terminus).

[0201] In some preferred embodiments, the alkene or alkyne group is an activated alkene or alkyne group that can spontaneously react with an azide (e.g., without the addition of a catalyst, such as a copper catalyst). For example, an activated alkyne group can be incorporated into a 7- or 8-membered ring, resulting in a strained species that spontaneously reacts with the azide group of an LPEI fragment. The activated alkene can include a maleimide moiety, where the alkene is activated by conjugation to an adjacent carbonyl group. In some preferred embodiments, the second end (i.e., terminus) of the PEG fragment is linked to a targeting fragment (e.g., hEGF) (e.g., a linking moiety "X" discussed herein). 2 " can be substituted).

[0202] PEG fragments can include a range of lengths (i.e., repeating units represented by the variable "m"). In other embodiments, PEG fragments can include a discrete number of repeating -O-CH-CH- units and are not defined in terms of average chain length. In preferred embodiments, the -(O-CH-CH) m - is a dispersed polymer moiety. In a preferred embodiment, the -(O-CH-CH) mThe - moiety comprises, and preferably consists of, a discrete number m of repeating units. In a preferred embodiment, the -(O-CH-CH) m The - moiety comprises, and preferably consists of, a discrete number of consecutive repeating units m.

[0203] In some preferred embodiments, the PEG fragment is a dispersed polymer moiety comprising from about 1 to about 200 repeating units, preferably from about 1 to about 200 repeating units. In some preferred embodiments, the PEG fragment can comprise from 1 to 100 repeating units (i.e., -O-CH2-CH2-). Preferably, the PEG fragment of the present invention comprises from about 1 to about 100 repeating units, from about 1 to about 90 repeating units, from about 1 to about 80 repeating units, from about 1 to about 70 repeating units, from about 1 to about 60 repeating units, from about 1 to about 50 repeating units, from about 1 to about 50 repeating units, from about 1 to about 40 repeating units, from about 1 to about 30 repeating units, or from about 1 to about 20 repeating units. In some other preferred embodiments, the PEG fragment comprises a discrete number of repeating units, m, preferably 12 repeating units or 24 repeating units. In some embodiments, the polyethylene glycol fragment is a dispersant polymer moiety having about 2 to about 80 repeating units and a dispersity of about 2.0 or less, preferably about 1.8 or less, and even about 1.5 or less; preferably about 2 to about 70 repeating units and a dispersity of about 1.8 or less, preferably about 1.5 or less; more preferably about 2 to about 50 repeating units and a dispersity of about 1.5 or less. In some embodiments, the -(O-CH2-CH2) m The - moiety is a disperse polymer moiety having from about 2 to about 80 repeating units and a dispersity of about 2.0 or less, preferably from about 2 to about 70 repeating units and a dispersity of about 1.8 or less; more preferably from about 2 to about 50 repeating units and a dispersity of about 1.5 or less.

[0204] In a preferred embodiment, the polyethylene glycol fragment PEG fragment comprises, and preferably consists of, a discrete number of repeating units m, preferably 12 or 24 repeating units. In a preferred embodiment, the -(O-CH-CH) m - the m moiety) comprises, preferably consists of, a discrete number of repeating units m, preferably 12 or 24 repeating units.

[0205] In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m between 2 and 100, preferably a discrete number of repeating units m between 4 and 60. In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m between 4 and 60, preferably a discrete number of repeating units m between 10 and 60. In preferred embodiments, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60. In preferred embodiments, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m of 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60. In preferred embodiments, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m of 4. In preferred embodiments, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m of 12. In preferred embodiments, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m of 24. In preferred embodiments, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m of 36.

[0206] In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of consecutive repeating units m between 2 and 100, preferably a discrete number of consecutive repeating units m between 4 and 60. In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of consecutive repeating units m between 4 and 60, preferably a discrete number of consecutive repeating units m between 10 and 60. In preferred embodiments, the PEG fragment comprises, or preferably consists of, a discrete number of consecutive repeating units m of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 4 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 12 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 24 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 36 discrete numbers of consecutive repeating units m.

[0207] In a preferred embodiment, the —(O—CH—CH) of formula I* or formula I m The - moiety comprises, or preferably consists of, a discrete number of repeating units m between 2 and 100, preferably between 4 and 60. In a preferred embodiment, the -(O-CH2-CH2) m The - moiety comprises, or preferably consists of, a discrete number of repeating units m between 4 and 60, preferably between 10 and 60. In a preferred embodiment, the -(O-CH2-CH2) mThe - moiety comprises, and preferably consists of, a discrete number of repeating units m of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60. In a preferred embodiment, the -(O-CH-CH) m The - moiety comprises, and preferably consists of, a discrete number of repeating units m of 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60. In a preferred embodiment, the -(O-CH-CH) m The - moiety comprises, and preferably consists of, a discrete number of repeating units m of 4. In a preferred embodiment, the -(O-CH-CH) m The - moiety comprises, and preferably consists of, a discrete number of repeating units m of 12. In a preferred embodiment, the -(O-CH-CH) m The - moiety comprises, and preferably consists of, a discrete number of repeating units m of 24. In a preferred embodiment, the -(O-CH-CH) m The moiety comprises, and preferably consists of, 36 discrete numbers of repeating units m.

[0208] In a preferred embodiment, the —(O—CH—CH) of formula I* or formula I m The - moiety comprises, or preferably consists of, a discrete number of consecutive repeating units m between 2 and 100, preferably a discrete number of consecutive repeating units m between 4 and 60. In a preferred embodiment, the -(O-CH2-CH2) m The - moiety comprises, or preferably consists of, a discrete number of consecutive repeating units m between 4 and 60, preferably a discrete number of consecutive repeating units m between 10 and 60. In a preferred embodiment, the -(O-CH2-CH2) mThe - moiety comprises, and preferably consists of, a discrete number of consecutive repeating units m of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60. In a preferred embodiment, the -(O-CH-CH) m The - moiety comprises, and preferably consists of, a discrete number of consecutive repeating units m of 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60. In a preferred embodiment, the -(O-CH-CH) m The - moiety comprises, and preferably consists of, a discrete number of consecutive repeating units m of 4. In a preferred embodiment, the -(O-CH-CH) m The - moiety comprises, and preferably consists of, a discrete number of consecutive repeating units m of 12. In a preferred embodiment, the -(O-CH-CH) m The - moiety comprises, and preferably consists of, a discrete number of consecutive repeating units m of 24. In a preferred embodiment, the -(O-CH-CH) m - the moiety comprises, and preferably consists of, 36 discrete numbers of consecutive repeating units m.

[0209] In preferred embodiments, the PEG fragments contained in the conjugates and compositions of the invention comprise, and preferably consist of, a discrete number m of repeating -(O-CH-CH)- units and are not defined in terms of average chain length. Thus, the PEG fragments contained in the conjugates and compositions of the invention comprise, and preferably consist of, a discrete number m of repeating -(O-CH-CH)- units and are not defined in terms of average chain length, but have a specifically defined, discrete molecular weight associated with the discrete number m of repeating -(O-CH-CH)- units. In preferred embodiments, the PEG fragments comprise, and preferably consist of, a discrete number m of repeating -(O-CH-CH)- units, typically and preferably, the discrete number (m) is between 25 and 100, more preferably between 25 and 60. In a preferred embodiment, the PEG fragment comprises, and preferably consists of, a discrete number m of consecutive repeating -(O-CH-CH) units, typically and preferably, the discrete number (m) is between 25 and 100, more preferably between 25 and 60.

[0210] The phrases "polyethylene glycol fragment comprising a discrete number (m) of repeating -(O-CH2-CH2)- units" or "PEG fragment comprising a discrete number (m) of repeating -(O-CH2-CH2)- units" are intended to refer to a fragment that comprises, and preferably consists of, a discrete number (typically referred to herein as a discrete number m) of repeating -(O-CH2-CH2)- units, wherein said discrete number (m) is a discrete, specific and single defined integer (m) between 25 and 100, preferably between 25 and 60. Thus, the phrases "polyethylene glycol fragments comprising a discrete number (m) of repeating -(O-CH-CH)- units" or "PEG fragments comprising a discrete number (m) of repeating -(O-CH-CH)- units" are intended to refer to fragments comprising, and preferably consisting of, a discrete number m of repeating -(O-CH-CH)- units, where the discrete number (m) is a specific, single, defined integer (m) between 25 and 100, preferably between 25 and 60. Thus, the defined PEG fragments comprise, and preferably consist of, a discrete number m of repeating -(O-CH-CH)- units, each having a specifically defined, discrete molecular weight, although not defined in terms of average chain length. Reference herein to a discrete number between 25 and 100 is intended to refer to any integer between 25 and 100, i.e., any integer between 25 and 100, including the integers and discrete numbers recited herein as boundaries, such as 25 and 100. As a further example, a PEG fragment comprising a discrete number (m) of repeating -(O-CH-CH)- units, where m is 36, refers to a PEG fragment comprising a chain of -(O-CH-CH)- units containing exactly 36 -(O-CH-CH)- units. Such a chain of exactly 36 -(O-CH-CH)- units is a PEG fragment. 36Such PEG fragments are contrasted with "polymeric PEG fragments," "polydisperse PEG fragments," or "disperse PEG fragments," which typically refer to a heterogeneous mixture of sizes and molecular weights resulting from a polymer reaction in a Poisson distribution (J. Herzberger et al.; Chem Rev, 2016, 116:2170-2243). The PEG fragments of the present invention, which contain a discrete number (m) of repeating -(O-CH-CH)- units, are not synthesized by a polymerization process. The PEG fragments of the present invention are single molecular fragments that contain a discrete number (m) of repeating -(O-CH-CH)- units and have a discrete, i.e., defined and specified, chain length. Thus, the PEG fragments of the present invention, which contain a discrete number (m) of repeating -(O-CH-CH)- units, are single molecular fragments that have a discrete, i.e., defined and specified, chain length. The PEG fragments of the present invention are not a mixture of molecular entities (such as those resulting from a random polymerization reaction). The discrete nature of the discrete PEG fragments of the present invention distinguishes them from polydisperse techniques.

[0211] The PEG fragments of the present invention can comprise, and preferably consist of, homogeneous discrete PEG fragments or heterogeneous discrete PEG fragments, typically and preferably homogeneous discrete PEG fragments. As used herein, the term "homogeneous discrete PEG fragments" refers to a discrete PEG structure whose entire chemical backbone is composed solely of a specific, discrete number of ethylene oxide units in a continuous sequence. In other words, no other functionality is present within the homogeneous discrete PEG fragments. However, the term "homogeneous discrete PEG fragments" refers to a discrete PEG structure whose basic ethylene oxide backbone, comprising a discrete number of ethylene oxide units, can, and typically does, have a functional group for conjugation with PEI fragments and targeting fragments. As used herein, the term "heterogeneous discrete PEG fragments" refers to a discrete PEG structure in which the basic ethylene oxide backbone, comprising a discrete number of ethylene oxide units, is divided or substituted by the inclusion of other functional groups or units within the structure, such as amide or ester linkages or other functional units. In a preferred embodiment of the present invention, the PEG fragments are homogeneous discrete PEG fragments.

[0212] The PEG fragments contained in the conjugates and compositions of the invention comprise, and preferably consist of, a discrete number m of repeating -O-CH-CH- units, and are not defined in terms of average chain length as is the case for polymeric PEG fragments. In a preferred embodiment, the -(O-CH-CH) m The - unit comprises, and preferably consists of, a discrete number of repeating units m. In a preferred embodiment, the -(O-CH-CH) m -unit comprises, and preferably consists of, a discrete number of consecutive repeating units m.

[0213] In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m between 25 and 100, preferably a discrete number of repeating units m between 25 and 60. In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m between 25 and 60, preferably a discrete number of repeating units m between 30 and 50. In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of repeating units m between 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60. Discrete number repeating -(O-CH2-CH2) mThe synthesis of such PEG fragments comprising or consisting of m-units, and thus discrete PEGs, is described in WO 2004 / 073620 and WO 2013 / 033476. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 28, 32, 36, 40, 44, 48, 52, 56, or 60 discrete numbers of repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 28 discrete numbers of repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 32 discrete numbers of repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 36 discrete numbers of repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 40 discrete numbers of repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 44 discrete numbers of repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 48 discrete numbers of repeating units m.

[0214] In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of consecutive repeating units m between 25 and 100, preferably a discrete number of consecutive repeating units m between 25 and 60. In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of consecutive repeating units m between 25 and 60, preferably a discrete number of consecutive repeating units m between 30 and 50. In a preferred embodiment, the PEG fragment comprises, or preferably consists of, a discrete number of consecutive repeating units m between 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 28, 32, 36, 40, 44, 48, 52, 56, or 60 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 28 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 32 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 36 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 40 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 44 discrete numbers of consecutive repeating units m. In preferred embodiments, the PEG fragment comprises, or preferably consists of, 48 discrete numbers of consecutive repeating units m.

[0215] In a preferred embodiment, the —(O—CH—CH) of formula I* or formula I m The - moiety consists of a discrete number of repeating units m between 25 and 100, preferably a discrete number of repeating units m between 25 and 60. In a preferred embodiment, the -(O-CH2-CH2) m The - moiety consists of a discrete number of repeating units m between 25 and 60, preferably a discrete number of repeating units m between 30 and 50. In a preferred embodiment, the -(O-CH2-CH2) mThe - moiety is comprised of a discrete number of repeating units m, which may be 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of a discrete number of repeating units m, which are 28, 32, 36, 40, 44, 48, 52, 56, or 60. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of 28 discrete repeating units m. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of 32 discrete repeating units m. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of 36 discrete repeating units m. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of 40 discrete repeating units m. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of a discrete number of repeating units m of 44. In a preferred embodiment, the -(O-CH-CH) m -The part consists of 48 discrete repeating units m.

[0216] In a preferred embodiment, the —(O—CH—CH) of formula I* or formula I m The - moiety consists of a discrete number of consecutive repeating units m between 25 and 100, preferably a discrete number of consecutive repeating units m between 25 and 60. In a preferred embodiment, the -(O-CH2-CH2) m The - moiety consists of a discrete number of consecutive repeating units m between 25 and 60, preferably a discrete number of consecutive repeating units m between 30 and 50. In a preferred embodiment, the -(O-CH2-CH2) m The - moiety consists of a discrete number of consecutive repeating units m, which is 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60. In a preferred embodiment, the -(O-CH-CH)m The - moiety consists of a discrete number of consecutive repeating units m, which are 28, 32, 36, 40, 44, 48, 52, 56, or 60. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of 28 discrete consecutive repeating units m. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of 32 discrete consecutive repeating units m. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of 36 discrete consecutive repeating units m. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of 40 discrete consecutive repeating units m. In a preferred embodiment, the -(O-CH-CH) m The - moiety consists of a discrete number of consecutive repeating units m of 44. In a preferred embodiment, the -(O-CH-CH) m - The part consists of 48 discrete consecutive repeating units m.

[0217] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor. 1 is —H. In a preferred embodiment, the R 1 is -CH3.

[0218] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of repeating units m from 2 to 100, preferably from 4 to 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor. 1 is —H. In a preferred embodiment, the R 1 is -CH3.

[0219] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is the repeating unit m of 36 discrete numbers; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor. 1 is —H. In a preferred embodiment, the R 1 is -CH3.

[0220] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is the repeating unit m of 36 discrete numbers; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor. 1 is —H. In a preferred embodiment, the R 1 is -CH3.

[0221] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of consecutive repeat units m between 2 and 100, preferably a discrete number of consecutive repeat units m between 4 and 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor. 1 is —H. In a preferred embodiment, the R 1 is -CH3.

[0222] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a discrete number of consecutive repeat units m between 2 and 100, preferably a discrete number of consecutive repeat units m between 4 and 60; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2 is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor. 1 is —H. In a preferred embodiment, the R 1 is -CH3.

[0223] In another aspect, the present invention provides a composition comprising a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500; m is a continuous repeating unit m of 36 discrete numbers; R 1 is the starting residue, preferably R 1 is -H or -CH3; R 2 are independently —H or an organic residue, and the —(NR 2 -CH2-CH2) n -The R in question 2 at least 80%, preferably at least 90% of is H; Ring A may have one or more R A1 R is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with A1 are independently selected from C1-C6 alkyl, C1-C6 alkoxy, oxo, or halogen; or two R A1 together with the atoms to which they are attached form one or more fused C6-C 10 can form an aryl, C5-C6 heteroaryl, or C3-C6 cycloalkyl ring, and each fused aryl, heteroaryl, or cycloalkyl can be joined by one or more R A2 optionally substituted with R A2 is independently selected from C1-C6 alkyl, C1-C6 alkoxy, halogen -SO3H or -OSO3H; X 1 is a divalent covalent linking moiety; X 2is a divalent covalent linking moiety; and L is a targeting fragment, preferably the targeting fragment is capable of binding to a cell, more preferably the targeting fragment is capable of binding to a cell surface receptor. 1 is —H. In a preferred embodiment, the R 1 is -CH3.

[0224] In another aspect, the present invention provides a conjugate of formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof: [ka] During the ceremony, [ka] is a single or double bond; n is any integer from 1 to 1500;...

Claims

1. 1. A composition comprising a conjugate, the conjugate comprising: a linear polyethyleneimine fragment comprising an alpha end and an omega end; A polyethylene glycol fragment comprising a first end and a second end, the fragment comprising a discrete number m of repeating -(O-CH ２ -CH ２ )- units, preferably consisting of repeating —(O—CH ２ -CH ２ the polyethylene glycol fragment, wherein the discrete number m of methyl-1-(2-methyl-2-methyl)-units is any discrete number between 25 and 100, preferably between 25 and 60; the alpha end of the polyethyleneimine fragment is the initiating residue; The omega end of the polyethyleneimine fragment is linked to the first terminal end of the polyethylene glycol fragment by a divalent covalent linking group -Z-X １ -Z-X １ - is not a single bond and -Z- is not an amide; The second end of the polyethylene glycol fragment is capable of binding to a targeting fragment, and preferably the second end of the polyethylene glycol fragment is capable of binding to a bivalent covalent linking moiety X ２ and more preferably, said targeting fragment is capable of binding to a cell.

2. A composition comprising a conjugate, said conjugate being of formula I* or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or enantiomer thereof; R １ -(NR ２ -CH ２ -CH ２ ) ｎ -Z-X １ -(O-CH ２ -CH ２ ) ｍ -X ２ -L (Formula I*); During the ceremony, n is any integer from 1 to 1500; m is a repeating group -(O-CH ２ -CH ２ )-units, and the repeating -(O-CH ２ -CH ２ said discrete number m of )-units is any discrete number between 25 and 100, preferably between 25 and 60; R １ is the initial residue, preferably R １ is -H or -CH ３ and R ２ are independently —H or an organic residue, and the —(NR ２ -CH ２ -CH ２ ) ｎ The R in - ２ at least 80%, preferably 90%, of is H; X １ and X ２ are independently divalent covalent linking moieties; Z is a divalent covalent linking moiety; Z-X １ - is not a single bond, -Z- is not -NHC(O)-; A composition wherein L is a targeting fragment, preferably said targeting fragment is capable of binding to a cell.

3. the conjugate is of Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof; 【Chemistry 1】 During the ceremony, 【Chemistry 2】 is a single or double bond; n is any integer from 1 to 1500; m is a repeating group -(O-CH ２ -CH ２ )-units, and the repeating -(O-CH ２ -CH ２ said discrete number m of )-units is any discrete number between 25 and 100, preferably between 25 and 60; R １ is the initial residue, preferably R １ is -H or -CH ３ and R ２ are independently —H or an organic residue, and the —(NR ２ -CH ２ -CH ２ ) ｎ The R in - ２ at least 80%, preferably at least 90% of is H; Ring A is one or more R Ａ１ is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with R Ａ１ is independent, C １ -C ６ Alkyl, C １ -C ６ alkoxy, oxo, or halogen; or two R Ａ１ together with the atoms to which they are attached, form one or more fused C ６ -C １０ Aryl, C ５ -C ６ Heteroaryl, or C ３ -C ６ Each fused aryl, heteroaryl, or cycloalkyl can form a cycloalkyl ring, and one or more R Ａ２ optionally substituted with R Ａ２ are independently １ -C ６ Alkyl, C １ -C ６ Alkoxy, halogen -SO ３ H or -OSO ３ H; X １ is a divalent covalent linking moiety; X ２ is a divalent covalent linking moiety; and 3. The composition of claim 1 or claim 2, wherein L is a targeting fragment, preferably said targeting fragment being capable of binding to a cell.

4. The —(O—CH ２ -CH ２ ) ｍ - The part is repeated with a discrete number m between 25 and 60 - (O-CH ２ -CH ２ )-unit, and preferably the —(O—CH ２ -CH ２ ) ｍ - The part is repeated with a discrete number m between 25 and 48 - (O-CH ２ -CH ２ )- units, more preferably repeating —(O—CH ２ -CH ２ 4. The composition of claim 1, wherein the discrete number m of hydroxy-1, hydroxy-2, hydroxy-3, hydroxy-4, hydroxy-5, hydroxy-6, hydroxy-7, hydroxy-8, hydroxy-9, hydroxy-11, hydroxy-12, hydroxy-13, hydroxy-14, hydroxy-15, hydroxy-16, hydroxy-17, hydroxy-18, hydroxy-19 ...

5. Ring A is an 8-membered cycloalkenyl, a 5-membered heterocycloalkyl, or a 7- to 8-membered heterocycloalkenyl, and each cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl may have one or more R Ａ１ The composition of any one of claims 3 to 4, optionally substituted with

6. Ring A is cyclooctene, succinimide, or a 7- to 8-membered heterocycloalkenyl, wherein the heterocycloalkenyl contains one or two heteroatoms selected from N, O, and S, and each cyclooctene or heterocycloalkenyl may contain one or more R Ａ１ is optionally substituted with, preferably R Ａ１ is oxo or fluorine, or two R Ａ１ are combined to form one or more fused phenyl rings, preferably one or two fused phenyl rings, each phenyl ring containing one or more -SO ３ H or -OSO ３ The composition of any one of claims 3 to 5, optionally substituted with H.

7. The conjugate of formula I is: 【Transformation 3】 【Chemistry 4】 【Transformation 5】 【Transformation 6】 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】 and 【Chemistry 11】 The composition according to any one of claims 3 to 6, selected from:

8. The conjugate of formula I is: 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】 【Chemistry 15】 【Chemistry 16】 【Chemistry 17】 and [Chemistry 18] The composition according to any one of claims 3 to 7, selected from:

9. The conjugate of formula I is: 【Chemistry 19】 and 【Chemistry 20】 The composition according to any one of claims 3 to 8, wherein the composition is selected from

10. The conjugate of formula I is: 【Chemistry 21】 The composition according to any one of claims 3 to 8, wherein the composition is selected from

11. The conjugate of formula I is: 【Chemistry 22】 and 【Chemistry 23】 The composition according to any one of claims 3 to 8, wherein the composition is selected from

12. X １ but the following: 【Chemistry 24】 wherein: r is independently, in each occurrence, 0 to 6, preferably 0, 1, 2, or 5; more preferably 0; s is independently, in each occurrence, 0 to 6, preferably 0, 2, 3, or 4; more preferably 2 or 3; t is independently, in each occurrence, 0 to 6, preferably 0, 1, 2, 4; more preferably 2; R １１ and R １２ is independently, in each occurrence, —H and —C １ -C ２ alkyl, preferably —H; and R １３ is —H; preferably, the wavy line closest to the integer “r” is the bond to Ring A and the wavy line closest to the integer “s” or “t” is —[OCH ２ -CH ２ ] ｍ The composition of any one of claims 1 to 11, wherein the bond is to -.

13. X １ but the following: 【Chemistry 25】 (In the formula, X Ａ is —NHC(O)— or —C(O)NH—; and 【Chemistry 26】 Preferably, the wavy line on the left is the bond to ring A and the wavy line on the right is —[OCH ２ -CH ２ ] ｍ The composition of any one of claims 1 to 12, wherein the bond is to -.

14. X １ but the following: 【Chemistry 27】 and 【Chemistry 28】 Preferably, the left wavy line is the bond to ring A and the right wavy line is —[OCH ２ -CH ２ ] ｍ The composition of any one of claims 1 to 13, wherein the bond is -.

15. X ２ but the following: 【Chemistry 29】 and 【Transformation 30】 wherein X is selected from Ｂ is —C(O)NH— or —NH—C(O)—, In the formula, Y ２ Each occurrence of ２１ R ２２ -, NR ２３ -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent carbocyclic moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R ２３ and each divalent heterocyclic moiety is optionally substituted with one or more R ２４ optionally substituted with; R ２１ , R ２２ and R ２３ are each independently, at each occurrence, —H, —SO ３ H, —NH ２ , -CO ２ H or C １ -C ６ alkyl, and each C １ -C ６ Alkyl is one or more of -OH, oxo, -CO ２ H, —NH ２ , C ６ -C １０ optionally substituted with aryl, or 5-8 membered heteroaryl; and R ２４ is independently, at each occurrence, —H, —CO ２ H, C １ -C ６ alkyl or oxo; preferably, the wavy line on the left is —[OCH ２ -CH ２ ] ｍ The composition of any one of claims 1 to 14, wherein the wavy line on the right is a bond to - and the wavy line on the right is a bond to L.

16. X ２ but the following: 【Chemistry 31】 【Chemistry 32】 and 【Transformation 33】 is selected from In the formula, Y ２ Each occurrence of ２１ R ２２ -, NR ２３ -, -O-, -S-, -C(O)-, an amino acid residue, a divalent phenyl moiety, a divalent carbocyclic moiety, a divalent heterocyclic moiety, and a divalent heteroaryl moiety, each divalent phenyl and divalent heteroaryl being independently selected from one or more R ２３ and each divalent heterocyclic moiety is optionally substituted with one or more R ２４ optionally substituted with; R ２１ , R ２２ and R ２３ are each independently, at each occurrence, —H, —SO ３ H, —NH ２ , -CO ２ H or C １ -C ６ alkyl, and each C １ -C ６ Alkyl is one or more of -OH, oxo, -CO ２ H, —NH ２ , C ６ -C １０ optionally substituted with aryl, or 5-8 membered heteroaryl; and R ２４ is independently, at each occurrence, —H, —CO ２ H, C １ -C ６ alkyl or oxo; preferably, the wavy line on the left is —[OCH ２ -CH ２ ] ｍ The composition of any one of claims 1 to 15, wherein the wavy line on the right is a bond to - and the wavy line on the right is a bond to L.

17. X ２ but the following: 【Transformation 34】 【Chemistry 35】 【Transformation 36】 【Chemistry 37】 【Transformation 38】 and 【Chemistry 39】 Preferably, the left wavy line is selected from -[OCH ２ -CH ２ ] ｍ The composition of any one of claims 1 to 16, wherein the wavy line on the right is a bond to - and the wavy line on the right is a bond to L.

18. X ２ but the following: 【Chemistry 40】 and preferably, the left wavy line is —[OCH ２ -CH ２ ] ｍ The composition of any one of claims 1 to 17, wherein the wavy line on the right is a bond to - and the wavy line on the right is a bond to L.

19. X ２ but the following: 【Chemistry 41】 The composition according to any one of claims 1 to 18, wherein

20. 20. The composition of any one of claims 1 to 19, wherein the targeting fragment L is capable of binding to a cell surface receptor, preferably wherein the targeting fragment is capable of specifically binding to a cell surface receptor.

21. 21. The composition of claim 20, wherein the cell surface receptor is selected from growth factor receptors, cytokine receptors, hormone receptors, extracellular matrix proteins, transmembrane proteins, glycosylphosphatidylinositol (GPI)-anchored membrane proteins, carbohydrate-binding integral membrane proteins, lectins, ion channels, G protein-coupled receptors, and enzyme-linked receptors such as tyrosine kinase-coupled receptors; preferably, the cell surface receptor is selected from epidermal growth factor receptor (EGFR), human epidermal growth factor receptor 2 (HER2), prostate-specific membrane antigen (PSMA), insulin-like growth factor 1 receptor (IGF1R), vascular endothelial growth factor receptor (VEGFR), platelet-derived growth factor receptor (PDGFR), asialoglycoprotein receptor (ASGPr), and fibroblast growth factor receptor (FGFR).

22. 22. The composition of any one of claims 1 to 21, wherein the targeting fragment L is capable of binding to a cell surface receptor, and the targeting fragment is a peptide, protein, small molecule ligand, sugar, oligosaccharide, oligonucleotide, lipid, amino acid, antibody, antibody fragment, aptamer, or affibody.

23. the targeting fragment L is an EGFR targeting fragment, preferably human EGF (hEGF); a PSMA targeting fragment, preferably a DUPA residue; an anti-HER2 peptide, preferably an anti-HER2 antibody or an affibody; folic acid; methotrexate; a somatostatin receptor targeting fragment, preferably somatostatin and / or octreotide; an integrin targeting fragment, preferably an arginine-glycine-aspartic acid (RGD)-containing fragment; a low pH insertion peptide; an ASGPr targeting fragment, preferably asialoorosomucoid; an insulin receptor targeting fragment, preferably insulin; a mannose-6-phosphate receptor targeting fragment, preferably mannose-6-phosphate; a mannose receptor targeting fragment, preferably mannose; a sialyl Lewis ｘ 23. The composition of any one of claims 1 to 22, selected from an antigen targeting fragment, preferably E-selectin; a sigma-2 receptor agonist, preferably N,N-dimethyltryptamine (DMT), a sphingolipid-derived amine and / or a steroid, more preferably progesterone; a p32 targeting ligand, preferably an anti-p32 antibody or a p32-binding LyP-1 tumor-homing peptide; a Trop-2 targeting fragment, preferably an anti-Trop-2 antibody and / or antibody fragment; insulin-like growth factor 1; vascular endothelial growth factor; platelet-derived growth factor; and fibroblast growth factor.

24. 24. The composition of any one of claims 1 to 23, wherein the targeting fragment L is an EGFR targeting fragment, preferably wherein the targeting fragment is capable of binding to a cell expressing EGFR, more preferably wherein the targeting fragment is capable of binding to a cell surface receptor, wherein the cell surface receptor is EGFR, and again more preferably wherein the targeting fragment L is human EGF (hEGF).

25. 25. The composition of any one of items 1 to 24, wherein the conjugate is selected from Compound 6a, Compound 6b, Compound 12a, Compound 12b, Compound 19a, Compound 19b, Compound 24, Compound 28a, Compound 28b, Compound 32a, Compound 32b, Compound 37a, Compound 37b, Compound 43, Compound 44a, Compound 44b, Compound 45, Compound 49a, Compound 49b, Compound 57a, Compound 57b, Compound 60a, Compound 60b, Compound 61a, Compound 61b, Compound 64a, Compound 64b, Compound 67a, Compound 67b, Compound 70a, and / or Compound 70b.

26. 26. The composition of any one of claims 1 to 25, wherein the composition further comprises a polyanion, preferably wherein the polyanion is a nucleic acid, and wherein the polyanion is preferably non-covalently bound to the conjugate, and wherein the polyanion and the conjugate form a polyplex.

27. 27. The composition of claim 26, wherein the polyanion is a nucleic acid, and the nucleic acid is dsRNA or ssRNA.

28. 28. The composition of claim 27, wherein the nucleic acid is a dsRNA.

29. 29. The composition of claim 28, wherein the dsRNA is polyinosinic:polycytidylic acid (poly(IC)).

30. 29. The composition of claim 28, wherein the nucleic acid is ssRNA.

31. 31. The composition of claim 30, wherein the ssRNA is mRNA.

32. 27. The composition of claim 26, wherein the polyanion is a nucleic acid, and the nucleic acid is DNA.

33. 33. The composition of claim 32, wherein the DNA is plasmid DNA.

34. 34. A polyplex of a conjugate and a polyanion according to any one of claims 1 to 33, wherein the polyanion is preferably non-covalently bound to the conjugate, and preferably the polyanion is a nucleic acid.

35. A polyplex comprising a conjugate of Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or enantiomer thereof, and a polyanion, preferably a nucleic acid, wherein said polyanion, preferably said nucleic acid, is preferably non-covalently bound to said conjugate; 【Chemistry 42】 During the ceremony, 【Chemistry 43】 is a single or double bond; n is any integer from 1 to 1500; m is a repeating group -(O-CH ２ -CH ２ )-units, and the repeating -(O-CH ２ -CH ２ said discrete number m of )-units is any discrete number between 25 and 100, preferably between 25 and 60; R １ is the initial residue, preferably R １ is -H or -CH ３ and R ２ are independently —H or an organic residue, and the —(NR ２ -CH ２ -CH ２ ) ｎ The R in - ２ at least 80%, preferably at least 90% of is H; Ring A may have one or more R Ａ１ is a 5-10 membered cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl optionally substituted with R Ａ１ is independent, C １ -C ６ Alkyl, C １ -C ６ alkoxy, oxo, or halogen; or two R Ａ１ together with the atoms to which they are attached, form one or more fused C ６ -C １０ Aryl, C ５ -C ６ Heteroaryl, or C ３ -C ６ Each fused aryl, heteroaryl, or cycloalkyl can form a cycloalkyl ring, and one or more R Ａ２ optionally substituted with R Ａ２ are independently １ -C ６ Alkyl, C １ -C ６ Alkoxy, halogen -SO ３ H or -OSO ３ H; X １ is a divalent covalent linking moiety; X ２ is a divalent covalent linking moiety; and A polyplex wherein L is a targeting fragment, preferably said targeting fragment is capable of binding to a cell.

36. 36. The polyplex of claim 34 or claim 35, wherein the polyanion is a nucleic acid, and the nucleic acid is RNA.

37. 37. The polyplex of claim 36, wherein the RNA is dsRNA or ssRNA.

38. 37. The polyplex of claim 36, wherein the RNA is dsRNA.

39. 39. The polyplex of claim 38, wherein the dsRNA is polyinosinic:polycytidylic acid (poly(IC)).

40. 37. The polyplex of claim 36, wherein the RNA is ssRNA.

41. 41. The polyplex of claim 40, wherein the ssRNA is mRNA.

42. 36. The polyplex of claim 34 or claim 35, wherein the polyanion is a nucleic acid, and the nucleic acid is DNA.

43. 43. The composition of claim 42, wherein the DNA is plasmid DNA.

44. A composition according to any one of claims 1 to 33 or a polyplex according to any one of claims 34 to 43 for use in the treatment of cancer, preferably head and neck cancer or melanoma.

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