Compounds comprising cleavable linkers and uses thereof

Abstract

The present invention relates to compounds comprising cleavable linkers and uses thereof. The present invention provides a compound comprising a cleavable linker, a use thereof, and an intermediate compound for preparing the compound, and more particularly, the compound comprising a cleavable linker of the present invention may comprise an active agent having a specific function or activity (e.g., a drug, a toxin, a ligand, a probe for detection, etc. S02 functional groups capable of selectively releasing the active agent, and functional groups that trigger chemical reactions, physicochemical reactions, and / or biological reactions by external stimuli, and may further include ligands (e.g., oligopeptides, polypeptides, antibodies, etc.) having binding specificity for the desired target receptor.

Description

[0001] This application is a divisional application of the PCT international application PCT / IB2018 / 000847 filed on July 3, 2018, which entered the Chinese national phase on February 28, 2020, and the invention patent application with Chinese patent number 201880056603.X and the invention name “Compounds containing cleavable linkers and their uses”.

[0002] Related applications

[0003] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 597,226, filed on December 11, 2017, and Korean Patent Application No. 10-2017-0084805, filed on July 4, 2017. The contents of each of these patents are hereby incorporated by reference in their entirety. Technical Field

[0004] The present application relates to compounds comprising a cleavable linker and uses thereof. Background Art

[0005] Antibody-drug conjugates (ADCs) are emerging as a powerful class of anti-tumor agents with efficacy against a range of cancers. ADCs are typically composed of three distinct components: a cell-binding or targeting moiety; a linker; and a cytotoxic agent. The linker component of an ADC is an important feature in the development of targeted anti-cancer agents with the desired target specificity, i.e., high activity in tumor cells but low activity in healthy cells.

[0006] Therefore, there is a need for improved linkers that can be used to prepare ADCs. Summary of the Invention

[0007] Provided herein are conjugates of formula (I'):

[0008] (DL) n -(CB) cb

[0009] (I')

[0010] or a pharmaceutically acceptable salt thereof,

[0011] in:

[0012] CB is the targeting moiety;

[0013] cb and n are each independently an integer having a value from 1 to about 20, preferably from 1 to about 10;

[0014] Each DL is independently a group having the structure of formula (I"):

[0015]

[0016] Q is an active agent linked to L' via a heteroatom, preferably O or N;

[0017] Z' is a linking group;

[0018] L' is a spacer moiety attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent;

[0019] X is -O-, -C(R b )2-or-N(R c )-, preferably -O-;

[0020] Ar represents a ring, such as aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0021] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0022] X and Y' are positioned on adjacent atoms of Ar;

[0023] TG is a trigger group that, when activated, generates a molecule capable of reacting with the SO2 to displace (Q) q -(L') w and N, O or S atoms forming a 5- or 6-membered ring containing the intervening atoms of X-SO2 and Ar;

[0024] q is an integer having a value from 1 to about 20, preferably from 1 to about 10;

[0025] w, x, and y are each independently an integer having a value of 0 or 1;

[0026] Each R a and R c are independently hydrogen or lower alkyl; and

[0027] Each R b are independently hydrogen or lower alkyl; or

[0028] Two R's b Together with the atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring;

[0029] The condition is that when w is 0, q is 1.

[0030] The present invention also relates to compositions (eg, pharmaceutical compositions) comprising a compound of Formula (I') and a carrier (eg, a pharmaceutically acceptable carrier).

[0031] In one aspect, the invention provides conjugates of formula (I') and compositions comprising such conjugates, eg, for use in therapy, imaging, as sensors, as molecular switches, as molecular machines and / or as nanomachines.

[0032] In another aspect, the present invention further provides a conjugate of formula (I') and a pharmaceutical composition thereof for use in a method of delivering an active agent to a cell, wherein the targeting moiety is selected to bind to a molecule associated with the target cell. In particular, the compounds, conjugates and compositions of the present invention can be used to inhibit abnormal cell growth or treat a proliferative disorder in a mammal (e.g., a human), such as when the target cell is a cancer cell and the targeting moiety is selected to bind to a molecule associated with the cancer cell (and not with healthy cells or at least preferentially associated with tumor cells rather than healthy cells).

[0033] The conjugates of formula (I') of the present invention and pharmaceutical compositions thereof can be used to treat diseases in mammals (e.g., humans), such as cancer, rheumatoid arthritis, multiple sclerosis, graft-versus-host disease (GVHD), transplant rejection, lupus, myositis, infection, immunodeficiency such as AIDS, and inflammatory diseases.

[0034] In some embodiments of the present invention, a conjugate of formula (I') is provided:

[0035] (DL) n -(CB) cb

[0036] (I')

[0037] or a pharmaceutically acceptable salt thereof,

[0038] in:

[0039] CB is the targeting moiety;

[0040] cb and n are each independently an integer having a value of 1 to about 20, preferably 1 to about 10;

[0041] Each DL is independently a group having the structure of formula (I"):

[0042]

[0043] Each Q is independently an active agent linked to L' through a heteroatom, preferably O or N;

[0044] Z' is a linking group;

[0045] L' is a spacer moiety attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent;

[0046] X is -O-, -C(R b )2-or-N(R c )-, preferably -O-;

[0047] Ar represents a ring, such as aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0048] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0049] X and Y' are positioned on adjacent atoms of Ar;

[0050] TG is a trigger group that, when activated, reacts with SO2 to displace (Q) q -(L') w And form a X-SO 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0051] q is an integer having a value from 1 to about 20, preferably 1 to about 10;

[0052] w, x, and y are each independently an integer having a value of 0 or 1;

[0053] Each R a and R c are independently hydrogen or lower alkyl; and

[0054] Each R b are independently hydrogen or lower alkyl; or

[0055] Two R's b Together with the atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring;

[0056] with the proviso that when w is 0, q is 1. In some embodiments of the present invention, X is -O-.

[0057] In some embodiments of the present invention, Ar is aryl.

[0058] In some embodiments of the present invention, Ar is phenyl or naphthyl.

[0059] In some embodiments of the present invention, Z' is a C comprising at least two of 10 -C 100 Straight or branched saturated or unsaturated alkylene moiety:

[0060] (i) at least one heteroatom selected from -NH-, -C(=O), -O-, -S- and -P-;

[0061] (ii) at least one heteroarylene group;

[0062] (iii) at least one amino acid moiety, sugar bond, peptide bond, or amide bond; and

[0063] (iv) one or more substituents selected from the group consisting of: C1-C 20 Alkyl, C6-C 20 Aryl C1-C8 alkyl, -(CH2) s COOH and -(CH2) p NH2, s is an integer with a value from 0 to 10, and p is an integer with a value from 1 to about 10. In some embodiments of the present invention, Z' comprises a functional group that can be generated by a click chemistry reaction, such as a triazole.

[0064] In some embodiments of the present invention, Z' comprises:

[0065]

[0066] in:

[0067] Each V is independently a single bond, -O-, -S-, -NR 21 -、-C(O)NR 22 -、-NR 23 C(O)-、-NR 24 SO2- or -SO2NR 25 -;

[0068] R 21 、R 22 、R 23 、R 24 and R 25 are independently hydrogen, (C1-C6)alkyl, (C1-C6)alkyl(C6-C 20 )aryl or (C1-C6)alkyl (C3-C 20 ) heteroaryl;

[0069] r is an integer with a value ranging from 1 to about 10;

[0070] p is an integer with a value ranging from 0 to about 10;

[0071] q is an integer having a value from 1 to about 10; and

[0072] L" is a single bond.

[0073] In some embodiments of the present invention, Z' is a linking group connecting CB and Ar, comprising (CH2) linked to each other by covalent bonds in a linear chain. b , L c 、(P 1 ) a 、W a1 、W a2 、W a3 、Y 1 and Y 2 Group, in which:

[0074] W a1 、W a2 and W a3 are each independently -NH-, -C(O)- or -CH2-;

[0075] W b1 is an amide bond or a triazole group;

[0076] P 1 is an amide bond, an amino acid residue, or a peptide;

[0077] L c is an alkylene group;

[0078] Y 1 Yes - (CH2) q -(CH2CH2X”) o -or-(CH2) q -(X"CH2CH2X) o -;

[0079] X" is -O-, -S-, -NH- or -CH2-;

[0080] Y 2 is a single bond or a group selected from the following:

[0081]

[0082] W b2 is an amide bond or a triazole group;

[0083] a is 0 to 10;

[0084] b, c, and d are each independently an integer with a value from 1 to about 10; and

[0085] o and q are each independently an integer having a value from 1 to about 10.

[0086] In some embodiments of the present invention, Z' is a linking group of formula (A):

[0087] **-L c -W b1 -(CH2) b -W a3 -(P 1 ) a -Y 2 -W a2 -Y 1 -W a1 -*

[0088] (A)

[0089] in:

[0090] * is the point of attachment to the CB; and

[0091] ** is the point of attachment to Ar.

[0092] In some embodiments of the present invention, wherein P 1 yes

[0093]

[0094] in:

[0095] R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s C(O)R 13 or -(CH2) p NR 14 R 15 ;

[0096] p is an integer with a value ranging from 1 to about 10;

[0097] s is an integer with a value ranging from 0 to about 10;

[0098] R 13 is OH or –NH(CH2) s’ (X''CH2CH2) s” Z"-(CB) m ;

[0099] R 14 and R 15 are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2) s" Z"-(CB) m ;

[0100] s" is an integer with a value ranging from 0 to about 10;

[0101] s' is an integer with a value ranging from 1 to about 10;

[0102] m is an integer with a value of 0 or 1;

[0103] X'' is -O-, -S-, -NH- or -CH2-; and

[0104] Z" is to connect CB and R 14 or R 15 or Z" is a linking group comprising a reactive group.

[0105] In some embodiments of the invention, Z' is a linking group of formula (F), (G), (H), (J), (K), (L), (M), or (N):

[0106]

[0107]

[0108] in:

[0109] R e is an alkyl group;

[0110] X" is -O-, -S-, -NH- or -CH2-;

[0111] X 4 is -NHC(O)-(CH2) g -NH- or -C(O)NH-(CH2) h -NH-;

[0112] W b1 and W b2 are independently -C(O)NH-, -NHC(O)-,

[0113] R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s C(O)R 13 or -(CH2) p NR 14 R 15 ;

[0114] R 13 is OH or –NH(CH2) s’ (X''CH2CH2) s” Z"-(CB) m ;

[0115] R 14 and R 15are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2) s" Z”-(CB) m ;

[0116] s and s" are each independently an integer with a value of 0 to about 10;

[0117] m is an integer with a value of 0 or 1;

[0118] X'' is -O-, -S-, -NH- or -CH2-; and

[0119] Z" is to connect CB and R 14 or R 15 or Z" is a linking group comprising a reactive group; and

[0120] b, c, d, e, g, h, o, and q are each independently an integer with a value from 1 to about 10; and

[0121] s' is an integer with a value of 1 to about 10.

[0122] In some embodiments of the invention, TG is a reactive chemical moiety or functional group that can be cleaved by nucleophilic conditions, basic reagent conditions, light irradiation, reducing agent conditions, acidic conditions, enzymatic conditions, or oxidative conditions.

[0123] In some embodiments of the invention, TG is selected from the group consisting of:

[0124]

[0125] in:

[0126] Each R 21 are independently hydrogen or acetyl; and

[0127] R 22 It is hydrogen or lower alkyl.

[0128] In some embodiments of the present invention, x is 0.

[0129] In some embodiments of the present invention, TG is selected from the group consisting of -NO2, -C(O)-(CH2)2C(O)-alkyl, and nitrobenzyl.

[0130] In some embodiments of the present invention, Q is a chemical factor, a biological factor, a hormone, an oligonucleotide, a drug, a toxin, an affinity ligand, a probe for detection, or a combination thereof.

[0131] In some embodiments of the present invention, Q is a drug selected from a cytokine, an immunomodulatory compound, an anticancer agent, an antiviral agent, an antibacterial agent, an antifungal agent, an anthelmintic, or a combination thereof.

[0132] In some embodiments of the present invention, (Q) q -(L') w -Selected from:

[0133]

[0134] in:

[0135] X 1 Is -O- or -NR a -;

[0136] X 2 and X 4 each independently absent or -C(O)- or -C(O)O-;

[0137] X 3 is -OC(=O)-;

[0138] w' is an integer with a value of 1, 2, 3, 4, or 5;

[0139] R 9 and R 10 are each independently hydrogen, alkyl, aryl or heteroaryl, wherein alkyl, aryl and heteroaryl are unsubstituted or substituted with one or more radicals selected from, for example, alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 and -(CH2) u SO2R u3 Substituents substituted;

[0140] R u1 、R u2 and R u3 are each independently hydrogen, alkyl, aryl, or heteroaryl; and

[0141] u is an integer having a value from 1 to about 10.

[0142] In some embodiments of the present invention, (Q) q -(L') w -Selected from:

[0143]

[0144]

[0145]

[0146]

[0147] Where * represents (Q)q -(L') w Attachment point to -SO2-.

[0148] In some embodiments of the invention, the targeting moiety is a nanoparticle, an immunoglobulin, a nucleic acid, a protein, an oligopeptide, a polypeptide, an antibody, a fragment or a repeat of an antigenic polypeptide.

[0149] In some embodiments of the invention, the targeting moiety is an antibody selected from the group consisting of an intact polyclonal antibody, an intact monoclonal antibody, an antibody fragment, a single-chain Fv (scFv) mutant, a multispecific antibody, a bispecific antibody, a chimeric antibody, a humanized antibody, a human antibody, a fusion protein comprising an antigenic determinant portion of an antibody, and other modified immunoglobulin molecules comprising an antigen recognition site.

[0150] In some embodiments of the invention, the antibody is selected from the group consisting of muromonab-CD3, abciximab, rituximab, daclizumab, palivizumab, infliximab, trastuzumab (Herceptin), etanercept, basiliximab, gemtuzumab ozogamicin, alemtuzumab, ibritumomab tentanate, adalimumab, alefacept, omalizumab, efalizumab, tositumomob-I 131 , cetuximab, bevacizumab, natalizumab, ranibizumab, panitumumab, eculizumab, rilonacept, becelimumab, romiplostim, AMG-531, CNTO-148, CNTO-1275, ABT-874, LEA-29Y, belimumab, TACI-Ig, second-generation anti-CD20, ACZ-885, tocilizumab, atezolizumab, mepolizumab, pertuzumab, Humax CD20, tremelimumab (CP-675 206), tesimumab, MDX-010, IDEC-114, itraconazole, HuMax EGFR, aflibercept, HuMax-CD4, Ala-Ala, ChAglyCD3, TRX4, catumaxomab, IGN101, MT-201, Pregovomab, CH-14.18, WX-G250, AMG-162, AAB-001, motavizumab, MEDI-524, efavirenz, Aurograb, rixibacumab, third-generation anti-CD20, LY2469298, and veltuzumab.

[0151] In some embodiments of the present invention, a compound of formula (Ia) is provided:

[0152]

[0153] or a pharmaceutically acceptable salt thereof, wherein:

[0154] Each Q is independently an active agent linked to L' through a heteroatom, preferably O or N;

[0155] Z' is absent or is a linking group;

[0156] L' is a linking group attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent;

[0157] X is -O-, -CR a 2- or -NR'-, preferably -O-;

[0158] Ar represents a ring, such as aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0159] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0160] X and Y' are positioned on adjacent atoms of Ar;

[0161] TG is a trigger group that, when activated, reacts with SO2 to displace (Q) q -(L') w And form a X-SO 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0162] q is an integer having a value from 1 to about 20, preferably from 1 to about 10;

[0163] w, x, and y are each independently an integer having a value of 0 or 1;

[0164] Each R a and R c are independently hydrogen or lower alkyl; and

[0165] Each R b are independently hydrogen or lower alkyl; or

[0166] Two R's b Together with the carbon atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring;

[0167] The condition is that when w is 0, q is 1.

[0168] In some embodiments of the present invention, a compound of formula (IIa), (IIb) or (IIc) is provided:

[0169]

[0170] or a pharmaceutically acceptable salt thereof, wherein:

[0171] G is halogen, imidazole or N-methylimidazolium;

[0172] Each R 11 are independently C1-C6-alkyl;

[0173] Ar represents a ring, such as aryl, heteroaryl, cycloalkyl or heterocycloalkyl;

[0174] TG is a trigger group that, when activated, generates a 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0175] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0176] O and Y' are positioned on adjacent atoms of Ar;

[0177] x and y are each independently an integer having a value of 0 or 1;

[0178] Z' is absent or is a linking group; and

[0179] Each R a are independently hydrogen or alkyl; and

[0180] Each R b are independently hydrogen or alkyl; or

[0181] Two R's b Together with the carbon atom to which they are attached, they form a 3-5 membered ring, such as a 3-membered ring.

[0182] In some embodiments of the present invention, there is provided a method for preparing a compound comprising making a compound of formula (IIc):

[0183]

[0184] or a pharmaceutically acceptable salt thereof with a sulfonyl halide:

[0185]

[0186] Reaction to provide compounds of formula (Iaa):

[0187]

[0188] or a pharmaceutically acceptable salt thereof, wherein:

[0189] X a It is a halogen;

[0190] Each Q is independently an active agent linked to L' through a heteroatom, preferably O or N;

[0191] Z is absent or is a linking group;

[0192] L' is a linking group attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent;

[0193] Ar represents a ring, such as aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0194] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, such that if y is 1, a N, O, or S atom is attached to TG;

[0195] O and Y' are positioned on adjacent atoms of Ar;

[0196] TG is a trigger group that, when activated, reacts with SO2 to displace (Q) q -(L') w And form a X-SO 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0197] q is an integer with a value of 1 to about 20, preferably 1 to about 10;

[0198] w, x, and y are each independently an integer having a value of 0 or 1;

[0199] Each R a and R c are independently hydrogen or lower alkyl; and

[0200] Each R bare independently hydrogen or lower alkyl; or

[0201] Two R's b Together with the carbon atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring;

[0202] The condition is that when w is 0, q is 1.

[0203] In some embodiments of the present invention, there is provided a method for preparing a compound comprising:

[0204] (a) making a compound of formula (IIa):

[0205]

[0206]

[0207] or a pharmaceutically acceptable salt thereof and 1,1'-sulfonylbis(1H-imidazole):

[0208]

[0209] Reaction to provide a compound of formula (IIbb):

[0210]

[0211] or a pharmaceutically acceptable salt thereof, wherein:

[0212] Z' is absent or is a linking group;

[0213] Ar represents a ring, such as aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0214] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0215] O and Y' are positioned on adjacent atoms of Ar;

[0216] TG is a triggering group that, when activated, produces an N, O, or S atom capable of reacting with SO2 to form a 5-6 membered ring containing an intervening atom of X-SO2 and Ar;

[0217] x is an integer with value 0 or 1;

[0218] Each R a and Rc are independently hydrogen or lower alkyl; and

[0219] Each R b are independently hydrogen or lower alkyl; or

[0220] Two R's b Together with the carbon atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring.

[0221] In some embodiments of the present invention, wherein the compound of formula (IIbb) is further reacted with formula (Ia'): (Q) q -(L') w -H or a pharmaceutically acceptable salt thereof to provide compound (Iaa):

[0222]

[0223] or a pharmaceutically acceptable salt thereof, wherein:

[0224] X is -O-, -C(R b )2-or-N(R c )-, preferably -O-;

[0225] Each Q is independently an active agent linked to L' through a heteroatom, preferably O or N, wherein Q is displaced upon activation of TG;

[0226] L' is a spacer moiety attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent; and

[0227] q is an integer with a value of 1 to about 20, preferably 1 to about 10;

[0228] w is an integer with a value of 0 or 1;

[0229] The condition is that when w is 0, q is 1.

[0230] In some embodiments of the present invention, wherein the compound of formula (Iaa) is further reacted with a targeting moiety to provide a conjugate of formula (I').

[0231] In some embodiments of the present invention, X is -O-.

[0232] In some embodiments of the present invention, Ar is aryl. In some embodiments of the present invention, Ar is phenyl or naphthyl.

[0233] In some embodiments of the present invention, Z' is a linking group comprising one or more groups selected from the group consisting of isocyanide, isothiocyanate, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-halo), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a ) and dihydrogen phosphate (-OP(=O)(OH)2).

[0234] In some embodiments of the present invention, x is 0.

[0235] In some embodiments of the present invention, TG is -NO2, -OC(O)(CH2) r C(O)R 1 、-NHNH2、-BR 2 R 3 or in:

[0236] R 1 is a C1-C6 alkyl group;

[0237] R 2 and R 3 are each independently hydrogen, C1-C6 alkyl, C1-C6 alkoxy or hydroxy;

[0238] R 4 、R 5 、R 6 and R 7 are each independently hydrogen or C1-C6 alkyl; and

[0239] r is an integer with a value of 1, 2, 3, 4, or 5.

[0240] In some embodiments of the present invention, TG is a trigger group, and the trigger group includes β-galactoside, β-glucuronide, or a combination of β-galactoside and β-glucuronide.

[0241] In some embodiments of the invention, TG is selected from the group consisting of:

[0242]

[0243] in:

[0244] Each R 21 are independently hydrogen or acetyl; and

[0245] R 22 It is hydrogen or lower alkyl.

[0246] In some embodiments of the present invention, TG is selected from the group consisting of -NO2, -C(O)-(CH2)2C(O)-alkyl, and nitrobenzyl.

[0247] In some embodiments of the present invention, (Q) q -(L') w -Selected from:

[0248]

[0249] in:

[0250] X 1 Is -O- or -NR a -;

[0251] X 2 and X 4 each independently absent or C(O)-, -C(O)O-, or -C(O)NH-;

[0252] X 3 is -OC(=O)-;

[0253] w' is an integer with a value of 1, 2, 3, 4, or 5;

[0254] R 9 and R 10 are each independently hydrogen, alkyl, aryl or heteroaryl, wherein alkyl, aryl and heteroaryl are optionally substituted by one or more groups selected from, for example, alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 and -(CH2) u SO2R u3 Substituents substituted;

[0255] R u1 、R u2 and R u3 each is independently hydrogen, alkyl, aryl, or heteroaryl; and u is an integer with a value of 1 to about 10.

[0256] In some embodiments of the present invention, (Q) q -(L') w -Selected from:

[0257]

[0258]

[0259]

[0260]

[0261] Where * represents (Q) q -(L') w -Attachment point to -SO2-.

[0262] In some embodiments of the invention, the compound is:

[0263]

[0264] or a pharmaceutically acceptable salt thereof.

[0265] In some embodiments of the present invention, there is provided a method of preparing a conjugate comprising reacting any of the compounds described above with a targeting moiety.

[0266] In some embodiments of the present invention, a pharmaceutical composition is provided, comprising any of the above conjugates and a pharmaceutically acceptable carrier or excipient.

[0267] In some embodiments of the present invention, an imaging composition is provided, comprising any of the conjugates described above.

[0268] In some embodiments of the present invention, a method for imaging is provided, comprising contacting a material (eg, a cell) with the imaging composition described above.

[0269] In some embodiments of the present invention, a sensor compound is provided, comprising any of the conjugates described above.

[0270] In some embodiments of the present invention, a detection method is provided, which comprises contacting a material with the above-mentioned sensor compound.

[0271] In some embodiments of the present invention, a molecular switch, molecular machine or nanomachine is provided, comprising any of the above conjugates.

[0272] In some embodiments of the present invention, a method for moving a portion of a molecular device is provided, comprising mixing in a solution:

[0273] (1) the molecular switches, molecular machines or nanomachines described above; and

[0274] (2) An activator that activates the trigger group.

[0275] In some embodiments of the present invention, there is provided a method for delivering an active agent to a cell comprising contacting the cell with any of the conjugates described above, wherein the targeting moiety is selected to bind to a molecule associated with the target cell.

[0276] In some embodiments of the invention, the cells are in a subject in need thereof, thereby treating the disease or disorder.

[0277] In some embodiments of the invention, the target cell is a cancer cell and the targeting moiety is selected to bind to a molecule that is associated with cancer cells and not with healthy cells or at least preferentially associated with tumor cells rather than healthy cells.

[0278] In some embodiments of the invention, the disease or disorder is an autoimmune disease, an infectious disease, or a tumor.

[0279] In some embodiments of the present invention, a method for treating a proliferative disease is provided, comprising administering any of the conjugates described above.

[0280] In some embodiments of the invention, the proliferative disease is selected from an autoimmune disorder (e.g., systemic lupus erythematosus, rheumatoid arthritis, graft-versus-host disease, myasthenia gravis, or Sjögren's syndrome), a chronic inflammatory disorder (e.g., psoriasis, asthma, or Crohn's disease), a hyperproliferative disorder (e.g., breast cancer, lung cancer), a viral infection (e.g., herpes, papilloma, or HIV), osteoarthritis, and atherosclerosis.

[0281] In some embodiments of the invention, the proliferative disease is a cancer selected from the group consisting of carcinoma, lymphoma, blastoma, sarcoma, leukemia, or lymphoid malignancy.

[0282] In some embodiments of the invention, the cancer is squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer (e.g., small cell lung cancer, non-small cell lung cancer ("NSCLC"), lung adenocarcinoma, and lung squamous carcinoma), peritoneal cancer, hepatocellular carcinoma, stomach or gastric cancer (e.g., gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, acute leukemia, and head / brain and neck cancer.

[0283] In some embodiments of the invention, the cancer is cervical cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0284] Figure 1 Shown are the results of an enzymatic cleavage assay of Compound A-1.

[0285] Figure 2 Shown are the results of an enzymatic cleavage assay of Compound A-2.

[0286] Figure 3 Shown are the results of an enzymatic cleavage assay of compound A-3.

[0287] Figure 4 Shown are the results of an enzymatic cleavage assay of compound A-4.

[0288] Figure 5 Shown are the results of an enzymatic cleavage assay of Compound B-1.

[0289] Figure 6 Shown are the results of an enzymatic cleavage assay of compound B-3.

[0290] Figure 7 Shown are the results of an enzymatic cleavage assay (pH 7.4) of Compound B-2.

[0291] Figure 8 Shown are the results of an enzymatic cleavage assay (pH 5.0) of Compound B-2.

[0292] Figure 9 Shown are the results of an enzymatic cleavage assay of compound B-4.

[0293] Figure 10 Shown are the results of an enzymatic cleavage assay of compound C-3.

[0294] Figure 11 The results of stability analysis of Compound A-1 are shown.

[0295] Figure 12 Shown are the results of in vitro analysis of compound D-1-AB.

[0296] Figure 13 Shown are the results of in vivo testing of compounds D-1-AB, D-2-AB, D-8-AB, and D-16-AB. DETAILED DESCRIPTION

[0297] The present invention relates to compounds and conjugates comprising cleavable linkers and uses thereof. Representative compounds disclosed herein and conjugates include activating agents (e.g., chemical factors, biological factors, hormones, oligonucleotides, drugs, toxins, ligands, probes for detection, etc.) with desired functions or activities, undergo chemical reactions (e.g., physicochemical reactions and / or biological reactions) under predetermined conditions to release functional groups and SO2 functional groups of nucleophilic heteroatoms, wherein the SO2 functional group is positioned adjacent to the nucleophilic heteroatoms so that it can react with the nucleophilic heteroatoms in an intramolecular cyclization reaction to release the activating agent. In some embodiments, compounds disclosed herein and conjugates further include targeting moieties (e.g., oligopeptides, polypeptides, antibodies, etc.) that are specific for desired target receptors or other molecules associated with target cells.

[0298] definition

[0299] The meaning of the term "alkyl" is as understood in the art. For example, "alkyl," used alone or as part of a larger moiety, such as "alkoxy," "haloalkyl," "cycloalkyl," "heterocycloalkyl," and the like, may refer to a fully saturated straight or branched chain hydrocarbon. Typically, a straight or branched chain alkyl is an acyclic group having from 1 to about 20 carbon atoms, preferably from 1 to about 10 carbon atoms, unless otherwise defined. Examples of straight and branched chain alkyls include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl. C1-C6 straight or branched chain alkyls are also referred to as "lower alkyl" groups. Alkyl groups having two open valences are sometimes referred to as having an "ene" suffix, as in alkylidene. Exemplary alkylidene groups include methylene, ethylene, propylene, and the like.

[0300] In addition, the term "alkyl" (or "lower alkyl") can include both "unsubstituted alkyl" and "substituted alkyl," wherein the latter refers to an alkyl moiety having a substituent replacing a hydrogen on one or more carbons of the hydrocarbon backbone. If not otherwise specified, such substituents may include, for example, halogen, hydroxy, carbonyl (such as carboxyl, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or an aromatic or heteroaromatic moiety. The skilled artisan will understand that the moiety substituted on the hydrocarbon chain may itself be substituted, if appropriate. For example, the substituents of the substituted alkyl group can include substituted and unsubstituted forms of alkyl, amino, azido, imino, amido, phosphoryl (including phosphonate and phosphinate), sulfonyl (including sulfate, sulfonamido, sulfamoyl and sulfonate) and silyl, as well as ether, alkylthio, carbonyl (including ketone, aldehyde, carboxylate and ester), -CF , -CN and the like. Exemplary substituted alkyl groups are described below. The cycloalkyl group can be further substituted with alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, alkyl substituted with carbonyl, -CF , -CN and the like.

[0301] When the term "C x -C y ", when used in conjunction with a chemical moiety such as, for example, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, can include groups containing from x to y carbons in the chain, where "x" and "y" are integers selected from 1 to about 20, and where x is an integer having a value less than y, and x and y are not the same value. For example, the term "C x -C y -alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group, including straight-chain alkyl and branched-chain alkyl groups containing from x to y carbon numbers in the chain, including halogenated alkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl. The term "C2-C y -alkenyl""C2-C y "-alkynyl" refers to a substituted or unsubstituted unsaturated aliphatic group similar in length and possible substitution to the alkyl groups described above, but containing at least one double or triple bond, respectively. When applied to heteroalkyl, "C x -C y " indicates that the group contains from x to y number of carbon and heteroatoms in the chain. When applied to carbocyclic structures such as aryl and cycloalkyl, "C x -C y " indicates that the ring contains from x to y number of carbon atoms in the ring.

[0302] The term "alkoxy" has a meaning understood in the art and, for example, may refer to an alkyl group, preferably a lower alkyl group, having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0303] The terms "hal," "halo," and "halogen" are used interchangeably throughout and refer to fluorine (or fluoro, F), chlorine (or chloro, Cl), bromine (or bromo, Br), or iodine (or iodo, I).

[0304] The term "cycloalkyl" is understood in the art and can refer, for example, to a fully saturated, substituted or unsubstituted cyclic hydrocarbon. Cycloalkyl groups include monocyclic and bicyclic rings. Typically, monocyclic cycloalkyl groups have from 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms, unless otherwise defined. The second ring of a bicyclic cycloalkyl group can be selected from saturated, unsaturated, and aromatic rings. Cycloalkyl groups include bicyclic molecules in which one, two, or three or more atoms are shared by the two rings. The term "fused cycloalkyl" refers to a bicyclic cycloalkyl group in which each ring shares two adjacent atoms with another ring. The second ring of a fused bicyclic cycloalkyl group can be selected from saturated, unsaturated, and aromatic rings. A "cycloalkenyl" group is a cyclic hydrocarbon group containing one or more double bonds.

[0305] The meaning of the term "aryl" is understood in the art, and for example can refer to a substituted or unsubstituted monocyclic aromatic group, each atom of the ring being carbon. Preferably, the ring is 5 to 7 membered, more preferably 6 membered. The term "aryl" also includes a polycyclic ring system with two or more cyclic rings, wherein two adjacent rings share two or more carbon atoms, wherein at least one ring is aromatic, for example, another cyclic ring can be a cycloalkyl, cycloalkenyl, cycloalkyl, aryl, heteroaryl and / or heterocyclic radical. Aryl includes benzene, naphthalene, phenanthrene, phenol, aniline etc.

[0306] The terms "heterocyclyl" and "heterocycle" are as understood in the art and may, for example, refer to a substituted or unsubstituted non-aromatic ring structure, preferably a 3- to 10-membered ring, more preferably a 3- to 7-membered ring, the ring structure of which contains at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. Such heterocycles also include polycyclic ring systems having two or more cyclic rings, wherein two adjacent rings share two or more carbon atoms, wherein at least one ring is heterocyclic, for example, the other cyclic ring may be a cycloalkyl, cycloalkenyl, cycloalkyl, aryl, heteroaryl and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

[0307] The term "heteroaryl" is understood in the art and, for example, may refer to a substituted or unsubstituted aromatic monocyclic structure, preferably a 5- to 7-membered ring, more preferably a 5- to 6-membered ring, the ring structure of which contains at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings, wherein two adjacent rings share two or more carbon atoms, wherein at least one ring is heteroaromatic, for example, the other cyclic ring can be a cycloalkyl, cycloalkenyl, cycloalkyl, aryl, heteroaryl and / or heterocyclyl. Heteroaryl includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine.

[0308] The term "substituted" refers to a moiety having a substituent that replaces a hydrogen on one or more carbon or heteroatom atoms of the moiety. It will be understood by those skilled in the art that "substituted" or "substituted with" includes the implicit proviso that such substitution complies with the permitted valences of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., the compound does not spontaneously undergo transformations such as rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds.

[0309] In some embodiments, the substituent group allowed includes the aromatic and non-aromatic substituents of acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic organic compounds.For suitable organic compounds, the substituent group allowed can be one or more and be identical or different.For purposes of the present invention, heteroatoms such as nitrogen can have the hydrogen substituent and / or any substituent group allowed of organic compounds described herein, and the substituent group satisfies the valence of heteroatoms.Substituent group can include any substituent group described herein, for example, as halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imines, cyano, nitro, azido, sulfydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclic radical, alkyl, aralkyl or aromatic or heteroaromatic part. The skilled artisan will appreciate that substituents themselves may be substituted, if appropriate. Unless specifically stated as "unsubstituted," references to chemical moieties herein are understood to include substituted variants. For example, reference to an "aryl" group or moiety implicitly includes both substituted and unsubstituted variants.

[0310] The term "subject" contemplated for administration includes, for example, humans (i.e., male or female, of any age group, such as pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or elderly adults)) and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys); mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds, including commercially relevant birds such as chickens, ducks, geese, and / or turkeys. Preferred subjects are humans.

[0311] As used herein, a therapeutic agent that "prevents" a condition or disorder can refer, for example, to a compound that reduces the occurrence of the condition or disorder in a treated sample relative to an untreated control sample, or delays the onset of or reduces the severity of one or more symptoms of the condition or disorder relative to an untreated control sample.

[0312] The term "treatment" includes preventative treatment and / or therapeutic treatment. The term "preventative" or "therapeutic" treatment is art-recognized and includes administering one or more of the following subject compositions to the host. If administered before the clinical manifestations of an undesirable disorder (e.g., a disease of a host animal or other undesirable disorder), treatment is preventative (i.e., it protects the host against the development of an undesirable disorder), and if administered after the manifestations of an undesirable disorder, treatment is therapeutic (i.e., it is intended to reduce, improve, or stabilize its undesirable disorder or its side effect).

[0313] In certain embodiments, compounds disclosed herein and conjugates can be used alone or in combination with another type of therapeutic compound or medicament. As used herein, the phrase "combined administration" refers to any form of administration of two or more different therapeutic compounds, so that the therapeutic compound previously administered is still effective in vivo when the second compound is administered (for example, two compounds are effective in the subject at the same time, which can include the synergistic effect of the two compounds). For example, different therapeutic compounds and conjugates can be administered concomitantly or sequentially in the same formulation or in separate formulations. In certain embodiments, different therapeutic compounds and conjugates can be administered to each other within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours or 1 week. Therefore, subjects receiving such treatments can benefit from the combined effects of different therapeutic compounds and conjugates.

[0314] The terms "abnormal cell growth" and "proliferative disorder" are used interchangeably in this application. Unless otherwise indicated, "abnormal cell growth" as used herein refers to cell growth independent of normal regulatory mechanisms (e.g., loss of contact inhibition). This includes, for example, the abnormal growth of: (1) tumor cells (tumors) that proliferate by expressing mutant tyrosine kinases or overexpressing receptor tyrosine kinases; (2) benign and malignant cells of other proliferative diseases in which abnormal tyrosine kinase activation occurs; (3) any tumor that proliferates through receptor tyrosine kinases; (4) any tumor that proliferates through abnormal serine / threonine kinase activation; and (5) benign and malignant cells of other proliferative diseases in which abnormal serine / threonine kinase activation occurs.

[0315] The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by uncontrolled cell growth. A "tumor" comprises one or more cancerous cells. Cancer includes, but is not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More specific examples of such cancers include squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer (including small cell lung cancer, non-small cell lung cancer ("NSCLC"), lung adenocarcinoma, and lung squamous carcinoma), peritoneal cancer, hepatocellular carcinoma, gastric cancer or stomach cancer (including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, acute leukemia, and head / brain and neck cancer.

[0316] Compounds and conjugates of the present invention

[0317] The present disclosure provides a conjugate of formula (I'):

[0318] (DL) n -(CB) cb

[0319] (I')

[0320] or a pharmaceutically acceptable salt thereof,

[0321] in:

[0322] CB is the targeting moiety;

[0323] cb and n are each independently an integer having a value from 1 to about 20, preferably from 1 to about 10;

[0324] Each DL is independently a group having the structure of formula (I"):

[0325]

[0326] Each Q is independently an active agent linked to L' through a heteroatom, preferably O or N;

[0327] Z' is a linking group;

[0328] L' is a spacer moiety attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent;

[0329] X is -O-, -C(R b )2-or-N(R c )-, preferably -O-;

[0330] Ar represents a ring, such as aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0331] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0332] X and Y' are positioned on adjacent atoms of Ar;

[0333] TG is a trigger group that, when activated, generates a molecule capable of reacting with the SO2 to displace (Q) q -(L') w And form a X-SO 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0334] q is an integer having a value from 1 to about 20, preferably from 1 to about 10;

[0335] w, x, and y are each independently an integer having a value of 0 or 1;

[0336] Each R a and R c are independently hydrogen or lower alkyl; and

[0337] Each R b are independently hydrogen or lower alkyl; or

[0338] Two R's b Together with the atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring,

[0339] The condition is that when w is 0, q is 1.

[0340] Each active agent can be any suitable active agent, as described in more detail below. Although many traditional conjugation methods require certain functional groups (such as amines or hydroxyls) to form stable bonds, the disclosure herein provides a strategy for forming linkages using functional groups (such as phenols and tertiary amines) to form stable bonds in the conjugates disclosed herein while still allowing release under predetermined conditions of activation of the trigger group.

[0341] Many suitable trigger groups are known in the art, and exemplary trigger groups and conditions for activating them, such as the moieties described below for Y, are discussed below. Some trigger groups include N, O, or S atoms, but in non-nucleophilic form. For example, the NO2 group is a trigger group that is reduced to an NH2 or NHOH group that can react with SO2 under reducing conditions, and the acetate group is a trigger group that is hydrolyzed to a hydroxyl group that can react with SO2 under hydrolysis conditions. Other trigger groups do not include N, O, or S atoms, but are converted into nucleophilic N, O, or S atoms after being activated. For example, a borate group is a trigger group that is converted into a hydroxyl group that can react with SO2 under oxidizing conditions (such as peroxides). Preferably, the trigger group is selected so that the conditions for activating it selectively activate in this way without cleaving or degrading other parts of the conjugate, such as the targeting moiety. Once nucleophilic N, O, or S atoms are generated, the atoms intramolecularly attack the SO2 moiety to form a ring, thereby expelling (Q) q -(L') w -H moiety, wherein H is bound to the heteroatom of Q or L' previously attached to the SO2 moiety.

[0342] In embodiments where w is 0, q is 1 and Q is attached directly to SO2 via a heteroatom. Thus, activation of the trigger group generates a nucleophilic heteroatom that intramolecularly attacks the SO2 moiety to form a ring, thereby expelling the active agent QH, wherein H is bound to the heteroatom previously attached to SO2.

[0343] In embodiments where w is 1, L' can be selected to allow attachment of multiple occurrences of Q (which can be the same or different). Thus, each instance of Q is indirectly attached to SO2 via the spacer moiety. In such embodiments, the activation trigger group generates a nucleophilic heteroatom that intramolecularly attacks the SO2 moiety to form a ring, thereby expelling (Q) q-L'-H moiety, wherein H binds to the heteroatom in L' that was previously attached to the SO2 moiety. In such embodiments, the released heteroatom triggers an intramolecular reaction that expels the one or more active agents Q (e.g., if Q has a tertiary amine attached to L' as a quaternary ammonium) or QH. For example, the heteroatom can undergo an intramolecular cyclization reaction with the ester moiety formed by the hydroxyl group of QH, thereby forming a ring and expelling the active agent QH. Alternatively, the heteroatom can undergo intramolecular tautomerization, which expels the active agent Q or QH.

[0344] Ar can be any suitable ring, including bicyclic or other polycyclic rings, so that the part undergoing intramolecular cyclization remains in close proximity to promote the reaction after activation of the triggering group. The planar characteristics of aromatic and heteroaromatic rings are preferred because the rigid geometry of the substituents on such rings ensures the favorable placement of the reactive part, although other types of rings (such as cycloalkenyl or heterocycloalkenyl) can achieve similar geometries. The number or identity of the heteroatoms in the five or six membered rings and / or the rings and / or the substituents on the other of the rings (e.g., electron donating substituents or electron withdrawing substituents) can be selected to regulate the cyclization rate based on the resulting bond angle of the ring. Similarly, when it is necessary to slow down the intramolecular cyclization rate, the more flexible conformations of the rings of cycloalkyl and heterocyclyl can be useful.

[0345] Z' can be any suitable linking group that connects Ar to one or more CB groups. Typically, the linking group should be sufficiently hydrophilic to promote water solubility and prevent the aggregation of the conjugate, such as by comprising parts such as polyethylene glycol moieties, peptide sequences, charged moieties (such as carboxylates, amines, nitrogen-containing rings, etc.) to balance the hydrophobic characteristics of any alkyl chains that may be included. Because it is generally advantageous to prepare conjugates in a modular manner, Z' can contain linking units that are functional groups generated by conjugating from one reactive part to another. Representative linking units are discussed in more detail below (e.g., in combination with variable Z), and common linking groups include amides, triazoles, oximes, carbamates, etc. Representative Z' groups include L 1’ -Z group, discussed in more detail below. In some embodiments, all DL groups attached to each CB are identical, while in other embodiments, each CB can be attached to two or more different DL groups. For example, some DL groups can have trigger groups that are activated under a first condition, while other DL groups can have trigger groups that are activated under a second condition, such that, for example, one active agent can be selectively released under the first condition, but a second active agent can be selectively released under the second condition.

[0346] The present disclosure also provides compounds useful as intermediates or reagents in the formation of the DL group in formula (I'), as described in formula (I"). Thus, in some embodiments, provided herein are compounds of formula (Ia):

[0347]

[0348] or a pharmaceutically acceptable salt thereof, wherein:

[0349] Each Q is independently an active agent linked to L' through a heteroatom, preferably O or N;

[0350] Z' is absent or is a linking group;

[0351] L' is a spacer moiety attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent;

[0352] X is -O-, -CR a 2- or -NR'-, preferably -O-;

[0353] Ar represents a ring, such as aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0354] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0355] X and Y' are positioned on adjacent atoms of Ar;

[0356] TG is a trigger group that, when activated, generates a molecule capable of reacting with the SO2 to displace (Q) q -(L') w And form a X-SO 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0357] q is an integer having a value from 1 to about 20, preferably from 1 to about 10;

[0358] w, x, and y are each independently an integer having a value of 0 or 1;

[0359] Each R a and R c are independently hydrogen or lower alkyl; and

[0360] Each R b are independently hydrogen or lower alkyl; or

[0361] Two R's b Together with the carbon atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring.

[0362] In certain embodiments of formula (I') and (Ia), -Y' is -(CH2) y NR"-、-(CH2) y O- or -(CH2) y S-, which is positioned such that if y is 1, a N, O, or S atom is attached to TG; R" is hydrogen or C1-C6-alkyl; and y is an integer having a value of 0 or 1. In some such embodiments, TG is β-galactoside, β-glucuronide, or a combination of β-galactoside and β-glucuronide.

[0363] In some embodiments of Formulas (I') and (Ia), (L') w Each Q is linked to -SO2-; and each Q is an active agent that is linked to one L' group via a heteroatom, preferably O or N, and forms an -O-, -OC(O)-, -OC(O)O-, or -OC(O)NH- bond containing the heteroatom of Q.

[0364] In other embodiments, (Q) q -(L') w -Selected from:

[0365]

[0366] in:

[0367] Q is an active agent linked to L' via a heteroatom, preferably O or N,

[0368] X 4 There is no -O-, -OC(O)-, -OC(O)O-, or -OC(O)NH- bond or bonds to the heteroatom containing Q;

[0369] X 1 Is -O- or -NR a -;

[0370] X 2 is -O-, -OC(O)-, -OC(O)O- or -OC(O)NH-;

[0371] X 3 is -OC(=O)-;

[0372] w' is an integer with a value of 1, 2, 3, 4, or 5;

[0373] R 9 and R 10 are each independently hydrogen, alkyl, aryl or heteroaryl, wherein alkyl, aryl and heteroaryl are unsubstituted or substituted with one or more radicals selected from, for example, alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 and -(CH2) u SO2R u3 Substituents substituted;

[0374] R u1 、R u2 and R u3 are each independently hydrogen, alkyl, aryl, or heteroaryl; and

[0375] u is an integer having a value from 1 to about 10.

[0376] In some such embodiments, (Q) q -(L') w -Selected from:

[0377]

[0378] Furthermore, the present invention provides intermediates for preparing conjugates according to formula (I') or compounds according to formula (Ia), wherein (Q) in these formulas q -(L') w is replaced by a leaving group such as a halogen (preferably fluorine) to allow (Q) q -(L') w Attachment.

[0379] In certain such embodiments, Z' comprises a reactive group (e.g., a precursor group, as discussed in more detail below with respect to Z) that can be used to attach the compound to a trigger, such as a CB (e.g., to prepare a compound of Formula (I') as discussed in more detail above), a solid surface (e.g., to form a solid-supported array or sensor particle), or any other molecule or support of interest. In certain preferred embodiments, the compound of Formula (I') is selected from:

[0380] in:

[0381] R 1 is a C1-C6 alkyl group; and

[0382] R 21 and R 22 Each is independently hydrogen or C1-C6-alkyl.

[0383] In other embodiments, the compound of formula (I') is selected from:

[0384]

[0385] In still other embodiments, the compound of formula (I') is selected from:

[0386]

[0387]

[0388] In other embodiments, the compound of formula (I') is selected from:

[0389]

[0390] In certain preferred embodiments, Z is a linking group having a structure of Formula (F), (G), (H), (J), (K), (L), (M), or (N):

[0391]

[0392] in:

[0393] * is the attachment point to CB; ** is the attachment point to Ar; R e is an alkyl group;

[0394] X" is -O-, -S-, -NH- or -CH2-;

[0395] X 4 is -NHC(O)-(CH2) g -NH- or -C(O)NH-(CH2) h -NH-;

[0396] W b1 and W b2 are independently -C(O)NH-, -NHC(O)-,

[0397] L 2 is an optional spacer moiety and may be further substituted with one or more substituents (such as C1-C6 alkyl, C5-C 14 alkyl, aryl and C3-C8 heteroaryl), wherein the alkyl, aryl and heteroaryl may be further substituted, for example, by one or more substituents selected from the group consisting of: C1-C 10 Alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 、-(CH2)u CO2H, -(CH2) u CO2R u1 and -(CH2) u SO2R u3 , where R u1 、R u2 and R u3 are independently hydrogen, C1-C 15 Alkyl, C6-C 20 Aryl or C3-C 10 heteroaryl; and u is an integer with a value of 1 to about 10;

[0398] R 12 is hydrogen, C1-C8 alkyl, or an amino acid moiety, such as a natural amino acid moiety;

[0399] b, c, d, e, g, h, o, and qq are each independently an integer with a value from 1 to about 10; and

[0400] s' is an integer with a value of 1 to about 10.

[0401] In other embodiments, Z is a linking group having a structure of Formula (F'), (G'), (H'), (J'), (K'), (L'), (M'), or (N'):

[0402]

[0403]

[0404] In certain preferred embodiments, CB is selected from the group consisting of:

[0405] In certain preferred embodiments, (Q) q -(L') w Selected from:

[0406]

[0407]

[0408]

[0409] Also provided herein are compounds of formula (I):

[0410]

[0411] or a pharmaceutically acceptable salt thereof, wherein:

[0412] X is -O-, -CH2- or -NR'-;

[0413] R' is hydrogen, C1-C6-alkyl, C6-C 14 -aryl or C2-C 20 -heteroaryl;

[0414] Ar is C5-C 20 -aromatic ring, C2-C 20 -heteroaromatic ring, C2-C 30 -fused ring or C5-C 20 -Aromatic ring-C2-C 20 -heteroaromatic ring;

[0415] R is a substituent on Ar or -L 1' -Z-(CB) cb , preferably -L 1' -Z-(CB) cb ;

[0416] L 1' It is C1-C 200 -alkylene or a C1-C1-C1-alkylene group further comprising at least one of a peptide bond, an amino bond, an ether bond, a triazole bond, a tetrazole bond, a sugar bond, a sulfonamide bond, a phosphonate bond, a sulfo bond or a dendrimer structure. 200 alkylene;

[0417] Z is connected to CB and L 1' a linking unit or reactive group (e.g., the reactive group enables connection to the CB);

[0418] The CB is a targeting moiety, such as a ligand, that has properties that bind it to the receptor;

[0419] cb is an integer with a value of 0, 1, or 2;

[0420] n is an integer with a value of 1, 2, 3, or 4;

[0421] Y is -NO2, -OC(O)(CH2) r C(O)R 1 、-O(CH2) r -Ar 1 -NO2, -NHOH, -NHNH2, -BR 2 R 3 、 Or -Y'-TG, preferably, Y is -NO2, -OC(O)(CH2) r C(O)R 1 、-O(CH2) r -Ar 1 -NO2, -NHNH2, -BR 2 R 3 、 or -Y'-TG;

[0422] R 1 is a C1-C6 alkyl group;

[0423] r is an integer with a value of 1, 2, 3, 4, or 5;

[0424] Ar 1 It is C6-C 20 arylene;

[0425] R 2 and R 3 are each independently hydrogen, C1-C6-alkyl, C1-C6-alkoxy or hydroxy;

[0426] R a 、R b 、R c and R d are each independently hydrogen or C1-C6 alkyl;

[0427] Y' is -(CH2) x NR"-、-(CH2) x O- or -(CH2) x S-;

[0428] R" is hydrogen or C1-C6 alkyl;

[0429] x is an integer with value 0 or 1;

[0430] TG is the trigger group;

[0431] Q is for -Q 1 or -L'-(Q 1 ) w ;

[0432] L' is a compound having -O- or -NR"'- at one end and -O-, -OC(O)-, -O(CO)O-, -OC(O)NR"'- or -OC(O)NR at the other end. 4 C7-C of CH2O- 30 -Hydrocarbon spacer, wherein -O-, -OC(O)-, -O(CO)O- or -OC(O)NR""- may further comprise a C7-C 30 In the hydrocarbon spacer, the C7-C 30 The hydrocarbon spacer is further substituted by one or more substituents (such as C1-C6 alkyl, C5-C 14 alkyl, aryl and C3-C8 heteroaryl), wherein the alkyl, aryl and heteroaryl may be further substituted, for example, by one or more substituents selected from the group consisting of: C1-C 10 Alkyl, -(CH2) u NH2, -(CH2)u NR u1 R u2 、-(CH2) u CO2H, -(CH2) u CO2R u1 and -(CH2) u SO2R u3 , where R u1 、R u2 and R u3 are independently hydrogen, C1-C 15 Alkyl, C6-C 20 Aryl or C3-C 10 heteroaryl; and u is an integer with a value of 1 to about 10;

[0433] Q 1 An active agent comprising at least one -OH, -NH-, -NR5R6, -SH, -SO2NH2 or -COOH functional group;

[0434] R 4 is hydrogen, C1-C6-alkyl, C5-C 14 -aryl or C3-C8-heteroaryl, wherein alkyl, aryl and heteroaryl are substituted or unsubstituted;

[0435] R 5 and R 6 are each independently hydrogen, C1-C6-alkyl, C3-C9-cycloalkyl or C5-C 10 -heteroaryl, wherein the heteroaryl group is substituted or unsubstituted;

[0436] R'" and R"" are each independently hydrogen or C1-C6-alkyl; and

[0437] w is an integer with a value of 1, 2, 3, 4, or 5.

[0438] In some embodiments, the compound of formula (I) comprises a functional group (eg, Y) capable of inducing intramolecular cyclization by external stimuli. In certain embodiments, the functional group is introduced at the ortho position relative to X.

[0439] In some embodiments, R' is C1-C6-alkyl, C6-C 14 -aryl or C2-C 20 -heteroaryl.

[0440] In some embodiments, Ar is C5-C 20 -aromatic ring, C2-C 20 -heteroaromatic ring, C2-C 30 -fused ring or C5-C 20 -Aromatic ring-C2-C20 - a heteroaromatic ring. For example, Ar can be a benzene ring, a naphthalene ring, a pyridine ring, or a quinolone ring. Preferably, Ar is a benzene ring or a naphthalene ring. In some embodiments, the compound of formula (I) is a compound having a structure according to formula (II):

[0441]

[0442] or a pharmaceutically acceptable salt thereof.

[0443] In other embodiments, the compound of formula (I) is a compound having a structure according to formula (III):

[0444]

[0445] or a pharmaceutically acceptable salt thereof.

[0446] In some embodiments, the compound is a compound of formula (I), (II) or (III), wherein R is selected from hydrogen, halogen (hal), aldehyde, acetal, ketal, -R*, -OR*, -SR*, -NR*R**, -C(hal)3, -CN, -OCN, -SCN, -N=C=O, -NCS, -NO, -NO2, -N3, -NC, -C(O)R*, -OC(O)R*, -OS(O)R*, -S(O)2R*, -S(O)2OR*, -OS(O)OR*, -OS(O)2OR*, -S(O)NR*R**, -S(O)2NR*R**, -S(O)R*, -OP(O)(OR*)2, -P(O) -(OR*)2, -OP(OR*)2, -OP(OR*)N(R**)2, -OP(O)(OR*)N(R**)2, -PR*, -P(O)2, -P(O)R*, -C(O)hal, -C(S)R*, -C02R*, -C(S)OR*, -C(O)SR*, -C(S)SR*, -C(O)NR*R**, -C(S)NR*R**, -C(=NR*)NR*R**, -NR*C(O)R**, -NR*S(O)2OR**, -NR*S(O)R**, -NR*C(O)NR**, -SS-R*, or -R*SSR**, wherein: R* and R** are each independently hydrogen, C1-C 18 Alkyl, C6-C 20 Aryl, C3-C 15 Heterocyclic or C3-C 20 Heteroaryl.

[0447] In some embodiments, the compound is of formula (I), (II) or (III), wherein R is hydrogen or *-(L a -A1-Lb -L c -Z) m -CB; where:

[0448] L a Is a single bond or C1-C 20 -alkylene;

[0449] A 1 is -C(O)NR*-, -NR*C(O)-, -NR*-, -O-, -PO3-, -OPO3-, -SO-, -SO2- or -SO3-;

[0450] L b is -(CH2CH2O) a -or-(CH2) a -;

[0451] R* is hydrogen, C1-C 18 -alkyl, C6-C 20 -aryl, C3-C 15 -heterocyclic or C3-C 20 heteroaryl;

[0452] a is an integer with a value ranging from 1 to about 20;

[0453] L c Is a single bond or C1-C 20 -alkylene;

[0454] n is an integer having a value of 1 or 2; and

[0455] Z is connected to CB and L c a connection unit; or

[0456] Z is a precursor selected from isocyanide, isothiocyanate, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where R a It is C1-C 10 -alkyl) and dihydrogen phosphate (-OP(=O)(OH)2);

[0457] CB is a targeting moiety, such as a ligand capable of binding to a receptor; and

[0458] m is an integer with a value of 0, 1, or 2.

[0459] In some embodiments, the compound is of formula (I), (II) or (III), wherein R is hydrogen or *-L a -A1-L b -L c -Z; where:

[0460] L a Is a single bond or C1-C 20 -alkylene;

[0461] A 1 is -C(O)NR*-, -NR*C(O)-, -NR*-, -O-, -PO3-, -PO4-, -SO-, -SO2- or -SO3-;

[0462] L b is -(CH2CH2O) a -or-(CH2) a -;

[0463] R* is hydrogen, C1-C 18 -alkyl, C6-C 20 -aryl, C3-C 15 -heterocyclic or C3-C 20 -heteroaryl;

[0464] a is an integer with a value ranging from 1 to about 20;

[0465] L c Is a single bond or C1-C 20 -alkylene;

[0466] n is an integer having a value of 1 or 2; and

[0467] Z is a precursor selected from isocyanide, isothiocyanate, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where Ra It is C1-C 10 -alkyl) and dihydrogen phosphate (-OP(=O)(OH)2).

[0468] In some embodiments, the compound is of formula (I), (II) or (III), wherein R is *(-L a -A1-L b -L c -Z) m -CB; where:

[0469] L a Is a single bond or C1-C 20 -alkylene;

[0470] A 1 is -C(O)NR*-, -NR*C(O)-, -NR*-, -O-, -PO3-, -PO4-, -SO-, -SO2- or -SO3-;

[0471] L b is -(CH2CH2O) a -or-(CH2) a -;

[0472] R* is hydrogen, C1-C 18 -alkyl, C6-C 20 -aryl, C3-C 15 -heterocyclic or C3-C 20 -heteroaryl;

[0473] a is an integer with a value ranging from 1 to about 20;

[0474] L c Is a single bond or C1-C 20 -alkylene;

[0475] n is an integer with a value of 1 or 2;

[0476] Z is connected to CB and L c connection unit;

[0477] CB is a targeting moiety, such as a ligand having properties that bind it to the receptor; and

[0478] m is an integer with a value of 1 to 2.

[0479] In some embodiments, the compound is of formula (I), (II) or (III), wherein L' is a C7-C ... 30 Hydrocarbon spacer.

[0480] In some embodiments, the compound is of formula (I), (II) or (III), wherein Q is selected from the group consisting of -L'-(Q 1 ) w :

[0481]

[0482] in:

[0483] Q 1 is an active agent comprising at least one functional group selected from: -OH, -NR 5 R 6 , -SH and -COOH;

[0484] Q 2 Yes Contains-NR 5 R 6 active agent;

[0485] X 1 is -O- or -NR"'-;

[0486] X 2 and X 4 each independently absent or selected from -O-, -OC(O)-, -OC(O)O-, and -OC(O)NH-;

[0487] X 3 is -OC(=O)-;

[0488] R 5 and R 6 is the same as defined above;

[0489] R 9 and R 10 are independently hydrogen, C1-C6 alkyl, C6-C 14 Aryl or C3-C9 heteroaryl, R9 and R 10 The alkyl, aryl and heteroaryl groups may be further substituted by one or more substituents selected from the group consisting of: C1-C 10 Alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 and -(CH2) u SO2R u3 , and R u1 、R u2 and R u3 are independently hydrogen, C1-C 15 Alkyl, C6-C 20 Aryl or C3-C 10heteroaryl; and u is an integer with a value of 1 to about 10;

[0490] R''' is hydrogen or C1-C6-alkyl; and

[0491] w is an integer with a value of 1, 2, 3, 4, or 5.

[0492] In certain embodiments, -L'-(Q 1 ) w Selected from

[0493]

[0494] Selected from -OH, -NR 5 R 6 , -SH and -COOH Q, Q 1 or Q 2 At least one functional group of serves as a point of attachment of the active agent to L'. The functional group may be present as part of an ester, thioester, carbonate, carbamate, amide, sulfonamide, sulfonate, sulfate, or other suitable linkage; i.e., when the active agent is part of a conjugate, -OH, -NR 5 R 6 , -SH and -COOH moieties themselves do not exist.

[0495] In some embodiments, Q 2 Yes Contains-NR 5 R 6 The active agent, wherein the active agent can be combined with a quaternary amine structure, for example, -NR 5 R 6 The moiety is capable of forming a quaternary amine bond with L'.

[0496] In some embodiments of Formula (I"), (Ia), (I), (II), and (III), R 4 is a substituted alkyl, aryl, or heteroaryl. In some such embodiments, R 4 Substituted by one or more substituents selected from the group consisting of: C1-C 10 Alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 、-(CH2) u CO2H, -(CH2) u CO2R u1 and -(CH2) u SO2R u3 , where R u1 、R u2 and R u3 are independently hydrogen, C1-C15 -alkyl, C6-C 20 -aryl or C3-C 10 -heteroaryl; and u is an integer with a value of 1 to about 10.

[0497] In some embodiments of Formula (I"), (Ia), (I), (II), and (III), R 5 and / or R 6 Is -NR 7 R 8 Substituted heteroaryl, wherein R 7 and R 8 are each independently hydrogen, C1-C6-alkyl, C3-C9-cycloalkyl or C5-C 14 -aryl.

[0498] In some embodiments of Formula (I"), (Ia), (I), (II), and (III), Q or -(L') w -(Q) q Selected from:

[0499]

[0500]

[0501]

[0502] In certain embodiments, provided herein are compounds of Formula (I), (II), or (III), wherein:

[0503] Y is -NO2, -OC(O)(CH2) r C(O)R 1 、-O(CH2) r -Ar 1 -NO2, -NHOH, -BR 2 R 3 Or -Y'-TG, preferably, Y is -NO2, -OC(O)(CH2) r C(O)R 1 、-O(CH2) r -Ar 1 -NO2, -BR 2 R 3 or -Y'-TG;

[0504] R 1 is a C1-C6 alkyl group;

[0505] r is an integer with a value of 1, 2, 3, 4, or 5;

[0506] Ar 1is phenylene, biphenylene or naphthylene;

[0507] R 2 and R 3 are each independently hydrogen, C1-C6-alkyl, C1-C6-alkoxy, or hydroxy;

[0508] Y' is -(CH2) x NR"-、-(CH2) x O- or -(CH2) x S-;

[0509] R" is hydrogen or C1-C6-alkyl;

[0510] x is an integer with value 0 or 1;

[0511] R" is hydrogen or C1-C6-alkyl; and

[0512] TG is a trigger group, such as β-galactoside, β-glucuronide, or a combination of β-galactoside and β-glucuronide.

[0513] In certain embodiments, the compound of Formula (I), (II) or (III) is selected from:

[0514]

[0515]

[0516] in:

[0517] R 1 is a C1-C6 alkyl group;

[0518] R 21 and R 22 are each independently hydrogen or acetyl;

[0519] R is hydrogen, *-L a -A1-L b -L c -Z, or a group having a structure of formula (F), (G), (H), (J), (K), (L), (M) or (N)

[0520]

[0521]

[0522] L a Is a single bond or C1-C 20 alkylene;

[0523] A 1is -C(O)NH-, -NHC(O)-, -NH-, -O-, -PO3-, -PO4-, -SO-, -SO2- or -SO3-;

[0524] L b is -(CH2CH2O) a -or-(CH2) a -;

[0525] a is an integer with a value ranging from 1 to about 20;

[0526] L c It is C1-C 20 alkylene;

[0527] X" is -O-, -S-, -NH- or -CH2-;

[0528] W b1 and W b2 are independently -C(O)NH-, -NHC(O)-,

[0529] R 12 is hydrogen, C1-C8 alkyl, amino acid moiety, -(CH2) s COR 13 or -(CH2) p NR 14 R 15 ;

[0530] R 13 is OH or -NH(CH2) s' (X"CH2CH2) s" Z;

[0531] R 14 and R 15 are each independently hydrogen or -(C(O)(CH2) s' (X"CH2CH2) s" Z) m -CB;

[0532] X" is -O-, -S-, -NH- or -CH2-;

[0533] R e is C1-C8-alkyl or -(L 1' -Z) m -CB;

[0534] X 4 is -NHC(O)-(CH2) g -NH- or -C(O)NH-(CH2) h -NH-;

[0535] b, c, d, e, g, h, o, and q are each independently an integer with a value ranging from 1 to about 10;

[0536] p is an integer with a value ranging from 1 to about 10;

[0537] s and s" are each independently an integer with a value of 0 to about 10;

[0538] s' is an integer with a value ranging from 1 to about 10;

[0539] m is an integer with a value of 0 or 1;

[0540] Z is isocyanide, isothiocyanate, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where R a It is C1-C 10 -alkyl) or dihydrogen phosphate (-OP(=O)(OH)2).

[0541] CB is a ligand selected from the group consisting of:

[0542]

[0543] And Q is selected from:

[0544]

[0545]

[0546]

[0547] Release of active agent

[0548] As described above, in certain embodiments, the compounds and conjugates disclosed herein are capable of cleaving one or more active agents (composed of Q, Q 1 , Q 2 In certain embodiments, the chemical reaction is a physicochemical reaction and / or a biochemical reaction.

[0549] In some embodiments, the compounds and conjugates disclosed herein comprise a nucleophilic functional group (Y or Y') introduced at an atom adjacent to X (e.g., O) on Ar. Typically, the nucleophilic functional group is masked by a trigger group (TG), as described in further detail below. Upon activation, the trigger group releases the nucleophilic functional group to react with a nearby SO2 moiety in an intramolecular cyclization, ultimately releasing one or more active agents (Q, Q 1 or Q 2 In some such embodiments, the one or more active agents are released by an intramolecular cyclization reaction following the chemical, physicochemical, and / or biochemical reaction (see, e.g., Reaction Scheme 1), or the active agent is released by a 1,6-elimination or 1,4-elimination following the intramolecular cyclization reaction (see, e.g., Reaction Scheme 2).

[0550] For example, the mechanism when Y is -Y'-TG is shown in Reaction Scheme 1:

[0551] Reaction Scheme 1:

[0552]

[0553] When Q is The mechanism is shown in Reaction Scheme 2:

[0554] Reaction Scheme 2:

[0555]

[0556] In some embodiments, Q 1 Upon release, the active agent comprises at least one functional group selected from: -OH, -NH-, -SH, and -COOH. According to these embodiments, as further described herein, Q 1 The compound as described herein is conjugated via -OH, -NH-, -SH and -COOH, for example, via a functional group selected from ester, amide, thioester, carbamate, urea, oxime, hydrazone, etc. In some such embodiments, the compound is conjugated using Q 2 Alternative Q 1 , and Q 2 is an amine-containing drug. In other embodiments, Q 2 is an active agent capable of binding to an ammonium unit. In still other embodiments, Q 2 Able to 2 The released active agent is released in its original form with amine groups, wherein the active agent can be a drug, a toxin, an affinity ligand, a probe for detection, or a combination thereof.

[0557] In some embodiments, the compounds and conjugates disclosed herein are chemically and physiologically stable. In some such embodiments, the compounds and conjugates disclosed herein reach the desired target cells with little dissociation of the active agent in the blood, thereby selectively releasing the drug.

[0558] Trigger group (TG)

[0559] In some embodiments, the conjugates of the present invention comprise a trigger group (TG). TG is a group that can be cleaved, preferably selectively cleaved, by a chemical reaction such as a biological reaction. Typically, the trigger group is used to mask the nucleophilic nature of the Y or Y' group, thereby providing stability to the compounds and conjugates disclosed herein (e.g., by preventing self-immolation or intramolecular cyclization before the conjugate reaches the target site or undergoes a predetermined trigger condition). Upon activation, the trigger group releases the nucleophilic Y or Y' group and allows self-immolation or intramolecular cyclization as described above to occur.

[0560] In some embodiments, TG comprises a sequence (such as a peptide sequence) or a portion that is recognized by TEV, trypsin, thrombin, cathepsin B, cathepsin D, cathepsin K, caspase 1, matrix metalloproteinase (MMP), etc., which can be hydrolyzed by an enzyme (e.g., oxidoreductase, transferase, hydrolase, lyase, isomerase, ligase, etc.) and / or can comprise a portion selected from phosphodiester, phospholipid, ester, β-galactose, β-glucose, fucose, oligosaccharide, etc.

[0561] In some embodiments, the TG comprises a reactive chemical moiety or functional group that is cleaved under nucleophilic conditions (e.g., a silyl ether, 2-N-acylnitrobenzenesulfonamide, an unsaturated vinyl thioether, an activated sulfonamide, a malondialdehyde-indole derivative, a levulinic acyl ester, a hydrazone, or an acylhydrazone).

[0562] In some embodiments, the TG may comprise a reactive chemical moiety or functional group that is cleavable under alkaline reagent conditions (eg, 2-cyanoethyl ester, ethylene glycol disuccinate, 2-sulfonylethyl ester, alkylthioester, or phenylthioester).

[0563] In some embodiments, the TG may comprise a reactive chemical moiety or functional group that is cleavable by photoirradiation (e.g., a 2-nitrobenzyl derivative, a benzoyl ester, 8-quinolinylbenzenesulfonate, a coumarin, a phosphotriester, a bis-arylhydrazone, or a bimane di-thiopropionic acid derivative).

[0564] In some embodiments, a TG may comprise a reactive chemical moiety or functional group (eg, a hydroxylamine, disulfide, levulinate, nitro, or 4-nitrobenzyl derivative) that can be cleaved by reducing agent conditions.

[0565] In some embodiments, the TG may comprise a reactive chemical moiety or functional group that can be cleaved using acidic conditions (e.g., saccharides, tert-butylcarbamate analogs, dialkyl or diaryl dialkoxysilanes, orthoesters, acetals, aconityl groups, hydrazones, β-thiopropionates, phosphoramidates, imines, trityl groups, vinyl ethers, polyketals, and alkyl 2-(diphenylphosphino)benzoate derivatives; alkyl esters, 8-hydroxyquinoline esters, and picolinates).

[0566] In some embodiments, the TG may comprise a reactive chemical moiety or functional group that can be cleaved under oxidative conditions (eg, a boronate ester, a vicinal diol, a p-methoxybenzyl derivative, or a selenium compound).

[0567] In certain preferred embodiments, the TG comprises a sugar that is cleavable under acidic or enzymatic conditions. In certain preferred embodiments, the trigger group is -NO2, which is cleavable under reducing conditions. In certain preferred embodiments, the trigger group is a boronate ester, which is cleavable under oxidative conditions. In certain preferred embodiments, the trigger group is an ester, which is cleavable under acidic, alkaline, or enzymatic conditions. In certain preferred embodiments, the trigger group is a hydrazone, which is cleavable under nucleophilic or acidic conditions. In certain preferred embodiments, the trigger group is a hydroxylamine, which is cleavable under reducing conditions.

[0568] Sugar trigger group

[0569] In some embodiments, the compounds and conjugates disclosed herein comprise a sugar trigger group, such as a trigger group selected from:

[0570]

[0571] Each R 21 are independently hydrogen or are selected such that OR 21 is a hydroxy protecting group (eg, acetyl); and R 22is hydrogen or lower alkyl (e.g., C1-C6-alkyl). In certain embodiments, the hydroxy protecting group can be used in organic synthesis, including but not limited to: methyl ether, methoxymethyl ether, methylthiomethyl ether, 2-methoxyethoxymethyl ether, bis(2-chloroethoxy)methyl ether, tetrahydropyranyl ether, tetrahydrothiopyranyl ether, 4-methoxytetrahydropyranyl ether, 4-methoxytetrahydrothiopyranyl ether, tetrahydrofuranyl ether, 1-ethoxyethyl ether, 1-methyl-1-methoxyethyl ether, 2-(phenylseleno)ethyl ether, tert-butyl ether, allyl ether, benzyl ether, o-nitrobenzyl ether, triphenylmethyl ether, α-naphthyldiphenylmethyl ether, p-methoxyphenyldiphenylmethyl ether, 9-(9-phenyl- The present invention also includes, but is not limited to, anthracene ether, trimethylsilyl ether, isopropyldimethylsilyl ether, tert-butyldimethylsilyl ether, tert-butyldiphenylsilyl ether, tribenzylsilyl ether, triisopropylsilyl ether, formates, acetates, trichloroacetates, phenoxyacetates, isobutyrates, pivalates, adamantates, benzoates, 2,4,6-trimethylbenzoate, methyl carbonate, 2,2,2-trichloroethyl carbonate, allyl carbonate, p-nitrophenyl carbonate, benzyl carbonate, p-nitrobenzyl carbonate, S-benzylthiocarbonate, N-phenylcarbamate, nitrates, 2,4-dinitrophenylsulfenate, and the like.

[0572] Protective groups as triggering groups

[0573] In some embodiments, TG is a group that can be cleaved by a chemical reaction, a physicochemical reaction, and / or a biological reaction. In certain embodiments, TG is a protecting group. In some such embodiments, the protecting group is an amine protecting group, an alcohol protecting group, or a thiol protecting group.

[0574] Amine protecting groups

[0575] In certain embodiments, the amine protecting group is a general protecting group that can be used in organic synthesis, including but not limited to: m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, phenyl (o-nitrophenyl) methyl carbamate, alkyl carbamate, 9-fluorenylmethyl carbamate, 2,2,2-trichloroethyl carbamate, 2-trimethylsilylethyl carbamate (Teoc), tert-butyl carbamate (Boc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), 8-quinolinyl carbamate, N-hydroxypiperidinyl carbamate, benzyl carbamate, p-methoxybenzyl carbamate, p-nitrobenzyl carbamate, diphenylmethylcarbamate, acetamide, chloroacetamide, trichloroacetamide, phenylacetamide, benzamide, N-phthalimide, N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, benzylsulfenamide, o-nitrobenzenesulfenamide, triphenylmethylsulfenamide, p-toluenesulfonamide, methanesulfonamide, etc., but are not limited to these.

[0576] Alcohol protecting group

[0577] In certain embodiments, the alcohol protecting group is a general protecting group that can be used in organic synthesis, including but not limited to: methyl ether, methoxymethyl ether (MOM ether), benzyloxymethyl ether (BOM ether), 2-(trimethylsilyl)ethoxymethyl ether (SEM ether), phenylthiomethyl ether (PTM ether), 2,2-dichloro-1,1-difluoroethyl ether, p-bromophenyl ether, chloropropyl methyl ether, isopropyl ether, cyclohexyl ether, 4-methoxybenzyl, 2,6-dichlorobenzyl ether, 4-(di ... methylaminocarbonyl)benzyl ether, 9-anthryl methyl ether, 4-picolyl ether, methylthiomethyl ether (MTM ether), 2-methoxyethoxymethyl ether (MEM ether), bis(2-chloroethoxy)methyl ether, tetrahydropyranyl ether (THP ether), tetrahydrothiopyranyl ether, 4-methoxytetrahydropyranyl ether, 4-methoxytetrahydrothiopyranyl ether, tetrahydrofuranyl ether, 1-ethoxyethyl ether, 1-methyl-1-methoxyethyl ether, 2-(phenylseleno)ethyl ether), tert-butyl ether, allyl Ether, benzyl ether, o-nitrobenzyl ether, triphenylmethyl ether, α-naphthyldiphenylmethyl ether, p-methoxyphenyldiphenylmethyl ether, 9-(9-phenyl-10-oxo)anthryl ether, trimethylsilyl ether (TMS ether), isopropyldimethylsilyl ether, tert-butyldimethylsilyl ether (TBDMS ether), tert-butyldiphenylsilyl ether, tribenzylsilyl ether, triisopropylsilyl ether, formates, acetates, trichloroacetates, phenoxyacetates, isobutyrates, pivalates , adamantate, benzoate, 2,4,6-trimethylbenzoate (Mesitoate), methyl carbonate, 2,2,2-trichloroethyl carbonate, allyl carbonate, p-nitrophenyl carbonate, benzyl carbonate, p-nitrobenzyl carbonate, S-benzyl thiocarbonate, N-phenylcarbamate, nitrate, 2,4-dinitrophenylsulfenate, dimethylphosphino ester (DMP ester), dimethylphosphinothioate (MPT ester), methanesulfonic acid aryl ester, toluenesulfonic acid aryl ester, etc., but are not limited to these.

[0578] Thiol protecting groups

[0579] In certain embodiments, the thiol protecting group can be used in organic synthesis, including but not limited to: S-benzyl sulfide, S-p-methoxybenzyl sulfide, S-o- or p-hydroxy or acetoxybenzyl sulfide, S-p-nitrobenzyl sulfide, S-4-picolyl sulfide, S-2-picolyl N-oxide sulfide, S-9-anthryl methyl sulfide, S-9-fluorenyl methyl sulfide, S-methoxymethyl monothioacetal, A-acetyl derivatives, S-benzoyl derivatives, S-(N-ethylcarbamate), S-(N-methoxymethylcarbamate), etc., but are not limited to these.

[0580] Linking group

[0581] In some embodiments, the compounds and conjugates disclosed herein comprise a linker group that covalently links each CB and Ar. Typical linkers are stable, non-hydrolyzable moieties such as C 10 -C 100 In certain embodiments, the linking unit satisfies at least two and more preferably at least three of the following four criteria:

[0582] (i) at least one -CH2- in the alkylene moiety is substituted (i.e., replaced) by a heteroatom selected from the group consisting of -NH-, -C(=O), -O-, -S-, and -P-;

[0583] (ii) the alkylene moiety comprises at least one heteroarylene group;

[0584] (iii) the alkylene moiety comprises at least one amino acid moiety, sugar bond, peptide bond, or amide bond; and

[0585] (iv) The alkylene group may be further substituted by one or more substituents selected from the group consisting of: C1-C 20 Alkyl, C6-C 20 Aryl C1-C8 alkyl, -(CH2) s COOH and -(CH2) p NH2, wherein s is an integer having a value from 0 to 10, and p is an integer having a value from 1 to about 10.

[0586] In certain embodiments, the linking unit comprises at least two, and more preferably at least three, of the following:

[0587] (i) at least one heteroatom selected from -NH-, -C(=O), -O-, -S- and -P-;

[0588] (ii) at least one heteroarylene group;

[0589] (iii) at least one amino acid moiety, sugar bond, peptide bond, or amide bond; and

[0590] (iv) The alkylene group may be further substituted by one or more substituents selected from the group consisting of: C1-C 20 Alkyl, C6-C 20 Aryl C1-C8 alkyl, -(CH2) s COOH and -(CH2) p NH2, wherein s is an integer having a value from 0 to 10, and p is an integer having a value from 1 to about 10.

[0591] In other embodiments, the linking group connecting each CB and Ar comprises a functional group generated by a click chemistry reaction.

[0592] In an alternative embodiment, the linking unit comprises a reactive functional group capable of participating in a click chemistry reaction.

[0593] Click chemistry is a reaction that can be performed under mild conditions and has extremely high selectivity for functional groups that are not commonly found in biomolecules (e.g., azide groups, acetylene groups, etc.). Therefore, the secondary reaction can be carried out in the presence of complex trigger groups, targeting moieties, etc. In addition, click chemistry has high reaction specificity. For example, the click chemistry reaction between the azide group and the acetylene group is carried out selectively without interference from other functional groups present in the molecule. For example, azide-acetylene click chemistry can obtain triazole moieties in high yield.

[0594] Thus, in some embodiments, the linking group connecting each CB and Ar comprises,

[0595]

[0596] V can be a single bond, -O-, -S-, -NR 21 -、-C(O)NR 22 -、-NR 23 C(O)-、-NR 24 SO2- or -SO2NR 25 -, R 21 to R 25 can be independently hydrogen, (C1-C6) alkyl, (C1-C6) alkyl (C6-C 20 )aryl or (C1-C6)alkyl (C3-C 20 ) heteroaryl, r can be an integer with a value of 1 to about 10, p can be an integer with a value of 0 to about 10, q can be an integer with a value of 1 to about 10, and L" can be a single bond.

[0597] In other embodiments, the linking unit connecting each CB and Ar is a linking group represented by formula (A):

[0598] **-L c -W b1 -(CH2) b -W a3 -(P 1 ) a -Y 2 -W a2 -Y 1 -W a1 -*

[0599] (A)

[0600] in:

[0601] * is the attachment point to the CB;

[0602] ** is the point of attachment to Ar;

[0603] W a1 、W a2 and W a3 are each independently -NH-, -C(=O)- or (-CH2-) b ;

[0604] W b1 is an amide bond or a triazole group;

[0605] P 1 Is connected to W a3 and Y 2 and is an amino acid moiety, a peptide bond, or an amide bond;

[0606] L c is an alkylene group;

[0607] Y 2 Is a single bond, -W a4 -(CH2) c -W b2 -(CH2) d -W a5 -or-W a6 -(CH2) e -CR e R f -X-;

[0608] R e Is C1-C8 alkyl or CB-W a7 -Y3-W c1 -(CH2) f -;

[0609] R f It's BW a7 -Y 3 -W c1 -(CH2) f -;

[0610] X is -NHC(=O)-(CH2) g -W a8 -or-C(=O)NH-(CH2) h -W a9 -;

[0611] W a4 、W a5 、W a6 、W a7 、W a8 and Wa9 are each independently -NH-, -C(=O)- or -CH2-;

[0612] W b2 is an amide bond or a triazole group;

[0613] W c1 is -NHC(=O)- or -C(=O)NH-;

[0614] Y 3 Yes - (CH2) i -(X'CH2CH2) j -(CH2) k -;

[0615] X' is -O-, -S-, -NH- or -CH2-;

[0616] CB is defined as above;

[0617] b, c, d, e, f, g, h, i, and j are each independently an integer with a value from 1 to about 10;

[0618] k and y are each independently an integer with a value ranging from 0 to about 10;

[0619] Y 1 Yes - (CH2) q -(CH2CH2X") o -or-(CH2) q -(X"CH2CH2) o -;

[0620] X" is -O-, -S-, -NH- or -CH2-; and

[0621] o and q are integers with values ​​ranging from 1 to about 10.

[0622] In some embodiments, P 1 Contains at least one unit represented by formula (B) or (C):

[0623]

[0624] in:

[0625] R 12 is hydrogen, C1-C8-alkyl, an amino acid side chain such as a natural amino acid side chain (e.g., H, methyl, isopropyl, isobutyl, sec-butyl, S-methyl sulfide, benzyl, indole, pyrrolidine, hydroxymethyl, amidoyl, lysyl, imidazole, glycyl, glutamyl, carbamoylbutyric acid, formamide, aspartic acid, 1-hydroxyethyl, and 2-hydroxyethyl)), -(CH2) s COR13 or -(CH2) p NR 14 R 15 ;

[0626] R 13 is OH or -NH(CH2) s' (X"CH2CH2) s" Z;

[0627] R 14 and R 15 are each independently hydrogen or -(C(O)(CH2) s' (X"CH2CH2) s" Z) m -CB;

[0628] X" is -O-, -S-, -NH- or -CH2-;

[0629] Z and CB are defined as above;

[0630] p is an integer with a value ranging from 1 to about 10;

[0631] s and s" are integers with a value ranging from 0 to about 10;

[0632] s' is an integer with a value ranging from 1 to about 10; and

[0633] m is an integer with a value of 0 or 1.

[0634] In some embodiments of Formula (B) or (C):

[0635] R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s C(O)R 13 or -(CH2) p NR 14 R 15 ;

[0636] p is an integer with a value ranging from 1 to about 10;

[0637] s is an integer with a value ranging from 0 to about 10;

[0638] R 13 is OH or –NH(CH2) s’ (X''CH2CH2) s” Z"-(CB) m ;

[0639] R 14 and R 15 are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2)s" Z”-(CB) m ;

[0640] s" is an integer with a value ranging from 0 to about 10;

[0641] s' is an integer with a value ranging from 1 to about 10;

[0642] m is an integer with a value of 0 or 1;

[0643] X'' is -O-, -S-, -NH- or -CH2-; and

[0644] Z" is to connect CB and R 14 or R 15 or Z" is a linking group comprising a reactive group.

[0645] In some such embodiments of Formula (B) or (C):

[0646] R 13 is OH or -NH(CH2) s' (X''CH2CH2) s" Z”;

[0647] R 14 and R 15 are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2) s" Z”; and

[0648] Z" is a reactive precursor of the linking unit, wherein the reactive precursor is selected from isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where R a It is C1-C 10 -alkyl) and dihydrogen phosphate (-OP(=O)(OH)2).

[0649] In other such embodiments of Formula (B) or (C):

[0650] R13 is OH or -NH(CH2) s' (X''CH2CH2) s" Z”CB;

[0651] R 14 and R 15 are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2) s" Z”CB; and

[0652] Z" is formed by the precursor CB and R 14 or R 15 The remaining part of the linker is connected to the precursor selected from isocyanide, isothiocyanate, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where R a It is C1-C 10 -alkyl) and dihydrogen phosphate (-OP(=O)(OH)2).

[0653] In some embodiments, Y 2 is a single bond or is selected from:

[0654]

[0655] in:

[0656] W b2 -C(O)NH-, -NHC(O)-,

[0657] R e is C1-C8-alkyl or -(L 1' -Z-) m CB;

[0658] R f It's BW b2' -(CH2) i -(X'CH2CH2) j -NH-C(=O)-(CH2) f-;

[0659] X a is -NHC(=O)-(CH2) g -NH- or -C(O)NH-(CH2) h -NH-;

[0660] W b2' is -C(O)NH- or -NHC(=O)-;

[0661] c, d, e, f, g, h, i, and j are each independently an integer with a value from 1 to about 10;

[0662] X" is -O-, -S-, -NH-, or -CH2-; and

[0663] L 1' , Z, m and B are the same as defined above.

[0664] In certain embodiments, the linking unit connecting each CB and Ar is a (CH2) b , L c 、(P 1 ) a 、W a1 、W a2 、W a3 、Y 1 and Y 2 A linker group of a group, wherein:

[0665] W a1 、W a2 and W a3 are each independently -NH-, -C(O)- or -CH2-;

[0666] W b1 is an amide bond or a triazole group;

[0667] P 1 is an amide bond, an amino acid residue, or a peptide;

[0668] L c is an alkylene group;

[0669] Y 1 Yes - (CH2) q -(CH2CH2X") o -or-(CH2) q -(X"CH2CH2X") o -;

[0670] X" is -O-, -S-, -NH- or -CH2-;

[0671] Y 2is a single bond or a group selected from the following:

[0672]

[0673] W b2 is an amide bond or a triazole group;

[0674] a is 0 to 10;

[0675] b, c, and d are each independently an integer with a value from 1 to about 10; and

[0676] o and q are each independently an integer having a value from 1 to about 10.

[0677] In some embodiments, R 12 is a natural amino acid side chain. In other embodiments, R 12 It is an unnatural amino acid side chain.

[0678] In some embodiments, the linking unit connecting each CB and Ar is a linking group represented by formula (A):

[0679] **-L c -W b1 -(CH2) b -W a3 -(P 1 ) a -Y 2 -W a2 -Y 1 -W a1 -*

[0680] (A)

[0681] in:

[0682] * is the point of attachment to the CB; and

[0683] ** is the point of attachment to Ar.

[0684] In some such embodiments, P 1 yes

[0685]

[0686] in:

[0687] R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s COOH or -(CH2) p NH2;

[0688] p is an integer with a value from 1 to about 10; and

[0689] s and s" are each independently an integer having a value from 0 to about 10.

[0690] In some embodiments, P 1 yes

[0691]

[0692] in:

[0693] R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s C(O)R 13 or -(CH2) p NR 14 R 15 ;

[0694] p is an integer with a value ranging from 1 to about 10;

[0695] s is an integer with a value ranging from 0 to about 10;

[0696] R 13 is OH or –NH(CH2) s’ (X''CH2CH2) s” Z"-(CB) m ;

[0697] R 14 and R 15 are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2) s" Z"-(CB) m ;

[0698] s" is an integer with a value ranging from 0 to about 10;

[0699] s' is an integer with a value ranging from 1 to about 10;

[0700] m is an integer with a value of 0 or 1;

[0701] X'' is -O-, -S-, -NH- or -CH2-; and

[0702] Z" is to connect CB and R 14 or R 15 or Z" is a linking group comprising a reactive group.

[0703] In P 1 In some such embodiments:

[0704] R 13 is OH or -NH(CH2) s' (X''CH2CH2) s" Z”;

[0705] R 14 and R 15 are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2) s" Z”; and

[0706] Z" is a reactive precursor of the linking unit, wherein the reactive precursor is selected from isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where R a It is C1-C 10 -alkyl) and dihydrogen phosphate (-OP(=O)(OH)2).

[0707] In P 1 In other such embodiments:

[0708] R 13 is OH or -NH(CH2) s' (X''CH2CH2) s" Z”CB;

[0709] R 14 and R 15 are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2) s" Z”CB; and

[0710] Z" is formed by the precursor CB and R 14 or R 15 The remaining part of the linker is connected to the precursor selected from isocyanide, isothiocyanate, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where R a It is C1-C 10 -alkyl) and dihydrogen phosphate (-OP(=O)(OH)2).

[0711] In alternative embodiments, the linking unit connecting CB and Ar is a linking group represented by formula (F), (G), (H), (J), (K), (L), (M), or (N):

[0712]

[0713]

[0714] in:

[0715] R e is an alkyl group;

[0716] X 4 is -NHC(O)-(CH2) g -NH- or -C(O)NH-(CH2) h -NH-;

[0717] e, g, and h are each independently an integer having a value of 1 to about 10; and s′ is an integer having a value of 1 to about 10.

[0718] In some embodiments of Formula (F), (G), (H), (I), (J), (K), (L), or (M): R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s C(O)R 13 or -(CH2) p NR 14 R 15 ; p is an integer with a value ranging from 1 to about 10;

[0719] s is an integer with a value ranging from 0 to about 10;

[0720] R 13 is OH or –NH(CH2) s’ (X''CH2CH2) s” Z”-(CB) m ;

[0721] R 14 and R 15 are each independently hydrogen or -C(O)(CH2)s' (X''CH2CH2) s" Z"-(CB) m ;

[0722] s" is an integer with a value ranging from 0 to about 10;

[0723] s' is an integer with a value ranging from 1 to about 10;

[0724] m is an integer with a value of 0 or 1;

[0725] X'' is -O-, -S-, -NH- or -CH2-; and

[0726] Z" is to connect CB and R 14 or R 15 or Z" is a linking group comprising a reactive group.

[0727] In some such embodiments of Formula (F), (G), (H), (I), (J), (K), (L), or (M):

[0728] R 13 is OH or -NH(CH2) s' (X''CH2CH2) s" Z”;

[0729] R 14 and R 15 are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2) s" Z”; and

[0730] Z" is a reactive precursor of the linking unit, wherein the reactive precursor is selected from isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where R a It is C1-C 10 -alkyl) and dihydrogen phosphate (-OP(=O)(OH)2).

[0731] In some such embodiments of Formula (F), (G), (H), (I), (J), (K), (L), or (M):

[0732] R 13 is OH or -NH(CH2) s' (X''CH2CH2) s" Z”CB;

[0733] R 14 and R 15 are each independently hydrogen or -C(O)(CH2) s' (X''CH2CH2) s" Z”CB; and

[0734] Z" is formed by the precursor CB and R 14 or R 15 The remaining part of the linker is connected to the precursor selected from isocyanide, isothiocyanate, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where R a It is C1-C 10 -alkyl) and dihydrogen phosphate (-OP(=O)(OH)2).

[0735] Targeting moiety

[0736] The compounds and conjugates of the present invention may further comprise a ligand or targeting moiety CB. In some embodiments, the ligand or targeting moiety is any molecular recognition element that can specifically interact with at least one other molecule through, for example, non-covalent bonds (such as hydrogen bonds, metal coordination, hydrophobic forces, van der Waals forces, π-π interactions, halogen bonds, electrostatic and / or electromagnetic effects). In certain embodiments, the CB is selected from nanoparticles, immunoglobulins, nucleic acids, proteins, oligopeptides, polypeptides, antibodies, fragments of antigenic polypeptides, repeats, and the like.

[0737] The compounds and conjugates of the present invention may comprise one or more targeting moieties. That is, the variable cb may have an integer value selected from 1, 2, 3, 4, 5, 1-10, or 1-20.

[0738] In some embodiments, the CB comprises two or more independently selected natural or unnatural amino acids conjugated by covalent bonds (e.g., peptide bonds), and can comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more natural or unnatural amino acids conjugated by peptide bonds. In some embodiments, the ligand comprises a shorter amino acid sequence (e.g., a natural protein fragment or a synthetic polypeptide fragment) as well as a full-length protein (e.g., a pre-engineered protein).

[0739] In some embodiments, the CB is selected from antibodies, hormones, drugs, antibody analogs (e.g., non-IgG), proteins, oligopeptides, polypeptides, etc. that bind to receptors. In certain embodiments, the CB selectively targets drugs in specific organs, tissues, or cells. In other embodiments, the CB specifically binds to receptors that are overexpressed in cancer cells, such as compared to normal cells, and can be classified as monoclonal antibodies (mAbs) or antibody fragments and low molecular weight non-antibodies. Preferably, the CB is selected from peptides identified in library screening, tumor cell-specific peptides, tumor cell-specific aptamers, tumor cell-specific carbohydrates, tumor cell-specific monoclonal antibodies, polyclonal antibodies, and antibody fragments.

[0740] Exemplary ligands or targeting moieties include, but are not limited to, carnitine, inositol, lipoic acid, pyridoxal, ascorbic acid, niacin, pantothenic acid, folic acid, riboflavin, thiamine, biotin, vitamin B 12, other water-soluble vitamins (vitamin B), fat-soluble vitamins (vitamins A, D, E, K), RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), transferrin, VIP (vasoactive intestinal peptide) receptor, APRPG (Ala-Pro-Arg-Pro-Gly) peptide, TRX-20 (thioredoxin-20), integrin, nucleolin, aminopeptidase N (CD13), endoglin, vascular epithelial growth factor receptor , low-density lipoprotein receptor, transferrin receptor, somatostatin receptor, bombesin, neuropeptide Y, luteinizing hormone-releasing hormone receptor, folate receptor, epidermal growth factor receptor, transforming growth factor, fibroblast growth factor receptor, asialoglycoprotein receptor, galectin-3 receptor, E-selectin receptor, hyaluronic acid receptor, prostate-specific membrane antigen (PSMA), cholecystokinin A receptor, cholecystokinin B receptor, lectin domain receptor, mucin receptor, opioid receptor, plasminogen receptor, bradykinin receptor , insulin receptor, insulin-like growth factor receptor, angiotensin AT1 receptor, angiotensin AT2 receptor, granulocyte macrophage colony-stimulating factor receptor (GM-CSF receptor), galactosamine receptor, sigma-2 receptor, delta-like 3 (DLL-3), aminopeptidase P, melanocyte transferrin, leptin, tetanus toxin Tet1, tetanus toxin G23, RVG (rabies virus glycoprotein) peptide, HER2 (human epidermal growth factor receptor 2), GPNMB (non-transfer glycoprotein b), Ley, CA6, C anAng, SLC44A4 (solute carrier family 44 member 4), CEACAM5 (carcinoembryonic antigen-related cell adhesion molecule 5), connexin-4, carbonic anhydrase 9, TNNB2, 5T4, CD30, CD37, CD74, CD70, PMEL17, EphA2 (ephrin A2 receptor), Trop-2, SC-16, tissue factor, ENPP-3 (AGS-16), SLITRK6 (SLIT and NTRK-like family member 6), CD27, LewisY antigen, LIV1, GPR161 (G protein-coupled receptor 161), PBR (peripheral benzodiazepine receptor), MERTK (Mer receptor tyrosine kinase) receptor, CD71, LLT1 (lectin-like transcript 1 or CLED2D), interleukin-22 receptor, σ1 receptor, peroxisome proliferator-activated receptor, DLL3, C4.4a, cKIT, ephrin A, CTLA4 (cytotoxic T-lymphocyte-associated protein 4), FGFR2b (fibroblast growth factor receptor 2b), N-acetylcholine receptor, gonadotropin Glandular hormone-releasing hormone receptor, gastrin-releasing peptide receptor, bone morphogenetic protein type 1B receptor (BMPR1B), E16 (LAT1, SLC7A5), STEAP1 (six-transmembrane epithelial antigen of the prostate), 0772P (CA125, MUC16), MPF (MSLN, mesothelin), Napi3b (SLC34A2), Sema5b (brain semaphorin 5b), ETBR (endothelin type B receptor), MSG783 (RNF124), STEAP2 (six-transmembrane epithelial antigen of the prostate 2), TrpM4 ( Transient receptor potential cation channel subfamily M member 4), CRIPTO (teratocarcinoma-derived growth factor), CD21, CD79b, FcRH2 (IFGP4), HER2 (ErbB2), NCA (CEACM6), MDP (DPEP1), IL20R-α (IN20Ra), Brevican (BCAN), EphB2R, ASLG659 (B7h), CD276, PSCA (prostate cancer stem cell antigen precursor), GEDA, BAFF-R (BR3), C D22 (BL-CAM), CD79a, CXCR5, HLA-DOB, P2X5, CD72, LY64, FcRH1, IRTA2, TENB2, SSTR2, SSTR5, SSTR1, SSTR3, SSTR4, ITGAV (integrin, α5), ITGB6 (integrin, β6), MET, MUC1, EGFRvIII, CD33, CD19, IL2RA (interleukin 2 receptor, α), AXL, BCMA, CTA (cancer testis antigen (CTA)antigen)), CD174, CLEC14A, GPR78, CD25, CD32, LGR5 (GPR49), CD133 (Prominin), ASG5, ENPP3 (extracellular nucleoside pyrophosphatase / phosphodiesterase 3), PRR4 (proline-rich protein 4), GCC (guanylate cyclase 2C), Liv-1 (SLC39A6), CD56, CanAg, TIM-1, RG-1, B7-H4, PTK7, CD138, claudin (Claudins), Her3 (ErbB3), RON (MST1R), CD20, TNC (tenascin C), FAP, DKK-1, CD52, CS1 (SLAMF7), annexin A1, V-CAM, gp100, MART-1, MAGE-1 (melanoma antigen encoding gene-1), MAGE-3 (melanoma-associated antigen 3), BAGE, GAGE-1, MUM-1 (multiple myeloma proto-oncogene 1), CDK4, TRP-1 (gp75), TAG-72 (tumor-associated glycoprotein-72), gangliosides GD2, GD3, GM2, GM3, VEP8, VEP9, My1, VIM-D5, D156-22, OX40, RNAK, PD-L1, TNFR1, TNFR2, etc.

[0741] target

[0742] In some embodiments, one or more targets of a molecular recognition element are specifically associated with one or more specific cells or tissue types. In some embodiments, a target is specifically associated with one or more specific disease states. In some embodiments, a target is specifically associated with one or more specific developmental stages. For example, the expression level of a cell type-specific marker in the cell type is typically at least 2 times greater than that in a reference cell population. In some embodiments, the presence level of a cell type-specific marker is at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 50 times, at least 100 times, or at least 1,000 times greater than its average expression in a reference population. The detection or measurement of a cell type-specific marker can distinguish one or more cell types of interest from many, most, or all other types of cells. In some embodiments, a target can comprise a protein, a carbohydrate, a lipid, and / or a nucleic acid, as described herein.

[0743] In some embodiments, a substance is considered "targeted" if it specifically binds to a targeting moiety, such as a nucleic acid targeting moiety. In some embodiments, a targeting moiety, such as a nucleic acid targeting moiety, specifically binds to a target under stringent conditions.

[0744] In certain embodiments, conjugates described herein and compounds include a targeting moiety that specifically binds to one or more targets (e.g., antigens) related to an organ, tissue, cell, extracellular matrix component and / or intracellular compartment. In some embodiments, conjugates described herein and compounds include a targeting moiety that specifically binds to a target related to a particular organ or organ system. In some embodiments, conjugates described herein and compounds include a targeting moiety that specifically binds to one or more intracellular targets (e.g., organelles, intracellular proteins). In some embodiments, conjugates described herein and compounds include a targeting moiety that specifically binds to a target related to a diseased organ, tissue, cell, extracellular matrix component and / or intracellular compartment. In some embodiments, conjugates described herein and compounds include a targeting moiety that specifically binds to a target related to a particular cell type (e.g., endothelial cells, cancer cells, malignant cells, prostate cancer cells, etc.).

[0745] In some embodiments, the conjugates and compounds described herein include a targeting moiety that is bound to a target that is specific for one or more specific tissue types (e.g., relative to prostate tissue, to liver tissue). In some embodiments, the conjugates and compounds described herein include a targeting moiety that is bound to a target that is specific for one or more specific cell types (e.g., relative to B cells, to T cells). In some embodiments, the conjugates and compounds described herein include a targeting moiety that is bound to a target that is specific for one or more specific disease states (e.g., relative to healthy cells, to tumor cells). In some embodiments, the conjugates and compounds described herein include a targeting moiety that is bound to a target that is specific for one or more specific developmental stages (e.g., relative to differentiated cells, to stem cells).

[0746] In some embodiments, target can be with one or more cell types, with one or more diseases and / or with one or more developmental stages exclusively or mainly relevant mark.The expression level of cell type specific marker in described cell type is typically greater than at least 2 times in reference cell colony, and described reference cell colony can be made up of approximately equal amounts of, for example, the mixture of the cell containing from multiple (for example, 5-10 or more) different tissues or organs.In some embodiments, the existence level of cell type specific marker is greater than at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 50 times, at least 100 times or at least 1000 times than its average expression in reference colony.Detection or measurement of cell type specific marker can make interested one or more cell types be distinguished from the cell of many, most or all other types.

[0747] In some embodiments, the target comprises a protein, a carbohydrate, a lipid, and / or a nucleic acid. In some embodiments, the target comprises a protein and / or a characteristic portion thereof, such as a tumor marker, an integrin, a cell surface receptor, a transmembrane protein, an intercellular protein, an ion channel, a membrane transport protein, an enzyme, an antibody, a chimeric protein, a glycoprotein, etc. In some embodiments, the target comprises a carbohydrate and / or a characteristic portion thereof, such as a glycoprotein, a sugar (e.g., a monosaccharide, a disaccharide, a polysaccharide), a glycocalyx (i.e., a carbohydrate-rich peripheral region on the outer surface of most eukaryotic cells), etc. In some embodiments, the target comprises a lipid and / or a characteristic portion thereof, such as an oil, a fatty acid, a glyceride, a hormone, a steroid (e.g., cholesterol, bile acid), a vitamin (e.g., vitamin E), a phospholipid, a sphingolipid, a lipoprotein, etc. In some embodiments, the target comprises a nucleic acid and / or a characteristic portion thereof, such as a DNA nucleic acid; an RNA nucleic acid; a modified DNA nucleic acid; a modified RNA nucleic acid; a nucleic acid comprising any combination of DNA, RNA, modified DNA, and modified RNA.

[0748] Many markers are known in the art. Typical markers include cell surface proteins, such as receptors. Exemplary receptors include, but are not limited to, transferrin receptors; LDL receptors; growth factor receptors, such as epidermal growth factor receptor family members (e.g., EGFR, Her2, Her3, Her4) or vascular endothelial growth factor receptors, cytokine receptors, cell adhesion molecules, integrins, selectins, and CD molecules. Markers can be molecules that are present only or in higher amounts on malignant cells (e.g., tumor antigens).

[0749] Nanoparticles

[0750] In some embodiments, the targeting moiety comprises a particle (e.g., a targeting particle), preferably a nanoparticle, optionally attached to a targeting molecule that can specifically or preferentially bind to a target. In some embodiments, the targeting particle itself directs the compounds of the invention (e.g., by accumulation in tumor cells or tissues), and there is no additional attached targeting molecule.

[0751] As used herein, "nanoparticle" means any particle having a diameter of less than 1000 nm. In some embodiments, the therapeutic agent and / or targeting molecule can be associated with the bulk of the particle, for example, within a polymer matrix. In some embodiments, the targeting molecule can be covalently associated with the surface of the polymer matrix. In some embodiments, the covalent association is mediated by a linker. In some embodiments, the therapeutic agent can be associated with the surface of, encapsulated within, surrounded by, and / or dispersed throughout the polymer matrix. See, for example, U.S. Patent No. 8,246,968, which is incorporated herein in its entirety.

[0752] Generally, the nanoparticles of the present invention comprise any type of particles. Any particles can be used according to the present invention. In some embodiments, the particles are biodegradable and biocompatible. Generally, biocompatible substances are non-toxic to cells. In some embodiments, if the substance is added to the cell and results in a certain threshold value less than cell death, then the substance is considered to be biocompatible. In some embodiments, if the substance is added to the cell and does not cause adverse effects, then the substance is considered to be biocompatible. Generally, biodegradable substances are substances that undergo decomposition under physiological conditions during the treatment-related time period (e.g., weeks, months, or years). In some embodiments, biodegradable substances are substances that can be decomposed by cellular machinery. In some embodiments, biodegradable substances are substances that can be decomposed by chemical processes. In some embodiments, particles are both biocompatible and biodegradable substances. In some embodiments, particles are biocompatible but non-biodegradable substances. In some embodiments, particles are biodegradable but non-biocompatible substances.

[0753] It is generally desirable to use a population of particles that is relatively uniform in size, shape, and / or composition so that each particle has similar properties. For example, the diameter or maximum dimension of at least 80%, at least 90%, or at least 95% of the particles may fall within 5%, 10%, or 20% of the average diameter or maximum dimension. In some embodiments, the population of particles may be heterogeneous in size, shape, and / or composition. According to the present invention, a variety of different particles may be used. In some embodiments, the particles are spheres or spheroids. In some embodiments, the particles are spheres or spheroids. In some embodiments, the particles are flat or plate-shaped. In some embodiments, the particles are cubes or cuboids. In some embodiments, the particles are ovoid or elliptical. In some embodiments, the particles are cylindrical, conical, or pyramidal.

[0754] In some embodiments, the particle is a microparticle (e.g., a microsphere). Typically, a "microparticle" refers to any particle with a diameter less than 1000 μm. In some embodiments, the particle is a picometer particle (e.g., a picometer sphere). Typically, a "picometer particle" refers to any particle with a diameter less than 1 nm. In some embodiments, the particle is a liposome. In some embodiments, the particle is a micelle.

[0755] The particles can be solid or hollow and can comprise one or more layers (e.g., nanoshells, nanorings). In some embodiments, each layer has a unique composition and unique properties relative to the other layer or layers. For example, the particles can have a core / shell structure, wherein the core is one layer and the shell is a second layer. The particles can include multiple different layers. In some embodiments, one layer can be substantially cross-linked, the second layer can not be substantially cross-linked, and so on. In some embodiments, one, several or all of the different layers can comprise one or more therapeutic agents or diagnostic agents to be delivered. In some embodiments, one layer comprises an agent to be delivered, the second layer does not comprise an agent to be delivered, and so on. In some embodiments, each individual layer comprises a different agent or group of agents to be delivered.

[0756] In some embodiments, the particles are porous, meaning that the particles contain holes or channels that are significantly smaller than the size of the particles. For example, the particles can be porous silica particles, such as mesoporous silica nanoparticles, or can have a mesoporous silica coating (Lin et al., 2005, J.Am.Chem.Soc, 17:4570). The particles can have a diameter ranging from about 1 nm to about 50 nm (e.g., a diameter between about 1 nm and 20 nm). Between about 10% and 95% of the particle volume can be composed of the voids in the holes or channels.

[0757] The particles can have a coating. For example, if the particles include a material that is toxic to cells, it may be advantageous to use a biocompatible coating. Suitable coating materials include, but are not limited to, natural proteins (such as bovine serum albumin (BSA)), biocompatible hydrophilic polymers (such as polyethylene glycol (PEG) or PEG derivatives), phospholipids-(PEG), silicon dioxide, lipids, polymers, carbohydrates (such as dextran), other nanoparticles that can be associated with the nanoparticles of the present invention, etc. Coatings can be applied or assembled in a variety of ways, such as by dipping, using layer-by-layer technology, by self-assembly, conjugation, etc. Self-assembly refers to the spontaneous assembly process of a higher-order structure, which depends on the natural attraction of the components (e.g., molecules) of the higher-order structure to each other. It typically occurs by the random motion of molecules and the formation of bonds based on size, shape, composition, or chemical properties.

[0758] Examples of polymers include polyalkylenes (e.g., polyethylene), polycarbonates (e.g., poly(1,3-dioxane-2-one)), polyanhydrides (e.g., poly(sebacic anhydride)), polyhydroxy acids (e.g., poly(3-hydroxyalkanoates)), polyfumarates, polycaprolactones, polyamides (e.g., polycaprolactam), polyacetals, polyethers, polyesters (e.g., polylactide, polyglycolide), poly(orthoesters), polyvinyl alcohol, polyurethanes, polyphosphazenes, polyacrylates, polymethacrylates, polycyanoacrylates, polyureas, polystyrenes, and polyamines. In some embodiments, polymers according to the present invention include polymers that have been approved for use in humans by the U.S. Food and Drug Administration (FDA) under 21 CFR §177.2600, including but not limited to polyesters (e.g., polylactic acid, polyglycolic acid, poly(lactic-co-glycolic acid), polycaprolactone, polyvalerolactone, poly(1,3-dioxane-2-one)); polyanhydrides (e.g., poly(sebacic anhydride)); polyethers (e.g., polyethylene glycol); polyurethanes; polymethacrylates; polyacrylates; and polycyanoacrylates.

[0759] In some embodiments, the particles can be non-polymer particles (e.g., metal particles, quantum dots, ceramic particles, polymers comprising inorganic materials, bone-derived materials, bone substitutes, viral particles, etc.). In some embodiments, the therapeutic agent or diagnostic agent to be delivered can be associated with the surface of such non-polymer particles. In some embodiments, the non-polymer particles are aggregates of non-polymer components, such as aggregates of metal atoms (e.g., gold atoms). In some embodiments, the therapeutic agent or diagnostic agent to be delivered can be associated with the surface of the non-polymer component aggregates and / or encapsulated therein, surrounded by it, and / or dispersed throughout it.

[0760] Particles (e.g., nanoparticles, microparticles) can be prepared using any method known in the art. For example, particle formulations can be formed by methods such as nanoprecipitation, flow focusing fluidic channels, spray drying, single and double emulsion solvent evaporation, solvent extraction, phase separation, grinding, microemulsion procedures, microfabrication, nanofabrication, sacrificial layers, simple and complex coacervation, and other suitable methods. Alternatively or additionally, aqueous and organic solvent syntheses for monodisperse semiconductor, conductive, magnetic, organic, and other nanoparticles have been described (Pellegrino et al., 2005, Small, 1:48; Murray et al., 2000, Ann. Rev. Mat. Sci., 30:545; and Trindade et al., 2001, Chem. Mat., 13:3843).

[0761] Methods for making microparticles for delivering encapsulated agents are described in the literature (see, e.g., Doubrow, ed., “Microcapsules and Nanoparticles in Medicine and Pharmacy,” CRC Press, Boca Raton, 1992; Mathiowitz et al., 1987, J. Control. Release, 5:13; Mathiowitz et al., 1987, Reactive Polymers, δ:275; and Mathiowitz et al., 1988, J. Appl. Polymer Sci., 35:755).

[0762] Nucleic acid targeting moiety

[0763] In some embodiments, the targeting moiety comprises a nucleic acid targeting moiety.

[0764] Generally, a nucleic acid targeting moiety is any polynucleotide that binds to a component associated with an organ, tissue, cell, extracellular matrix component, and / or intracellular compartment (target).

[0765] In some embodiments, the nucleic acid targeting moiety is an aptamer. An aptamer is typically a polynucleotide that is attached to a specific target structure related to a specific organ, tissue, cell, extracellular matrix component and / or intracellular compartment. Generally, the targeting function of an aptamer is based on the three-dimensional structure of an aptamer. In some embodiments, the combination of an aptamer and a target is typically mediated by the interaction between the two-dimensional and / or three-dimensional structures of an aptamer and a target. In some embodiments, the combination of an aptamer and a target is not only based on the primary sequence of an aptamer, but also depends on one or more three-dimensional structures of an aptamer and / or a target. In some embodiments, an aptamer is combined with their target by complementary Watson-Crick (Watson-Crick) base pairing, and the complementary Watson-Crick base pairing is interrupted by the structure (for example, hairpin loop) that destroys base pairing.

[0766] In some embodiments, the nucleic acid targeting moiety is a spiegelmer (PCT publications WO 98 / 08856, WO 02 / 100442, and WO 06 / 117217). Typically, a spiegelmer is a synthetic mirror image nucleic acid (i.e., a mirror image aptamer) that can specifically bind to a target. A spiegelmer is characterized by structural features that make it insensitive to exonucleases and endonucleases.

[0767] Those of ordinary skill in the art will recognize that any nucleic acid targeting moiety (e.g., an aptamer or spiegelmer) that is capable of specifically binding to a target can be used in accordance with the present invention. In some embodiments, the nucleic acid targeting moiety used in accordance with the present invention can target a marker associated with a disease, condition, and / or illness. In some embodiments, the nucleic acid targeting moiety to be used in accordance with the present invention can target a cancer-related target. In some embodiments, the nucleic acid targeting moiety used in accordance with the present invention can target a tumor marker. Any type of cancer and / or any tumor marker can be targeted using the nucleic acid targeting moiety in accordance with the present invention. To give just a few examples, the nucleic acid targeting moiety can target a marker associated with prostate cancer, lung cancer, breast cancer, colorectal cancer, bladder cancer, pancreatic cancer, endometrial cancer, ovarian cancer, bone cancer, esophageal cancer, liver cancer, gastric cancer, brain tumor, skin melanoma, and / or leukemia.

[0768] Nucleic acids of the invention (including nucleic acid-targeting nucleic acid moieties and / or functional RNAs to be delivered, such as RNAi-inducing entities, ribozymes, tRNAs, etc., as described in further detail below) can be prepared according to any available technique, including but not limited to chemical synthesis, enzymatic synthesis, enzymatic or chemical synthesis of longer precursors, and the like.

[0769] Methods for synthesizing RNA are known in the art (see, e.g., Gait, MJ (ed.) Oligonucleotide synthesis: a practical approach, Oxford [Oxfordshire], Washington, DC: IRL Press, 1984; and Herdewijn, P. (ed.) Oligonucleotide synthesis: methods and applications, Methods in molecular biology, v. 288 (Clifton, NJ.) Totowa, NJ: Humana Press, 2005).

[0770] The nucleic acids forming the nucleic acid targeting moiety can comprise naturally occurring nucleosides, modified nucleosides, naturally occurring nucleosides wherein hydrocarbon linkers (e.g., alkylene) or polyether linkers (e.g., PEG linkers) are inserted between one or more nucleosides, modified nucleosides wherein hydrocarbon or PEG linkers are inserted between one or more nucleosides, or combinations thereof. In some embodiments, the nucleotides or modified nucleotides of the nucleic acid targeting moiety can be replaced with hydrocarbon linkers or polyether linkers, provided that the binding affinity and selectivity of the nucleic acid targeting moiety are not significantly reduced by the substitution (e.g., the dissociation constant of the nucleic acid targeting moiety for the target should not be greater than about 1 x 10 -3 M).

[0771] It will be understood by those of ordinary skill in the art that nucleic acids according to the present invention may comprise nucleotides of the type found entirely in naturally occurring nucleic acids, or may alternatively comprise one or more nucleotide analogs, or have a structure that differs in other respects from the structure of naturally occurring nucleic acids. U.S. Patent Nos. 6,403,779; 6,399,754; 6,225,460; 6,127,533; 6,031,086; 6,005,087; 5,977,089; and references therein disclose a wide variety of specific nucleotide analogs and modifications that can be used. See Crooke, S. (ed.) Antisense Drug Technology: Principles, Strategies, and Applications (1st ed.), Marcel Dekker; ISBN: 0824705661; 1st ed. (2001) and references therein. For example, 2'-modifications include halogen, alkoxy, and allyloxy. In some embodiments, the 2'-OH group is replaced by a group selected from H, OR, R, halogen, SH, SR, NH2, NHR, NR2, or CN, wherein R is C1-C6 alkyl, alkenyl, or alkynyl, and halogen is F, CI, Br, or I. Examples of modified bonds include phosphorothioate and 5'-N-phosphoramidite bonds.

[0772] According to the present invention, nucleic acids comprising a plurality of different nucleotide analogs, modified backbones or non-natural internucleoside bonds can be used. Nucleic acids of the present invention can include natural nucleosides (i.e., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine and deoxycytidine) or modified nucleosides. Examples of modified nucleotides include base-modified nucleosides (e.g., cytarabine, inosine, isoguanosine, muscimol, pseudouridine, 2,6-diaminopurine, 2-aminopurine, 2-thiothymidine, 3-deaza-5-azacytidine, 2'-deoxyuridine, 3-nitropyrrole, 4-methylindole, 4-thiouridine, 4-thiouridine, 2-aminoadenosine, 2-thithymidine, 2-thiouridine, 5-bromocytidine, 5-iodouridine, inosine, 6-azauridine, 6-chloropurine, 7-deazaadenosine, 7-deazaguanosine, 8-azaadenosine, 8-azidoadenosine, benzimidazole, M1-methyladenosine, pyrrolo-pyrimidine, The invention also provides a kind of nucleic acid synthesis method for the preparation of nucleic acid. The nucleic acid synthesis method comprises the following steps: 1. The nucleic acid synthesis method comprises the following steps: 2-amino-6-chloropurine, 3-methyladenosine, 5-propynylcytidine, 5-propynyluridine, 5-bromouridine, 5-fluorouridine, 5-methylcytidine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine and 2-thiocytidine), chemically or biologically modified bases (e.g., methylated bases), modified sugars (e.g., 2'-fluororibose, 2'-aminoribose, 2'-azidoribose, 2'-O-methylribose, L-enantiomeric nucleoside arabinose and hexose), modified phosphate groups (e.g., thiophosphate and 5'-N-phosphoramidite bond), and combinations thereof. Natural nucleotides and modified nucleotide monomers for chemical synthesis of nucleic acids are readily available. In some cases, nucleic acids comprising such modifications exhibit improved properties relative to nucleic acids consisting only of naturally occurring nucleotides. In some embodiments, nucleic acid modifications described herein are used to reduce and / or prevent digestion by nucleases (e.g., exonucleases, endonucleases, etc.). For example, the structure of the nucleic acid can be stabilized by including nucleotide analogs at the 3' end of one or both strands to reduce digestion.

[0773] Modified nucleic acids do not need to be uniformly modified along the entire length of the molecule. Different nucleotide modifications and / or backbone structures can be present at various positions in the nucleic acid. It will be understood by those skilled in the art that nucleotide analogs or other one or more modifications can be located at any one or more positions in the nucleic acid so that the function of the nucleic acid is substantially unaffected. As just one example, modifications can be located at any position in the nucleic acid targeting moiety so that the ability of the nucleic acid targeting moiety to specifically bind to the target is substantially unaffected. The modified region can be at the 5' end and / or the 3' end of one or both strands. For example, modified nucleic acid targeting moieties have been used in which approximately 1-5 residues at the 5' end and / or the 3' end of either strand are nucleotide analogs and / or have backbone modifications. Modifications can be 5' end or 3' end modifications. One or both nucleic acid strands can contain at least 50% unmodified nucleotides, at least 80% unmodified nucleotides, at least 90% unmodified nucleotides, or 100% unmodified nucleotides.

[0774] Nucleic acid according to the present invention can for example comprise the modification of sugar, nucleoside or internucleoside bond, such as those described in U.S. Patent Application Publications 2003 / 0175950, 2004 / 0192626, 2004 / 0092470, 2005 / 0020525 and 2005 / 0032733. The present invention includes any nucleic acid using any one or more modifications described therein. For example, many end conjugates have been reported, for example lipids, such as cholesterol, lithocholic acid, aluric acid or long alkyl side chains, to improve cellular uptake. For example, any suitable assay known in the art can be used to test analogs and modifications, for example, to select the delivery, improved nucleic acid targeting moiety and target specific binding that cause improved therapeutic agent or diagnostic agent. In some embodiments, nucleic acid according to the present invention can comprise one or more non-natural nucleoside bonds. In some embodiments, one or more internal nucleotides at the 3'-end, 5'-end, or both the 3'-end and 5'-end of the nucleic acid targeting moiety are inverted to create a linkage, such as a 3'-3' linkage or a 5'-5' linkage.

[0775] In some embodiments, the nucleic acids according to the present invention are not synthetic, but are naturally occurring entities that have been isolated from their natural environment.

[0776] Any method can be used to design novel nucleic acid targeting moieties (see, e.g., U.S. Patent Nos. 6,716,583; 6,465,189; 6,482,594; 6,458,543; 6,458,539; 6,376,190; 6,344,318; 6,242,246; 6,184,364; 6,001,577; 5,958,691; 5,874,218; 5,853,984; 5,843,732; 5,843 ,653; 5,817,785; 5,789,163; 5,763,177; 5,696,249; 5,660,985; 5,595,877; 5,567,588; and 5,270,163; and U.S. patent application publications 2005 / 0069910, 2004 / 0072234, 2004 / 0043923, 2003 / 0087301, 2003 / 0054360, and 2002 / 0064780).

[0777] Nucleic acid targeting moieties can be designed and identified that bind to proteins, carbohydrates, lipids, and / or nucleic acids. In some embodiments, nucleic acid targeting moieties can be designed and identified for use in complexes of the invention that bind to proteins and / or characteristic portions thereof, such as tumor markers, integrins, cell surface receptors, transmembrane proteins, intercellular proteins, ion channels, membrane transporters, enzymes, antibodies, chimeric proteins, and the like. In some embodiments, nucleic acid targeting moieties can be designed and identified for use in complexes of the invention that bind to carbohydrates and / or characteristic portions thereof, such as glycoproteins, sugars (e.g., monosaccharides, disaccharides, and polysaccharides), glycocalyx (i.e., the carbohydrate-rich peripheral region on the outer surface of most eukaryotic cells), and the like. In some embodiments, nucleic acid targeting moieties can be designed and identified for use in complexes of the invention that bind to lipids and / or characteristic portions thereof, such as oils, saturated fatty acids, unsaturated fatty acids, glycerides, hormones, steroids (e.g., cholesterol, bile acids), vitamins (e.g., vitamin E), phospholipids, sphingolipids, lipoproteins, and the like. In some embodiments, nucleic acid targeting moieties can be designed and / or identified for use in complexes of the invention that bind to nucleic acids and / or characteristic portions thereof, such as DNA nucleic acids; RNA nucleic acids; modified DNA nucleic acids; modified RNA nucleic acids; and nucleic acids comprising any combination of DNA, RNA, modified DNA, and modified RNA; etc.

[0778] Any available method can be used to design and / or identify nucleic acid targeting moieties (eg, aptamers or spiegelmers). In some embodiments, nucleic acid targeting moieties are designed and / or identified by identifying nucleic acid targeting moieties from a mixture of candidate nucleic acids.

[0779] Methods for preparing the compounds of the present invention

[0780] The compounds and conjugates disclosed herein can be prepared by simple preparative methods (see, e.g., Examples 1-78). Such preparative methods allow for easy purification.

[0781] Therefore, the present invention also provides a method for preparing the compounds of the present invention. For example, the compounds of the present invention can be prepared as shown in any one of reaction schemes 3, 4, 5, 6 or 7:

[0782] Reaction Scheme 3:

[0783]

[0784] Reaction Scheme 4:

[0785]

[0786] Reaction Scheme 5:

[0787]

[0788] Reaction Scheme 6:

[0789]

[0790] Reaction Scheme 7:

[0791]

[0792] wherein X, Y, Ar, R and n are the same as defined above.

[0793] Also provided herein is a method for preparing a compound comprising making a compound of formula (IIc):

[0794]

[0795] or a pharmaceutically acceptable salt thereof with a sulfonyl halide:

[0796]

[0797] Reaction to provide compounds of formula (Iaa):

[0798]

[0799] or a pharmaceutically acceptable salt thereof, wherein:

[0800] X a is halogen (preferably fluorine), each R 11independently represents an alkyl, aryl, aralkyl, or alkoxy substituent, and the remaining groups may be selected according to any of the definitions provided herein.

[0801] For example, in certain such embodiments,

[0802] Q is an active agent linked to L' via a heteroatom, preferably O or N;

[0803] Z' is absent or is a linking group comprising at least one reactive group (such as a precursor described above for Z);

[0804] L' is a spacer moiety attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent;

[0805] Ar is aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0806] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-, or -(CR b 2) y S-, such that if y is 1, the N, O or S atom is attached to TG;

[0807] O and Y' are positioned on adjacent atoms of Ar;

[0808] TG is a trigger group which, when activated, results in the formation of a molecule capable of reacting with the SO2 to displace (Q) q -(L') w And form a X-SO 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0809] w, x, and y are each independently an integer having a value of 0 or 1;

[0810] Each R a and R c are independently hydrogen or lower alkyl; and

[0811] Each R b are independently hydrogen or lower alkyl; or

[0812] Two R's b Together with the carbon atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring.

[0813] The reactive group of the linking group can be a moiety capable of participating in a 1,3-dipolar cycloaddition reaction, a hetero-Diels-Alder reaction, a nucleophilic substitution reaction, a non-aldol carbonyl reaction, an addition to a carbon-carbon multiple bond, an oxidation reaction, a click reaction, or any other intermolecular coupling reaction. Preferably, the reactive group is selected to participate in selective reactions with reaction partners that are not common in biomolecules, such as a 1,3-dipolar cycloaddition, a hetero-Diels-Alder reaction, an oxime / hydrazone condensation, or a click reaction.

[0814] In certain preferred embodiments, the linking group may comprise an alkyne or azide (which reacts to form a triazole), an alkyne and nitrile oxide (which reacts to form an isoxazole), or a carbonyl (e.g., an aldehyde or ketone), or a hydrazine or hydroxylamine (which reacts to form an oxime or hydrazone).

[0815] In other embodiments, provided herein are methods for preparing a compound comprising:

[0816] (a) making a compound of formula (IIa):

[0817]

[0818] or a pharmaceutically acceptable salt thereof and 1,1'-sulfonylbis(1H-imidazole):

[0819]

[0820] Reaction to provide a compound of formula (IIbb):

[0821]

[0822]

[0823] or a pharmaceutically acceptable salt thereof, wherein the variables may be selected according to any of the definitions provided herein.

[0824] For example, in certain such embodiments,

[0825] Z' is absent or is a linking group comprising a reactive group as discussed in more detail above;

[0826] Ar is aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0827] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) yS-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0828] -SO2- and Y' are positioned on the adjacent atoms of Ar;

[0829] TG is a trigger group which, when activated, results in the formation of a molecule capable of reacting with the SO2 to displace (Q) q -(L') w And form a X-SO 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0830] w, x, and y are each independently an integer having a value of 0 or 1;

[0831] Each R a and R c are independently hydrogen or lower alkyl; and

[0832] Each R b are independently hydrogen or lower alkyl; or

[0833] Two R's b Together with the carbon atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring.

[0834] The compound of formula (IIbb) can then be further reacted with a compound of formula (Ia'): Q-(L') w -H or a pharmaceutically acceptable salt thereof to provide a compound of (Ia):

[0835]

[0836] or a pharmaceutically acceptable salt thereof, wherein the variables may be selected according to any of the definitions provided herein.

[0837] For example, in certain embodiments,

[0838] X is O;

[0839] Q is an active agent linked to L' via a heteroatom, preferably O or N;

[0840] L' is a linking group attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent; and

[0841] w is 0 or 1.

[0842] In some embodiments, the method uses an intermediate compound of Formula (IIa), (IIb), or (IIc) to provide a compound of Formula (Iaa), wherein Ar, TG, Y'Z', and R a is as defined above for the conjugate of formula (I') or the compound of formula (Ia).

[0843] The invention further provides compounds described above that are useful in these methods.

[0844] Intermediate compounds

[0845] In some embodiments, the compounds and conjugates disclosed herein can be prepared by methods using intermediate compounds having structures according to Formula (IV):

[0846]

[0847] or a pharmaceutically acceptable salt thereof, wherein:

[0848] W is hydrogen, -SiR 16 R 17 R 18 or -SO2-G;

[0849] R 16 、R 17 and R18 are each independently C1-C6-alkyl;

[0850] G is halogen (preferably fluorine), imidazole or N-methylimidazolium;

[0851] R is a substituent or -L 1' -Z;

[0852] L 1' is a C1-C ... 200 -alkylene;

[0853] Z is a precursor selected from isocyanide, isothiocyanate, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10Cycloalkyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a , where R a It is C1-C 10 -alkyl) and dihydrogen phosphate (-OP(=O)(OH)2);

[0854] n is an integer with a value from 1 to 4;

[0855] Y is -NO2, -OC(O)(CH2) r C(O)R1, -O(CH2) r -Ar1-NO2, -NHOH, -NHNH2, -BR2R3, or -Y'-TG, such as -NO2, -OC(O)(CH2) r C(O)R1, -O(CH2) r -Ar1-NO2, -NHNH2, -BR2R3, or -Y'-TG;

[0856] R 1 is a C1-C6 alkyl group;

[0857] r is an integer from 1 to 5;

[0858] Ar 1 It is C6-C 20 -arylene;

[0859] R 2 and R 3 are each independently hydrogen, C1-C6-alkyl, C1-C6-alkoxy, or hydroxy;

[0860] R a 、R b 、R c and R d are each independently hydrogen or C1-C6 alkyl;

[0861] Y' is -(CH2) x NR"-、-(CH2) x O- or -(CH2) x S-;

[0862] R" is hydrogen or C1-C6 alkyl;

[0863] x is an integer of 0 or 1; and

[0864] TG is a trigger group.

[0865] In some embodiments, the compounds and conjugates disclosed herein can be prepared by methods using intermediate compounds having structures according to Formula (V):

[0866]

[0867] or a pharmaceutically acceptable salt thereof, wherein:

[0868] W, L 1’ and Z are the same as defined for formula (IV); and

[0869] TG is a trigger group, such as β-galactoside, β-glucuronide, or a combination of β-galactoside and β-glucuronide.

[0870] In other embodiments, the compounds and conjugates disclosed herein can be prepared by methods using intermediate compounds having structures according to Formula (VI):

[0871]

[0872] or a pharmaceutically acceptable salt thereof, wherein:

[0873] W is defined as for formula (IV);

[0874] Y is -NO2, -OC(O)(CH2) r C(O)R 1 、-O(CH2) r -Ar 1 -NO2, -NHOH, -NHNH2, -BR 2 R 3 or -O-TG;

[0875] R 1 is C1-C6-alkyl, such as NO2, -OC(O)(CH2) r C(O)R 1 、-O(CH2) r -Ar 1 -NO2, -NHOH, -NHNH2, -BR 2 R 3 or -O-TG;

[0876] R 1 is C1-C6-alkyl;

[0877] r is an integer from 1 to 5;

[0878] Ar 1 is phenylene, biphenylene or naphthalene;

[0879] R2 and R 3 are each independently hydrogen, C1-C6-alkyl, C1-C6-alkoxy, or hydroxy;

[0880] R a 、R b 、R c and R d are each independently hydrogen or C1-C6-alkyl; and

[0881] TG is a trigger group, β-galactoside, β-glucuronide, or a combination of β-galactoside and β-glucuronide.

[0882] Also provided herein are intermediate compounds of formula (IIa), (IIb), or (IIc):

[0883]

[0884] or a pharmaceutically acceptable salt thereof, wherein:

[0885] G is halogen, imidazole or N-methylimidazolium;

[0886] Each R 11 are independently C1-C6-alkyl;

[0887] Ar is aryl, heteroaryl, cycloalkyl or heterocycloalkyl;

[0888] TG is a trigger group that, when activated, leads to the formation of a molecule capable of forming a 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0889] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0890] O and Y' are positioned on adjacent atoms of Ar;

[0891] x and y are each independently an integer having a value of 0 or 1;

[0892] Z' is absent or is a linking unit comprising, for example, a reactive unit or a binding unit; and

[0893] Each R a are independently hydrogen or alkyl; or

[0894] Two R'sa Together with the carbon atom to which they are attached, they form a three-membered ring.

[0895] In some embodiments, the intermediate compound is a compound of formula (IIa), (IIb) or (IIc), wherein Ar, TG, Y'Z' and R a is as defined above for the conjugate of formula (I') or the compound of formula (Ia).

[0896] In preferred embodiments, the intermediate compound is a compound of formula (IIa), (IIb) or (IIc), wherein Ar is aryl (eg, phenyl or naphthyl).

[0897] In some embodiments, provided herein is an intermediate compound, the intermediate compound being a compound of formula (IIa), (IIb) or (IIc),

[0898] wherein Z' is a linking group comprising one or more groups selected from the group consisting of isocyanide, isothiocyanate, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-halo), maleimide, diene, olefin, halide, p-toluenesulfonate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivatives (-C(O)C≡CR a ) and dihydrogen phosphate (-OP(=O)(OH)2.

[0899] In other embodiments, the intermediate compound is a compound of formula (IIa), (IIb) or (IIc), wherein x is 0. In some such embodiments, TG is -NO2, -OC(O)(CH2) r C(O)R 1 、-NHOH、-NHNH2、-BR 2 R 3 、 Such as NO2, -OC(O)(CH2) r C(O)R 1 、-NHNH2、-BR 2 R 3 、 in:

[0900] R 1 is a C1-C6 alkyl group;

[0901] R 2 and R 3 are each independently hydrogen, C1-C6 alkyl, C1-C6 alkoxy or hydroxy;

[0902] R 4 、R 5 、R 6 and R 7 are each independently hydrogen or C1-C6 alkyl; and

[0903] r is an integer with a value of 1, 2, 3, 4, or 5.

[0904] In alternative embodiments, the intermediate compound is a compound of formula (IIa), (IIb) or (IIc), wherein TG is a trigger group comprising β-galactoside, β-glucuronide or a combination of β-galactoside and β-glucuronide.

[0905] In certain embodiments, the intermediate compound is:

[0906]

[0907] or a pharmaceutically acceptable salt thereof.

[0908] In certain other embodiments, the intermediate compound is:

[0909]

[0910] or a pharmaceutically acceptable salt thereof.

[0911] Antibody-drug conjugates (ADCs)

[0912] In some embodiments, CB is an antibody and Q is a drug. Thus, the compounds and conjugates disclosed herein can be used to conjugate an antibody to a drug moiety to form an antibody-drug conjugate (ADC). Due to the ability of ADC to selectively deliver one or more drug moieties to target tissues (such as tumor-associated antigens), antibody-drug conjugates (ADCs) can improve the therapeutic efficacy of treating diseases (e.g., cancer). Therefore, in certain embodiments, the present invention provides ADCs for therapeutic uses (e.g., treating cancer).

[0913] The ADC of the present invention comprises an antibody linked to one or more drug moieties. The specificity of the ADC is defined by the specificity of the antibody. In one embodiment, the antibody is linked to one or more cytotoxic drugs that are delivered internally to cancer cells.

[0914] Examples of drugs that can be used in the ADCs of the present invention are provided below. The terms "drug," "agent," and "drug moiety" are used interchangeably herein. The terms "linked" and "conjugated" are also used interchangeably herein and indicate that the antibody and moiety are covalently linked.

[0915] In some embodiments, the ADC has the following formula (Formula VII):

[0916] (DL) n -Ab(VII)

[0917] wherein Ab is an antibody and (DL) is a linker-drug moiety. The linker-drug moiety is made up of a linker L and a drug moiety D. The drug moiety can have, for example, cytostatic, cytotoxic or other therapeutic activity against target cells. n is an integer with a value from 1 to about 20, preferably from 1 to about 10. Preferably, DL has the structure of formula (I"):

[0918]

[0919] Q is an active agent linked to L' via a heteroatom, preferably O or N;

[0920] Z' is a linking group;

[0921] L' is a spacer moiety attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent;

[0922] X is -O-, -C(R b )2-or-N(R c )-, preferably -O-;

[0923] Ar is aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl;

[0924] Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG;

[0925] X and Y' are positioned on adjacent atoms of Ar;

[0926] TG is a trigger group which, when activated, results in the formation of a molecule capable of reacting with the SO2 to displace (Q) q -(L')w And form a X-SO 2 and Ar intervening atom of the 5-6 membered ring of N, O or S atom;

[0927] w, x, and y are each independently an integer having a value of 0 or 1;

[0928] Each R a and R c are independently hydrogen or lower alkyl; and

[0929] Each R b are independently hydrogen or lower alkyl; or

[0930] Two R's b Together with the atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring.

[0931] In some embodiments, n has a value ranging from 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or is an integer having a value of 1. When cb is 1 and n is 1, the drug to antibody ratio (DAR) of the ADC is equal to the number of drugs present in (DL). When cb is not 1, the drug to antibody ratio (DAR) of the ADC is equal to the ratio of the number of drugs present in (DL) to the number of antibodies present in the conjugate.

[0932] Exemplary drugs for conjugation

[0933] The ADCs of the present invention provide targeted therapies that can reduce side effects often seen with anti-cancer therapies, for example when one or more active agents or one or more drugs are delivered to specific cells.

[0934] For example, the drug can be selected from the group consisting of: erlotinib (TARCEVA; Genentech / OSI Pharm.); bortezomib (VELCADE; Millenium Pharm.); fulvestrant (FASLODEX; AstraZeneca); sutent (SU11248; Pfizer); letrozole (FEMARA; Novartis); imatinib mesylate (Gleevec; Novartis); PTK787 / ZK 222584 (Novartis); oxaliplatin (Eloxatin; Sanofi); 5-fluorouracil (5-FU); folinic acid; rapamycin (Sirolimus, RAPAMUNE; Wyeth); lapatinib (TYKERB, GSK572016; GlaxoSmithKline); lonafarnib (SCH 66336); sorafenib (BAY43-9006; Bayer Labs.); gefitinib (IRESSA; AstraZeneca); AG1478, AG1571 (SU 5271; Sugen); alkylating agents (e.g., thiotepa or cyclophosphamide); alkyl sulfonates (e.g., busulfan, improsulfan, or piposulfan); aziridines (e.g., benzodopa, carboquinone, meturedopa, or uredopa); ethyleneimine, methylmelamine, hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, trishydroxymethylmelamine; acetogenins (e.g., bullatacin or bullatacinone); camptothecins, including the synthetic analog topotecan; bryostatin; sponge callystatin; CC-1065 (including adolesin, carzelesin, or biszelesin, or synthetic analogs thereof); candidin (e.g., candidin 1 or candidin 8); caudate; duocarmycin (including synthetic analogs, KW-2189, and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; sponge toxins; nitrogen mustards (e.g., chlorambucil, naphthyl mustard, cholophosphamide, estramustine, ifosfamide, dichloromethyldiethylamine, mechlorethamine hydrochloride, oxide hydrochloride), melphalan, nembixin, phenylephrine, prednimustine, trofosfamide, or uramustine); nitrosoureas (e.g., carmustine, chlorozotocin, fotemustine, lomustine, nimustine, or ranimnustine); antibiotics (e.g., geldanamycin, selected from the group consisting of calicheamicin gamma 1I and calicheamicin omega 1 1, or a dynemicin as an enediyne antibiotic, including dynemicin A); a bisphosphonate (e.g., clodronate); esperamicin, a new tumor suppressor protein chromoprotein or a related chromoprotein enediyne antibiotic chromoprotein, aclarubicin, actinomycin, antramycin, azaserine, bleomycin, cactinomycin, carabicin, carninomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubucin, 6-diazo-5-oxo-L-norleucine, Doxorubicin ( doxorubicin (e.g., morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrroline-doxorubicin, liposomal doxorubicin, or deoxydoxorubicin), epirubicin, esorubicin, maseromycin, mitomycin (e.g., mitomycin C, mycophenolic acid, nogamycin, olivomycin, peplomycin, potfiromycin, puromycin, triferon-doxorubicin, rhodorubicin, streptomigrin, streptozotocin, tuberculin, ubenimex, zolpidem, or daunorubicin); antimetabolites (e.g., 5-fluorouracil (5-FU)); folic acid analogs (e.g., leucovorin, methotrexate, pteropterin, or trimetrexate); purine analogs (e.g., fludarabine, 6-mercaptopurine, thiabendine, or thiguanine); pyrimidine analogs (e.g., ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, or floxuridine); androgens (e.g., calutosterone, drostanolone propionate, cyclothiocarbamate, melastane, or testolactone); antiadrenal drugs (e.g., aminoglutethimide, mitotane, or trilostane); folic acid supplements Fillers (e.g., folinic acid); aceglucuronolide; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate, defofamine; colcemid; diacrazone; elfornithine; elliptinium acetate acetate); epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansine alkaloids (e.g., maytansine or ansamitocin); trichothecenes (e.g., T-2 toxin, verracurin A, myrosin A, or serpentin); mitoxantrone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; 2-ethylhydrazide; procarbazine; Polysaccharides; razoxane; rhizoxin; sizolan; spirogermanamine; tricholomanic acid; triamin; 2,2',2"-trichlorotriethylamine; trichothecenes (particularly T-2 toxin, verrucosporin A, mylocin A, or serpentin); ethyl carbamates; vindesine; dacarbazine; mannitol mustard; dibromomannitol; dibromodulcitol; pipobroman; gacytosine; arabinoside ('Ara-C'); cyclophosphamide; thiotepa; taxanes (e.g., Paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE TM Cremophor-free albumin-engineered paclitaxel nanoparticle formulation, American Pharmaceutical Partners, Schaumber, 111. Docetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; platinum analogs (e.g., cisplatin or carboplatin); vinblastine; platinum; etoposide, ifosfamide; mitoxantrone; vincristine; Vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS2000; difluoromethylornithine (DFMO); retinoids (e.g., retinoic acid); capecitabine; and pharmaceutically acceptable salts, solvates, acids, or derivatives thereof.

[0935] Mitotic inhibitors

[0936] In some embodiments, the linkers of the present invention can be used to conjugate antibodies to one or more mitotic inhibitors to form ADCs for the treatment of cancer. As used herein, the term "mitotic inhibitor" refers to a cytotoxic and / or therapeutic agent that blocks mitosis or cell division, which is particularly important for cancer cells. Mitotic inhibitors disrupt microtubules, thereby preventing cell division, generally by affecting microtubule polymerization or microtubule depolymerization. Therefore, in certain embodiments, antibodies are conjugated to one or more mitotic inhibitors that disrupt microtubule formation by inhibiting tubulin polymerization. In one embodiment, the mitotic inhibitor used in the ADC of the present invention is (paclitaxel), (docetaxel) or (Ixabepilone). The following provides examples of mitotic inhibitors that can be used in the ADCs disclosed herein. The genus of mitotic inhibitors includes the auristatins described above.

[0937] Auristatin

[0938] The linker of the present invention can be used to conjugate an antibody to at least one auristatin. Auristatin represents a group of Aplysia analogs, which have generally been shown to have anticancer activity by interfering with microtubule dynamics and GTP hydrolysis to inhibit the effect of cell division. For example, auristatin E (U.S. Patent number 5,635,483) is a synthetic analog of the marine natural product Aplysia 10, which is a compound (GRPettit, Prog. Chem. Org. Nat. Prod, 70: 1-79 (1997)) that inhibits tubulin polymerization by binding to the same site on tubulin (same as the anticancer drug vincristine). Aplysia 10, auristatin PE and auristatin E are linear peptides with four amino acids, three of which are unique to the compound of the Aplysia class. Exemplary embodiments of mitotic inhibitors of the auristatin subclass include, but are not limited to, monomethyl auristatin D (MMAD or auristatin D derivatives), monomethyl auristatin E (MMAE or auristatin E derivatives), monomethyl auristatin F (MMAF or auristatin F derivatives), auristatin F phenylenediamine (AFP), auristatin EB (AEB), auristatin EFP (AEFP) and 5-benzoylvaleric acid-AE ester (AEVB). The synthesis and structure of auristatin derivatives are described in U.S. Patent Application Publication Nos. 2003-0083263, 2005-0238649 and 2005-0009751; International Patent Publication Nos. WO 04 / 010957, WO 04 / 010958 and WO 04 / 010959. 02 / 088172, and U.S. Patent Nos. 6,323,315; 6,239,104; 6,034,065; 5,780,588; 5,665,860; 5,663,149; 5,635,483; 5,599,902; 5,554,725; 5,530,097; 5,521,284; 5,504,191; 5,410,024; 5,138,036; 5,076,973; 4,986,988; 4,978,744; 4,879,278; 4,816,444; and 4,486,414, each of which is incorporated herein by reference.

[0939] Aplysia

[0940] The linker of the present invention can be used to conjugate an antibody to at least one Aplysia to form an ADC. Aplysia is a short peptide compound isolated from the Indian Ocean sea hare Dolabella auricularia (see Pettit et al., J.Am.Chem.Soc., 1976, 98, 4677). Examples of Aplysia include Aplysia 10 and Aplysia 15. Aplysia 15 is a seven-subunit depsipeptide derived from Dolabella auricularia and is an effective antimitotic agent structurally related to the anti-tubulin agent Aplysia 10 (a five-subunit peptide obtained from the same organism). Therefore, in one embodiment, the ADC of the present invention comprises an antibody, a linker as described herein, and at least one Aplysia. The auristatin described above is a synthetic derivative of Aplysia 10.

[0941] Maytansinoids

[0942] The linker of the present invention can be used to conjugate an antibody to at least one maytansinoid to form an ADC. Maytansinoids are effective antitumor agents originally isolated from members of the higher plant families of Celastraceae, Rhamnaceae and Euphorbiaceae, as well as some moss species (Kupchan et al., J.Am.Chem.Soc.94:1354-1356

[1972] ; Wani et al., J.Chem.Soc.Chem.Commun 390:

[1973] ; Powell et al., J.Nat.Prod.46:660-666

[1983] ; Sakai et al., J.Nat.Prod.51:845-850

[1988] ; and Suwanborirux et al., Experientia 46:117-120

[1990] ). There is evidence that maytansinoids inhibit mitosis by inhibiting the polymerization of tubulin (tubulin protein tubulin) to prevent the formation of microtubules (see, for example, U.S. Patent No. 6,441,163 and Remillard et al., Science, 189, 1002-1005 (1975)). Using cell culture models in vitro and using laboratory animal systems in vivo, it has been shown that maytansinoids inhibit tumor cell growth. In addition, the cytotoxicity of maytansinoids is 1,000 times larger than conventional chemotherapeutic agents (such as methotrexate, daunomycin and vincristine) (see, for example, U.S. Patent No. 5,208,020).

[0943] Maytansinoids include C-3 esters of maytansine, maytansinol, maytansinol and other maytansinol analogs and derivatives (see, for example, U.S. Patent number 5,208,020 and 6,441,163, each of which is incorporated herein by reference). The C-3 ester of maytansinol can be naturally occurring or synthetically derived. In addition, naturally occurring and synthetic C-3 maytansinol esters can be classified as C-3 esters with simple carboxylic acids or with the C-3 esters of derivatives of N- methyl-L- alanine, the latter's cytotoxicity being greater than the former. Synthetic maytansinoid analogs are described in, for example, Kupchan et al., J.Med.Chem., 21, 31-37 (1978).

[0944] Suitable maytansinoids for ADCs of the present invention can be isolated from natural sources, produced synthetically or semi-synthetically. In addition, maytansinoids can be modified in any suitable manner, as long as enough cytotoxicity is retained in the final conjugate molecule. The structure of exemplary maytansinoids (mertansine, DM1) is provided below.

[0945]

[0946] Representative examples of maytansinoids include, but are not limited to, DM1 (N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-maytansine; also known as mertansine, drug maytansinoid 1; ImmunoGen, Inc.; see also Chari et al. (1992) Cancer Res 52:127), DM2, DM3 (N2'-deacetyl-N2'-(4-mercapto-1-oxopentyl)-maytansine), DM4 (4-methyl-4-mercapto-1-oxopentyl)-maytansine), and maytansinol (a synthetic maytansinoid analog). Other examples of maytansinoids are described in U.S. Pat. No. 8,142,784, which is incorporated herein by reference for all purposes.

[0947] Ansamitocin is a group of maytansinoid antibiotics that have been isolated from various bacterial sources. These compounds have potent antitumor activity. Representative examples include, but are not limited to, ansamitocin P1, ansamitocin P2, ansamitocin P3, and ansamitocin P4.

[0948] plant alkaloids

[0949] The linkers of the present invention can be used to conjugate antibodies to at least one plant alkaloid, such as a taxane or a vinca alkaloid. Plant alkaloids are chemotherapeutic agents derived from certain types of plants. Vinca alkaloids are made from the periwinkle plant, catharanthus rosea, while taxanes are made from the bark of the Pacific Yewtree taxus. Both vinca alkaloids and taxanes are also known as antimicrotubule agents and are described in more detail below.

[0950] Taxanes

[0951] The linker of the present invention can be used to conjugate an antibody to at least one taxane. As used herein, the term "taxane" refers to a class of antitumor agents that have a microtubule mechanism of action and have a structure comprising a taxane ring structure and stereospecific side chains required for cell inhibition activity. The term "taxane" also includes a variety of known derivatives, including both hydrophilic and hydrophobic derivatives. Taxane derivatives include, but are not limited to, galactose and mannose derivatives described in International Patent Application No. WO 99 / 18113; piperazine and other derivatives described in WO 99 / 14209; taxane derivatives described in WO 99 / 09021, WO 98 / 22451 and U.S. Patent No. 5,869,680; 6-thio derivatives described in WO 98 / 28288; sulfenamide derivatives described in U.S. Patent No. 5,821,263; and paclitaxel derivatives described in U.S. Patent No. 5,415,869, each of which is incorporated herein by reference. Taxane compounds have also been previously described in U.S. Pat. Nos. 5,641,803, 5,665,671, 5,380,751, 5,728,687, 5,415,869, 5,407,683, 5,399,363, 5,424,073, 5,157,049, 5,773,464, 5,821,263, 5,840,929, 4,814,470, 5,438,072, 5,403,858, 4,960,790, 5,433,364, 4,942,184, 5,362,831, 5,705,503, and 5,278,324, all of which are expressly incorporated by reference. Additional examples of taxanes include, but are not limited to, docetaxel ( Sanofi Aventis), paclitaxel ( or Abraxis Oncology and nanoparticle paclitaxel (ABI-007 / Abraxis Bioscience.

[0952] In one embodiment, the linkers of the present invention can be used to conjugate an antibody to at least one docetaxel. In one embodiment, the linkers of the present invention can be used to conjugate an antibody to at least one paclitaxel.

[0953] Vinca alkaloids

[0954] In one embodiment, the linker of the present invention can be used to conjugate an antibody to at least one vinca alkaloid. Vinca alkaloids are a class of cell cycle-specific drugs that work by acting on tubulin and preventing microtubule formation, thereby inhibiting the ability of cancer cells to divide. Examples of vinca alkaloids that can be used in the ADCs of the present invention include, but are not limited to, vindesine sulfate, vincristine, vinblastine, and vinorelbine.

[0955] antitumor antibiotics

[0956] The linker of the present invention can be used to conjugate an antibody to one or more antitumor antibiotics for treating cancer. As used herein, the term "antitumor antibiotic" means an antitumor drug that blocks cell growth by interfering with DNA and is manufactured by microorganisms. Typically, antitumor antibiotics damage DNA chains or slow down or stop DNA synthesis. Examples of antitumor antibiotics that may be included in the ADC disclosed herein include, but are not limited to, actinomycin (e.g., pyrrolo[2,1-c][1,4]benzodiazepines), anthracyclines, calicheamicins, and duocarmycins, which are described in more detail below.

[0957] Actinomycin

[0958] The linker of the present invention can be used to conjugate an antibody to at least one actinomycin. Actinomycin is a subclass of antitumor antibiotics isolated from bacteria of the genus Streptomyces. Representative examples of actinomycin include, but are not limited to, actinomycin D (Cosmegen [also known as actinomycin, dactinomycin, actinomycin IV, actinomycin C1], Lundbeck, Inc.), anthramycin, chicamycin A, DC-81, mazethramycin, neothramycin A, neothramycin B, porothramycin, prothracarcin B, SG2285, sibanomicin, sibiromycin, and tomaymycin. In one embodiment, D is a pyrrolobenzodiazepine (PBD). Examples of PBDs include, but are not limited to, anthramycin, cheikamycin A, DC-81, methylanthramycin, neothramycin A, neothramycin B, porothramycin, prothracarcin B, SG2000 (SJG-136), SG2202 (ZC-207), SG2285 (ZC-423), sibamicin, sibiricomycin, and tomamycin. Thus, in one embodiment, D is an actinomycin, such as actinomycin D, or a PBD, such as a pyrrolobenzodiazepine (PBD) dimer.

[0959] The structure of a PBD can be found, for example, in US Patent Application Publication Nos. 2013 / 0028917 and 2013 / 0028919 and WO 2011 / 130598 A1, each of which is incorporated herein by reference in its entirety. The general structure of a PBD is provided below.

[0960]

[0961] PBDs differ in the number, type, and position of substituents, both their aromatic A ring and pyrrolo C ring, and the degree of saturation of the C ring. In the B ring, an imine (N=C), carbinolamine (NH—CH(OH)), or carbinolamine methyl ether (NH—CH(OMe)) is typically present at the N10-C11 position, which serves as the electrophilic center responsible for alkylating the DNA. All known natural products have an (S)-configuration at the chiral C11α position, which provides them with a right-handed twist when viewed from the C ring to the A ring. Additional examples of PBDs that can be conjugated to antibodies via the linkers disclosed herein can be found, for example, in U.S. Patent Application Publication Nos. 2013 / 0028917A1 and 2013 / 0028919A1, U.S. Patent No. 7,741,319B2, and WO 2011 / 130598A1 and WO 2006 / 111759 A1, each of which is incorporated herein by reference in its entirety.

[0962] anthracycline

[0963] The linker of the present invention can be used to conjugate an antibody to at least one anthracycline. Anthracyclines are a subclass of antitumor antibiotics isolated from bacteria of the genus Streptomyces. Representative examples include, but are not limited to, daunorubicin (Cerubidine, Bedford Laboratories), doxorubicin (Adriamycin, Bedford Laboratories; also known as doxorubicin hydrochloride, hydroxydaunorubicin, and Rubex), epirubicin (Ellence, Pfizer), and idarubicin (Idamycin; Pfizer Inc.). Therefore, in one embodiment, D is an anthracycline, such as doxorubicin.

[0964] Calcheamicin

[0965] The linkers of the present invention can be used to conjugate antibodies to at least one calicheamicin. The calicheamicins are a family of enediyne antibiotics derived from the soil organism Micromonospora echinospora. The calicheamicins bind to the minor groove of DNA and induce double-stranded DNA breaks, leading to cell death, which is increased 100-fold relative to other chemotherapeutic agents (Damle et al. (2003) Curr Opin Pharmacol 3:386). The preparation of calicheamicins that can be used as drug conjugates in the present invention has been described, see U.S. Patent Nos. 5,712,374; 5,714,586; 5,739,116; 5,767,285; 5,770,701; 5,770,710; 5,773,001; and 5,877,296. Structural analogs of calicheamicin that can be used include, but are not limited to, γ 1 I, α 2 I, α 3 I, N-acetyl-γ 1 I, PSAG, and θ 1 I (Hinman et al., Cancer Research 53:3336-3342 (1993), Lode et al., Cancer Research 58:2925-2928 (1998), and aforementioned U.S. Patent Nos. 5,712,374; 5,714,586; 5,739,116; 5,767,285; 5,770,701; 5,770,710; 5,773,001; and 5,877,296). Thus, in one embodiment, D is calicheamicin.

[0966] Duocarmycin

[0967] The linker of the present invention can be used to conjugate an antibody to at least one duocarmycin. Duocarmycins are a subclass of antitumor antibiotics isolated from bacteria of the genus Streptomyces. (See Nagamura and Saito (1998) Chemistry of Heterocyclic Compounds, Vol. 34, No. 12). Duocarmycins bind to the minor groove of DNA and alkylate the nucleobase adenine at the N3 position (Boger (1993) Pure and Appl Chem 65(6):1123; and Boger and Johnson (1995) PNAS USA 92:3642). Synthetic analogs of duocarmycins include, but are not limited to, adolesin, biszelesin, and carzelesin. Thus, in one embodiment, D is a duocarmycin.

[0968] Other antitumor antibiotics

[0969] In addition to the foregoing, additional antitumor antibiotics that can be used in the ADCs of the present invention include bleomycin (Blenoxane, Bristol-Myers Squibb), mitomycin, and plicamycin (also known as mithramycin).

[0970] Immunomodulators

[0971] In some embodiments, the linker of the present invention can be used to conjugate an antibody to at least one immunomodulator. As used herein, the term "immunomodulator" refers to an agent that can stimulate or change an immune response. In one embodiment, an immunomodulator is an immunostimulator that enhances the immune response of a subject. In some embodiments, an immunomodulator is an immunosuppressant that prevents or reduces the immune response of a subject. Immunomodulators can regulate myeloid cells (monocytes, macrophages, dendritic cells, megakaryocytes, and granulocytes) or lymphoid cells (T cells, B cells, and natural killer (NK) cells) and any further differentiated cells thereof. Representative examples include, but are not limited to, Bacillus Calmette-Guérin (BCG) and levamisole (Ergamisol). Other examples of immunomodulators that can be used for the ADC of the present invention include, but are not limited to, cancer vaccines, cytokines, and immunomodulatory gene therapy.

[0972] cancer vaccines

[0973] The linker of the present invention can be used to conjugate antibodies to cancer vaccines. As used herein, the term "cancer vaccine" refers to a composition (e.g., tumor antigens and cytokines) that causes a tumor-specific immune response. By administering a cancer vaccine or, in the case of the present invention, administering an ADC comprising an antibody and a cancer vaccine, a reaction is elicited from the subject's own immune system. In a preferred embodiment, the immune response leads to the eradication of tumor cells (e.g., primary or metastatic tumor cells) in vivo. The use of cancer vaccines generally involves the administration of a specific antigen or a group of antigens, which, for example, are present on the surface of specific cancer cells or on the surface of a specific infectious agent that is shown to contribute to cancer formation. In some embodiments, the use of cancer vaccines is for preventive purposes, while in other embodiments, the use is for therapeutic purposes. Non-limiting examples of cancer vaccines that can be used with the ADCs disclosed herein include recombinant bivalent human papillomavirus (HPV) vaccine types 16 and 18 (Cervarix, GlaxoSmithKline), recombinant quadrivalent human papillomavirus (HPV) vaccine types 6, 11, 16, and 18 (Gardasil, Merck & Company), and sipuleucel-T (Provenge, Dendreon). Thus, in one embodiment, D is a cancer vaccine that is an immunostimulatory agent or an immunosuppressive agent.

[0974] cytokines

[0975] The linker of the present invention can be used to conjugate an antibody to at least one cytokine. The term "cytokine" generally refers to a protein released by a cell population that acts on another cell as an intercellular medium. Cytokines directly stimulate immune effector cells and stromal cells at the tumor site and enhance tumor cell recognition by cytotoxic effector cells (Lee and Margolin (2011) Cancers 3:3856). A large number of animal tumor model studies have shown that cytokines have a wide range of anti-tumor activity, and this has been converted into a variety of cytokine-based methods for cancer therapy (Lee and Margoli, supra). In recent years, many cytokines have been seen, including GM-CSF, IL-7, IL-12, IL-15, IL-18 and IL-21, and have entered clinical trials for patients with advanced cancer (Lee and Margoli, supra).

[0976] Examples of cytokines that can be used in the ADCs of the present invention include, but are not limited to, parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; prorelaxin; glycoprotein hormones such as follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), and luteinizing hormone (LH); liver growth factor; fibroblast growth factor; prolactin; placental lactogen; tumor necrosis factor; Müllerian inhibitory substance; mouse gonadotropin-related peptide; inhibin; activin; vascular endothelial growth factor; integrin; thrombopoietin (TPO); nerve growth factors such as NGF; platelet-derived growth factor; transforming growth factor (TGF- ); insulin-like growth factor-I and insulin-like growth factor-II; erythropoietin (EPO); osteoinductive factors; interferons, such as interferon α, β and γ, colony stimulating factor (CSF); granulocyte-macrophage-C-SF (GM-CSF); and granulocyte-CSF (G-CSF); interleukins (IL), such as IL-1, IL-1a, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12; tumor necrosis factor; and other polypeptide factors, including LIF and kit ligand (KL). As used herein, the term cytokine includes proteins from natural sources or from recombinant cell culture and biologically active equivalents of native sequence cytokines. Therefore, in one embodiment, D is a cytokine.

[0977] colony-stimulating factor (CSF)

[0978] The linkers of the present invention can be used to conjugate antibodies to at least one colony stimulating factor (CSF). Colony stimulating factor (CSF) is a growth factor that helps the bone marrow produce red blood cells. Because some cancer treatments (e.g., chemotherapy) can affect white blood cells (which help fight infection), colony stimulating factors may be introduced to help support white blood cell levels and enhance the immune system. Colony stimulating factors can also be used after a bone marrow transplant to help the new bone marrow begin to produce white blood cells. Representative examples of CSFs that can be used in the ADCs disclosed herein include, but are not limited to, erythropoietin (Epoetin), filgrastim (Neopogen (also known as granulocyte colony-stimulating factor (G-CSF); Amgen, Inc.), sargramostim (leukine (granulocyte-macrophage colony-stimulating factor and GM-CSF); Genzyme Corporation), promegapoietin, and oprelvekin (recombinant IL-11; Pfizer, Inc.). Thus, in one embodiment, D is CSF.

[0979] Gene therapy

[0980] The linker of the present invention can be used to conjugate an antibody to at least one nucleic acid (directly or indirectly via a vector) for gene therapy. Gene therapy generally refers to the introduction of genetic material into cells, whereby the genetic material is designed to treat a disease. Because it is related to immunomodulators, gene therapy is used to stimulate a subject's natural ability to inhibit cancer cell proliferation or kill cancer cells. In one embodiment, the ADC of the present invention comprises a nucleic acid encoding a functional therapeutic gene that is used to replace a mutated or dysfunctional (e.g., truncated) gene associated with cancer. In other embodiments, the ADC of the present invention comprises a nucleic acid encoding a therapeutic protein or providing the production of a therapeutic protein to treat cancer in other aspects. The nucleic acid encoding the therapeutic gene can be conjugated directly to the antibody, or alternatively, it can be conjugated to the antibody via a vector. Examples of vectors that can be used to deliver nucleic acids for gene therapy include, but are not limited to, viral vectors or liposomes.

[0981] Alkylating agents

[0982] The linkers of the present invention can be used to conjugate antibodies to one or more alkylating agents. Alkylating agents are a class of anti-tumor compounds in which an alkyl group is attached to DNA. Examples of alkylating agents that can be used in the ADCs of the present invention include, but are not limited to, alkyl sulfonates, ethylenimine, methylamine derivatives, epoxides, nitrogen mustards, nitrosoureas, triazines, and hydrazines.

[0983] Alkyl sulfonate

[0984] The linker of the present invention can be used to conjugate an antibody to at least one alkyl sulfonate. Alkyl sulfonate is a compound having the general formula: R-SO2-OR 1 A subclass of alkylating agents, where R and R 1 Typically, it is an alkyl or aryl group. A representative example of an alkyl sulfonate is busulfan ( GlaxoSmithKline; Busulfex PDL BioPharma, Inc.

[0985] Nitrogen mustard

[0986] The linker of the present invention can be used to conjugate an antibody to at least one nitrogen mustard. Representative examples of this subclass of anticancer compounds include, but are not limited to, chlorambucil ( GlaxoSmithKline), cyclophosphamide ( Bristol-Myers Squibb; Neosar, Pfizer), estramustine (estramustine sodium phosphate or ), ifosfamide ( Bristol-Myers Squibb Company), dichloromethyl diethylamine ( Lundbeck Inc.) and Melphalan or L- or phenylalanine mustard; GlaxoSmithKline).

[0987] Nitrosourea

[0988] The linkers of the present invention can be used to conjugate an antibody to at least one nitrosourea. Nitrosoureas are a subclass of lipid-soluble alkylating agents. Representative examples include, but are not limited to, carmustine (BCNU [also known as BiCNU, N,N-bis(2-chloroethyl)-N-nitrosourea or 1,3-bis(2-chloroethyl)-1-nitrosourea], Bristol-Myers Squibb Company), fotemustine (also known as ), lomustine (CCNU or 1-(2-chloro-ethyl)-3-cyclohexyl-1-nitrosourea, Bristol-Myers Squibb), nimustine (also known as ACNU), and streptozotocin ( Teva Pharmaceuticals).

[0989] Triazine and hydrazine

[0990] The linkers of the present invention can be used to conjugate antibodies to at least one triazine or hydrazine. Triazines and hydrazines are subclasses of nitrogen-containing alkylating agents. In some embodiments, these compounds decompose spontaneously or can be metabolized to produce alkyldiazo intermediates that promote the transfer of alkyl groups to nucleic acids, peptides and / or polypeptides, thereby causing mutagenic, carcinogenic or cytotoxic effects. Representative examples include, but are not limited to, dacarbazine (DTIC-Dome, Bayer Healthcare Pharmaceuticals Inc.), procarbazine ( Sigma-Tau Pharmaceuticals, Inc.) and temozolomide ( Schering Plough.

[0991] Other alkylating agents

[0992] The linkers of the present invention can be used to conjugate antibodies to at least one ethyleneimine, methylamine derivative, or epoxide. Ethylenimines are a subclass of alkylating agents that typically contain at least one aziridine ring. Epoxides represent a subclass of alkylating agents that are characterized as cyclic ethers with only three ring atoms.

[0993] Representative examples of ethyleneimines include, but are not limited to, thiopeta (Thioplex, Amgen), diaziridinylbenzoquinone (also known as aziridinylbenzoquinone (AZQ)), and mitomycin C. Mitomycin C is a natural product containing an aziridine ring and appears to cross-link DNA to induce cytotoxicity (Dorr RT, et al. Cancer Res. 1985; 45: 3510; Kennedy KA, et al. Cancer Res. 1985; 45: 3541). Representative examples of methylamine derivatives and their analogs include, but are not limited to, hexamethylmelamine (Hexalen, MGI Pharma, Inc.), also known as hexamethylamine and hexastat. Representative examples of epoxides of this class of anticancer compounds include, but are not limited to, dianhydrogalactitol. Dianhydrogalactitol (1,2:5,6-dianhydrodulcitol) is chemically related to aziridines and generally promotes the transfer of alkyl groups by a similar mechanism as described above. Dibromodulcitol is hydrolyzed to dianhydrogalactitol and is therefore a prodrug of the epoxide (Sellei C et al. Cancer Chemother Rep. 1969; 53: 377).

[0994] Anti-angiogenic agents

[0995] In some embodiments, the linkers of the present invention can be used to conjugate antibodies to at least one anti-angiogenic agent. Anti-angiogenic agents inhibit the growth of new blood vessels. Anti-angiogenic agents exert their effects in a variety of ways. In some embodiments, these agents interfere with the ability of growth factors to reach their targets. For example, vascular endothelial growth factor (VEGF) is one of the main proteins that triggers angiogenesis by binding to specific receptors on the cell surface. Therefore, certain anti-angiogenic agents that block the interaction of VEGF with its cognate receptor prevent VEGF from triggering angiogenesis. In other embodiments, these agents interfere with intracellular signaling cascades. For example, once a specific receptor on the cell surface is triggered, a cascade of other chemical signals is triggered to promote blood vessel growth. Therefore, certain enzymes (e.g., some tyrosine kinases) that are known to promote certain intracellular signaling cascades that contribute to, for example, cell proliferation are targets for cancer treatment. In other embodiments, these agents interfere with intercellular signaling cascades. However, in other embodiments, these agents inactivate specific targets that activate and promote cell growth or inactivate them by directly interfering with vascular cell growth. Angiogenesis-inhibiting properties have been found in more than 300 substances, with a variety of direct and indirect inhibitory effects.

[0996] Representative examples of anti-angiogenic agents that can be used in the ADCs of the present invention include, but are not limited to, angiostatin, ABXEGF, C1-1033, PKI-166, EGF vaccine, EKB-569, GW2016, ICR-62, EMD 55900, CP358, PD153035, AG1478, IMC-C225 (Erbitux, ZD1839 (Iressa), OSI-774, erlotinib (tarceva), angiostatin, arrestin, endostatin, BAY 12-9566 and with fluorouracil or doxorubicin, angiostatin, carboxyamidotriazole and with paclitaxel, EMD121974, S-24, vitamin, dimethylxanthenone acetic acid, IM862, interleukin-12, interleukin-2, NM-3, HuMV833, PTK787, RhuMab, angiostatin (ribozyme), IMC-1C11, neovastat, marimstat, prinostat mastat), BMS-275291, COL-3, MM1270, SU101, SU6668, SU11248, SU5416 (with paclitaxel, with gemcitabine and cisplatin, and with irinotecan and cisplatin, and with radiation), tecogalan, temozolomide and PEG interferon alfa 2b, tetrathiomolybdate, TNP-470, thalidomide, CC-5013 and with taxotere, tumstatin, 2-methoxyestradiol, VEGF trap, mTOR inhibitors (rapamycin, everolimus (Afinitor, Novartis Pharmaceutical Corporation), and temsirolimus (Torisel, Pfizer)), tyrosine kinase inhibitors (e.g., erlotinib (Tarceva, Genentech, Inc.), imatinib (Gleevec, Novartis Pharmaceuticals), gefitinib (Iressa, AstraZeneca Pharmaceuticals), dasatinib (Sprycel, Brystol-Myers Squibb), sunitinib (Sutent, Pfizer), nilotinib (Tasigna, Novartis Pharmaceuticals), lapatinib (Tykerb, GlaxoSmithKline Pharmaceuticals), sorafenib (Nexavar, Bayer and Onyx), phosphoinositide 3-kinase (PI3K).

[0997] Antimetabolites

[0998] The linker of the present invention can be used to conjugate an antibody to at least one antimetabolite. Antimetabolites are a type of chemotherapy treatment that is very similar to normal substances in cells. When cells incorporate antimetabolites into cellular metabolism, the result is negative for the cells, for example, the cells cannot divide. Antimetabolites are classified according to the substances they interfere with. Examples of antimetabolites that can be used for the ADC of the present invention include, but are not limited to, folic acid antagonists (e.g., methotrexate), pyrimidine antagonists (e.g., 5-fluorouracil, Foxuridine, cytarabine, capecitabine and gemcitabine), purine antagonists (e.g., 6-mercaptopurine and 6-thioguanine) and adenosine deaminase inhibitors (e.g., cladribine, fludarabine, nelarabine and pentostatin), as described in more detail below.

[0999] Antifolates

[1000] The linker of the present invention can be used to conjugate the antibody to at least one antifolate. Antifolates are a subclass of antimetabolites that are structurally similar to folate. Representative examples include, but are not limited to, methotrexate, 4-amino-folic acid (also known as aminopterin and 4-aminopteroic acid), lometrexol (LMTX), pemetrexed (Alimpta, Eli Lilly and Company), and trimetrexate (Neutrexin, Ben Venue Laboratories, Inc.).

[1001] Purine antagonists

[1002] The linker of the present invention can be used to conjugate an antibody to at least one purine antagonist. Purine analogs are a subclass of antimetabolites that are structurally similar to the group of compounds called purines. Representative examples of purine antagonists include, but are not limited to, azathioprine (Azasan, Salix; Imuran, GlaxoSmithKline), cladribine (Leustatin [also known as 2-CdA], Janssen Biotech, Inc.), mercaptopurine (Purinethol [also known as 6-mercaptoethanol], GlaxoSmithKline), fludarabine (Fludara, Genzyme), pentostatin (Nipent, also known as 2'-deoxycoformycin (DCF)), 6-thioguanine (Lanvis [also known as thioguanine], GlaxoSmithKline).

[1003] Pyrimidine antagonists

[1004] The linker of the present invention can be used to conjugate an antibody to at least one pyrimidine antagonist. Pyrimidine antagonists are a subclass of antimetabolites structurally similar to the group of compounds called purines. Representative examples of pyrimidine antagonists include but are not limited to azacitidine (Vidaza, Celgene Corporation), capecitabine (Xeloda, Roche Laboratories), cytarabine (also known as cytosine arabinoside and arabinosylcytosine, Bedford Laboratories), decitabine (Dacogen, Eisai Pharmaceuticals), 5-fluorouracil (Adrucil, Teva Pharmaceuticals; Efudex, Valeant Pharmaceuticals, Inc), 5-fluoro-2'-deoxyuridine 5'-phosphate (FdUMP), 5-fluorouridine triphosphate and gemcitabine (Gemzar, Eli Lilly and Company).

[1005] Boron-containing agents

[1006] The linkers of the present invention can be used to conjugate antibodies to at least one boron-containing agent. Boron-containing agents include a class of cancer therapeutic compounds that interfere with cell proliferation. Representative examples of boron-containing agents include, but are not limited to, borophycin and bortezomib (Velcade, Millenium Pharmaceuticals).

[1007] Chemical protective agents

[1008] The linker of the present invention can be used to conjugate an antibody to at least one chemoprotectant. Chemoprotectants are compounds that help protect the body from the specific toxic effects of chemotherapy. Chemoprotectants can be administered together with a variety of chemotherapy to protect healthy cells from the toxic effects of chemotherapy drugs while allowing the administered chemotherapeutic agents to treat cancer cells. Representative chemoprotectants include, but are not limited to, amifostine (Ethyol, Medimmune, Inc.) (for alleviating the nephrotoxicity associated with the cumulative dose of cisplatin), dexrazoxane (Totect, Apricus Pharma; Zinecard) (for treating the overflow caused by administering anthracyclines (Totect) and for treating the heart-related complications caused by administering antitumor antibiotic doxorubicin (Zinecard)) and mesna (Mesnex, Bristol-Myers Squibb) (for preventing hemorrhagic cystitis during chemotherapy with ifocfamide).

[1009] Hormonal agents

[1010] The linkers of the present invention can be used to conjugate an antibody to at least one hormone agent. Hormones (including synthetic hormones) are compounds that interfere with the production or activity of hormones produced endogenously by the endocrine system. In some embodiments, these compounds interfere with cell growth or produce cytotoxic effects. Non-limiting examples include androgens, estrogens, medroxyprogesterone acetate (Provera, Pfizer), and progestogens.

[1011] Antihormonal agents

[1012] The linkers of the present invention can be used to conjugate an antibody to at least one anti-hormonal agent. An "anti-hormonal" agent is an agent that inhibits the production of certain endogenous hormones and / or prevents their function. In one embodiment, the anti-hormonal agent interferes with the activity of a hormone selected from the group consisting of androgens, estrogens, progesterone, and goanadotropin-releasing hormone, thereby interfering with the growth of various cancer cells. Representative examples of anti-hormonal agents include, but are not limited to, aminoglutethimide, anastrozole (Arimidex, AstraZeneca), bicalutamide (Casodex, AstraZeneca), cyproterone acetate (Cyprostat, Bayer PLC), degarelix (Firmagon, Ferring Pharmaceuticals), exemestane (Aromasin, Pfizer), flutamide (Drogenil, Schering-Plough Ltd), fulvestrant (Faslodex, AstraZeneca), goserelin (Zolodex, AstraZeneca), letrozole (Femara, Novartis), leuprorelin (Prostap), lupron, medroxyprogesterone acetate (Provera, Pfizer), megestrol acetate (Megace, Bristol-Myers Squibb), tamoxifen (Nolvadex, AstraZeneca), and triptorelin (Decapetyl, Ferring).

[1013] corticosteroids

[1014] The linker of the present invention can be used to conjugate an antibody to at least one corticosteroid. Corticosteroids can be used in the ADC of the present invention to reduce inflammation. Examples of corticosteroids include, but are not limited to, glucocorticoids, such as prednisone (Deltasone, a branch of Pfizer, Pharmacia Pharmaceuticals).

[1015] Photoactive therapeutic agents

[1016] The linkers of the present invention can be used to conjugate antibodies to at least one photoactive therapeutic agent. Photoactive therapeutic agents include compounds that can be deployed to kill the treated cells upon exposure to electromagnetic radiation of a specific wavelength. Therapeutically relevant compounds absorb electromagnetic radiation of a wavelength that penetrates tissue. In preferred embodiments, the compounds are administered in a non-toxic form that, upon sufficient activation, is capable of producing a photochemical effect that is toxic to cells or tissues. In other preferred embodiments, these compounds are retained by cancerous tissue and readily cleared from normal tissue. Non-limiting examples include various chromogens and dyes.

[1017] Oligonucleotides

[1018] The joint of the present invention can be used for antibody and at least one oligonucleotide being put together.Oligonucleotide is made up of short nucleic acid chain, and described short nucleic acid chain works by interfering with the processing of genetic information.In some embodiments, the oligonucleotide for ADC is unmodified single-stranded and / or double-stranded DNA or RNA molecule, and in other embodiments, these therapeutic oligonucleotides are chemically modified single-stranded and / or double-stranded DNA or RNA molecule.In one embodiment, the oligonucleotide used in ADC is relatively short (19-25 nucleotides), and hybridizes with the unique nucleic acid sequence in the pool of the nucleic acid target present in the cell.Some important oligonucleotide technologies include antisense oligonucleotide (including RNA interference (RNAi)), aptamer, CpG oligonucleotide and ribozyme.

[1019] antisense oligonucleotides

[1020] The joint of the present invention can be used for antibody and at least one antisense oligonucleotide conjugated.Antisense oligonucleotide is designed to be combined with RNA by Watson-Crick hybridization.In some embodiments, the antisense oligonucleotide is complementary to the nucleotides of the region, domain, part or segment of the coding conjugated antibody.In some embodiments, the antisense oligonucleotide comprises from about 5 to about 100 nucleotides, from about 10 to about 50 nucleotides, from about 12 to about 35 and from about 18 to about 25 nucleotides.

[1021] Once an oligonucleotide binds to a target RNA, a variety of mechanisms can be used to inhibit RNA function. (Crooke ST. (1999). Biochim. Biophys. Acta, 1489, 30-42). The best characterized antisense mechanism results in cleavage of the target RNA by endogenous cellular nucleases (such as RNase H or nucleases associated with the RNA interference mechanism). However, oligonucleotides that inhibit the expression of a target gene through non-catalytic mechanisms (such as regulation of splicing or translational arrest) can also be effective and selective regulators of gene function.

[1022] Another RNA enzyme-dependent antisense mechanism that has recently received widespread attention is RNAi (Fire et al. (1998). Nature, 391, 806-811; Zamore PD. (2002). Science, 296, 1265-1269.). RNA interference (RNAi) is a post-transcriptional process in which double-stranded RNA inhibits gene expression in a sequence-specific manner. In some embodiments, RNAi is achieved by introducing relatively long double-stranded RNA (dsRNA), while in preferred embodiments, this RNAi is achieved by introducing shorter double-stranded RNA (e.g., small interfering RNA (siRNA) and / or microRNA (miRNA)). In yet another embodiment, RNAi can also be achieved by introducing a plasmid that produces a dsRNA complementary to the target gene. In each of the aforementioned embodiments, the double-stranded RNA is designed to interfere with the gene expression of a specific target sequence in the cell. Typically, the mechanism involves conversion of dsRNA into short RNAs that guide ribonucleases to their cognate mRNA targets (for an overview, Ruvkun, Science 2294:797 (2001)). The ribonucleases then degrade the corresponding endogenous mRNA, leading to regulation of gene expression. Notably, dsRNAs are reported to have antiproliferative properties, which also lead to the envision of therapeutic applications (Aubel et al., Proc. Natl. Acad. Sci., USA 88:906 (1991)). For example, synthetic dsRNA has been shown to inhibit tumor growth in mice (Levy et al. Proc. Nat. Acad. Sci. USA, 62: 357-361 (1969)), is active in the treatment of leukemic mice (Zeleznick et al., Proc. Soc. Exp. Biol. Med. 130: 126-128 (1969)), and inhibits chemically induced tumorigenesis in mouse skin (Gelboin et al., Science 167: 205-207 (1970)). Therefore, in a preferred embodiment, the present invention provides the use of antisense oligonucleotides in ADCs for the treatment of breast cancer. In other embodiments, the present invention provides compositions and methods for initiating antisense oligonucleotide therapy, wherein the dsRNA interferes with target cell expression of EGFR at the mRNA level. As used above, dsRNA refers to naturally occurring RNA, partially purified RNA, recombinantly produced RNA, synthetic RNA, and RNA that is modified differently by including non-standard nucleotides, non-nucleotide materials, nucleotide analogs (e.g., locked nucleic acids (LNA)), deoxyribonucleotides, and any combination thereof. The RNA of the present invention need only be sufficiently similar to natural RNA so that it has the ability to mediate the antisense oligonucleotide-based regulation described herein.

[1023] Aptamer

[1024] The linkers of the present invention can be used to conjugate antibodies to at least one aptamer. Aptamers are nucleic acid molecules selected from a random pool based on their ability to bind other molecules. Like antibodies, aptamers can bind to target molecules with remarkable affinity and specificity. In many embodiments, aptamers exhibit complex, sequence-dependent three-dimensional shapes that allow them to interact with target proteins, creating tightly bound complexes similar to antibody-antigen interactions, thereby interfering with the protein's function. The unique ability of aptamers to tightly and specifically bind to their target proteins highlights their potential as targeted molecular therapies.

[1025] CpG oligonucleotides

[1026] The linker of the present invention can be used to conjugate an antibody to at least one CpG oligonucleotide. Bacterial and viral DNA are known to be strong activators of both human innate and specific immunity. These immunological characteristics are associated with unmethylated CpG dinucleotide motifs found in bacterial DNA. Because these motifs are rare in humans, the human immune system has developed the ability to recognize these motifs as early signs of infection and subsequently trigger an immune response. Therefore, oligonucleotides containing such CpG motifs can be used to trigger an anti-tumor immune response.

[1027] Ribozyme

[1028] The linkers of the present invention can be used to conjugate antibodies to at least one ribozyme. Ribozymes are catalytic RNA molecules ranging in length from about 40 to 155 nucleotides. The ability of ribozymes to recognize and cleave specific RNA molecules makes them potential candidates for therapeutic agents. Representative examples include angiotensin.

[1029] Radionuclide agents (radioisotopes)

[1030] The linker of the present invention can be used to conjugate an antibody to at least one radionuclide agent. Radionuclide agents include agents characterized by unstable nuclei that can undergo radioactive decay. The basis of successful radionuclide therapy depends on sufficient concentrations of the radionuclide and the retention of cancer cells for an extended period of time. Other factors considered include the radionuclide half-life, the energy of the emitted particles, and the maximum range to which the emitted particles can travel. In a preferred embodiment, the therapeutic agent is a radionuclide selected from the group consisting of: 111In, 177Lu, 212Bi, 213Bi, 211At, 62Cu, 64Cu, 67Cu, 90Y, 125I, 131I, 32P, 33P, 47Sc, 111Ag, 67Ga, 142Pr, 153Sm, 161Tb, 166Dy, 166Ho, 186Re, 188Re, 189Re, 212Pb, 223Ra, 225Ac, 59Fe, 75Se, 77As, 89Sr, 99Mo, 105Rh, 109Pd, 143Pr, 149Pm, 169Er, 194Ir, 198Au, 199Au, and 211Pb. Also preferred are radionuclides that decay significantly with Auger-emitting particles. Examples include Co-58, Ga-67, Br-80m, Tc-99m, Rh-103m, Pt-109, In-111, Sb-119, I-125, Ho-161, Os-189m, and Ir-192. Useful beta-emitting nuclides with decay energies preferably include Dy-152, At-211, Bi-212, Ra-223, Rn-219, Po-215, Bi-211, Ac-225, Fr-221, At-217, Bi-213, and Fm-255. The decay energy of useful alpha-particle-emitting radionuclides is preferably 2,000-10,000 keV, more preferably 3,000-8,000 keV, and most preferably 4,000-7,000 keV. Additional potential radioisotopes used include 11C, 13N, 150, 75Br, 198Au, 95Ru, 97Ru, 103Ru, 105Ru, 107Hg, 203Hg, 121mTe, 122mTe, 125mTe, 165Tm, 167Tm, 168Tm, 197Pt, 109Pd, 105Rh, 142Pr, 143Pr, 161Tb, 166Ho, 199Au, 57Co, 58Co, 51Cr, 59Fe, 75Se, 201Tl, 225Ac, 76Br, 169Yb, etc.

[1031] Radiosensitizers

[1032] The linker of the present invention can be used to conjugate an antibody to at least one radiosensitizer. As used herein, the term "radiosensitizer" is defined as a molecule, preferably a low molecular weight molecule, that increases the sensitivity of cells to electromagnetic radiation and / or promotes the treatment of diseases that can be treated with electromagnetic radiation. Radiosensitizers are agents that make cancer cells more sensitive to radiotherapy while typically having a much smaller effect on normal cells. Therefore, radiosensitizers can be used in combination with radiolabeled antibodies or ADCs. Compared to treatment with radiolabeled antibodies or antibody fragments alone, the addition of radiosensitizers can lead to enhanced efficacy. Radiosensitizers are described in DM Goldberg (ed.), Cancer Therapy with Radiolabeled Antibodies, CRC Press (1995). Examples of radiosensitizers include gemcitabine, 5-fluorouracil, taxanes, and cisplatin.

[1033] Radiosensitizers can be activated by electromagnetic radiation of X-rays. Representative examples of X-ray activated radiosensitizers include, but are not limited to, the following: metronidazole, misonidazole, desmethylmisonidazole, pimonidazole, etanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233, E09, RB 6145, nicotinamide, 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR), bromodeoxycytidine, fluorodeoxyuridine (FUdR), hydroxyurea, cisplatin, and therapeutically effective analogs and derivatives thereof. Alternatively, photodynamic therapy (PDT) can be used to activate the radiosensitizer. Representative examples of photodynamic radiosensitizers include, but are not limited to, hematoporphyrin derivatives, photoporphyrin (r), benzoporphyrin derivatives, NPe6, tin etioporphyrin (SnET2), pheoborbide a, bacteriochlorophyll a, naphthalocyanine, phthalocyanine, zinc phthalocyanine, and therapeutically effective analogs and derivatives thereof.

[1034] Topoisomerase inhibitors

[1035] The linker of the present invention can be used to conjugate an antibody to at least one topoisomerase inhibitor.Topoisomerase inhibitors are chemotherapy agents designed to interfere with the effects of topoisomerases (topoisomerases I and II), which are enzymes that control DNA structural changes by catalysis, then breaking and recombining the phosphodiester backbone of the DNA chain during the normal cell cycle. Representative examples of DNA topoisomerase I inhibitors include, but are not limited to, camptothecin and its derivatives irinotecan (CPT-11, Camptosar, Pfizer) and topotecan (Hycamtin, GlaxoSmithKline Pharmaceuticals). Representative examples of DNA topoisomerase II inhibitors include, but are not limited to, amsacrine, daunorubicin, doxorubicin, epipodophyllotoxin, ellipticine, epirubicin, etoposide, razoxane, and teniposide.

[1036] Tyrosine kinase inhibitors

[1037] The linkers of the present invention can be used to conjugate antibodies to at least one tyrosine kinase inhibitor. Tyrosine kinases are enzymes within cells that attach phosphate groups to the amino acid tyrosine. By blocking the ability of protein tyrosine kinases to function, tumor growth can be inhibited. Examples of tyrosine kinases that can be used in the ADCs of the present invention include, but are not limited to, axitinib, bosutinib, cediranib, dasatinib, erlotinib, gefitinib, imatinib, lapatinib, lestaurtinib, nilotinib, semaxanib, sunitinib, and vandetanib.

[1038] Other medicines

[1039] Examples of other agents that can be used in the ADCs of the present invention include, but are not limited to, abrin (e.g., abrin A chain), alpha toxin, Aleurites fordii protein, amatoxin, crotin, curcin, dianthin protein, diphtheria toxin (e.g., diphtheria A chain and inactive fragments of diphtheria toxin), deoxyribonuclease (DNase), gelonin, mitogellin, modeccin A chain, momordica charantia inhibitor, streptothricin, onconase, phenomycin, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), pokeweed antiviral proteins, Pseudomonas exotoxin, Pseudomonas exotoxin (e.g., exotoxin A chain (from Pseudomonas aeruginosa), and phenotypes thereof). aeruginosa), restrictocin, ricin A chain, ribonuclease (RNase), sapaonaria officinalis inhibitors, saporin, α-sarcin, Staphylococcal enterotoxin-A, tetanus toxin, cisplatin, carboplatin, and oxaliplatin (Eloxatin, Sanofi Aventis), proteasome inhibitors (e.g., PS-341 [bortezomib or Velcade]), HDAC inhibitors (vorinostat (Zolinza, Merck), belinostat, entinostat, motistat, and panobinostat), COX-2 inhibitors, substituted ureas, heat shock protein inhibitors (e.g., geldanamycin and its various analogs), adrenocortical suppressants, and tricothecenes. (See, e.g., WO 93 / 21232). Other agents include asparaginase (Espar, Lundbeck), hydroxyurea, levamisole, mitotane (Lysodren, Bristol-Myers Squibb), and tretinoin (Renova, Valeant Pharmaceuticals).

[1040] It should be noted that the foregoing group of drug moieties useful in the ADCs of the present invention is not exclusive, as some examples of drugs can be found in more than one class, e.g., ansamitocin is both a mitotic inhibitor and an antitumor antibiotic.

[1041] All stereoisomers of the above drug moieties are contemplated for use in the compounds of the present invention, ie, any combination of R and S configurations at the chiral carbons of D.

[1042] A "detectable moiety" or "marker" refers to a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, radioactive, or chemical means. For example, useful labels include 32 P. 35 The detectable moiety can be a fluorescent marker, a fluorescent dye, an electron-dense reagent, an enzyme (e.g., an enzyme commonly used in ELISA), a biotin-streptavidin, a digoxigenin, a hapten, and a protein or nucleic acid molecule with a sequence complementary to the target, such as a fluorescent marker, a colorimetric marker, or a fluorescent marker. The detectable moiety can typically produce a measurable signal, such as a radioactive signal, a colorimetric marker, or a fluorescent signal, which can be used to quantitatively measure the amount of the detectable moiety bound to the sample. Quantification of the signal can be accomplished by, for example, scintillation counting, a densitometer, flow cell analysis, ELISA, or by mass spectrometry of a cyclic or subsequently digested peptide (one or more peptides can be determined). Those skilled in the art are familiar with the techniques and detection methods for labeled compounds of interest. These techniques and methods are conventional and well-known in the art.

[1043] The probe used for detection refers to (i) a material that can provide a detectable signal, (ii) a material that can interact with the first probe or the second probe to change the detectable signal provided by the first probe or the second probe (such as fluorescence resonance energy transfer (FRET)), (iii) a material that can stabilize the interaction with the antigen or ligand or increase the binding affinity, (iv) a material that can affect electromigration or cell invasion through physical parameters such as charge, hydrophobicity, etc., or (v) a material that can regulate ligand affinity, antigen-antibody binding or ion complex formation.

[1044] Antibody

[1045] The antibody of the ADC can be any antibody that typically, but not necessarily specifically, binds to an antigen expressed on the surface of a target cell of interest. The antigen is not required, but in some embodiments, the ADC bound thereto can be internalized into the cell. The target cell of interest may include cells in which apoptosis is desired to be induced. The target antigen can be any protein, glycoprotein, polysaccharide, lipoprotein, etc. expressed on the target cell of interest, but will typically be a protein that is uniquely expressed on the target cell rather than on a normal or healthy cell or overexpressed on the target cell compared to a normal or healthy cell, so that the ADC selectively targets a specific cell of interest, such as a tumor cell. As will be understood by the skilled person, the specific antigen selected and therefore the antibody will depend on the identity of the desired target cell of interest. In a specific embodiment, the antibody of the ADC is an antibody suitable for administration to a human.

[1046] Antibodies (Ab) and immunoglobulins (Ig) are glycoproteins with identical structural features. Although antibodies exhibit binding specificity for specific targets, immunoglobulins include antibodies and other antibody-like molecules that lack target specificity. Natural antibodies and immunoglobulins are typically heterotetrameric glycoproteins of about 150,000 daltons, which are composed of two identical light (L) chains and two identical heavy (H) chains. Each heavy chain has a variable domain (VH) at one end, followed by multiple constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end.

[1047] References to "VH" refer to the variable region of an immunoglobulin heavy chain (including the heavy chain of an Fv, scFv, or Fab) of an antibody. References to "VL" refer to the variable region of an immunoglobulin light chain (including the light chain of an Fv, scFv, dsFv, or Fab).

[1048] The term "antibody" as used herein is used in the broadest sense and refers to an immunoglobulin molecule that specifically binds to or is immunoreactive with a particular antigen, and includes polyclonal, monoclonal, genetically engineered, and otherwise modified forms of antibodies (including, but not limited to, murine chimeric antibodies, humanized antibodies, heteroconjugate antibodies (e.g., bispecific antibodies, diabodies, triabodies, and tetrabodies)), as well as antigen-binding fragments of antibodies (including, for example, Fab', F(ab')2, Fab, Fv, rIgG, and scFv fragments). The term "scFv" refers to a single-chain Fv antibody in which the variable domains of the heavy and light chains of a traditional antibody have been linked to form a single chain.

[1049] Antibodies can be murine, human, humanized, chimeric or derived from other species. Antibodies are proteins produced by the immune system that can recognize and bind to specific antigens. (Janeway, C., Travers, P., Walport, M., Shlomchik (2001) Immuno Biology, 5th edition, Garland Publishing, New York). The target antigen typically has many binding sites, also known as epitopes, that are recognized by the CDRs on multiple antibodies. Each antibody that specifically binds to a different epitope has a different structure. Therefore, an antigen can have more than one corresponding antibody. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immunospecifically binds to an antigen or a portion of a target of interest, such targets including but not limited to cancer cells or cells that produce autoimmune antibodies associated with autoimmune diseases. The immunoglobulins disclosed herein can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecules. The immunoglobulins can be derived from any species. However, in one aspect, the immunoglobulins are of human, mouse, or rabbit origin.

[1050] The term "antibody fragment" refers to a portion of a full-length antibody, typically the target binding or variable region. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments. The "Fv" fragment is the smallest antibody fragment that contains a complete target recognition and binding site. This region consists of a dimer of one heavy-chain variable domain and one light-chain variable domain in a tight, non-covalent association (VH-VL dimer). It is in this configuration that the three CDRs of each variable domain interact to define a target binding site on the surface of the VH-VL dimer. Typically, the six CDRs confer target binding specificity to the antibody. However, in some cases, even a single variable domain (or half of an Fv, containing only three CDRs specific for a target) can have the ability to recognize and bind a target. A "single-chain Fv" or "scFv" antibody fragment contains the VH and VL domains of an antibody in a single polypeptide chain. Typically, the Fv polypeptide further comprises a polypeptide linker between the VH domain and the VL domain that enables the scFv to form the desired structure for antigen binding. A "single-domain antibody" is composed of a single VH or VL domain that exhibits sufficient affinity for the target. In specific embodiments, the single-domain antibody is a camelized antibody (see, e.g., Riechmann, 1999, Journal of Immunological Methods 231: 25-38).

[1051] Fab fragments contain the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab' fragments differ from Fab fragments by the addition of several residues at the carboxyl terminus of the heavy chain CH1 domain (including one or more cysteines from the antibody hinge region). F(ab') fragments are produced by cleavage of disulfide bonds at the hinge cysteines of the F(ab')2 pepsin digest. Additional chemical couplings of antibody fragments are known to those of ordinary skill in the art.

[1052] The light chain variable domain and the heavy chain variable domain both have complementarity determining regions (CDRs), also referred to as hypervariable regions. The more highly conserved portion of the variable domain is referred to as framework region (FR). As known in the art, the amino acid position / boundary of the hypervariable region of an antibody can vary, depending on the context and various definitions known in the art. Some positions within the variable domain can be considered as hybrid hypervariable positions, because these positions can be considered to be within the hypervariable region under a set of standards, and are considered to be outside the hypervariable region under a set of different standards. One or more of these positions can also be found in the hypervariable region of extension. The CDRs in each chain are closely held together by the FR region and together with the CDRs from another chain, contribute to the formation of the target binding site of the antibody (see Kabat et al., Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md. 1987). As used herein, unless otherwise noted, the numbering of immunoglobulin amino acid residues is performed according to the immunoglobulin amino acid residue numbering system of Kabat et al.

[1053] In certain embodiments, the antibody of the ADC of the present disclosure is a monoclonal antibody. The term "monoclonal antibody" (mAb) refers to an antibody that is derived from a single copy or clone, including, for example, any eukaryotic, prokaryotic, or phage clone, rather than the method of producing it. Preferably, the monoclonal antibodies of the present disclosure are present in a homogeneous or substantially homogeneous population. Monoclonal antibodies include both complete molecules and antibody fragments (such as Fab fragments and F(ab')2 fragments) that can specifically bind to proteins. Fab fragments and F(ab')2 fragments lack the Fc fragment of complete antibodies (cleared faster from animal circulation) and can have less non-specific tissue binding than complete antibodies (Wahl et al., 1983, J.Nucl.Med 24:316). Monoclonal antibodies that can be used in the present disclosure can be prepared using a variety of techniques known in the art, including the use of hybridomas, recombinants, and phage display technology or a combination thereof. Antibodies of the present disclosure include chimeric, primatogenic, humanized, or human antibodies.

[1054] Although in most cases antibodies are composed only of genetically encoded amino acids, in some embodiments, non-encoded amino acids can be specifically incorporated. Examples of non-encoded amino acids that can be incorporated into antibodies for controlling stoichiometry and attachment position, as well as methods for making such modified antibodies, are discussed in Tian et al., 2014, Proc Nat'l Acad Sci USA 111(5):1766-1771 and Axup et al., 2012, Proc Nat'l Acad Sci USA 109(40):16101-16106 (incorporated herein by reference in their entireties).

[1055] In certain embodiments, the antibody of the ADC described herein is a chimeric antibody. As used herein, the term "chimeric" antibody refers to an antibody having a variable sequence derived from a non-human immunoglobulin (such as a rat or mouse antibody) and a human immunoglobulin constant region (typically selected from a human immunoglobulin template). Methods for producing chimeric antibodies are known in the art. See, for example, Morrison, 1985, Science 229 (4719): 1202-7; Oi et al., 1986, BioTechniques 4: 214-221; Gillies et al., 1985, J. Immunol. Methods 125: 191-202; U.S. Patent Nos. 5,807,715; 4,816,567; and 4,816397, which are incorporated herein by reference in their entirety.

[1056] In certain embodiments, the antibody of ADC described herein is a humanized antibody. " Humanized " form of non-human (for example, mouse) antibody is a chimeric immunoglobulin, immunoglobulin chain or its fragment (such as Fv, Fab, Fab', F(ab')2 or other target binding subdomains of an antibody) containing the minimum sequence derived from a non-human immunoglobulin. Generally, a humanized antibody will comprise substantially all of at least one and typically two variable domains, all or substantially all of the CDR regions corresponding to those of a non-human immunoglobulin, and all or substantially all of those humanized antibodies in the FR regions that are human immunoglobulin sequences can also comprise at least a portion of an immunoglobulin constant region (Fc), typically a portion of a human immunoglobulin consensus sequence. Methods for antibody humanization are known in the art. See, e.g., Riechmann et al., 1988, Nature 332:323-7; U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,761; 5,693,762; and U.S. Pat. No. 6,180,370, Queen et al.; EP 239400; PCT Publication WO 91 / 09967; U.S. Pat. No. 5,225,539; EP 592106; EP 519596; Padlan, 1991, Mol. Immunol., 28:489-498; Studnicka et al., 1994, Prot. Eng. 7:805-814; Roguska et al., 1994, Proc. Natl. Acad. Sci. USA 91:969-973; and US Patent No. 5,565,332, all of which are hereby incorporated by reference in their entirety.

[1057] In certain embodiments, the antibody of ADC described herein is a human antibody. Completely "human" antibodies may be desirable for therapeutic treatment of human patients. As used herein, "human antibodies" include antibodies with the amino acid sequence of human immunoglobulin, and include antibodies separated from human immunoglobulin libraries or from animals for one or more human immunoglobulin transgenics and not expressing endogenous immunoglobulins. Human antibodies can be manufactured by a variety of methods known in the art, including phage display methods using antibody libraries derived from human immunoglobulin sequences. See U.S. Patent Nos. 4,444,887, 4,716,111, 6,114,598, 6,207,418, 6,235,883, 7,227,002, 8,809,151 and U.S. Publication Application No. 2013 / 189218, the contents of which are incorporated herein by reference in their entirety. Human antibodies can also be produced using transgenic mice that are unable to express functional endogenous immunoglobulins but can express human immunoglobulin genes. See, for example, U.S. Patent Nos. 5,413,923; 5,625,126; 5,633,425; 5,569,825; 5,661,016; 5,545,806; 5,814,318; 5,885,793; 5,916,771; 5,939,598; 7,723,270; 8,809,051 and U.S. Published Application No. 2013 / 117871, which are incorporated herein by reference in their entirety. In addition, companies such as Medarex (Princeton, New Jersey), Astellas Pharma (Astellas Pharma, Deerfield, Illinois) and Regeneron (Regeneron, Tarrytown, New York) can be hired to provide human antibodies for the selected antigen using techniques similar to those described above. A technique called "guided selection" can be used to generate fully human antibodies that recognize the selected epitope. In this method, a selected non-human monoclonal antibody (e.g., mouse antibody) is used to guide the selection of fully human antibodies that recognize the same epitope (Jespers et al., 1988, Biotechnology 12:899-903).

[1058] In certain embodiments, the antibodies of the ADCs described herein are primatized antibodies. The term "primatized antibody" refers to an antibody comprising a monkey variable region and a human constant region. Methods for producing primatized antibodies are known in the art. See, for example, U.S. Patent Nos. 5,658,570; 5,681,722; and 5,693,780, which are incorporated herein by reference in their entireties.

[1059] In certain embodiments, the antibody of ADC described herein is a bispecific antibody or a dual variable domain antibody (DVD). Bispecific and DVD antibodies are monoclonal, generally human or humanized antibodies that have binding specificity for at least two different antigens. DVD is described in, for example, U.S. Patent No. 7,612,181, the disclosure of which is incorporated herein by reference.

[1060] In certain embodiments, the antibodies of the ADCs described herein are derivative antibodies. For example, but not limited to, derivative antibodies are typically modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting / blocking groups, proteolytic cleavage, connection to cellular ligands or other proteins, etc. Any of a variety of chemical modifications can be performed by known techniques, including but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. In addition, the derivatives can contain one or more non-natural amino acids, for example, using ambrx technology (see, for example, Wolfson, 2006, Chem.Biol.13(10):1011-2).

[1061] In certain embodiments, the antibodies of the ADCs described herein have sequences that have been modified to alter at least one constant region-mediated biological effector function relative to the corresponding wild-type sequence. For example, in some embodiments, the antibodies can be modified to reduce at least one constant region-mediated biological effector function relative to an unmodified antibody, such as reducing binding to an Fc receptor (FcR). FcR binding can be reduced by mutating segments of the immunoglobulin constant region of the antibody in specific regions necessary for FcR interaction (see, e.g., Canfield and Morrison, 1991, J. Exp. Med 173: 1483-1491; and Lund et al., 1991, J. Immunol. 147: 2657-2662).

[1062] In certain embodiments, the antibodies of the ADCs described herein are modified to obtain or improve at least one constant region-mediated biological effector function relative to an unmodified antibody, such as to enhance FcγR interactions (see, e.g., US 2006 / 0134709). For example, antibodies having a constant region that binds to FcγRIIA, FcγRIIB, and / or FcγRIIIA with an affinity greater than that of the corresponding wild-type constant region can be produced according to the methods described herein.

[1063] In certain specific embodiments, the antibody of ADC described herein is an antibody that binds to tumor cells, such as an antibody to a cell surface receptor or a tumor associated antigen (TAA). In an attempt to find effective cell targets for cancer diagnosis and therapy, researchers seek to identify transmembrane polypeptides or other tumor-associated polypeptides that are specifically expressed on the surface of one or more specific types of cancer cells compared to one or more normal non-cancerous cells. Typically, such tumor-associated polypeptides are more abundantly expressed on the surface of cancer cells compared to the surface of non-cancerous cells. Such cell surface receptors and tumor associated antigens are known in the art and can be prepared for the generation of antibodies using methods and information well known in the art.

[1064] Exemplary Cell Surface Receptors and TAAs

[1065] Examples of cell surface receptors and TAAs that the antibodies of the ADCs described herein can target include, but are not limited to, the various receptors and TAAs listed in Table 1 below. For convenience, information related to these antigens (all of which are known in the art) is listed below and follows the National Center for Biotechnology Information (NCBI) National Center for Nucleic Acid and Protein Sequence Identification Conventions, including names, alternative names, Genbank accession numbers, and one or more primary references. Nucleic acid and protein sequences corresponding to the listed cell surface receptors and TAAs are available in public databases such as GenBank.

[1066] Table 1.

[1067]

[1068]

[1069]

[1070]

[1071] Exemplary antibodies

[1072] Exemplary antibodies to be used in the ADCs of the present disclosure include, but are not limited to, 3F8 (GD2), Abagovomab (CA-125 (generic)), Adecatumumab (EpCAM), Afutuzumab (CD20), Alacizumab pegol (VEGFR2), ALD518 (IL-6), Alemtuzumab (CD52), Altumomab pentetate (CEA), Amatuximab (mesothelin), Anatumomnab (C-10), and 4-Hydroxyprogesterone (Hydroxyprogesterone). Mafenatox (TAG-72), Apolizumab (HLA-DR), Arcitumomab (CEA), Bavituximab (phosphatidylserine), Betumomab (CD22), Belimumab (BAFF), Besilesomab (CEA-related antigen), Bevacizumab (VEGF-A), Bivatuzumab mertansine (CD44v6), Blinatumomab (CD19), Brentuximab vedotin (CD30 (TNFRSF8)), Cantuzumab mertansine (CanAg), Cantuzumab ravtansine (MUC1), Capromab pendetide (Centrolide A). Pendetide (prostate cancer cells), Carlumab (MCP-1), Catumaxomab (EpCAM, CD3), CC49 (Tag-72), cBR96-DOX ADC (Lewis-Y antigen), Cetuximab (EGFR), Citatuzumab bogatox (EpCAM), Cixutumumab (IGF-1 receptor), Clivatuzumab tetraxetan (MUC1), Conatumumab (TRAIL-E2), Dacetuzumab (CD40), Dalotuzumab (Insulin-like growth factor 1 receptor), Deratumumab ((CD38 (cyclic ADP ribose hydrolase)), Demcizumab (DLL4), Denosumab (RANKL), Detumomab (B-lymphoma cells), Drozitumab (DR5), Dusigitumab (ILGF2), Ecromeximab (D3 ganglioside), Eculizumab (C5), Edrecolomab (EpCAM), Elotuzumab (SLAMF7), Elsilimomab (IL-6), Enavatuzumab (TWEAK receptor), Enoticumab (DLL4), Ensituximab (5AC), Epitumomab cituxetan (Episialin), Epratuzumab (CD22), Ertumaxomab (HER2 / neu, CD3), Etancizumab (Integrin αvβ3), Farletuzumab (Folate Receptor 1), FBTA05 (CD20), Ficlatuzumab (HGF), Figitumumab (IGF-1 Receptor), Flanvotumab (TYRP1 (Glycoprotein 75)), Fresolimumab (TGF-1), Galiximab (CD80), Ganitumab (IGF-1), Gemtuzumab Ozogamicin (CD33), Girentuximab (Carbonic Anhydrase 9 (CA-IX)), Glembatumumab Vedotin (GPNMB), Icrucimab (CD20), Icrucumab (VEGFR-1), Igovomab (CA-125), IMAB362 (CLDN18.2), Imgatuzumab (EGFR), Indatuximab ravtansine (SDC1), Intetumumab (CD51), Inotuzumab ozogamicin (CD22), Ipilimumab (CD152), Iratumumab (CD30 (TNFRSF8)), Labetuzumab (CEA), Lambrolizumab (PDCD1), Lexatumumab (TRAIL-R2), Lintuzumab (CD33), Lorvotuzumab mertansine (CD56), Lucatumumab (CD40), Lumiliximab (CD23 (IgE receptor)), Mapatumumab (TRAIL-R1), Margetuximab (ch4DS), Matuzumab (EGFR), Milatuzumab (CD74), Mitumomab (GD3 ganglioside), Mogamulizumab (CCR4), Moxetumomab pasudotox (CD22), Nacolomabtafenatox (C2-42 antigen), Naptumomab estafenatox, 5T4), Narnatumab (RON), Natalizumab (integrin α4), Necitumumab (EGFR), Nesvacumab (angiopoietin 2), Nimotuzumab (EGFR), Nivolumab (IgG4), Ocaratuzumab (CD20), Ofatumumab (CD20), Olaratumab (PDGF-Rα), Onartuzumab (human discrete factor receptor kinase), Ontuxizumab (TEM1), Monto-Oportuzumab monato, EpCAM), Oregovomab (CA-125), Otlertuzumab (CD37), Panitumumab (EGFR), Pankomab (tumor-specific glycosylation of MUC1), Parsatuzumab (EGFL7), Patritumab (HER3), Pemtumomab (MUC1), Pertuzumab (HER2 / neu), Pidilizumab (PD-1), Vitin-Pinatuzumab Vedotin (CD22), Pritumumab (vimentin), Racotumomab (N-glycolylneuraminic acid), Radretumab (fibronectin extra domain-B), Ramucirumab (VEGFR2), Rilotumumab (HGF), Rituximab (CD20), Robatumumab (IGF-1 receptor), Samalizumab (CD200), Satumomab pendetide (TAG-72), Seribantumab (ERBB3), Sibrotuzumab (FAP), SGN-CD19A (CD19), SGN-CD33A (CD33), Siltuximab (IL-6), Solitomab (EpCAM), Sonepcizumab (sphingosine-1-phosphate), Tabalumb (BAFF), Tacatuzumab tetraxetan (α-fetoprotein), Taplitumomab paptox (CD19), tenatumomab (tenascin C), teprotumumab (CD221), TGN1412 (CD28), tesitumomab (CTLA-4), tigatuzumab (TRAIL-R2), TNX-650 (IL-13), tovetumab (CD40a), trastuzumab (HER2 / neu), TRBS07 (GD2), tremelimumab (CTLA-4), tucotuzumab (interleukin-6,1-dT), trastuzumab (HER2 / neu), TRBS07 (GD2), tremelimumab (CTLA-4), tucotuzumab (interleukin-7,1-dT ...7,1-dT), trastuzumab ( celmoleukin (EpCAM), Ublituximab (MS4A), Urelumab (4-1BB), Vandetanib (VEGF), Vantictumab (Frizzled receptor), Volociximab (Integrin α5β1), Vorsetuzumab mafodotin (CD70), Votumumab (Tumor antigen CTAA16.88), Zalutumumab (EGFR), Zanolimumab (CD4), and Zatuximab (HER1).

[1073] Methods for producing antibodies

[1074] The antibody of ADC can be prepared by recombinantly expressing immunoglobulin light chain and heavy chain genes in host cells. For example, in order to recombinantly express antibodies, host cells are transfected with recombinant expression vectors carrying one or more DNA fragments of immunoglobulin light chain and heavy chain encoding antibodies so that light chain and heavy chain are expressed in host cells, and optionally, secreted into the culture medium of the host cells, from which antibodies can be recovered. Standard recombinant DNA methods are used to obtain antibody heavy chain and light chain genes, these genes are incorporated into recombinant expression vectors and the vectors are introduced into host cells, such as those described in Molecular Cloning; A Laboratory Manual, Second Edition (Sambrook, Fritsch and Maniatis (eds.), Cold Spring Harbor, NY, 1989), Current Protocols in Molecular Biology (Ausubel, FM et al., eds., Greene Publishing Associates, 1989) and U.S. Patent No. 4,816,397.

[1075] In one embodiment, Fc variant antibodies are similar to their wild-type equivalents, but their Fc domains are altered. To generate nucleic acids encoding such Fc variant antibodies, a DNA fragment encoding the Fc domain of a wild-type antibody (referred to as a "wild-type Fc domain") or a portion of the Fc domain can be synthesized and used as a mutagenesis template to produce antibodies described herein using conventional mutagenesis techniques; alternatively, a DNA fragment encoding the antibody can be directly synthesized.

[1076] Once a DNA fragment encoding a wild-type Fc domain is obtained, these DNA fragments can be further manipulated by standard recombinant DNA techniques, for example to convert constant region genes into full-length antibody chain genes. In these manipulations, the DNA fragment encoding CH is operably linked to another DNA fragment encoding another protein, such as an antibody variable region or a flexible linker. As used in this context, the term "operably linked" is intended to mean connecting two DNA fragments so that the amino acid sequences encoded by the two DNA fragments remain in frame.

[1077] In order to express Fc variant antibodies, the DNA encoding part or full-length light chain and heavy chain obtained as described above is inserted into an expression vector so that the gene is operably linked to transcription and translation control sequences. In this context, the term "operably linked" is intended to mean that the antibody gene is connected to a vector so that the transcription and translation control sequences in the vector play the function of its expected regulation of the transcription and translation of the antibody gene. Expression vector and expression control sequences are selected to be compatible with the expression host cell used. Variant antibody light chain gene and antibody heavy chain gene can be inserted into a separate vector, or more typically, two genes are inserted into the same expression vector.

[1078] The antibody gene is inserted into an expression vector by standard methods (for example, by connecting the complementary restriction sites on the antibody gene fragment and the vector, or by flat-end connection if there is no restriction site). Before inserting the variant Fc domain sequence, the expression vector may have carried the antibody variable region sequence. Additionally or alternatively, the recombinant expression vector may encode a signal peptide that promotes secretion of the antibody chain from the host cell. The antibody chain gene may be cloned into a vector so that the signal peptide is connected to the amino terminus of the antibody chain gene in frame. The signal peptide may be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., a signal peptide from a non-immunoglobulin protein).

[1079] In addition to the antibody chain genes, the recombinant expression vector also carries regulatory sequences that control the expression of the antibody chain genes in the host cell. The term "regulatory sequence" is intended to include promoters, enhancers and other expression control elements (e.g., polyadenylation signals) that control the transcription or translation of the antibody chain genes. Such regulatory sequences are described in, for example, Goeddel, Gene Expression Technology: Methods in Enzymology 185 (Academic Press, San Diego, Calif., 1990). It will be appreciated by those skilled in the art that the design of the expression vector (including the selection of regulatory sequences) can depend on factors such as the selection of the host cell to be transformed, the expression level of the desired protein, etc. Suitable regulatory sequences for mammalian host cell expression include viral elements that direct high-level protein expression in mammalian cells, such as promoters and / or enhancers derived from: cytomegalovirus (CMV) (such as CMV promoter / enhancer), simian virus 40 (SV40) (such as SV40 promoter / enhancer), adenovirus, (e.g., adenovirus major late promoter (AdMLP)) and polyomavirus. For further description of viral regulatory elements and their sequences, see, eg, US Patent No. 5,168,062 to Stinski, US Patent No. 4,510,245 to Bell et al., and US Patent No. 4,968,615 to Schaffner et al.

[1080] In addition to antibody chain genes and regulatory sequences, the recombinant expression vector carries and can also carry other sequences, such as sequences (for example, replication origin) and selective marker genes for regulating vector replication in host cells. The selective marker gene helps to select the host cell (see, for example, U.S. Patent numbers 4,399,216, 4,634,665 and 5,179,017, all by Axel et al.) into which vectors have been introduced. For example, typically, the selective marker gene gives resistance to drugs (such as G418, puromycin, blasticidin, hygromycin or methotrexate) to the host cell into which vectors have been introduced. Suitable selective marker genes include dihydrofolate reductase (DHFR) gene (for use in DHFR host cells selected / amplified with methotrexate) and neo gene (selected for G418). For the expression of light chain and heavy chain, one or more expression vectors encoding heavy chain and light chain are transfected into host cells by standard techniques. The various forms of the term "transfection" are intended to encompass a wide variety of techniques commonly used to introduce exogenous DNA into prokaryotic or eukaryotic host cells, such as electroporation, lipofection, calcium phosphate precipitation, DEAE-dextran transfection, and the like.

[1081] Can express antibody in prokaryotic or eukaryotic host cell.In certain embodiments, antibody is expressed in eukaryotic cell (for example, mammalian host cell), to secrete correctly folded and immunocompetent antibody optimally.Exemplary mammalian host cell for expressing recombinant antibodies comprises Chinese hamster ovary (Chinese Hamster Ovary, CHO cell) (comprising DHFR-CHO cell, is described in Urlaub and Chasin, 1980, Proc.Natl.Acad.Sci.USA77:4216-4220, use together with DHFR selective marker, for example, as described in Kaufman and Sharp, 1982, Mol.Biol.159:601-621), NSO myeloma cell, COS cell, 293 cell and SP2 / 0 cell.When the recombinant expression vector of encoding antibody gene is introduced into mammalian host cell, by described host cell culture continuing to be enough to allow described antibody to be expressed in host cell or described antibody is secreted into the time period in the culture medium of wherein growth host cell to produce antibody.Standard protein purification method can be used to reclaim antibody from culture medium. The host cells can also be used to produce portions of intact antibodies, such as Fab fragments or scFv molecules.

[1082] In some embodiments, the antibody of the ADC can be a bifunctional antibody. Such antibodies, in which one heavy chain and one light chain are specific for one antigen and the other heavy chain and light chain are specific for a second antigen, can be produced by cross-linking one antibody with a second antibody using standard chemical cross-linking methods. Bifunctional antibodies can also be made by expressing a nucleic acid engineered to encode the bifunctional antibody.

[1083] In certain embodiments, bispecific antibodies can be produced by mutating the amino acid residues in the light chain and / or heavy chain CDR, that is, antibodies that combine an antigen with a second unrelated antigen using the same binding site. Exemplary second antigens include proinflammatory cytokines (such as lymphotoxin, interferon-γ or interleukin-1). Bispecific antibodies can be produced, for example, by mutating the amino acid residues in the periphery of the antigen binding site (see, for example, Bostrom et al., 2009, Science 323: 1610-1614). Bifunctional antibodies can be manufactured by expressing a nucleic acid engineered to encode a bispecific antibody.

[1084] Antibodies can also be produced by chemical synthesis (e.g., by the methods described in Solid Phase Peptide Synthesis, 2nd Edition, 1984 The Pierce Chemical Co., Rockford, 111.) Antibodies can also be produced using a cell-free platform (see, e.g., Chu et al., Biochemia No. 2, 2001 (Roche Molecular Biologicals)).

[1085] Methods for recombinant expression of Fc fusion proteins are described in Flanagan et al., Methods in Molecular Biology, Vol. 378: Monoclonal Antibodies: Methods and Protocols.

[1086] Once antibodies have been produced by recombinant expression, they can be purified by any method known in the art for purifying immunoglobulin molecules, such as chromatography (e.g., ion exchange, affinity, particularly for the antigen after protein A or protein G selection, and size column chromatography), centrifugation, differential solubility, or by any other standard technique for protein purification.

[1087] Once isolated, the antibodies can be further purified, if desired, for example, by high performance liquid chromatography (see, e.g., Fisher, Laboratory Techniques In Biochemistry And Molecular Biology (Work and Burdon, eds., Elsevier, 1980)) or by chromatography on a Superdex TM Further purification was performed by gel filtration chromatography on a 75 column (Pharmacia Biotech AB, Uppsala, Sweden).

[1088] Imaging compounds and sensors

[1089] In certain embodiments, provided herein are uses of the disclosed compounds in imaging compositions and as sensors.

[1090] The sensor may be a biosensor, a chemical sensor or a molecular switch. Biosensors can identify the presence or amount of a specific material (e.g., cancer cells, viruses, various chemicals, etc.) by reacting a specific material with a bioreceptor (a portion designed to adsorb and react with a biomaterial such as DNA, RNA, antibodies, enzyme proteins, cells, biomembranes, hormone receptors, etc.) having selective specificity and performing measurement using a signal converter (a device that converts the reaction between the specific material and the bioreceptor into an electrical signal using various methods). They can be used in medicine, the environment, process industry, research, food, etc. (see, for example, Biosensors and Bioelectronics, 2016, 32-45; Pol. J. Environ. Stud. 2015, 19-25; Analytica Chimica Acta 568 (2006) 200-210; Biosensors and Bioelectronics 2017, 217-231; ACS Appl. Mater. Interfaces 2015, 7, 20190-20199; Journal of Coastal Life Medicine). 2016;4(3):200-202;ArtificialCells,Blood Substitutes,and Biotechnology,39:281-288;Journal of ControlledRelease 159(2012)154-163).

[1091] Chemical sensors are used to quickly and accurately monitor specific materials in many fields such as clinical diagnosis, medical research, chemical material measurement, and environmental measurement by using electrical properties such as electricity, resistance, and potential difference, and optical properties such as color and fluorescence. They include gas sensors (hydrogen, oxygen, carbon monoxide), ion sensors (cations, anions, gas-sensitive ions), component sensors (gas phase, liquid phase, luminescent components), humidity sensors (relative humidity, absolute humidity, condensation), dust / soot sensors (floating dust, dirty dust, soot, turbidity), etc. (see, for example, Chem. Soc. Rev., 2015, 44, 3358; Journal of the Korean Chemical Society, 2010, 451-459; Chem. Sci., 2015, 6, 1150-1158; KR 10-1549347; J. Phys. Chem. B 2016, 120, 7053-7061; ACS Appl. Mater. Interfaces). 2015,7,704-712; J.Am.Chem.Soc.2011,133,10960-10965; J.Am.Chem.Soc.2012,134,20412-20420; O rg.Lett.2014,16,1680-1683; J.Org.Chem.2013,78,702-705; J.Org.Chem.2015,80,12129-12136; ACS Macro Lett. 2014, 3, 1191-1195; New J. Chem., 2012, 36, 386-393; Chem. Commun., 2010, 46, 6575-6577; 2013).

[1092] A molecular switch is a molecule that can be reversibly switched between two or more stable states. The molecule can switch between multiple states in response to environmental stimuli, such as changes in the chemical environment (such as pH), light irradiation (such as light of a specific wavelength), temperature, current, microenvironment, or the presence of a ligand. In some cases, the switching between multiple states can depend on a combination of stimuli. The oldest form of synthetic molecular switches is a pH indicator, which displays different colors depending on pH. Synthetic molecular switches can be applied to molecular computers or responsive drug delivery systems. Molecular switches are also important in biology because many biological functions are based on them, such as allosteric regulation and vision.

[1093] Such biosensors, chemical sensors and molecular switches may further comprise additional photoreactive moieties such as rhodamine, phenol red, orange azo dyes, papa red, non-sulfonated cyanines, sulfonated cyanines, chemiluminescent fluoride sensors (1,2-dioxetane derivatives) and D2A dyes (NIR fluorescent dyes). Alternatively, the photoreactive moiety may be selected from compounds having functional groups and structures similar to the following:

[1094]

[1095]

[1096] in:

[1097] R 100 is H or C1-C6-alkyl;

[1098] R 101 is H or SO3H; R 102 is C1-C6-alkyl or -(CH2) z COOH;

[1099] z is an integer from 3 to 8;

[1100] R 103 and R 104 are each independently H or C1-C6 alkyl; and

[1101] R 105 and R 106 Each is independently hydrogen, COOH or SO3H.

[1102] Additional photoreactive moieties are known in the art. See, for example, Org. Lett. 2014, 16, 1680-1683; J. Am. Chem. Soc. 2011, 133, 10960-10965; Dye Lasers, 3rd ed. (Springer-Verlag, Berlin, 1990); J. Am. Chem. Soc. 2012, 134, 20412-20420).

[1103] Treatment

[1104] Target-directed therapy

[1105] The targeting portion of the conjugate can be recognized by cells, thereby providing so-called target-directed therapy.

[1106] In some embodiments, the conjugate comprises an active agent for use in target-directed therapy to treat autoimmune diseases. In some such embodiments, the active agent is selected from the group consisting of cyclosporine, cyclosporine A, mycophenylate mofetil, sirolimus, tacrolimus, enanercept, prednisone, azathioprine, methotrexate cyclophosphamide, aminocaproic acid, chloroquine, hydroxychloroquine, hydrocortisone, dexamethasone, clomiphene, DHEA, danazol, bromocriptine, meloxicam, infliximab, and the like.

[1107] In some embodiments, the compound comprises an active agent Q for use in target-directed therapy for the treatment of infectious diseases. In some such embodiments, Q is selected from the group consisting of: β-lactam series (penicillin G, penicillin V, cloxacillin, dicloxacillin, methicillin, nafcillin, oxacillin, ampicillin, amoxicillin, bacampicillin, azlocillin, carbenicillin, mezlocillin, piperacillin, ticarcillin), aminoglycoside series (amikacin, gentamicin, kanamycin, neostrepton, netilmicin, streptomycin, tobramycin), macrolide series (azithromycin, Clarithromycin, erythromycin, lincomycin, clindamycin), tetracycline series (demeclocycline, doxycycline, minocycline, tetracycline), quinolone series (cinoxacin, nalidixic acid), fluoroquinolone series (ciprofloxacin, enoxacin, grefloxacin, levofloxacin, lomefloxacin, norfloxacin, ofloxacin, sparfloxacin, trovafloxicin), polypeptide series (bacitracin, colistin, polymyxin B), sulfonamide series ( Sulfisoxazole, sulfamethoxazole, sulfadiazine, sulfamethoxazole, sulfacetamide), other antibiotics (trimethoprim, sulfamethoxazole, chloramphenicol, vancomycin, metronidazole, quinupristin, dalfopristin, rifampicin, spectinomycin, nitrofurantoin), general antiviral agents (iodine glucoside, vidarabine, acyclovir, famciclovir, pencicyclovir, valacyclovir, gancicyclovir), Foscarnet, ribavirin, amantadine, rimantadine, cidofovir, antisense oligonucleotides, immunoglobulins, interferons), HIV infection therapeutic agents (tenofovir, emtricitabine, zidovudine, didanosine, zalcitabine, stavudine, lamivudine, nevirapine, delaviridine, saquinavir, ritonavir, indinavir, nelfinavir), etc.

[1108] In some embodiments, the compounds and conjugates disclosed herein comprise an active agent Q for use in a method for delivering an active agent to a cell to treat a tumor, wherein the targeting moiety is selected to bind to the target cell (i.e., a cancer cell). In particular, the compounds, conjugates, and compositions of the invention can be used to inhibit abnormal cell growth or treat a proliferative disorder in a mammal (e.g., a human), such as when the target cell is a cancer cell and the targeting moiety is selected to bind to a molecule that is associated with the cancer cell (and not with healthy cells or at least preferentially associated with tumor cells rather than healthy cells).

[1109] In some such embodiments, the active agent is selected from the group consisting of: a cytotoxic or immunomodulatory agent, an anticancer agent, an anti-tubulin agent, a cytotoxic agent, and the like. Preferably, the cytotoxic or immunomodulatory agent includes an anti-tubulin agent, auristatin, a DNA minor groove binder, a DNA transcription inhibitor, an alkylating agent, an anthracycline, an antibiotic (antibitiotic), an antifolate, an antimetabolite, a calmodulin inhibitor, a chemotherapy sensitizer, a duocarmycin, an etoposide, a fluorinated pyrimidine, an ionophore, a lecithin, a maytansinoid alkaloid, a nitrosourea, a cisplatin, a pore-forming compound, a purine antimetabolite, puromycin, a radiosensitizer, a rapamycin, a steroid, a taxane, a topoisomerase inhibitor, a vinca alkaloid, etc.; and the anticancer agent includes methotrexate, taxol, L-asparaginase, mercaptopurine, thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosourea, cisplatin, carboplatin, mitomycin, dacarbazine, proocarbizine, topotecan, nitrogen mustard, cytoxan ), etoposide, 5-fluorouracil, BCNU, irinotecan, camptothecin, bleomycin, doxorubicin, idarubicin, daunorubicin, dactinomycin, plicamycin, mitoxantrone, asparaginase, vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel, etc.; anti-tubulin agents include taxanes (e.g., paclitaxel, docetaxel), T67, vinca alkaloids (e.g., vincristine, vinblastine, vindesine, vinorelbine), baccatin ( baccatin) derivatives, taxane derivatives, epothilones (e.g., epothilone A, epothilone B), nocodazole, colchicine, colcemid, estramustine, carboxylic acid peptides, cemadotin, maytansinoids, combrestatin, discodermolide, sclerocortin, auristatin derivatives (AFP, MMAF, MMAE), etc.;Cytotoxic agents include androgens, anthromycin (AMC), asparaginase, 5-azacytidine, azathioprine, bleomycin, busulfan, buthionine sulfoxide, sulfoximine), calicheamicin, calicheamicin derivatives, camptothecin, carboplatin, carmustine (BSNU), CC-1065, chlorambucin, cisplatin, colchicine, cyclophosphamide, cytarabine, cytosine arabinoside, cytochalasin B, dacarbazine, dactinomycin (actinomycin), daunomycin, decarbazine, DM1, DM4, docetaxel, doxorubicin, etoposide, estrogen, 5-fluorodeoxyuridine, 5-fluorouracil, gemcitabine, gramicidin D, hydroxyurea, idarubicin, ifosfamide, irinotecan, lomustine (CCNU), maytansine, bischloromethyl diethylamide, melphalan, 6-mercaptopurine, methotrexate, mithramycin, mitomycin C, mitoxantrone, nitroimidazoles, paclitaxel, palytoxin, plicamycin , procarbizine, rhizotomycin, streptozotocin, tenoposide, 6-thioguanine, thio-TEPA, topotecan, vinblastine, vincristine, vinorelbine, VP-16, VM-26; DNA minor groove binders (e.g., enediynes, lecithin, CBI compounds), duocarmycins, taxanes (e.g., paclitaxel, docetaxel), puromycin, vinca alkaloids, CC-1065, SN-38, topotecan, morpholino-doxorubicin, rhizotomycin, cyanomorpholino-doxorubicin, echinomycin, combretastatin, fusobactin, epothilone A, epothilone B, estramustine, scutellarin, cemadotin, maytansinoids, scutellarin, mitoxantrone, etc.;

[1110] Cell proliferation and apoptosis

[1111] The compounds and conjugates disclosed herein can be used in methods of inducing apoptosis.

[1112] Apoptosis dysregulation is implicated in a variety of diseases, including, for example, autoimmune disorders (e.g., systemic lupus erythematosus, rheumatoid arthritis, graft-versus-host disease, myasthenia gravis, or Sjögren's syndrome), chronic inflammatory conditions (e.g., psoriasis, asthma, or Crohn's disease), hyperproliferative disorders (e.g., breast cancer, lung cancer), viral infections (e.g., herpes, papilloma, or HIV), and other conditions such as osteoarthritis and atherosclerosis. The compounds, conjugates, and compositions described herein can be used to treat or alleviate any of these diseases. Such treatments typically involve administering to a subject suffering from the disease an amount of a compound, conjugate, or composition described herein sufficient to provide a therapeutic benefit. The identity of the antibody to the compound, conjugate, or composition administered will depend on the disease being treated—thus, the antibody should bind to a cell surface antigen expressed in a cell type where inhibition would be beneficial. The therapeutic benefit obtained will also depend on the specific disease being treated. In some cases, when administered as a monotherapy, the compounds and compositions disclosed herein can treat or alleviate the disease itself or the symptoms of the disease. In other cases, the compounds and compositions disclosed herein may be part of an overall treatment regimen that includes other agents that, together with the inhibitors or compounds and compositions disclosed herein, treat or alleviate the disease or symptoms of the disease being treated. Agents useful for treating or alleviating a particular disease that can be administered in conjunction with or in conjunction with the compounds and compositions disclosed herein will be readily apparent to those skilled in the art.

[1113] Although an absolute cure is always ideal in any treatment regimen, a cure need not be achieved to provide a therapeutic benefit. A therapeutic benefit can include halting or slowing the progression of a disease, causing regression of the disease without a cure, and / or alleviating or slowing the progression of symptoms of the disease. Prolonged survival and / or improved quality of life compared to the statistical mean can also be considered a therapeutic benefit.

[1114] A specific class of diseases involving apoptosis disorders and as a significant health burden in the global context is cancer. In a specific embodiment, the compounds and compositions disclosed herein can be used to treat cancer. Cancer can be, for example, a solid tumor or a hematological tumor. Cancers that can be treated with the compounds and compositions disclosed herein include, but are not limited to, bladder cancer, brain cancer, breast cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T cell or B cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, myeloma, prostate cancer, small cell lung cancer and spleen cancer. The compounds and compositions disclosed herein may be particularly beneficial in the treatment of cancer because the antibodies can be used to specifically target tumor cells, thereby potentially avoiding or alleviating the undesirable side effects and / or toxicity that may be associated with systemic administration of unconjugated inhibitors. One embodiment relates to a method of treating a disease involving dysregulation of intrinsic apoptosis, comprising administering to a subject suffering from a disease involving dysregulation of apoptosis an amount of the compounds and compositions disclosed herein effective to provide a therapeutic benefit, wherein the ligand of the compounds and compositions disclosed herein binds to a cell surface receptor on a cell in which intrinsic apoptosis is dysregulated. One embodiment relates to a method of treating cancer, comprising administering to a subject suffering from cancer an amount of the compounds and compositions disclosed herein effective to provide a therapeutic benefit, wherein the ligand is capable of binding to a cell surface receptor or tumor-associated antigen expressed on the surface of a cancer cell.

[1115] In the context of tumorigenic cancers, therapeutic benefit may specifically include, in addition to the effects discussed above, arresting or slowing the progression of tumor growth, causing regression of tumor growth, eradicating one or more tumors, and / or increasing patient survival, compared to statistical means for the type and stage of cancer being treated. In one embodiment, the cancer being treated is a tumorigenic cancer.

[1116] The compounds and conjugates disclosed herein can be administered as a monotherapy to provide therapeutic benefit, or can be administered in conjunction with or in addition to other chemotherapeutic agents and / or radiation therapy. The chemotherapeutic agents with which the compounds and compositions disclosed herein can be used as adjuvant therapy can be targeted (e.g., ADCs, protein kinase inhibitors, etc.) or non-targeted (e.g., non-specific cytotoxic agents such as radionucleotides, alkylating agents, and intercalating agents). Non-targeted chemotherapeutic agents that can be administered adjuvantly with the compounds and compositions disclosed herein include, but are not limited to, methotrexate, tacrolimus, L-asparaginase, mercaptopurine, thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosoureas, cisplatin, carboplatin, mitomycin, dacarbazine, procarbazine, topotecan, mechlorethamine, carbazine, etoposide, 5-fluorouracil, BCNU, irinotecan, camptothecin, bleomycin, doxorubicin, idarubicin, daunorubicin, dactinomycin, plicamycin, mitoxantrone, asperaginase, vinblastine, vincristine, vinorelbine, paclitaxel, calicheamicin, and docetaxel.

[1117] The compounds and conjugates disclosed herein, which may not be effective as a monotherapy for the treatment of cancer, may be administered in adjunct to or in conjunction with other chemotherapeutic agents or radiation therapy to provide therapeutic benefit. One embodiment relates to a method wherein the compounds or compositions disclosed herein are administered in an amount effective to sensitize tumor cells to standard chemotherapy and / or radiation therapy. Thus, in the context of treating cancer, "therapeutic benefit" includes administering the compounds and compositions disclosed herein in adjunct to or in conjunction with other chemotherapeutic agents and / or radiation therapy in patients who have not yet started such therapy, or who have started but have not yet shown signs of resistance, or in patients who have begun to show signs of resistance, as a means of sensitizing the tumor to chemotherapy and / or radiation therapy.

[1118] Pharmaceutical compositions and their administration

[1119] The compounds and conjugates disclosed herein can be used to treat individuals in need thereof. In certain embodiments, the individual is a mammal, such as a human or a non-human mammal. When administered to an animal, such as a human, the composition or compound is preferably administered in the form of a pharmaceutical composition comprising, for example, a disclosed compound and a pharmaceutically acceptable carrier.

[1120] Pharmaceutically acceptable carrier is well known in the art, and includes, for example, aqueous solution, such as water or physiologically buffered saline or other solvents or vehicles, such as ethylene glycol, glycerol, oil such as olive oil or injectable organic ester. In a preferred embodiment, when such pharmaceutical composition is used for people, particularly for invasive route of administration (that is, circumventing the route of transporting or diffusion through epithelial barrier, such as injection or implantation), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to realize the delayed release of medicament or selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form, such as tablets, capsules (including sprinkle capsules (sprinkle capsules) and gelatin capsules), granules, lyophilized agents for reconstruction, powders, solutions, syrups, suppositories, injections etc. The composition can also be present in transdermal delivery systems (for example, skin patches). The composition can also be present in solutions (such as ointments or creams) suitable for topical application.

[1121] Pharmaceutically acceptable carriers can contain physiologically acceptable agents, which, for example, act to stabilize compounds (such as the compounds of the present invention), increase their solubility, or increase their absorption. Such physiologically acceptable agents include, for example, carbohydrates (such as glucose, sucrose, or dextran), antioxidants (such as ascorbic acid or glutathione), chelating agents, low molecular weight proteins, or other stabilizers or excipients. The selection of pharmaceutically acceptable carriers (including physiologically acceptable agents) depends on, for example, the route of administration of the composition. The preparation of the pharmaceutical composition can be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) can also be a liposome or other polymer matrix, into which, for example, the compounds of the present invention may be incorporated. For example, the liposome comprising phospholipids or other lipids is a nontoxic, physiologically acceptable, and metabolizable carrier that is relatively easy to manufacture and administer.

[1122] The phrase "pharmaceutically acceptable" is used herein to refer to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[1123] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other nontoxic compatible substances used in pharmaceutical formulations.

[1124] The pharmaceutical composition (formulation) can be administered to a subject in any of a variety of routes of administration, including, for example, oral administration (e.g., as a drench in an aqueous or non-aqueous solution or suspension, a tablet, a capsule (including a sprinkle capsule and a gelatin capsule), a bolus, a powder, a granule, a paste for application to the tongue); absorption through the oral mucosa (e.g., sublingually); anally, rectally, or vaginally (e.g., as a pessary, cream, or foam); parenterally (including intramuscularly, intravenously, subcutaneously, or intrathecally, as, for example, a sterile solution or suspension); nasally; intraperitoneally; subcutaneously; transdermally (e.g., as a patch applied to the skin); and topically (e.g., as a cream, ointment, or spray applied to the skin, or as eye drops). The compound can also be formulated for inhalation. In certain embodiments, the compound can be simply dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable therefor can be found, for example, in US Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and patents cited therein.

[1125] These preparations can exist in unit dosage form conveniently and can be prepared by any method well-known in the pharmaceutical field.The amount of the active ingredient that can be combined with carrier materials to produce single dosage form will depend on the host to be treated and the specific mode of administration and change.The amount of the active ingredient that can be combined with carrier materials to produce single dosage form will be the amount of the compound that produces therapeutic effect generally.In general, in one hundred percent, the scope of this amount will be from about 1% to about 99% active ingredient, preferably from about 5% to about 70%, most preferably from about 10% to about 30%.

[1126] The method for preparing these preparations or compositions comprises the step of combining an active compound such as a compound of the present invention with a carrier and optionally one or more auxiliary ingredients. Generally, the preparation is prepared in the following manner: uniformly and intimately combining a compound of the present invention with a liquid carrier or a finely divided solid carrier or both, and then (if necessary) shaping the product.

[1127] Formulations of the present invention suitable for oral administration can be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), lyophilized forms, powders, granules; or as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil liquid emulsion; or as an elixir or syrup; or as a pastille (using an inert base, such as gelatin and glycerin or sucrose and acacia) and / or as a mouthwash, etc., each containing a predetermined amount of the compound of the present invention as the active ingredient. The compound, conjugate, or composition thereof can also be administered as a bolus, suppository, or paste.

[1128] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers (such as sodium citrate or dicalcium phosphate) and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) solution retardants. retarding agent), such as paraffin; (6) absorption promoters, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glyceryl monostearate; (8) adsorbents, such as kaolin and bentonite; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also contain a buffer. Similar types of solid compositions may also be used as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar and high molecular weight polyethylene glycols and the like.

[1129] Tablet can be prepared by optionally compressing or molding with one or more auxiliary components. Compressed tablets can be prepared using a binder (e.g., gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), a surfactant or a dispersant. Molded tablets can be prepared by molding a mixture of a powdered compound moistened with an inert liquid diluent in a suitable machine.

[1130] Tablets and other solid dosage forms of the pharmaceutical composition (such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules) can optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the art of pharmaceutical formulation. They can also be formulated using, for example, hydroxypropyl methylcellulose, other polymer matrices, liposomes and / or microspheres in varying proportions to provide the desired release characteristics in order to provide slow or controlled release of the active ingredient therein. They can be sterilized by, for example, filtering through a bacteria-retaining filter or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or some other injectable sterile medium just before use. Optionally, these compositions may also contain opacifiers and may be compositions that release one or more active ingredients only or preferentially in a certain part of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, if appropriate, containing one or more of the excipients described above.

[1131] Liquid dosage forms that can be used for oral administration include pharmaceutically acceptable emulsions, lyophilized agents for reconstitution, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage form may also contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and their derivatives, solubilizers, and emulsifiers, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuran alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof.

[1132] Besides inert diluents, the oral compositions may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[1133] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[1134] Formulations of pharmaceutical compositions for rectal, vaginal or urethral administration can be presented as suppositories, which can be prepared by mixing one or more active compounds with one or more suitable non-irritating excipients or carriers (including, for example, cocoa butter, polyethylene glycol, suppository wax or salicylates), and which are solid at room temperature but liquid at body temperature and therefore will melt in the rectum or vaginal cavity and release the active compound.

[1135] Formulations of pharmaceutical compositions for administration to the mouth may be in the form of a mouthwash, oral spray, or oral ointment.

[1136] Alternatively or additionally, the composition can be formulated for delivery via a catheter, stent, wire or other intraluminal device. Delivery via such a device may be particularly useful for delivery to the bladder, urethra, ureter, rectum or intestine.

[1137] Formulations suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

[1138] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound can be mixed under sterile conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers, or propellants that may be required.

[1139] Ointments, pastes, creams and gels may contain, in addition to the active compounds, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[1140] In addition to the active compound, powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. Sprays may additionally contain customary propellants, such as chlorofluorocarbons and unsubstituted volatile hydrocarbons, such as butane and propane.

[1141] Transdermal patches have the additional advantage of providing controlled delivery of the compounds of the invention to the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in an appropriate medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of this flux can be controlled by providing a rate-controlling membrane or dispersing the active compound in a polymer matrix or gel.

[1142] Ophthalmic formulations, eye ointments, powders, solutions, and the like are also contemplated as being within the scope of the present invention. Exemplary ophthalmic formulations are described in U.S. Publication Nos. 2005 / 0080056, 2005 / 0059744, 2005 / 0031697, and 2005 / 004074; and U.S. Patent No. 6,583,124, the contents of which are incorporated herein by reference. If desired, the liquid ophthalmic formulation has properties similar to those of tears, aqueous humor, or vitreous humor, or is compatible with such fluids.

[1143] As used herein, the phrases "parenteral administration" and "administered parenterally" mean modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[1144] Pharmaceutical compositions suitable for parenteral administration may comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders to be reconstituted into sterile injectable solutions or dispersions just before use, which may contain antioxidants, buffers, bacteriostats, solutes (to render the formulation isotonic with the blood of the intended recipient), or suspending or thickening agents.

[1145] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Suitable fluidity can be maintained, for example, by the use of coating materials (such as lecithin), by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[1146] These compositions can also contain adjuvants, such as preservatives, wetting agents, emulsifiers and dispersants.Can ensure to prevent the effect of microorganisms by comprising various antibacterials and antifungals (for example, parabens, chlorobutanol, phenol sorbic acid etc.).Also can desirably comprise isotonic agents, such as sugar, sodium chloride etc. in the compositions.In addition, can realize the prolongation absorption of injectable drug form by comprising the reagent (such as aluminum monostearate and gelatin) that delays absorption.

[1147] In some cases, in order to prolong the effect of the drug, it is desirable to slow down the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or non-crystalline material with poor water solubility. The absorption rate of the drug then depends on its dissolution rate, which in turn may depend on crystal size and crystalline form. Alternatively, delayed absorption of parenteral drug forms is achieved by dissolving or suspending the drug in an oil vehicle.

[1148] Injectable depot forms are manufactured by forming a microencapsulated matrix of the subject compound in a biodegradable polymer (such as polylactide-polyglycolide). Depending on the ratio of the drug to the polymer and the properties of the specific polymer used, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot formulations can also be prepared by embedding the drug into liposomes or microemulsions compatible with body tissues.

[1149] For use in the methods of the present invention, the active compound can be administered per se or as a pharmaceutical composition containing, for example, 0.1% to about 99.5% (more preferably about 0.5% to about 90.0%) active ingredient in combination with a pharmaceutically acceptable carrier.

[1150] In some embodiments of the present invention, the compounds of the present invention are administered in combination with one or more additional compounds / agents.

[1151] In certain such embodiments, the co-administration is simultaneous. In certain such embodiments, the compounds of the invention are co-formulated with one or more additional compounds. In certain other such embodiments, the compounds of the invention and one or more additional compounds are administered separately but simultaneously. In certain such embodiments, the co-administration is sequential, with administration of the compounds of the invention, or minutes or hours before or after administration of the one or more additional compounds.

[1152] Methods for introducing the compounds of the present invention can also be provided by rechargeable or biodegradable devices. In recent years, various slow-release polymer devices have been developed and tested in vivo for the controlled delivery of drugs (including protein-like biopharmaceuticals). A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form implants for sustained release of compounds at specific target sites.

[1153] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without unacceptable toxicity to the patient.

[1154] The selected dosage level will depend upon a variety of factors, including the activity of the specific compound, conjugate or combination of compounds and / or conjugates employed, or the esters, salts or amides thereof, the route of administration, the time of administration, the rate of excretion of the specific compound or compounds employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the specific compound or compounds employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[1155] A physician or veterinarian with ordinary skill in the art can easily determine and prescribe a therapeutically effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian can start with a dose of the pharmaceutical composition or compound that is lower than the level required to achieve the desired therapeutic effect, and gradually increase the dose until the desired effect is achieved. A "therapeutically effective amount" refers to a concentration of the compound sufficient to cause the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the subject's weight, sex, age, and medical history. Other factors affecting the effective amount may include, but are not limited to, the severity of the patient's illness, the condition being treated, the stability of the compound, and (if desired) another type of therapeutic agent administered with the compounds of the present invention. A larger total dose can be delivered by multiple administrations of the agent. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13th edition, 1814-1882, incorporated herein by reference).

[1156] In general, a suitable daily dose of the active compound used in the compositions and methods of the invention will be the amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[1157] If desired, the effective daily dose of the active compound or conjugate may be administered in unit dosage form as one, two, three, four, five, six or more sub-doses administered at appropriate intervals throughout the day. In certain embodiments of the invention, the active compound may be administered two or three times daily. In preferred embodiments, the active compound will be administered once daily.

[1158] The patient receiving such treatment is any animal in need thereof, including primates, particularly humans, and other mammals such as horses, cattle, pigs, and sheep; and poultry and pets in general.

[1159] In certain embodiments, compound disclosed herein or conjugate can be used alone or in combination with another type of therapeutic agent.As used herein, phrase "combined administration" refers to any form of administration of two or more different therapeutic compounds or conjugates, so that the therapeutic compound or conjugate previously administered is still effective in vivo when the second compound or conjugate is administered (for example, two compounds or conjugates are effective in the patient at the same time, which can include the synergistic effect of two compounds or conjugates). For example, different therapeutic compounds or conjugates can be administered concomitantly or sequentially in the same formulation or in a separate formulation. In certain embodiments, different therapeutic compounds or conjugates can be administered to each other within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours or 1 week or more weeks. Therefore, individuals receiving such treatments can benefit from the combined effects of different therapeutic compounds or conjugates.

[1160] The present invention includes the purposes of the pharmaceutically acceptable salt of compound disclosed herein or conjugate.In certain embodiments, the salt of consideration of the present invention includes but is not limited to alkyl, dialkyl, trialkyl or tetraalkyl ammonium salt.In certain embodiments, the salt of consideration of the present invention includes but is not limited to L- arginine, phenethylbenzylamine (benenthamine), benzophenone, betaine, calcium hydroxide, choline, deanol (deanol), diethanolamine, diethylamine, 2- (diethylamino) ethanol, ethanolamine, ethylenediamine, N- methylglucosamine, hydrabamine (hydrabamine), 1H-imidazole, lithium, L- lysine, magnesium, 4- (2- hydroxyethyl) morpholine, piperazine, potassium, 1- (2- hydroxyethyl) pyrrolidine, sodium, triethanolamine, tromethamine (tromethamine) and zinc salt.In certain embodiments, the salt of consideration of the present invention includes but is not limited to the salt of Na, Ca, K, Mg, Zn or other metals.

[1161] Pharmaceutically acceptable acid addition salts may also exist as various solvates (such as with water, methanol, ethanol, dimethylformamide, etc.). Mixtures of such solvates may also be prepared. The source of such solvates may be derived from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or exogenous to such solvent.

[1162] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[1163] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[1164] Having now generally described the invention, the invention will be more readily understood by reference to the following examples, which are included merely for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention.

[1165] Examples

[1166] Synthesis scheme

[1167] abbreviation

[1168] AcO: acetyl

[1169] AcOH: acetic acid

[1170] EA: ethyl acetate

[1171] DCM: dichloromethane

[1172] m-CPBA: meta-chloroperbenzoic acid

[1173] TBDMSOTf: tert-Butyldimethylsilyl trifluoromethanesulfonate

[1174] TBDMS: tert-butyldimethylsilyl

[1175] DMF: dimethylformamide

[1176] EDCI: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide

[1177] HOBt: 1-Hydroxybenzotriazole hydrate

[1178] ACN: acetonitrile

[1179] TBDMS-Cl: tert-Butyldimethylsilyl chloride

[1180] DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene

[1181] THF: Tetrahydrofuran

[1182] DCC: N,N'-dicyclohexylcarbodiimide

[1183] DMAP: 4-dimethylaminopyridine

[1184] NHS: N-hydroxysuccinimide DIPEA: diisopropylethylamine

[1185] TEA: triethylamine

[1186] DEAD: diethyl azodicarboxylate

[1187] Boc: tert-Butoxycarbonyl

[1188] LAH: lithium aluminum hydride

[1189] CDI: 1,1'-carbonyldiimidazole

[1190] BEMP: 2-tert-butylimino-2-diethylamino-1,3-dimethylperhydro-1,3,2-diazaphosphine TPSCl: triphenylsilyl chloride TFA: trifluoroacetyl

[1191] PyBop: Benzotriazol-1-yl-oxytripyrrolophosphonium hexafluorophosphate

[1192] HBTU: N,N,N',N'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate

[1193] TFA: trifluoroacetic acid

[1194] DIC: N,N'-diisopropylcarbodiimide

[1195] DMPA: 2,2-dimethoxy-2-phenylacetophenone

[1196] TBAF: Tetra-n-butylammonium fluoride AgOTf: Silver trifluoromethanesulfonate (BimC4A)3: 5,5',5"-[2,2',2"-nitrilotris(methylene)tris(1H-benzimidazole-2,1-diyl)]tripalkenate tripotassium hydrate

[1197] [Example 1] Preparation of BGal-Br (hereinafter referred to as 'Int-TG')

[1198]

[1199] Under N2 atmosphere at 0 ℃, beta-D-galactose pentaacetate (Alfa, CAS 4163-60-4, 5.0g, 12.81mmol) is dissolved in 33% HBr in AcOH (20mL).The mixture is warmed to room temperature.After stirring at room temperature for 4 hours, the mixture is concentrated under reduced pressure, and then EA (1000mL) and saturated sodium bicarbonate (1000mL) are added.The organic layer is through anhydrous Na2SO4 drying, filtered, and concentrated under reduced pressure.Residue is purified by column chromatography to obtain compound Int-TG (5.2g, 99%).

[1200] 1 H NMR (400MHz, CDCl3) δ6.70(d,J=4.0Hz,1H),5.52(d,J=2.4Hz,1H),5.41(dd,J=7.6,2.8Hz,1H ), 5.05 (dd, J = 6.4, 4.0Hz, 1H), 4.49 (t, J = 6.4Hz, 1H), 4.22-4.09 (m, 2H), 2.16-2.01 (m, 12H).

[1201] [Example 2] Preparation of compound Int-TG1

[1202]

[1203] Preparation of compound Int-TG1-1

[1204] Under N2 atmosphere, to a solution of salicyl aldehyde (Aldrich, CAS 90-02-8, 148 mg, 1.22 mmol) and compound Int-TG (0.5 g, 1.22 mmol) in acetonitrile (10 mL), dry molecular sieves (2.5 g) and Ag2O (845 mg, 3.65 mmol) were added. After stirring at room temperature for 1 hour, distilled water (50 mL) and EA (50 mL X2) were added. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound Int-TG1-1 (441 mg, 81%).

[1205] 1H NMR (400MHz, CDCl3) δ10.37(s,1H),7.88(t,J=7.6Hz,1H),7.21(t,J=7.4Hz,1H),7.14(t,J=8.4 Hz,1H),5.62(m,1H),5.48(m,1H),5.16(d,J=7.2Hz,2H),4.27-4.23(m,1H),4.18-4.09(m,2H)m 2.21(s,3H),2.07(s,6H),2.03(s,3H).

[1206] Preparation of compound Int-TG1-2

[1207] At 0 ℃ under N2 atmosphere, to a solution of compound Int-TG1-1 (260mg, 0.575mmol) in DCM (3mL) was added m-CPBA (283mg, 1.149mmol). After 5 hours, the mixture was concentrated under reduced pressure. EA (50mL X2) and aqueous sodium bicarbonate solution (30mL) were then added. The organic layer obtained was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain compound Int-TG1-2 (270mg, quantitative). Compound Int-TG1-2 was used directly in the next reaction without purification.

[1208] 1 H NMR (400MHz, CDCl3) δ8.18(s,1H),7.90(d,J=8.0Hz,1H),7.64(d,J=8.0Hz,1H),7.46(t,J=7.6Hz,1H),7.15(d,J=8.4Hz,1H),5.51(m,2H) ,5.11(d,J=8.8Hz,1H),5.04(d,J=8.0Hz,1H),4.24(m,1H),4.16(m,1H),4.08(m,1H),2.18(s,3H),2.09(s,3H),2.07(s,3H),2.02(s,3H). EI-MSm / z:491(M + +Na).

[1209] Preparation of compound Int-TG1-3

[1210] To a solution of compound Int-TG1-2 in CHCl (3 mL) was added hydrazine monohydrate (21 μL, 0.427 mmol) at 0° C. under N atmosphere. After stirring at 0° C. for 0.5 hours, EA (30 mL × 2) and 1 M aqueous HCl (10 mL) were added. The obtained organic layer was dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to obtain compound Int-TG1-3 (161 mg, 86%).

[1211] 1 H NMR (400MHz, CDCl3) δ7.03(t,J=8.0Hz,1H),6.98-6.95(m,2H),6.83(t,J=7.6Hz,1H),6.02(s,1H),5.47(d,J=3.2Hz,2H),5.13(dd,J=10 .8,2.8Hz,1H),4.93(d,J=7.6Hz,1H),4.26(m,1H),4.19-4.09(m,2H),4.06(m,1H),2.21(s,3H),2.13(s,3H),2.08(s,3H),2.03(s,3H). EI-MSm / z:463(M + +Na).

[1212] Preparation of compound Int-TG1

[1213] At 0 ℃ under N2 atmosphere, to a solution of compound Int-TG1-3 (161 mg, 0.366 mmol) in DCM (3 mL) was added Et3N (102 μ L, 0.732 mmol) and TBDMS-OTf (126 μ L, 0.549 mmol). The mixture was stirred at room temperature for 2 hours. Then DCM (30 mL × 2) and 1M HCl aqueous solution (10 mL) were added. The organic layer obtained was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound Int-TG1 (147 mg, 91%).

[1214] 1 H NMR (400MHz, CDCl3) δ7.02(d,J=7.6Hz,1H),6.95-6.84(m,3H),5.48-5.43(m,2H),5.15(d,J=8.0Hz,1H),5.10(d,J=10.4Hz,1H), 4.21-4.11(m,2H),4.03-3.99(m,1H),2.19(s,3H),2.04(s,3H),2.02(s,3H),2.00(s,3H),0.99(s,9H),0.20(s,3H),0.16(s,3H). EI-MSm / z:555(M + ).

[1215] [Example 3] Preparation of compound Int-TG2

[1216]

[1217] Preparation of compound Int-TG2-1

[1218] To a solution of 3-formyl-4-hydroxybenzoic acid (3 g, 18.06 mmol) and 11-azido-3,6,9-trioxa-1-amine (Aldrich, CAS134179-38-7, 5.98 g, 23.48 mmol) in DMF (20 mL) was added EDCI (5.19 g, 27.09 mmol), HOBt (4.15 g, 27.09 mmol) and EtN (10.1 mL, 72.24 mmol) at 0 ° C. under N2 atmosphere. The mixture was stirred at room temperature under N2 atmosphere overnight. The reactant was quenched with EA (60 mL × 2) and citric acid (60 mL). The organic layer was extracted with sodium bicarbonate aqueous solution (80 mL). The organic layer obtained was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound Int-TG2-1 (2.56 g, 39%).

[1219] 1 H NMR (400MHz, CDCl3) δ11.26(s,1H),9.96(s,1H),8.16(s,1H),7.98-7.96(d,J=8.4Hz,1H),7.0 4-7.02(d,J=9.2Hz,1H),6.91(s,1H),3.68-3.61(m,14H),3.37-3.34(m,2H),EI-MSm / z:367(M + ).

[1220] Preparation of compound Int-TG2-2

[1221] At room temperature under N2 atmosphere, molecular sieves (8 g) and Ag2O (2.68 g, 11.55 mmol) were added to a solution of compound Int-TG2-1 (1.41 g, 3.85 mmol) and compound Int-TG (1.74 g, 4.24 mmol) in anhydrous ADC (20 mL). The mixture was stirred at room temperature for 3 hours and then filtered through diatomaceous earth. The organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound Int-TG2-2 (1.88 g, 70%).

[1222] 1H NMR (400MHz, CDCl3) δ10.35(s,1H),8.20-8.17(m,2H),7.26(s,1H),7.20-7.18(d,J=9.2Hz,1H),6.96(s,1H),5.63-5.58(m,1H),5.50-5.49(m,1 H),5.23-5.21(m,1H),5.18-5.14(m,1H),4.24-4.14(m,3H),3.69-3.64( m,14H),3.37-3.35(m,2H),2.21(s,3H),2.08-2.07(m,6H),2.03(s,3H). EI-MSm / z:697(M + ).

[1223] Preparation of compound Int-TG2-3

[1224] To a solution of compound Int-TG2-2 (1.69 g, 2.42 mmol) in DCM (15 mL) was added m-CPBA (2.4 g, 9.70 mmol) at 0°C under N2 atmosphere. After stirring for 7 hours at 0°C, the mixture was quenched by adding saturated sodium bicarbonate (40 mL x 2). The mixture was separated and the organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound Int-TG2-3 (1.25 g, 76%).

[1225] 1 H NMR (400MHz, CDCl3) δ7.36-7.33(m,2H),7.01-6.99(d,J=8.4Hz,1H),6.71(m,1H),6.06(s,1H),5.49-5.44(m,2H),5.15-5.12(m,1H),4.99-4.97 (d,J=8.0Hz,1H),4.24-4.09(m,3H),3.69-3.63(m,14H),3.37-3.34(m,2 H),2.20(s,3H),2.13(s,3H),2.12(s,3H),2.03(s,3H),EI-MSm / z:685(M + ).

[1226] Preparation of compound Int-TG2

[1227] To a solution of compound Int-TG2-3 (750 mg, 1.09 mmol) in DCM (10 mL) was added TBDMS-OTf (504 μL, 2.19 mmol) and EtN (458 μL, 3.29 mmol) at 0 ° C under N atmosphere. The mixture was stirred at room temperature overnight and then quenched by adding citric acid (20 ml). The organic layer was washed with brine (20 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound Int-TG2 (799 mg, 91%).

[1228] 1 H NMR (400MHz, CDCl3) δ7.35 (d, J=2.4Hz, 1H), 7.30 (dd, J=8.4, 2.0Hz, 1H), 7.02 (d, J=8.0Hz, 1H),6.65(t,J=5.2Hz,1H),5.49-5.44(m,2H),5.20(d,J=7.6Hz,1H),5.12(dd,J=10.0,3.6 Hz,1H),4.20-4.11(m,2H),4.06-4.03(m,1H),3.69-3.62(m,15H),3.37(t,J=5.2Hz,2H),2 .19(s,3H),2.05(s,3H),2.02(s,3H),2.01(s,3H),1.01(s,9H),0.22(s,3H),0.18(s,3H). EI-MSm / z:799(M + ).

[1229] [Example 4] Preparation of compound Int-TG3

[1230]

[1231] Preparation of compound Int-TG3-1

[1232] At room temperature, to a solution of salicyl aldehyde (Aldrich, 200 mg, 1.64 mmol) and compound Bg-Br (813 mg, 1.64 mmol) in acetonitrile (12 mL), dry molecular sieves (1.0 g) and Ag o (1.42 g, 4.92 mmol) were added. The mixture was stirred overnight and distilled water (50 mL) and EA (50 mL × 2) were added. The organic layer was dried over anhydrous Na sO , filtered, and concentrated under reduced pressure. Residue was purified by column chromatography to obtain compound Int-TG-3-1 (218 mg, 30%).

[1233] 1H NMR (400MHz, CDCl3) δ10.35(s,1H),7.86(dd,J=6.0,1.6Hz,1H),7.56(td,J=7.6,1.6Hz,1H),7.20(t,J=7.6Hz,1H ),7.13(d,J=8.8Hz,1H),5.39-5.31(m,3H),5.27-5.25(m,1H),5.24-4.19(m,1H),3.75(s,3H),2.07-2.04(m,9H). EI-MSm / z:461(M + +Na).

[1234] Preparation of compound Int-TG3-2

[1235] To a solution of compound Int-TG3-1 (217.6 mg, 0.50 mmol) in DCM (10 mL) was added m-CPBA (367.1 mg, 1.50 mmol) at 0°C under atmosphere. The mixture was stirred at room temperature overnight and 40 mL of DCM was added. The organic layer was washed with saturated sodium bicarbonate (10 mL) and dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure.

[1236] The residue was dissolved in CHCl (5 ml) again. Hydrazine (36.2 μL, 0.74 mmol) was then added. After 30 minutes, DCM (20 mL × 2) and water (10 mL) were added. The obtained organic layer was dried over anhydrous NaSO, filtered and concentrated under reduced pressure, and the residue was purified by column chromatography to obtain compound Int-TG1-3-2 (195 mg, 92%).

[1237] 1 H N...

Claims

1. A compound comprising the following structural formula: or a pharmaceutically acceptable salt thereof, in: X is -O-, -C(R b )2-or-N(R c )-, preferably -O-; Ar represents a ring, such as aryl, heteroaryl, cycloalkyl or heterocycloalkyl, preferably aryl or heteroaryl; Y' is -(CR b 2) y N(R a )-、-(CR b 2) y O-or-(CR b 2) y S-, positioned such that if y is 1, then the N, O, or S atom is attached to TG; X and Y' are positioned on adjacent atoms of Ar; TG is a triggering group that, when activated, generates an N, O, or S atom capable of reacting with the SO2 to form a 5-6 membered ring containing an intervening atom of X-SO2 and Ar; x is 0 or 1; y is 0 or 1; Each R a and R c are independently hydrogen or lower alkyl; and Each R b are independently hydrogen or lower alkyl; or two R b Together with the atoms to which they are attached, they form a 3-5 membered ring, preferably a 3-4 membered ring.

2. The compound of claim 1, wherein the compound has the structure of formula (Ia): or a pharmaceutically acceptable salt thereof, wherein: Each Q is independently an active agent linked to L' through a heteroatom, preferably O or N; Z' is a solubilizing group, a reactive group, or a linking group that connects the structure of Formula (Ia) to an antibody, a solid surface, a stabilizing group, a chelator, a biopolymer, or a detectable moiety; L' is a linking group attached to SO2 via a heteroatom selected from O, S and N, preferably O or N, and is selected such that cleavage of the bond between L' and SO2 promotes cleavage of the bond between L' and Q to release the active agent; q is an integer having a value from 1 to about 20, preferably from 1 to about 10; w is 0 or 1; The condition is that when w is 0, q is 1.

3. The compound of any one of claims 1-2, wherein X is -O-.

4. The compound of any one of claims 1 to 3, wherein Ar is aryl or heteroaryl.

5. The compound of claim 4, wherein Ar is phenyl, naphthyl, pyridyl or quinolone. The compound of claim 5 , wherein Ar is phenyl or naphthyl.

7. The compound of any one of claims 2 to 6, wherein Z' is a linking group connecting the antibody and Ar, comprising (CH2) linked to each other by covalent bonds in a linear chain. b , L c 、(P 1 ) a 、W a1 、W a2 、W a3 、Y 1 and Y 2 Group, in which: W a1 、W a2 and W a3 are each independently -NH-, -C(O)- or -CH2-; W b1 is an amide bond or a triazole group; P 1 is an amide bond, an amino acid residue, or a peptide; L c is an alkylene group; Y 1 Yes - (CH2) q -(CH2CH2X”) o -or-(CH2) q -(X'CH2CH2X) o -; X" is -O-, -S-, -NH- or -CH2-; Y 2 is a single bond or a group selected from the following: W b2 is an amide bond or a triazole group; a is an integer with a value from 0 to 10; b, c, and d are each independently an integer with a value from 1 to about 10; and o and q are each independently an integer having a value from 1 to about 10.

8. The compound of claim 7, wherein Z' is a linking group of formula (A): **-L c -W b1 -(CH2) b -W a3 -(P 1 ) a -Y 2 -W a2 -Y 1 -W a1 -* (A) in: * is the point of attachment to the antibody; and ** is the point of attachment to Ar.

9. The compound of any one of claims 1 to 8, wherein Z' comprises:

10. The compound of any one of claims 1 to 9, wherein TG is a reactive chemical moiety or functional group that can be cleaved by nucleophilic reagent conditions, alkaline reagent conditions, light irradiation, reducing agent conditions, acidic conditions, enzymatic conditions, or oxidative conditions.

11. The compound of any one of claims 1 to 10, wherein x is 0.

12. The compound of claim 11, wherein TG is -NO2, -OC(O)(CH2) r C(O)R 1 、-NHNH2、-BR 2 R 3 or in: R 1 is a C1-C6 alkyl group; R 2 and R 3 are each independently hydrogen, C1-C6 alkyl, C1-C6 alkoxy or hydroxy; R 4 、R 5 、R 6 and R 7 are each independently hydrogen or C1-C6 alkyl; and r is an integer with a value of 1, 2, 3, 4, or 5.

13. The compound of any one of claims 1 to 10, wherein TG is selected from: in: Each R 21 are independently hydrogen or acetyl; and R 22 It is hydrogen or lower alkyl.

14. The compound of any one of claims 1 to 11, wherein TG is selected from: -NO2, -C(O)-(CH2)2C(O)-alkyl and nitrobenzyl.

15. The compound of any one of claims 1 to 14, wherein Q is a chemical factor, a biological factor, a hormone, an oligonucleotide, a drug, a toxin, an affinity ligand, a probe for detection, or a combination thereof.

16. The compound of claim 15, wherein Q is a drug selected from a cytokine, an immunomodulatory compound, an anticancer agent, an antiviral agent, an antibacterial agent, an antifungal agent, an anthelmintic, or a combination thereof.

17. The compound of any one of claims 1 to 14, wherein (Q) q -(L') w -Selected from: in: X 1 Is -O- or -NR a -; X 2 and X 4 each independently absent or C(O)- or -C(O)O-; X 3 is -OC(=O)-; w' is an integer with a value of 1, 2, 3, 4, or 5; R 9 and R 10 are each independently hydrogen, alkyl, aryl or heteroaryl, wherein alkyl, aryl and heteroaryl are optionally substituted by one or more groups selected from, for example, alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 and -(CH2) u SO2R u3 Substituents substituted; R u1 、R u2 and R u3 each is independently hydrogen, alkyl, aryl, or heteroaryl; and u is an integer with a value of 1 to about 10.

18. The compound of claim 17, wherein (Q) q -(L') w -Selected from: Where * represents (Q) q -(L') w -Attachment point to -SO2-.

19. The compound of any one of claims 1 to 18, wherein Z' links the structure of Formula (Ia) to an antibody selected from the group consisting of an intact polyclonal antibody, an intact monoclonal antibody, an antibody fragment, a single-chain Fv (scFv) mutant, a multispecific antibody, a bispecific antibody, a chimeric antibody, a humanized antibody, a human antibody, a fusion protein comprising an antigenic determinant portion of an antibody, and other modified immunoglobulin molecules comprising an antigen recognition site.

20. The compound of claim 19, wherein the antibody is selected from the group consisting of muromonab-CD3, abciximab, rituximab, daclizumab, palivizumab, infliximab, trastuzumab (Herceptin), etanercept, basiliximab, gemtuzumab ozogamicin, alemtuzumab, ibritumomab tentanycin, adalimumab, alefacept, omalizumab, efalizumab, tositumomob-I 131 , cetuximab, bevacizumab, natalizumab, ranibizumab, panitumumab, eculizumab, rilonacept, becelimumab, romiplostim, AMG-531, CNTO-148, CNTO-1275, ABT-874, LEA-29Y, belimumab, TACI-Ig, second-generation anti-CD20, ACZ-885, tocilizumab, atezolizumab, mepolizumab, pertuzumab, Humax CD20, tremelimumab (CP-675 206), tesimumab, MDX-010, IDEC-114, itraconazole, HuMax EGFR, aflibercept, HuMax-CD4, Ala-Ala, ChAglyCD3, TRX4, catumaxomab, IGN101, MT-201, Pregovomab, CH-14.18, WX-G250, AMG-162, AAB-001, motavizumab, MEDI-524, efavirenz, Aurograb, rixibacumab, third-generation anti-CD20, LY2469298, and veltuzumab.

21. A pharmaceutical composition comprising the compound according to any one of claims 1 to 20 and a pharmaceutically acceptable carrier or excipient.

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