Drug antibody conjugates

Abstract

having the formula [D-(X) b -(AA) w -(T) g -(L)] n a drug conjugate of Ab, wherein: D is a drug moiety having the following formula (I), or a pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer thereof, wherein D is covalently attached to (X) b (if present), or (AA) w (if present), or to (T) g (if present), or (L); which is useful for treating cancer.

Description

TECHNICAL FIELD

[0001] The present invention relates to novel drug conjugates, drugs, drug linker compounds, methods for their preparation, pharmaceutical compositions containing said drug conjugates, and their use as antitumor agents. BACKGROUND

[0002] Ecteinascidins are extremely potent antitumor agents isolated from the marine tunicate Ecteinascidia turbinata. One of these compounds, trabectedin, is used to treat patients with advanced and metastatic soft tissue sarcoma (STS) after failure of, or unsuitable for, anthracyclines and ifosfamide, and in combination with pegylated liposomal doxorubicin for the treatment of platinum-sensitive recurrent ovarian cancer.

[0003] U.S. Patent (Patent No. 5,149,804) describes Ecteinascidin 722 (ET-722) isolated from the marine tunicate Ecteinascidia turbinata and its structure. ET-722 protects mice from P388 lymphoma, B16 melanoma and Lewis lung carcinoma at very low concentrations in vivo.

[0004]

[0005] WO03066638 describes several synthetic analogs of ET-722 and their cytotoxic activity against tumor cells. Specifically, WO03066638 describes compounds 1 to 3 and their cytotoxic activity against a panel of cancer cell lines.

[0006]

[0007] Another compound described in WO 03 / 014127, lurbinectedin, is currently in clinical trials for the treatment of cancer. Lurbinectedin has the following chemical structure

[0008]

[0009] WO2018197663 relates to a novel ecteinascidin derivative that exhibits very promising antitumor activity. One of the compounds disclosed in this patent application is currently in phase I clinical trials for the prevention and treatment of solid tumors.

[0010] In recent years, the treatment of cancer has made significant progress with the development of drug entities that can more effectively target and kill cancer cells. Researchers have exploited cell surface receptors and antigens selectively expressed by target cells, such as cancer cells, to develop antibody-based drug entities that, for example, bind to tumor-specific or tumor-associated antigens in tumors. To achieve this goal, cytotoxic molecules such as chemotherapeutic drugs, bacterial and plant toxins, and radionuclides have been chemically linked to monoclonal antibodies that bind to tumor-specific or tumor-associated cell surface antigens.

[0011] Accordingly, ADCs represent a challenging area of development given the complex payload, linker, and antibody structures, but there remains a need to develop further ADCs. SUMMARY

[0012] There is a need for novel active drug conjugates. The present invention addresses this need. The present invention further provides novel drugs and drug linker compounds for use in the preparation of the drug conjugates of the present invention, methods of preparing the novel drug conjugates of the present invention, pharmaceutical compositions containing the drug conjugates and their use as antitumor agents, and kits comprising the drug conjugates of the present invention for use in the treatment of cancer.

[0013] In a first aspect of the present invention, there is provided a drug conjugate comprising a drug moiety covalently linked to the remainder of the drug conjugate, the drug conjugate having the formula [D-(X) b -(AA) w -(T) g -(L)-] n -Ab, wherein:

[0014] D is a drug moiety having the following formula (I), or a pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer thereof,

[0015]

[0016] wherein:

[0017] D is covalently linked to (X) b (if present), or (AA) w (if present), or (T) g (if present), or (L);

[0018] Y is -NH- or -O-;

[0019] R1is -OH or -CN;

[0020] R2is -C(=O)R a group;

[0021] R3is hydrogen or -OR b group;

[0022] R4is selected from hydrogen, -CH2OH, -CH2OC(=0)R c , -CH2NH2, and -CH2NHProt NH ;

[0023] R a is selected from hydrogen, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl;

[0024] R b is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl;

[0025] R c is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl; and

[0026] Prot NH is a protecting group for an amino group;

[0027] X and T are extension groups which can be the same or different;

[0028] each AA is independently an amino acid unit;

[0029] L is a linker group;

[0030] w is an integer in the range of 0 to 12;

[0031] b is an integer of 0 or 1 ;

[0032] g is an integer of 0 or 1 ;

[0033] Ab is a moiety comprising at least one antigen binding site; and

[0034] n is the ratio of the group [D-(X) b -(AA) w -(T) g -(L)-] to the moiety comprising at least one antigen binding site and is in the range of 1 to 20.

[0035] In another aspect of the application, a drug conjugate is provided comprising a drug moiety covalently linked to the remainder of the drug conjugate, the drug conjugate having the formula [D-(X) b -(AA) w -(T) g -(L)-] n -Ab, wherein:

[0036] D is a drug moiety having the following formula (I), or a pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer thereof,

[0037]

[0038] wherein:

[0039] D is covalently linked to (X) b (if present), or (AA) w (if present), or (T) g (if present), or (L);

[0040] Y is -NH- or -O-;

[0041] R1is -OH or -CN;

[0042] R2is -C(=O)R a group;

[0043] R3is hydrogen or -OR b group;

[0044] R4is selected from the group consisting of hydrogen, -CH2OH, -CH2OC(=O)R c , -CH2NH2, and -CH2NHProt NH ;

[0045] R a is selected from the group consisting of hydrogen, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl;

[0046] R b is selected from the group consisting of substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl;

[0047] R c is selected from the group consisting of substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12alkenyl, substituted or unsubstituted C2-C 12 alkynyl; and

[0048] Prot NH is a protecting group for an amino group;

[0049] with the proviso that when R4is hydrogen, Y is -O-;

[0050] X and T are extension groups which can be the same or different;

[0051] each AA is independently an amino acid unit;

[0052] L is a linker group;

[0053] w is an integer in the range of 0 to 12;

[0054] b is an integer of 0 or 1 ;

[0055] g is an integer of 0 or 1 ;

[0056] Ab is a moiety comprising at least one antigen binding site; and

[0057] n is a group [D-(X) b -(AA) w -(T) g -(L)-] to a moiety comprising at least one antigen binding site and is in the range of 1 to 20.

[0058] In another aspect of the application, there is provided a drug conjugate comprising a drug moiety covalently linked to the remainder of the drug conjugate, the drug conjugate having the formula [D-(X) b -(AA) w -(T) g -(L)-] n -Ab, wherein:

[0059] D is a drug moiety having the following formula (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer thereof,

[0060]

[0061] wherein:

[0062] the wavy line indicates the point of covalent attachment to (X) b (if present), or (AA) w (if present), or to (T) g (if present), or to (L);

[0063] Y is -NH- or -O-;

[0064] R1is -OH or -CN;

[0065] R2is -C(=O)R a ;

[0066] R3is hydrogen or -OR b ;

[0067] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c , -CH2NH2, and -CH2NHProt NH ;

[0068] R a is selected from hydrogen, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl;

[0069] R b is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl;

[0070] R c is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl; and

[0071] Prot NH is a protecting group for an amino group;

[0072] X and T are extension groups, which can be the same or different;

[0073] each AA is independently an amino acid unit;

[0074] L is a linker group;

[0075] w is an integer ranging from 0 to 12;

[0076] b is an integer of 0 or 1;

[0077] g is an integer of 0 or 1;

[0078] Ab is a moiety comprising at least one antigen binding site; and

[0079] n is a group [D-(X) b -(AA) w -(T) g- (L) -] to the moiety comprising at least one antigen binding site and is in the range of 1 to 20.

[0080] In another aspect of the application, a drug conjugate is provided comprising a drug moiety covalently linked to the remainder of the drug conjugate, the compound having the formula [D-(X) b - (AA) w - (T) g - (L) -] n - Ab, wherein:

[0081] D is a drug moiety having the following formula (I), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof,

[0082]

[0083] wherein:

[0084] D is covalently linked to (X) through a hydroxyl or an amino group b (if present), or (AA) w (if present), or (T) g (if present), or (L);

[0085] Y is selected from -NH- and -O-;

[0086] R1is -OH or -CN;

[0087] R2is -C(=O)R a group;

[0088] R3is hydrogen or -OR b group;

[0089] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c , -CH2NH2and -CH2NHProt NH ;

[0090] R a is selected from hydrogen, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl, wherein the optional substituents are one or more substituents R x ;

[0091] R b is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12alkynyl, wherein the optional substituent is one or more substituents R x ;

[0092] R c is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl, wherein the optional substituent is one or more substituents R x ;

[0093] Prot NH is a protecting group for an amino group;

[0094] substituents R x is selected from the group consisting of C1-C y alkyl optionally substituted by at least one group R 12 , C2-C y alkenyl optionally substituted by at least one group R 12 , C2-C y alkynyl optionally substituted by at least one group R 12 , a halogen atom, an oxyl group, a thioxy group, a cyano group, a nitro group, OR y , OCOR y , OCOOR y , COR y , COOR y , OCONR y R z , CONR y R z , S(O)R y , SO2R y , P(O)(R y )OR z , NR y R z , NR y COR z , NR y C(=O)NR y R z , NR y C(=NR y )NR y R z , aryl having 6 to 18 carbon atoms in one or more rings (the aryl being optionally substituted by one or more groups R y which can be the same or different, OR y , OCOR y , OCOOR y , NR y R z , NRy COR z and NR y C(=NR y )NR y R z substituted by one or more substituents R y , and in the case of more than one optional substituent on any given group, the optional substituents R y may be the same or different;

[0095] each R y and R z is independently selected from the group consisting of hydrogen, C1-C 12 alkyl, C1-C 12 alkyl substituted by at least one halogen atom, aralkyl comprising C1-C 12 alkyl substituted by aryl having 6 to 18 carbon atoms in one or more rings, and heterocycloalkyl comprising C1-C 12 alkyl substituted by 5- to 14-membered saturated or unsaturated heterocyclyl having one or more rings and comprising at least one oxygen, nitrogen, or sulfur atom in the one or more rings;

[0096] X and T are the same or different extending groups;

[0097] each AA is independently an amino acid unit;

[0098] L is a linker group;

[0099] w is an integer in the range of 0 to 12;

[0100] b is an integer of 0 or 1;

[0101] g is an integer of 0 or 1;

[0102] wherein b + g + w is optionally other than 0;

[0103] Ab is a moiety comprising at least one antigen binding site; and

[0104] n is a group [D-(X b -(AA) w -(T) gthe ratio of (L) to the moiety comprising at least one antigen binding site and is in the range of 1 to 20.

[0105] In another aspect of the application, a drug conjugate is provided comprising a drug moiety covalently linked to the remainder of the drug conjugate, the drug conjugate having the formula [D-(X) b -(AA) w -(T) g -(L)-] n -Ab, wherein:

[0106] D is a drug moiety having the following formula (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer thereof,

[0107]

[0108] wherein:

[0109] the wavy line indicates the point of covalent attachment to (X) b (if present), or (AA) w (if present), or to (T) g (if present), or to (L);

[0110] Y is -NH- or -O-;

[0111] R1is -OH or -CN;

[0112] R2is -C(=O)R a group;

[0113] R3is hydrogen or -OR b group;

[0114] R4is selected from the group consisting of hydrogen, -CH2OH, -CH2OC(=O)R c , -CH2NH2, and -CH2NHProt NH ;

[0115] R a is selected from the group consisting of hydrogen, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl, wherein the optional substituent is one or more substituents R x ;

[0116] R b is selected from the group consisting of substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12alkynyl, wherein the optional substituent is one or more substituents R x ;

[0117] R c is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl, wherein the optional substituent is one or more substituents R x ;

[0118] Prot NH is a protecting group for an amino group;

[0119] substituents R x is selected from the group consisting of C1-C y alkyl optionally substituted by at least one group R 12 , C2-C y alkenyl optionally substituted by at least one group R 12 , C2-C y alkynyl optionally substituted by at least one group R 12 , a halogen atom, an oxyl group, a thioxyl group, a cyano group, a nitro group, OR y , OCOR y , OCOOR y , COR y , COOR y , OCONR y R z , CONR y R z , S(O)R y , SO2R y , P(O)(R y )OR z , NR y R z , NR y COR z , NR y C(=O)NR y R z , NR y C(=NR y )NR y R z , aryl having 6 to 18 carbon atoms in one or more rings (the aryl being optionally substituted by one or more groups R y , OR y , OCOR y , OCOOR y , NR y R z , NRy COR z and NR y C(=NR y )NR y R z substituted by one or more substituents R y , which can be the same or different, on any given group, and in the case of more than one optional substituent on any given group, the optional substituents R y may be the same or different;

[0120] each R y and R z is independently selected from the group consisting of hydrogen, C1-C 12 alkyl, C1-C 12 alkyl substituted by at least one halogen atom, aralkyl comprising C1-C 12 alkyl substituted by aryl having 6 to 18 carbon atoms in one or more rings, and heterocycloalkyl comprising C1-C 12 alkyl substituted by 5- to 14-membered saturated or unsaturated heterocyclyl having one or more rings and comprising at least one oxygen, nitrogen, or sulfur atom in the one or more rings;

[0121] X and T are the same or different extended groups;

[0122] each AA is independently an amino acid unit;

[0123] L is a linker group;

[0124] w is an integer ranging from 0 to 12;

[0125] b is an integer of 0 or 1;

[0126] g is an integer of 0 or 1;

[0127] wherein b + g + w is optionally other than 0;

[0128] Ab is a moiety comprising at least one antigen binding site; and

[0129] n is a group [D-(X b -(AA) w -(T) gthe ratio of (L) to the moiety comprising at least one antigen binding site and is in the range of 1 to 20.

[0130] In another aspect of the application, a drug conjugate is provided comprising a drug moiety covalently linked to the remainder of the drug conjugate, the drug conjugate having the formula [D-(X) b (AA) w (T) g (L) n -Ab, wherein:

[0131] D is a drug moiety having the following formula (I), or a pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer thereof,

[0132]

[0133] wherein:

[0134] D is covalently linked to (X) through a hydroxyl or an amino group b (AA), if present, or (T) w (AA), if present, or (T) g (L), if present;

[0135] Y is selected from -NH- and -O-;

[0136] R1is -OH or -CN;

[0137] R2is -C(=O)R a group;

[0138] R3is hydrogen or -OR b group;

[0139] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c , -CH2NH2, and -CH2NHProt NH ;

[0140] R a is selected from hydrogen, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl;

[0141] R b is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl;

[0142] Rc Selected from substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl group; and

[0143] Prot NH It is a protecting group of amino groups;

[0144] X and T are extension groups that can be the same or different;

[0145] Each AA is an independent amino acid unit;

[0146] L is a linker group;

[0147] w is an integer in the range of 0 to 12;

[0148] b is 1;

[0149] g is an integer that is either 0 or 1;

[0150] Ab is a moiety containing at least one antigen-binding site; and

[0151] n represents the group [D-(X)] b -(AA) w -(T) g The ratio of -(L)-] to the portion containing at least one antigen binding site and in the range of 1 to 20.

[0152] In another aspect of the invention, a pharmaceutical conjugate is provided comprising a pharmaceutical portion covalently linked to the remainder of the pharmaceutical conjugate, the compound having the formula [D-(X)]. b -(AA) w -(T) g -(L)-] n -Ab, where:

[0153] D is a pharmaceutical moiety having the following formula (I), or a pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer thereof.

[0154]

[0155] in:

[0156] D is covalently linked to (X) via a hydroxyl or amino group. b (If it exists), or (AA) w (If it exists), or (T) g (if it exists), or (L);

[0157] Y is selected from -NH- and -O-;

[0158] R1is -OH or -CN;

[0159] R2is -C(=O)R a ;

[0160] R3is hydrogen or -OR b ;

[0161] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c , -CH2NH2, and -CH2NHProt NH ;

[0162] R a is selected from hydrogen, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl;

[0163] R b is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl;

[0164] R c is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl; and

[0165] Prot NH is a protecting group for an amino group;

[0166] X and T are extension groups, which can be the same or different;

[0167] each AA is independently an amino acid unit;

[0168] L is a linker group;

[0169] w is 2;

[0170] b is 1;

[0171] g is an integer of 0 or 1;

[0172] Ab is a moiety comprising at least one antigen binding site; and

[0173] n is a group [D-(X) b -(AA) w -(T) g- (L) -] to a moiety comprising at least one antigen binding site and in the range of 1 to 20.

[0174] As we will explain and illustrate in more detail below, the compounds of formula [D-(X) b - (AA) w - (T) g - (L) -] n - Ab represent a breakthrough to solve the above problems (in addition to the need for further drug conjugates exhibiting superior anti-tumor activity in addition to the three major families of drug conjugates based on cytotoxic drugs that have been used so far as payloads).

[0175] In another aspect of the application, there is provided a compound of formula D-(X) b - (AA) w - (T) g - L1or formula D-(X) b - (AA) w - (T) g - H, wherein:

[0176] L1is a linker selected from the group consisting of the following formulae:

[0177]

[0178] each wavy line represents a covalent attachment to (T) g (if present), or (AA) w (if present), or to (X) b (if present), or to D;

[0179] G is selected from halogen, -O-methanesulfonyl, and -O-toluenesulfonyl;

[0180] J is selected from halogen, hydroxyl, -N-succinimidoxy, -O-(4-nitrophenyl), -O-pentafluorophenyl, -O-tetrafluorophenyl, and -O-C(O)-OR 20 ;

[0181] R 19 is selected from -C1-C 12 alkylene-, -C3-C8carbocyclyl, -O-(C1-C 12 alkylene), optionally substituted -C6-C x arylene, -C1-C 18 alkylene-C6-C 12 arylene, -C1-C 18Ar1is arylene (wherein said arylene is monocyclic or polycyclic, which is optionally substituted by one or more substituents R x substituted), -C6-C 18 substituted), -C6-C 12 substituted), -C6-C x substituted), -C6-C 12 substituted), -C6-C 12 substituted), -C6-C 14 substituted), -C6-C x substituted), -C6-C 12 substituted), -C6-C 14 substituted), -C6-C x substituted), -C6-C 14 substituted), -C6-C 12 substituted), -C6-C x substituted), -C6-C r substituted), -C6-C r substituted), -C6-C x substituted), -C6-C

[0182] R 20 substituted), -C6-C 12 substituted), -C6-C x substituted), -C6-C

[0183] r is an integer in the range of 1 to 10;

[0184] g is an integer of 0 or 1;

[0185] b is an integer of 0 or 1;

[0186] w is an integer in the range of 0 to 12; and

[0187] D, R xeach of X, T and AA is as defined in the first aspect of the application.

[0188] In preferred embodiments of the application, b + g + w is not 0. In other embodiments, b + w is not 0. In yet other embodiments, b is 1 when w is not 0. In another embodiment, b is 1 when w is 0.

[0189] In another aspect of the application, there is provided a compound of formula D-(X) b -(AA) w -(T) g -L1or formula D-(X) b -(AA) w -(T) g -H, or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof; wherein each of D, X, AA, T, L1, b, g and w is as defined herein; but further wherein if the compound is of formula D-(X) b -(AA) w -(T) g -H, then b + w + g ≠ 0.

[0190] In preferred embodiments according to the aspects of the application, n is the ratio of the moiety [D-(X) b -(AA) w -(T) g -(L)-] to the moiety comprising at least one antigen binding site and is in the range of 1 to 20. In other embodiments, n is in the range of 1-12, 1-8, 3-8, 3-6, 3-5 or is 1, 2, 3, 4, 5 or 6, preferably 3, 4 or 5 or 4.

[0191] In another aspect of the application, there is provided a drug moiety D for use in an antibody drug conjugate. In another aspect of the application, there is provided a drug moiety D for use as a payload in an antibody drug conjugate. In another aspect of the application, there is provided the use of a drug moiety D as described herein for the manufacture of an antibody drug conjugate.

[0192] In another aspect of the application, there is provided a drug moiety of formula (IA)

[0193]

[0194] wherein:

[0195] Y is -NH- or -O-;

[0196] R1is -OH or -CN;

[0197] R2is -C(=O)R agroup;

[0198] R3is hydrogen or -OR b group;

[0199] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c , -CH2NH2and -CH2NHProt NH ;

[0200] R a is selected from hydrogen, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl;

[0201] R b is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl;

[0202] R c is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl; and

[0203] Prot NH is a protecting group for an amino group;

[0204] with the proviso that when R4is hydrogen, Y is -O-.

[0205] In another aspect of the application, there is provided a pharmaceutical conjugate according to the application for use as a medicament.

[0206] In another aspect of the application, there is provided a pharmaceutical composition comprising a pharmaceutical conjugate according to the application and a pharmaceutically acceptable carrier.

[0207] In another aspect of the application, there is provided a pharmaceutical conjugate according to the application for use in the treatment of cancer.

[0208] In another aspect of the application, there is provided a method for preventing or treating cancer, comprising administering to a patient in need thereof an effective amount of a pharmaceutical conjugate according to the application.

[0209] In another aspect of the application, there is provided the use of a pharmaceutical conjugate according to the application in the manufacture of a medicament for the treatment of cancer.

[0210] In another aspect of the application, a kit is provided comprising a therapeutically effective amount of a drug conjugate according to the application and a pharmaceutically acceptable carrier.

[0211] In the above aspects of the application, the cancer can be selected from lung cancer (including NSCLC), gastric cancer, colorectal cancer, breast cancer, pancreatic cancer, endometrial cancer, bladder cancer, cervical cancer, esophageal cancer, gall bladder cancer, uterine cancer, salivary ductal carcinoma, ovarian cancer, renal cancer, leukemia, multiple myeloma and lymphoma. In a preferred embodiment, the cancer is a HER2 positive cancer. Preferred HER2 positive cancers include HER2 positive lung cancer (including HER2 positive NSCLC), HER2 positive gastric cancer, HER2 positive colorectal cancer, HER2 positive breast cancer, HER2 positive pancreatic cancer, HER2 positive endometrial cancer, HER2 positive bladder cancer, HER2 positive cervical cancer, HER2 positive esophageal cancer, HER2 positive gall bladder cancer, HER2 positive uterine cancer, HER2 positive salivary ductal carcinoma and HER2 positive ovarian cancer. More preferred cancers are HER2 positive breast cancer, HER2 positive ovarian cancer and HER2 positive gastric cancer. Most preferred cancer is HER2 positive breast cancer.

[0212] In another aspect of the application, a method of preparing a drug conjugate according to the application is provided, comprising conjugating a moiety Ab comprising at least one antigen binding site to a drug D, Ab and D being as defined herein. DETAILED DESCRIPTION

[0213] The following applies to all aspects of the application:

[0214] In the compounds of the application, alkyl groups can be branched or unbranched and preferably have from 1 to about 12 carbon atoms. A more preferred class of alkyl groups has from 1 to about 6 carbon atoms. Even more preferred are alkyl groups having 1, 2, 3 or 4 carbon atoms. Methyl, ethyl, n-propyl, isopropyl and butyl (including n-butyl, isobutyl, sec-butyl and t-butyl) are particularly preferred alkyl groups in the compounds of the application.

[0215] In the compounds of the application, alkenyl groups can be branched or unbranched, have one or more double bonds and have from 2 to about 12 carbon atoms. A more preferred class of alkenyl groups has from 2 to about 6 carbon atoms. Even more preferred are alkenyl groups having 2, 3 or 4 carbon atoms. Ethenyl, 1-propenyl, 2-propenyl, 1-methylethenyl, 1-butenyl, 2-butenyl and 3-butenyl are particularly preferred alkenyl groups in the compounds of the application.

[0216] In the compounds of the application, alkynyl groups can be branched or unbranched, have one or more triple bonds and have from 2 to about 12 carbon atoms. A more preferred class of alkynyl groups has from 2 to about 6 carbon atoms. Even more preferred are alkynyl groups having 2, 3 or 4 carbon atoms. In the compounds of the application, alkynyl groups can be branched or unbranched, have one or more triple bonds and have from 2 to about 12 carbon atoms. A more preferred class of alkynyl groups has from 2 to about 6 carbon atoms. Even more preferred are alkynyl groups having 2, 3 or 4 carbon atoms.

[0217] Suitable aryl groups in the compounds of the application include monocyclic and polycyclic compounds, including polycyclic compounds containing separate and / or fused aryl groups. Typical aryl groups contain one to three separate and / or fused rings and from six to about eighteen carbon ring atoms. The aryl groups preferably contain from six to about ten carbon ring atoms. Particularly preferred aryl groups include substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted biphenyl, substituted or unsubstituted phenanthryl, and substituted or unsubstituted anthracenyl.

[0218] Suitable heterocyclic groups include heteroaromatic and heteroaliphatic groups containing one to three separate and / or fused rings and from five to about eighteen ring atoms. Preferably, the heteroaromatic and heteroaliphatic groups contain from five to about ten ring atoms, most preferably five, six, or seven ring atoms. Suitable heteroaromatic groups in the compounds of the application contain one, two, or three heteroatoms selected from N, O, or S, and include, for example, coumarinyl (including 8-coumarinyl), quinolinyl (including 8-quinolinyl), isoquinolinyl, pyridyl, pyrazinyl, pyrazolyl, pyrimidinyl, furanyl, pyrrolyl, thienyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazolyl, indolyl, isoindolyl, indazolyl, indolizinyl, phtalazinyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furopyridinyl, pyridazinyl, triazinyl, cinnolinyl, benzimidazolyl, benzofuranyl, benzofuropyridinyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. Suitable heteroaliphatic groups in the compounds of the application contain one, two, or three heteroatoms selected from N, O, or S, and include, for example, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazapanyl, diazepanyl, thiazepanyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolizinyl, and quinolizinyl.

[0219] The above-mentioned groups can be substituted at one or more available positions with one or more suitable groups such as OR', =0, SR', SOR', SO2R', NO2, NHR', NR'R', =N-R', NHCOR', N(COR')2, NHSO2R', NR'C(=NR')NR'R', CN, halogen, COR', COOR', OCOR', OCONHR', OCONR'R', CONHR', CONR'R', protected OH, protected amino, protected SH, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclyl, wherein each R' group is independently selected from the group consisting of hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, COalkyl, CO2H, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclyl. In the case that such groups are themselves substituted, the substituents can be selected from those listed above. Additionally, in the case that there is more than one R' group on a substituent, each R' can be the same or different.

[0220] In the compounds of the application, halogen substituents include F, CI, Br, and I.

[0221] More particularly, in the compounds of the application, R 20 , Ra, Rb, Rc, R x , Ry, and Rz can be a linear or branched alkyl chain radical having 1 to 12 carbon atoms, and it is preferably an alkyl radical having 1 to 6 carbon atoms, more preferably methyl, ethyl, or isopropyl, and most preferably methyl. In the definitions of M and Q, it can be a linear or branched alkyl chain radical having 1 to 6 carbon atoms. Methyl, ethyl, n-propyl, isopropyl, and butyl (including n-butyl, isobutyl, sec-butyl, and t-butyl) are especially preferred alkyl radicals in the compounds of the application.

[0222] In the compounds of the application, Ra, Rb, Rc, and R xThe alkenyl group defined as branched or unbranched may have one or more double bonds and 2 to 12 carbon atoms. Preferably, it has 2 to 6 carbon atoms, and more preferably, it is a branched or unbranched alkenyl group having 2, 3, or 4 carbon atoms. Vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 2-butenyl, and 3-butenyl are particularly preferred alkenyl groups in the compounds of the present invention.

[0223] In the compounds of this invention, Ra, Rb, Rc, and R x The alkynyl group is defined as branched or unbranched, and may have one or more triple bonds and 2 to 12 carbon atoms. Preferably, it has 2 to 6 carbon atoms, and more preferably, it is a branched or unbranched alkynyl group having 2, 3 or 4 carbon atoms.

[0224] In the compounds of this invention, R x R y and R z The halogen substituents in the definition include F, Cl, Br and I, with Cl being preferred.

[0225] In the compounds of this invention, R xThe 5- to 14-membered saturated or unsaturated heterocyclic group in the definition is a heterocyclic group having one or more rings containing at least one oxygen, nitrogen, or sulfur atom in said one or more rings. The heterocyclic group can be a heteroaromatic group or a heteroalicyclic group, the latter of which can be partially unsaturated. Both aromatic and alicyclic heterocyclic groups contain 1 to 3 independent or fused rings. Preferably, the heteroaromatic group and the heteroalicyclic group contain 5 to 10 ring atoms. The suitable heteroaromatic groups in the compounds of the present invention contain one, two, or three heteroatoms selected from N, O, or S atoms, and include, for example, quinolinyl (including 8-quinolinyl), isoquinolinyl, coumarinyl (including 8-coumarinyl), pyridyl, pyrazinyl, pyrazolyl, pyrimidinyl, furanyl, pyrroleyl, thiophenyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazoleyl, indolyl, isoyndolyl, indazoleyl, indolazinyl, phthalazinyl, pteridinyl, purineyl, oxadiazolyl, thiadiazolyl, furazolidyl, pyridazinyl, triazinyl, cenolinyl, benzimidazolyl, benzofuranyl, benzofuranyl, benzothiophenyl, benzothiazolyl, benzooxazolyl, quinazolinyl, quinoxolinyl, naphthidyl, and furanopyridyl. The suitable heterocyclic groups in the compounds of this invention contain one, two, or three heteroatoms selected from N, O, or S atoms, and include, for example, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiopheneyl, tetrahydrothiopiperanyl, piperidinyl, linyl, thiolinyl, thiooxacyclohexyl, piperazine, azacyclic butyl, oxacyclobutyl, thiooxacyclobutyl, high-piperidinyl, oxacycloheptyl, thiooxacycloheptyl, oxazine, diazine, thiazopine, 1, 2,3,6-Tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-piperanyl, 4H-piperanyl, dioxane, 1,3-dioxolanecycloyl, pyrazolinyl, dithiaalkyl, dithiocyclopentaneyl, dihydropiperanyl, dihydrothiopheneyl, dihydrofuranyl, pyrazolidine, imidazolinyl, imidazodinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolyl and quinazinyl.

[0226] In the compounds of this invention, R x and R 20 The aryl group, as defined, is a monocyclic or polycyclic compound containing independent and / or fused aryl groups and having 6 to 18 ring atoms, which may be selectively substituted. A typical aryl group contains 1 to 3 independent or fused rings. The aryl group preferably contains 6 to 12 carbon ring atoms. Particularly preferred aryl groups include substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted biphenyl, substituted or unsubstituted phenanthyl, and substituted or unsubstituted anthraceneyl, with substituted or unsubstituted phenyl being the most preferred, wherein the substituents are as indicated above.

[0227] In the compounds of this invention, R x R y and R zArylalkyl in the definition of R

[0228] In the compounds of the application, R x Aryloxy in the definition of R

[0229] In the compounds of the application, R y Heteroaryl in the definition of R z Heterocycloalkyl in the definition of R

[0230] In the compounds of the application, R 19 In the definition of R 30 In the definition of R 19 In the definition of R 30 In the definition of R

[0231] In the compounds of the application, R19 The carbocyclic group in the definition of M is a cycloalkyl group having 3 to 8 carbon atoms, which has two covalent bonds at any position on the cycloalkyl ring that connect the cycloalkyl group to the remainder of the drug conjugate. Preferably, R 19 The carbocyclic group in the definition of M is a cycloalkyl group having 3 to 7 carbon atoms, and more preferably a carbocyclic group having 5 to 7 carbon atoms.

[0232] In the compounds of this invention, R 19 The aryl group in the definition is an aryl group having 6 to 18 carbon atoms in one or more rings, having two covalent bonds at any position on the aromatic ring system that connect the aryl group to the remainder of the drug conjugate. Preferably, R 19 The aryl group defined in the definition is an aryl group having 6 to 12 carbon atoms in one or more rings, having two covalent bonds at any position on the aromatic ring system, and most preferably a phenylene group.

[0233] In the compounds of this invention, R 19 The heterocyclic group defined herein is a heterocyclic group containing 1 to 3 independent or fused rings, having 5 to 14 ring atoms, and including at least one oxygen, nitrogen, or sulfur atom in one or more of the rings, wherein two covalent bonds are present at any position on the ring system of the heterocyclic group. The heterocyclic group can be a heteroaromatic group or a heteroalicyclic group (the latter may be partially unsaturated). Preferably, R 19 The definition of a heterocyclic group is a heterocyclic group containing 1 to 3 independent or fused rings, having 5 to 12 ring atoms and including at least one oxygen, nitrogen or sulfur atom in one or more rings, wherein two covalent bonds are present at any position on the ring system of the heterocyclic group.

[0234] There is more than one selectable substituent R on the substituent. x R y or R z In the case of each substituent R x They can be the same or different, each substituent R y They can be the same or different, and each R z They can be the same or different.

[0235] In one embodiment, D may be a pharmaceutical portion of formula (I) or a pharmaceutically acceptable salt or ester thereof:

[0236]

[0237] in:

[0238] D is covalently linked to (X) via a hydroxyl or amino group. b (If it exists), or (AA) w (If it exists), or (T)g (if it exists), or (L);

[0239] Y is selected from -NH- and -O-;

[0240] R1 is -OH or -CN;

[0241] R2 is -C(=O)R a Group;

[0242] R3 is hydrogen or -OR b Group;

[0243] R4 is selected from hydrogen, -CH2OH, -CH2OC (=O)R c -CH2NH2 and -CH2NHProt NH ;

[0244] R a Selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl group;

[0245] R b Selected from substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl group;

[0246] R c Selected from substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 alkenyl, substituted or unsubstituted C2-C 12 alkynyl group; and

[0247] Prot NH It is a protecting group of amino groups.

[0248] In embodiments according to all aspects of the invention, the substituent group is replaced by one or more substituents R. x Instead, the R x Independently selected from the group consisting of: optionally coated with at least one group R y Replacement C1-C 12 Alkyl group, optionally with at least one R group y Replacement C2-C 12 Alkenyl, optionally with at least one R group y Replacement C2-C 12 Alkyne, halogen atom, oxygen, thio, cyano, nitro, OR y OCORy OCOOR y COR y COOR y OCONR y R z CONR y R z S(O)R y SO2R y 、P(O)(R y OR z NR y R z NR y COR z NR y C(=O)NR y R z NR y C(=NR y )NR y R z aryl groups having 6 to 18 carbon atoms in one or more rings (the aryl groups may be selectively composed of one or more identical or different R groups). y OR y OCOR y OCOOR y NR y R z NR y COR z and NR y C(=NR y )NR y R z The group comprises: aralkyl groups having 1 to 12 carbon atoms, including alkyl groups having 1 to 12 carbon atoms that are selectively substituted with aryl groups as defined above; aralkoxy groups having 1 to 12 carbon atoms that are selectively substituted with aryl groups as defined above; and 5 to 14 member saturated or unsaturated heterocyclic groups having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, wherein the heterocyclic group may be selectively substituted with one or more substituents R. y Substitution, and in the case where more than one alternative substituent is present on any given group, the alternative substituent R y They can be the same or different;

[0249] Each R y and R z Independently select from the following groups: hydrogen, C1-C 12 Alkyl groups, C1-C atoms substituted with at least one halogen atom 12 Alkyl groups, including C1-C groups substituted with aryl groups having 6 to 18 carbon atoms in one or more rings.12 aralkyl of an alkyl group, and C1-C6alkyl substituted with a 5- to 14-membered saturated or unsaturated heterocyclyl group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings 12 heterocycloalkyl of an alkyl group.

[0250] In another embodiment, D can be a drug moiety of Formula (IH) or a pharmaceutically acceptable salt or ester thereof:

[0251]

[0252] wherein the wavy line indicates a covalent attachment to (X) b (AA) w (BB) g (CC)

[0253] Y is selected from -NH- and -O-;

[0254] R1is -OH or -CN;

[0255] R2is -C(=O)R a group;

[0256] R3is hydrogen or -OR b group;

[0257] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c , -CH2NH2, and -CH2NHProt NH ;

[0258] R a is selected from hydrogen, substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl, wherein the optional substituent is one or more substituents R x ;

[0259] R b is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12 alkenyl, and substituted or unsubstituted C2-C 12 alkynyl, wherein the optional substituent is one or more substituents R x ;

[0260] R c is selected from substituted or unsubstituted C1-C 12 alkyl, substituted or unsubstituted C2-C 12alkenyl, substituted or unsubstituted C2-C 12 Alkyne group, wherein the optional substituent is one or more substituents R x ;

[0261] Prot NH It is a protecting group of amino groups;

[0262] Where the substituent group is replaced by one or more substituents R x Instead, the R x Independently selected from the group consisting of: optionally coated with at least one group R y Replacement C1-C 12 Alkyl group, optionally with at least one R group y Replacement C2-C 12 Alkenyl, optionally with at least one R group y Replacement C2-C 12 Alkyne, halogen atom, oxygen, thio, cyano, nitro, OR y OCOR y OCOOR y COR y COOR y OCONR y R z CONR y R z S(O)R y SO2R y 、P(O)(R y OR z NR y R z NR y COR z NR y C(=O)NR y R z NR y C(=NR y )NR y R z aryl groups having 6 to 18 carbon atoms in one or more rings (the aryl groups may be selectively composed of one or more identical or different R groups). y OR y OCOR y OCOOR y NR y R z NR y COR z and NR y C(=NR y )NR y R zsubstituted with one or more substituents R y substituted with one or more substituents R y which can be the same or different;

[0263] each R y and R z is independently selected from the group consisting of hydrogen, C1-C 12 alkyl, C1-C 12 alkyl substituted with at least one halogen atom, C1-C 12 alkyl substituted with one or more substituents R 12 alkyl substituted with one or more substituents R

[0264] Preferred compounds of the compounds of general formula (I) or (IH) and the pharmaceuticals of general formula (IA) are those having general formula a or b or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof:

[0265]

[0266] Note that in the case of compounds having general formula a or b, R4may not be hydrogen.

[0267] Preferred prodrugs and pharmaceuticals include those of general formula (I) or (IH) and the pharmaceuticals of general formula (IA) wherein:

[0268] Y is -NH-;

[0269] and R1, R2, R3, R4, R a , R b , R c and Prot NH are as described above.

[0270] Preferred prodrugs and pharmaceuticals include those of general formula (I) or (IH) and the pharmaceuticals of general formula (IA) wherein:

[0271] Y is -O-;

[0272] and R1, R2, R3, R4, R a , R b , R c and Prot NH are as described above.

[0273] Further preferred drug moieties and drugs comprise moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0274] R1is -OH;

[0275] and Y, R2, R3, R4, R a , R b , R c and Prot NH are as described above.

[0276] Further preferred drug moieties and drugs comprise moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0277] R1is -CN;

[0278] and Y, R2, R3, R4, R a , R b , R c and Prot NH are as described above.

[0279] Further preferred drug moieties and drugs comprise moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0280] R2is a -C(=O)R a group, wherein R a is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R a is selected from the group consisting of a substituted or unsubstituted methyl group, a substituted or unsubstituted ethyl group, a substituted or unsubstituted n-propyl group, a substituted or unsubstituted i-propyl group, a substituted or unsubstituted n-butyl group, a substituted or unsubstituted i-butyl group, a substituted or unsubstituted sec-butyl group, a substituted or unsubstituted tert-butyl group. Most preferred R2is an acetyl group;

[0281] and Y, R1, R3, R4, R b , R c and Prot NH are as described above.

[0282] Further preferred drug moieties and drugs comprise moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0283] R3is hydrogen or a -OR b group; wherein R bis selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy; b is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy;

[0284] and Y, R1, R2, R4, R a , R c and Prot NH are as described above.

[0285] Further preferred drug moieties and drugs include moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0286] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy; c is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy; c is methyl. More preferred R4is selected from hydrogen, -CH2OH and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferred R4is hydrogen;

[0287] and Y, R1, R2, R3, R a , and R b are as described above.

[0288] Further preferred drug moieties and drugs include moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0289] Y is -NH-;

[0290] R1is -OH;

[0291] and R2, R3, R4, R a , R b , R c , and Prot NH are as described above.

[0292] Further preferred drug moieties and drugs include moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0293] Y is -NH-;

[0294] R2is -C(=O)R a ; wherein R a is substituted or unsubstituted C1-C6alkyl. Particularly preferred R a groups are selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl.

[0295] Most preferred R2is acetyl;

[0296] and R1, R3, R4, R b , R c , and Prot NH are as described above.

[0297] Further preferred drug moieties and drugs include moieties of general formula (I) or (IH) and drugs of general formula (IA), wherein:

[0298] Y is -NH-;

[0299] R3is hydrogen or an -OR b group; wherein R b is substituted or unsubstituted C1-C6alkyl. Particularly preferred R b groups are selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy;

[0300] and R1, R2, R4, R a , R c , and Prot NH are as described above.

[0301] Further preferred drug moieties include moieties of general formula (I) or (IH), wherein:

[0302] Y is -NH-;

[0303] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c , and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R csubstituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted tert-butyl. Most preferred R c is methyl. More preferred R4is selected from hydrogen, -CH2OH and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferred R4is hydrogen;

[0304] and R1, R2, R3, R a , and R b are as described above.

[0305] Further preferred are the drugs of general formula (IA), wherein:

[0306] Y is -NH-;

[0307] R4is selected from -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R c is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted tert-butyl. Most preferred R c is methyl. More preferred R4is -CH2OH or -CH2NH2. Most preferred R4is -CH2OH;

[0308] and R1, R2, R3, R a , and R b are as described above.

[0309] Further preferred are the drugs of general formula (IA), wherein:

[0310] Y is -NH-;

[0311] R1is -OH;

[0312] R2is a -C(=O)R a group, wherein R a is substituted or unsubstituted C1-C6alkyl. Particularly preferred R a is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted tert-butyl. Most preferred R2is acetyl;

[0313] and R3, R4, R b , R c and Prot NH are as described above.

[0314] Further preferred drug moieties and drugs include moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0315] Y is -NH-;

[0316] R1is -OH;

[0317] R3is hydrogen or -OR b ; wherein R b is substituted or unsubstituted C1-C6alkyl. Particularly preferred R b is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy;

[0318] and R2, R4, R a , R c and Prot NH are as described above.

[0319] Further preferred drug moieties include moieties of the general formula (I) or (IH), wherein:

[0320] Y is -NH-;

[0321] R1is -OH;

[0322] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R c is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4is selected from hydrogen, -CH2OH and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferred R4is hydrogen;

[0323] and R2, R3, R a , and R b are as described above.

[0324] Further preferred are the drugs of general formula (IA), wherein:

[0325] Y is -NH-;

[0326] R1is -OH;

[0327] R4is selected from the group consisting of -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R c is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4is -CH2OH and -CH2NH2. Most preferred R4is -CH2OH;

[0328] and R2, R3, R a , and R b are as described above.

[0329] Further preferred are the drug moieties and drugs comprising a moiety of general formula (I) or (IH) and a drug of general formula (IA), wherein:

[0330] Y is -NH-;

[0331] R2is a -C(=O)R a group, wherein R a is substituted or unsubstituted C1-C6alkyl. Particularly preferred R a is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R2is acetyl;

[0332] R3is hydrogen or a -OR b group; wherein R b is substituted or unsubstituted C1-C6alkyl. Particularly preferred R b is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy;

[0333] and R1, R4, Rc and Prot NH as described above.

[0334] Further preferred drug moieties include moieties of the general formula (I) or (IH), wherein:

[0335] Y is -NH-;

[0336] R2is a -C(=O)R a group, wherein R a is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred R a is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R2is acetyl;

[0337] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred R c is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4is selected from hydrogen, -CH2OH and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferred R4is hydrogen;

[0338] and R1, R3, and R b are as described above.

[0339] Further preferred drugs include drugs of the general formula (IA), wherein:

[0340] Y is -NH-;

[0341] R2is a -C(=O)R a group, wherein R a is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred R a is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R2is acetyl;

[0342] R4is selected from -CH2OH, -CH2OC(=O)Rc And -CH2NH2; where R c It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. c Selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. The most preferred R... c It is a methyl group. More preferably, R4 is selected from -CH2OH and -CH2NH2. The most preferred R4 is -CH2OH;

[0343] And R1, R3, and R b As stated above.

[0344] Further preferred drug portions include portions of general formula (I) or (IH), wherein:

[0345] Y is -NH-;

[0346] R3 is hydrogen or -OR b Group; wherein R b It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. b The R3 group is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. More preferably, R3 is hydrogen and methoxy, and most preferably, R3 is methoxy.

[0347] R4 is selected from hydrogen, -CH2OH, -CH2OC (=O)R c And -CH2NH2; where R c It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. c Selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. The most preferred R... c It is a methyl group. More preferably, R4 is selected from hydrogen, -CH2OH, and -CH2NH2. More preferably, R4 can be hydrogen or -CH2OH. Most preferably, R4 is hydrogen.

[0348] And R1, R2, and R a As stated above.

[0349] Further preferred drugs include drugs of general formula (IA), wherein:

[0350] Y is -NH-;

[0351] R3is hydrogen or -OR b is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R b is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R b is hydrogen and methoxy, and most preferred R3groups are methoxy;

[0352] R4is selected from -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R c is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4is selected from -CH2OH and -CH2NH2. Most preferred R4is -CH2OH;

[0353] and R1, R2, and R a are as described above.

[0354] Further preferred drug moieties and drugs include moieties of general formula (I) or (IH) and drugs of general formula (IA), wherein:

[0355] Y is -NH-;

[0356] R1is -OH;

[0357] R2is -C(=O)R a group, wherein R a is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R a is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R2is acetyl;

[0358] R3is hydrogen or -OR b group; wherein R b is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R bselected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, and most preferred R3group is methoxy;

[0359] and R4, R c and Prot NH as described above.

[0360] Further preferred drug moieties include moieties of the general formula (I) or (IH), wherein:

[0361] Y is -NH-;

[0362] R1is -OH;

[0363] R2is -C(=O)R a wherein R a is substituted or unsubstituted C1-C6alkyl. Particularly preferred R a is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R2is acetyl;

[0364] R4is selected from the group consisting of hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R c is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4is selected from the group consisting of hydrogen, -CH2OH and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferred R4is hydrogen;

[0365] and R3, and R b as described above.

[0366] Further preferred drugs include drugs of the general formula (IA), wherein:

[0367] Y is -NH-;

[0368] R1is -OH;

[0369] R2is -C(=O)Ra R2is a -C(=O)R a group, wherein R a is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R c groups are selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4groups are selected from -CH2OH and -CH2NH2. Most preferred R4is -CH2OH;

[0370] R4is selected from -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R c groups are selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4groups are selected from -CH2OH and -CH2NH2. Most preferred R4is -CH2OH;

[0371] and R3, and R b are as described above.

[0372] Further preferred drug moieties include moieties of general formula (I) or (IH), wherein:

[0373] Y is -NH-;

[0374] R2is a -C(=O)R a group, wherein R a is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R a groups are selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R b is methyl. More preferred R2groups are selected from -CH2OH and -CH2NH2. Most preferred R2is -CH2OH;

[0375] R3is hydrogen or -OR b group; wherein R b is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R b groups are selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3groups are hydrogen and methoxy, most preferred R3groups are methoxy;

[0376] R4is selected from the group consisting of hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R c is selected from the group consisting of a substituted or unsubstituted methyl group, a substituted or unsubstituted ethyl group, a substituted or unsubstituted n-propyl group, a substituted or unsubstituted iso-propyl group, a substituted or unsubstituted n-butyl group, a substituted or unsubstituted iso-butyl group, a substituted or unsubstituted sec-butyl group, a substituted or unsubstituted t-butyl group. Most preferred R c is a methyl group. More preferred R4is selected from the group consisting of hydrogen, -CH2OH and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferred R4is hydrogen;

[0377] R1is as described above.

[0378] Further preferred pharmaceuticals include those of general formula (IA), wherein:

[0379] Y is -NH-;

[0380] R2is a -C(=O)R a group, wherein R a is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R a is selected from the group consisting of a substituted or unsubstituted methyl group, a substituted or unsubstituted ethyl group, a substituted or unsubstituted n-propyl group, a substituted or unsubstituted iso-propyl group, a substituted or unsubstituted n-butyl group, a substituted or unsubstituted iso-butyl group, a substituted or unsubstituted sec-butyl group, a substituted or unsubstituted t-butyl group. Most preferred R2is an acetyl group;

[0381] R3is hydrogen or a -OR b group; wherein R b is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R b is selected from the group consisting of a substituted or unsubstituted methyl group, a substituted or unsubstituted ethyl group, a substituted or unsubstituted n-propyl group, a substituted or unsubstituted iso-propyl group, a substituted or unsubstituted n-butyl group, a substituted or unsubstituted iso-butyl group, a substituted or unsubstituted sec-butyl group, a substituted or unsubstituted t-butyl group. More preferred R3is hydrogen and a methoxy group, most preferred R3group is a methoxy group;

[0382] R4is selected from the group consisting of -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R csubstituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4is selected from -CH2OH and -CH2NH2. Most preferred R4is -CH2OH;

[0383] R1is as described above.

[0384] Further preferred drug moieties include moieties of general formula (I) or (IH) wherein:

[0385] Y is -NH-;

[0386] R1is -OH;

[0387] R2is -C(=O)R a wherein R a is substituted or unsubstituted C1-C6alkyl. Particularly preferred R a is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R2is acetyl;

[0388] R3is hydrogen or -OR b wherein R b is substituted or unsubstituted C1-C6alkyl. Particularly preferred R b is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy;

[0389] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R c is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R cis hydrogen. More preferably, R4is selected from the group consisting of -CH2OH, -CH2OC(=O)R5and -CH2NH2. Most preferably, R4is -CH2OH.

[0390] Further preferred drugs include drugs of general formula (IA), wherein:

[0391] Y is -NH-;

[0392] R1is -OH;

[0393] R2is -C(=O)R a group, wherein R a is substituted or unsubstituted C1-C6alkyl. Particularly preferred R a is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R2is acetyl;

[0394] R3is hydrogen or -OR b group; wherein R b is substituted or unsubstituted C1-C6alkyl. Particularly preferred R b is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy;

[0395] R4is selected from the group consisting of -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R c is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4is selected from the group consisting of -CH2OH and -CH2NH2. Most preferred R4is -CH2OH.

[0396] Further preferred drug moieties and drugs include moieties of general formula (I) or (IH) and drugs of general formula (IA), wherein:

[0397] Y is -O-;

[0398] R1is -OH;

[0399] and R2, R3, R4, R a , R b , R c , and Prot NH are as described above.

[0400] Further preferred drug moieties and drugs comprise moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0401] Y is -O-;

[0402] R2is a -C(=O)R a group, wherein R a is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred R a is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R2is acetyl;

[0403] and R1, R3, R4, R b , R c , and Prot NH are as described above.

[0404] Further preferred drug moieties and drugs comprise moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0405] Y is -O-;

[0406] R3is hydrogen or a -OR b group; wherein R b is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred R b is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy;

[0407] and R1, R2, R4, R a , R c , and Prot NH are as described above.

[0408] Further preferred drug moieties and drugs comprise moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0409] Y is -O-;

[0410] R4is selected from the group consisting of hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R c is selected from the group consisting of a substituted or unsubstituted methyl group, a substituted or unsubstituted ethyl group, a substituted or unsubstituted n-propyl group, a substituted or unsubstituted i-propyl group, a substituted or unsubstituted n-butyl group, a substituted or unsubstituted i-butyl group, a substituted or unsubstituted sec-butyl group, a substituted or unsubstituted t-butyl group. Most preferred R c is a methyl group. More preferred R4is selected from the group consisting of hydrogen, -CH2OH and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferred R4is hydrogen;

[0411] and R1, R2, R3, R a , and R b are as described above.

[0412] Further preferred drug moieties and drugs comprise moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0413] Y is -O-;

[0414] R1is -OH;

[0415] R2is a -C(=O)R a group, wherein R a is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R a is selected from the group consisting of a substituted or unsubstituted methyl group, a substituted or unsubstituted ethyl group, a substituted or unsubstituted n-propyl group, a substituted or unsubstituted i-propyl group, a substituted or unsubstituted n-butyl group, a substituted or unsubstituted i-butyl group, a substituted or unsubstituted sec-butyl group, a substituted or unsubstituted t-butyl group. Most preferred R2is an acetyl group;

[0416] and R3, R4, R b , R c , and Prot NH are as described above.

[0417] Further preferred drug moieties and drugs comprise moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0418] Y is -O-;

[0419] R1is -OH;

[0420] R3is hydrogen or a -OR b group; wherein R b is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred Rb The R3 group is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. More preferably, R3 is hydrogen and methoxy, and most preferably, R3 is methoxy.

[0421] And R2, R4, R a R c and Prot NH As stated above.

[0422] Further preferred pharmaceutical portions and pharmaceuticals include portions of general formula (I) or (IH) and pharmaceuticals of general formula (IA), wherein:

[0423] Y is -O-;

[0424] R1 is -OH;

[0425] R4 is selected from hydrogen, -CH2OH, -CH2OC (=O)R c And -CH2NH2; where R c It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. c Selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. The most preferred R... c It is a methyl group. More preferably, R4 is selected from hydrogen, -CH2OH, and -CH2NH2. More preferably, R4 can be hydrogen or -CH2OH. Most preferably, R4 is hydrogen.

[0426] And R2, R3, R a and R b As stated above.

[0427] Further preferred pharmaceutical portions and pharmaceuticals include portions of general formula (I) or (IH) and pharmaceuticals of general formula (IA), wherein:

[0428] Y is -O-;

[0429] R2 is -C(=O)R a Group, wherein R a It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. aThe R2 is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. The most preferred R2 is acetyl.

[0430] R3 is hydrogen or -OR b Group; wherein R b It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. b The R3 group is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. More preferably, R3 is hydrogen and methoxy, and most preferably, R3 is methoxy.

[0431] And R1, R4, R c and Prot NH As stated above.

[0432] Further preferred pharmaceutical portions and pharmaceuticals include portions of general formula (I) or (IH) and pharmaceuticals of general formula (IA), wherein:

[0433] Y is -O-;

[0434] R2 is -C(=O)R a Group, wherein R a It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. a The R2 is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. The most preferred R2 is acetyl.

[0435] R4 is selected from hydrogen, -CH2OH, -CH2OC (=O)R c And -CH2NH2; where R c It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. c Selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. The most preferred R... c It is a methyl group. More preferably, R4 is selected from hydrogen, -CH2OH, and -CH2NH2. More preferably, R4 can be hydrogen or -CH2OH. Most preferably, R4 is hydrogen.

[0436] and R1, R3and R b As described above.

[0437] Further preferred drug moieties and drugs include moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0438] Y is -O-;

[0439] R3is hydrogen or -OR b ; wherein R b is substituted or unsubstituted C1-C6alkyl. Particularly preferred R b is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy;

[0440] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R c is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4is selected from hydrogen, -CH2OH and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferred R4is hydrogen;

[0441] and R1, R2and R a As described above.

[0442] Further preferred drug moieties and drugs include moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0443] Y is -O-;

[0444] R1is -OH;

[0445] R2is -C(=O)R a ; wherein R a is substituted or unsubstituted C1-C6alkyl. Particularly preferred R aR2is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R2groups are hydrogen and methoxy, most preferred R2groups are methoxy;

[0446] R3is hydrogen or -OR b wherein R b is substituted or unsubstituted C1-C6alkyl. Particularly preferred R b groups are selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3groups are hydrogen and methoxy, most preferred R3groups are methoxy;

[0447] and R4, R c , and Prot NH are as described above.

[0448] Further preferred drug moieties and drugs include moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0449] Y is -O-;

[0450] R1is -OH;

[0451] R2is -C(=O)R a wherein R a is substituted or unsubstituted C1-C6alkyl. Particularly preferred R a groups are selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R2groups are hydrogen and methoxy, most preferred R2groups are methoxy;

[0452] R4is selected from the group consisting of hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R c groups are selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R4groups are hydrogen and methoxy, most preferred R4groups are methoxy; cis hydrogen. More preferably, R4is selected from the group consisting of hydrogen, -CH2OH, and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferably, R4is hydrogen;

[0453] and R3and R b As described above.

[0454] Further preferred drug moieties and drugs include moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0455] Y is -O-;

[0456] R2is -C(=O)R a wherein R a is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R a is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferably, R2is acetyl;

[0457] R3is hydrogen or -OR b wherein R b is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R b is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferably, R3is hydrogen and methoxy, most preferably the R3group is methoxy;

[0458] R4is selected from the group consisting of hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is a substituted or unsubstituted C1-C6alkyl group. Particularly preferred R c is selected from the group consisting of substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferably, R c is methyl. More preferably, R4is selected from the group consisting of hydrogen, -CH2OH, and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferably, R4is hydrogen;

[0459] R1is as described above.

[0460] Further preferred drug moieties and drugs include moieties of the general formula (I) or (IH) and drugs of the general formula (IA), wherein:

[0461] Y is -O-;

[0462] R1is -OH;

[0463] R2is -C(=O)R a group, wherein R a is substituted or unsubstituted C1-C6alkyl. Particularly preferred R a is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R2is acetyl;

[0464] R3is hydrogen or -OR b group; wherein R b is substituted or unsubstituted C1-C6alkyl. Particularly preferred R b is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. More preferred R3is hydrogen and methoxy, most preferred R3group is methoxy;

[0465] R4is selected from hydrogen, -CH2OH, -CH2OC(=O)R c and -CH2NH2; wherein R c is substituted or unsubstituted C1-C6alkyl. Particularly preferred R c is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted i-propyl, substituted or unsubstituted n-butyl, substituted or unsubstituted i-butyl, substituted or unsubstituted sec-butyl, substituted or unsubstituted t-butyl. Most preferred R c is methyl. More preferred R4is selected from hydrogen, -CH2OH and -CH2NH2. More preferably, R4may be hydrogen or -CH2OH. Most preferred R4is hydrogen.

[0466] The following preferred substituents (where possible substituents are allowed) apply to the drug moieties of the general formula (I) or (IH) and the drugs of the general formula (IA):

[0467] Particularly preferred R1is -OH.

[0468] Particularly preferred R2is -C(=O)R a group, wherein R aIt is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. a The R2 is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. The most preferred R2 is acetyl.

[0469] The particularly preferred R3 is hydrogen or -OR b Group; wherein R b It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. b The R3 group is selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. More preferably, R3 is hydrogen and methoxy, and most preferably, R3 is methoxy.

[0470] The particularly preferred R4 is selected from hydrogen, -CH2OH, and -CH2OC(=O)R. c And -CH2NH2; where R c It is a substituted or unsubstituted C1-C6 alkyl group. Particularly preferred is R. c Selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, substituted or unsubstituted sec-butyl, and substituted or unsubstituted tert-butyl. The most preferred R... c It is a methyl group. More preferably, R4 is selected from hydrogen, -CH2OH, and -CH2NH2. More preferably, R4 can be hydrogen or -CH2OH. Most preferably, R4 is hydrogen.

[0471] The particularly preferred pharmaceutical portion and the pharmaceutical according to the present invention comprise:

[0472] • Part of formula (I) or (IH), where

[0473] Y is -NH-; and

[0474] R4 is selected from hydrogen, -CH2OH, and -CH2NH2.

[0475] Drugs of formula (IA), in which

[0476] Y is -NH-; and

[0477] R4 is selected from -CH2OH and -CH2NH2.

[0478] • A drug of formula (I) or (IH) or formula (IA), wherein

[0479] Y is -O-;

[0480] R4is selected from hydrogen, -CH2OH, and -CH2NH2.

[0481] • a pharmaceutical of the partial formula (I) or (IH) or of the formula (IA), wherein

[0482] R2is -C(=O)R a group;

[0483] R3is hydrogen or -OR b group;

[0484] R4is selected from -CH2OH, and -CH2NH2.

[0485] R a is selected from hydrogen, substituted or unsubstituted C1-C6alkyl; and

[0486] R b is substituted or unsubstituted C1-C6alkyl.

[0487] More preferred pharmaceuticals of the partial formula (I) or (IH) according to the present application include

[0488] • a pharmaceutical of the partial formula (I) or (IH), wherein

[0489] Y is -NH-;

[0490] R2is -C(=O)R a group;

[0491] R3is hydrogen or -OR b group;

[0492] R4is hydrogen or -CH2OH;

[0493] R a is selected from hydrogen, substituted or unsubstituted C1-C6alkyl; and

[0494] R b is substituted or unsubstituted C1-C6alkyl.

[0495] • a pharmaceutical of the formula (IA), wherein

[0496] Y is -NH-;

[0497] R2is -C(=O)R a group;

[0498] R3is hydrogen or -OR b group;

[0499] R4is -CH2OH;

[0500] Ra is selected from hydrogen, substituted or unsubstituted Ci-C6alkyl; and

[0501] R b is substituted or unsubstituted Ci-C6alkyl.

[0502] • a moiety of formula (I) or (IH) or a drug of formula (IA), wherein Y is -0-;

[0503] R2is -C(=0)R a group;

[0504] R3is hydrogen or -OR b group;

[0505] R4is hydrogen or -CH2OH;

[0506] R a is selected from hydrogen, substituted or unsubstituted Ci-C6alkyl; and

[0507] R b is substituted or unsubstituted Ci-C6alkyl.

[0508] • a moiety of formula (I) or (IH) or a drug of formula (IA), wherein

[0509] R2is -C(=0)R a group;

[0510] R3is hydrogen or -OR b group;

[0511] R4is hydrogen or -CH2OH;

[0512] R a is selected from hydrogen, substituted or unsubstituted Ci-C6alkyl; and

[0513] R b is substituted or unsubstituted Ci-C6alkyl.

[0514] Particularly preferred preferred drug moieties according to the present application include:

[0515] • a moiety of formula (I) or (IH), wherein

[0516] Y is -NH-;

[0517] R2is -C(=0)R a group;

[0518] R3is hydrogen or methoxy;

[0519] R4is hydrogen or -CH2OH; and

[0520] R ais substituted or unsubstituted C1-C6alkyl.

[0521] • a drug of formula (IA), wherein

[0522] Y is -NH-;

[0523] R2is -C(=O)R a group;

[0524] R3is hydrogen or methoxy;

[0525] R4is -CH2OH; and

[0526] R a is substituted or unsubstituted C1-C6alkyl.

[0527] • a drug of formula (I) or (IH) or of formula (IA), wherein

[0528] Y is -O-;

[0529] R2is -C(=O)R a group;

[0530] R3is hydrogen or methoxy;

[0531] R4is hydrogen or -CH2OH; and

[0532] R a is substituted or unsubstituted C1-C6alkyl.

[0533] • a drug of formula (I) or (IH) or of formula (IA), wherein

[0534] R2is -C(=O)R a group;

[0535] R3is hydrogen or methoxy;

[0536] R4is hydrogen or -CH2OH; and

[0537] R a is selected from the group consisting of methyl, ethyl, n-propyl, i-propyl and butyl (including n-butyl, sec-butyl, i-butyl and t-butyl).

[0538] Even more preferably, the drug moiety according to the application comprises:

[0539] • a drug of formula (I) or (IH), wherein

[0540] Y is -NH-;

[0541] R2is acetyl;

[0542] R3is methoxy; and

[0543] R4 is hydrogen.

[0544] • a drug of formula (IA) wherein Y is -NH-;

[0545] R2 is acetyl;

[0546] R3 is methoxy; and R4 is -CH2OH.

[0547] • a moiety of formula (I) or (IH) or a drug of formula (IA) wherein Y is -O-;

[0548] R2 is acetyl;

[0549] R3 is methoxy; and R4 is hydrogen.

[0550] • a moiety of formula (I) or (IH) or a drug of formula (IA) wherein R2 is acetyl;

[0551] R3 is methoxy; and R4 is hydrogen.

[0552] • a moiety of formula (I) or (IH) or a drug of formula (IA) wherein R1 is -OH;

[0553] R2 is acetyl;

[0554] R3 is methoxy; and R4 is hydrogen.

[0555] • a moiety according to the present application of the following formula:

[0556]

[0557] or a pharmaceutically acceptable salt or ester thereof; wherein D is covalently attached to (X) through a hydroxyl or amino group b (if present), or (AA) w (if present), or (T) g (if present), or (L).

[0558] Particularly preferred is a moiety of the following formula:

[0559]

[0560] or a pharmaceutically acceptable salt or ester thereof; wherein D is covalently attached to (X) through a hydroxyl or amino group b (if present), or (AA) w (if present), or (T) g (if present), or (L).

[0561] • a moiety of the following formula:

[0562]

[0563] In another preferred embodiment, the preferred cases described above for different substituents are combined. The invention also relates to such combinations of preferred substituents in a pharmaceutical portion of formula (I) or (IH) according to the invention and in a pharmaceutical product of formula (IA) (where possible substituents are permitted).

[0564] To avoid any doubt, the above compounds may be pharmaceutical moiety D and covalently linked to (X) via a hydroxyl or amino group. b (If it exists) or (AA) w (If it exists) or to (T) g (If present) or (L). Therefore, when conjugated, the covalent bond replaces the proton on the hydroxyl group or amino group of the compound.

[0565] Preferred pharmaceutical conjugates according to the present invention are given below. As described below (X) b (AA) w (T) g The preferred definition of (L) applies to all of the above-described pharmaceutical fractions, specifically compound D. Preferred pharmaceutical conjugates according to the invention include:

[0566] ●According to the formula [D-(X)] of the present invention b -(AA) w -(T) g -(L)-] n -Ab drug conjugates, where L is a linker group selected from the group consisting of:

[0567]

[0568] in

[0569] The wavy line indicates a covalent connection to Ab (the wavy line on the right) and to (T). g (If it exists), or (AA) w (If it exists), or to (X) b (If it exists), or to point D (the wavy line on the left);

[0570] R 19 Selected from -C1-C 12 Alkylene, -C3-C8 carbocyclic, -O-(C1-C 12 Alkylene, monocyclic or polycyclic, optionally substituted with one or more R groups. x Replacement -C6-C 18 Alpha-aryl, -C1-C 12 Alkylene-C6-C 18 arylene-(wherein the arylene group is monocyclic or polycyclic, and may be selectively replaced by one or more substituents R) x Replacement), -C6-C 18Aspartic-C1-C 12 alkylene-(wherein the arylene group is monocyclic or polycyclic, and may be selectively substituted by one or more substituents R) x Replacement, -C1-C 12 Alkylene-(C3-C8 carbocyclic)-, -(C3-C8 carbocyclic)-C1-C 12 Alkylene-, -C5-C 14 Heterocyclic group - (wherein the heterocyclic group may be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, the group may be selectively substituented by one or more R groups) x Replacement), -C1-C 12 Alkylene-(C5-C 14 Heterocyclic group) - (wherein the heterocyclic group may be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, the group may be selectively substituented by one or more R groups. x Replace), -(C5-C 14 (heterocyclic group)-C1-C 12 Alkylene (wherein the heterocyclic group may be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, the group may be selectively substituented by one or more R groups) x (substitute), -(OCH2CH2) r - and -CH2-(OCH2CH2) r - wherein, whether each of the above alkylene substituents is alone or attached to another part, the carbon chain can be selectively substituented by one or more R groups. x replace;

[0571] R 30 It is a -C1-C6 alkylene group;

[0572] M is selected from the following groups: -C1-C6 alkylene-, -C1-C6 alkylene-(C3-C8 carbocyclic)-, -(CH2CH2O) s -, -C1-C6 alkylene-(C3-C8 carbocycloyl)-CON(H or C1-C6 alkyl)-C1-C6 alkylene-, optionally with one or more substituents R x Substituted phenylene, phenylene-C1-C6 alkylene- (wherein the phenylene moiety may be selectively replaced by one or more substituents R) x Substitution), and -C1-C6 alkylene-CON(H or C1-C6 alkyl)C1-C6 alkylene-;

[0573] Q is selected from the group consisting of -N(H or C1-C6alkyl)phenylene- and -N(H or C1-C6alkyl)-(CH2) s ;

[0574] r is an integer ranging from 1 to 10; and

[0575] s is an integer ranging from 1 to 10.

[0576] • a pharmaceutical conjugate of Ab according to the application of formula [D-(X) b -(AA) w -(T) g -(L)-] n -Ab, wherein L is selected from the group consisting of:

[0577]

[0578] wherein:

[0579] the wavy line represents the covalent attachment to Ab (wavy line on the right) and to (T) g (if present), or (AA) w (if present), or to (X) b (if present), or to D (wavy line on the left);

[0580] R 19 is selected from -C1-C 12 alkylene-, -O-(C1-C 12 alkylene), -C6-C x aryl, -C1-C 12 alkylene-C6-C 12 aryl- (wherein said aryl is a monocyclic or polycyclic ring, optionally substituted by one or more substituents R 12 ), -C6-C x aryl-C1-C 12 alkylene- (wherein said aryl is a monocyclic or polycyclic ring, optionally substituted by one or more substituents R 12 ), -C5-C x heterocyclyl- (wherein said heterocyclyl can be a saturated or unsaturated group having one or more rings and comprising at least one oxygen, nitrogen or sulfur atom in said one or more rings, said group being optionally substituted by one or more substituents R 12 ), -C1-C x alkylene- (C5-C 12 heterocyclyl- (wherein said heterocyclyl can be a saturated or unsaturated group having one or more rings and comprising at least one oxygen, nitrogen or sulfur atom in said one or more rings, said group being optionally substituted by one or more substituents R 12heterocyclyl)-(wherein said heterocyclyl can be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen or sulfur atom in said one or more rings, said group being optionally substituted by one or more substituents R x substituted), -(C5-C 12 substituted), -(C5-C 12 heterocyclyl)-(wherein said heterocyclyl can be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen or sulfur atom in said one or more rings, said group being optionally substituted by one or more substituents R x substituted), -(OCH2CH2) r - and -CH2-(OCH2CH2) r - wherein each of the above alkylene substituents, whether alone or attached to another moiety, the carbon chain can be optionally substituted by one or more substituents R x substituted;

[0581] R 30 is a -C1-C6 alkylene- group;

[0582] M is selected from the group consisting of -C1-C6 alkylene-, -C1-C6 alkylene-(C3-C8 carbocyclyl)- and phenylene optionally substituted by one or more substituents R x substituted; and

[0583] r is an integer ranging from 1 to 6.

[0584] • a pharmaceutical conjugate of Ab of formula [D-(X b -(AA) w -(T) g -(L)-] n selected from formulae (IV), (V) and (VI):

[0585]

[0586] wherein:

[0587] X and T are extension groups as defined herein;

[0588] each AA is independently an amino acid unit as defined herein;

[0589] w is an integer ranging from 0 to 12;

[0590] b is an integer of 0 or 1 ;

[0591] g is an integer of 0 or 1 ;

[0592] wherein b + g + w is optionally other than 0;

[0593] D is a drug moiety;

[0594] Ab is a moiety comprising at least one antigen binding site;

[0595] n is a group [D-(X) b -(AA) w -(T) g -(L)-] (where L is defined as in formula (IV), (V) or (VI)) to a moiety comprising at least one antigen binding site in a ratio and in the range of 1 to 20;

[0596] R 19 is selected from -Ci-C8alkylene-, -0-(Ci-C8alkylene), -Ci-C8alkylene-C6-C 12 arylene- (wherein said arylene is monocyclic or polycyclic, optionally substituted with one or more substituents R x ), and -C6-C 12 arylene-Ci-C8alkylene- (wherein said arylene is monocyclic or polycyclic, optionally substituted with one or more substituents R x ), wherein each of the above alkylene substituents, whether alone or connected to another moiety, is optionally substituted with one or more substituents R x ;

[0597] R 30 is a -C2-C4alkylene- group; and

[0598] M is selected from the group consisting of -Ci-C3alkylene- and -Ci-C3alkylene-(C5-C7carbocyclyl)-.

[0599] • a pharmaceutical conjugate of formula [D-(X) b -(AA) w -(T) g -(L)-] n -Ab selected from formula (IV), (V) and (VI):

[0600]

[0601] wherein:

[0602] X and T are extension groups which can be the same or different;

[0603] each AA is independently an amino acid unit;

[0604] w is an integer in the range of 0 to 12;

[0605] b is an integer of 0 or 1 ;

[0606] g is an integer of 0 or 1 ;

[0607] wherein b+g+w are optionally not 0;

[0608] D is a drug moiety;

[0609] Ab is a moiety comprising at least one antigen binding site;

[0610] n is a group [D-(X) b -(AA) w -(T) g -(L)-] (wherein L is as defined in formula (IV), (V) or (VI)) to a moiety comprising at least one antigen binding site in a ratio and in the range of 1 to 20;

[0611] R 19 is selected from -C1-C6alkylene-, phenylene-C1-C6alkylene- (wherein the phenylene is optionally substituted by one or more substituents R x selected from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, aryl having 6 to 12 carbon atoms, a halogen atom, nitro and cyano), wherein each of the above alkylene substituents, whether alone or in conjunction with another moiety in the carbon chain, is optionally substituted by one or more substituents R x selected from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, aryl having 6 to 12 carbon atoms, a halogen atom, nitro and cyano), wherein each of the above alkylene substituents, whether alone or in conjunction with another moiety in the carbon chain, is optionally substituted by one or more substituents R 19 is preferably a -C1-C6alkylene- group;

[0612] R 30 is a -C2-C4alkylene- group; and

[0613] M is -C1-C3alkylene-(C5-C7carbocyclyl)-.

[0614] • preferably, in the definition of the drug conjugate of formula [D-(X) b -(AA) w -(T) g -(L)-] n (AA) w has the formula (II):

[0615]

[0616] wherein the wavy line indicates the point of covalent attachment to (X) b (if present), or to a drug moiety (wavy line on the left) and to (T) g (if present), or to a linker (wavy line on the right); and

[0617] R21 at each occurrence is selected from the group consisting of hydrogen, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, -CH2OH, -CH(OH)CH3, -CH2CH2SCH3, -CH2CONH2, -CH2COOH, -CH2CH2CONH2, -CH2CH2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)3NH2, -(CH2)3NHCOCH3, -(CH2)3NHCHO, -(CH2)4NHC(=NH)NH2, -(CH2)4NH2, -(CH2)4NHCOCH3, -(CH2)4NHCHO, -(CH2)3NHCONH2, -(CH2)4NHCONH2, -CH2CH2CH(OH)CH2NH2, 2-pyridylmethyl-, 3-pyridylmethyl-, 4-pyridylmethyl-, phenyl, cyclohexyl,

[0618]

[0619] and w is an integer in the range of 0 to 12.

[0620] • a compound of formula [D-(X b -(AA) w -(T) g -(L)-] n a drug conjugate of -Ab, wherein L is defined as above in the preferred definition of said group and (AA) w of formula (II), wherein:

[0621] R 21 at each occurrence is selected from the group consisting of hydrogen, methyl, isopropyl, sec-butyl, benzyl, indolylmethyl, -(CH2)3NHCONH2, -(CH2)4NH2, -(CH2)3NHC(=NH)NH2, and -(CH2)4NHC(=NH)NH2; and

[0622] w is an integer in the range of 0 to 6.

[0623] • a compound of formula [D-(X b -(AA) w -(T) g -(L)-] n a drug conjugate of -Ab, wherein L is defined as above in the preferred definition of said group, wherein w is 0 or 2 and when w is 2, (AA) w of formula (III), wherein:

[0624]

[0625] wavy line indicates covalent attachment to (X) b (if present), or to a drug moiety (left wavy line) and to (T) g (if present), or to a linker (right wavy line);

[0626] R 22 is selected from the group consisting of methyl, benzyl, isopropyl, sec-butyl and indolylmethyl; and

[0627] R 23 is selected from the group consisting of methyl, -(CH2)4NH2, -(CH2)3NHCONH2 and -(CH2)3NHC(=NH)NH2.

[0628] • in embodiments of the application, b + g + w is not 0. In other embodiments, b + w is not 0. In other embodiments, b is 1 when w is not 0. Furthermore, it is preferred that in formula [D-(X) b -(AA) w -(T) g -(L)-] n in the definition of the drug conjugate of Ab, L and (AA) w as defined above in the preferred definition of the group and X is an extension group selected from the group consisting of:

[0629] in case of conjugation of D via an amino group:

[0630] -COO-(C1-C6alkylene)NH-;

[0631] -COO-CH2-(optionally substituted phenylene)-NH-; x -COO-CH2-(optionally substituted phenylene)-NH-;

[0632] -COO-CH2-(optionally substituted phenylene)-NH-; x -COO-CH2-(optionally substituted phenylene)-NH-;

[0633] -COCH2NH-COCH2-NH-;

[0634] -COCH2NH-;

[0635] -COO-(C1-C6alkylene)S-;

[0636] -COO-(C1-C6alkylene)S-;

[0637] in case of conjugation of D via a hydroxyl group:

[0638] -CONH-(C1-C6alkylene)NH-;

[0639] -COO-CH2-(optionally substituted by one or more substituents R x substituted phenylene)-NH-;

[0640] -COO-CH2-(optionally substituted by one or more substituents R x substituted phenylene)-NH-;

[0641] -COCH2NH-COCH2-NH-;

[0642] -COCH2NH-;

[0643] -CONH-(C1-C6alkylene)S-;

[0644] -CONH-(C1-C6alkylene)S-;

[0645] b is 0 or 1, preferably 1.

[0646] • a pharmaceutical conjugate of Ab, wherein L and (AA) b -(AA) w -(T) g -(L)-] n -Ab, wherein L and (AA) w as defined above in the preferred definition of the group and X is an extension group selected from the group consisting of:

[0647] wherein D is in the case of a conjugation via an amino group:

[0648] -COO-(C2-C4alkylene)NH-;

[0649] -COO-CH2-phenylene-NH-, wherein the phenylene is optionally substituted by one to four substituents R x substituted;

[0650] -COO-(C2-C4alkylene)NH-COO-CH2-(optionally substituted by one to four substituents R x substituted phenylene)-NH-;

[0651] -COCH2NH-COCH2-NH-;

[0652] -COO-(C2-C4alkylene)S-;

[0653] -COO-(C2-C4alkylene)NHCO(C1-C3alkylene)S-; or

[0654] in case D is conjugated via a hydroxyl group:

[0655] -CONH-(C2-C4alkylene)NH-;

[0656] -COO-CH2-phenylene-NH-, wherein the phenylene is optionally substituted by one to four substituents R selected from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, halogen atoms, nitro and cyano x substituted;

[0657] -CONH-(C2-C4alkylene)NH-COO-CH2-(phenylene optionally substituted by one to four substituents R selected from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, halogen atoms, nitro and cyano)-NH-; x substituted;

[0658] -COCH2NH-COCH2-NH-;

[0659] -CONH-(C2-C4alkylene)S-;

[0660] -CONH-(C2-C4alkylene)NHCO(C1-C3alkylene)S-; and

[0661] b is 0 or 1, preferably 1.

[0662] • a pharmaceutical conjugate of Ab, wherein L and (AA) b -(T) w -(T) g -(L)-] n -Ab, wherein L and (AA) w as defined above in the preferred definition of the group and X is an extension group selected from the group consisting of:

[0663] in case D is conjugated via an amino group:

[0664] -COO-CH2-phenylene-NH-

[0665] -COO(CH2)3NHCOOCH2-phenylene-NH-;

[0666] -COO(CH2)3NH-;

[0667] -COO(CH2)3-S-;

[0668] -COO(CH2)3NHCO(CH2)2S-; or

[0669] in case of a conjugation of D via a hydroxyl group:

[0670] -COO-CH2-phenylene-NH-

[0671] -CONH(CH2)3NHCOOCH2-phenylene-NH-;

[0672] -CONH(CH2)3NH-;

[0673] -CONH(CH2)3-S-;

[0674] -CONH(CH2)3NHCO(CH2)2S-; and b is 0 or 1, preferably 1.

[0675] • a pharmaceutical conjugate of the formula [D-(X b -(AA) w -(T) g -(L)-] n Ab, wherein L, (AA) w and (X) b as

[0676] are defined above in the preferred definitions of said groups and T is an extension group selected from the group consisting of:

[0677] -CO-(Ci-C6-alkylene)-NH-;

[0678] -CO-(Ci-C6-alkylene)-[O-(C2-C6-alkylene)] j -NH-;

[0679] -COO-(Ci-C6-alkylene)-[O-(C2-C6-alkylene)] j -NH-;

[0680] wherein j is an integer from 1 to 25 and g is 0 or 1.

[0681] • a pharmaceutical conjugate of the formula [D-(X b -(AA) w -(T) g -(L)-] n Ab, wherein L, (AA) w and (X) b as

[0682] are defined above in the preferred definitions of said groups and T is an extension group selected from the group consisting of:

[0683] -CO-(Ci-C4alkylene)NH-

[0684] -CO-(Ci-C4alkylene)-[O-(C2-C4alkylene)] j -NH-;

[0685] -COO-(Ci-C4alkylene)-[O-(C2-C4alkylene)] j -NH-;

[0686] wherein j is an integer from 1 to 10; and g is 0 or 1.

[0687] • a preferred pharmaceutical conjugate according to the application is a pharmaceutical conjugate of Ab, wherein L, (AA) b -(AA) w -(T) g -(L)-] n - Ab, wherein L, (AA) w and (X) b as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein

[0688] as defined above in the preferred definition of said group and T is an extension group selected from the group consisting of:

[0689] -CO-(Ci-C4alkylene)NH-

[0690] -CO-(Ci-C4alkylene)-[O-(C2-C4alkylene)] j -NH-;

[0691] -COO-(Ci-C4alkylene)-[O-(C2-C4alkylene)] j -NH-;

[0692] wherein j is an integer from 1 to 5; and g is 0 or 1.

[0693] • a preferred pharmaceutical conjugate according to the application is a pharmaceutical conjugate of Ab, wherein L, (AA) b -(AA) w -(T) g -(L)-] n - Ab, wherein L, (AA) w , (X) b , and (T) g as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein

[0694] R1is CN or OH, and more preferably R1is CN.

[0695] • a preferred pharmaceutical conjugate according to the application is a pharmaceutical conjugate of Ab, wherein L, (AA) b -(AA) w -(T) g -(L)-] n - Ab, wherein L, (AA) w , (X) b , and (T) g as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, whereinb -(AA) w -(T) g -(L)-] n Another preferred drug conjugate of Ab is a drug conjugate of the formula: wherein L, (AA) w , (X) b , and (T) g as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein R2is C(=O)R a wherein R a is selected from hydrogen and substituted or unsubstituted C1-C6alkyl, wherein the optional substituent is one or more substituents R x More preferably R2is acetyl.

[0696] • a compound of formula [D-(X) b -(AA) w -(T) g -(L)-] n Another preferred drug conjugate of Ab is a drug conjugate of the formula: wherein L, (AA) w , (X) b , and (T) g as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein R3is hydrogen or -OR b wherein R b is substituted or unsubstituted C1-C6alkyl, wherein the optional substituent is one or more substituents R x More preferably R3is hydrogen or methoxy. Most preferably R3is methoxy.

[0697] • a compound of formula [D-(X) b -(AA) w -(T) g -(L)-] n Another preferred drug conjugate of Ab is a drug conjugate of the formula: wherein L, (AA) w , (X) b , and (T) g as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein R4is selected from hydrogen, -CH2OH and -CH2NH2, more preferably R4is hydrogen or -CH2OH. Most preferably R4is hydrogen.

[0698] • a compound of formula [D-(X) b -(AA)w -(T) g -(L)-] n A further preferred drug conjugate of Ab is a drug conjugate of the following formula: wherein L, (AA) w , (X) b , and (T) g are as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein Y is -NH- or -O-.

[0699] • a compound of formula [D-(X) b -(AA) w -(T) g -(L)-] n A further preferred drug conjugate of Ab is a drug conjugate of the following formula: wherein L, (AA) w , (X) b , and (T) g are as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein:

[0700] R1is -CN or -OH;

[0701] R2is -C(=O)R a , wherein R a is selected from hydrogen and substituted or unsubstituted C1-C6alkyl, wherein the optional substituents are one or more substituents R

[0702] ; x ;

[0703] R3is hydrogen or -OR b , wherein R b is substituted or unsubstituted C1-C6alkyl, wherein the optional substituents are one or more substituents R

[0704] ; x ;

[0705] R4is hydrogen, -CH2OH or -CH2NH2; and

[0706] Y is -NH- or -O.

[0707] • a compound of formula [D-(X) b -(AA) w -(T) g -(L)-] n A further preferred drug conjugate of Ab is a drug conjugate of the following formula: wherein L, (AA) w , (X)b , and (T) g as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein:

[0708] R1is -CN or -OH;

[0709] R2is acetyl;

[0710] R3is hydrogen or methoxy, more preferably methoxy;

[0711] R4is hydrogen or -CH2OH; and

[0712] Y is -NH- or -O-.

[0713] • a further preferred drug conjugate of formula [D-(X) b -(AA) w -(T) g -(L)-] n -Ab according to the application is a drug conjugate of formula: wherein L, (AA) w , (X) b , and (T) g as defined above, wherein D is a compound of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein:

[0714] R1is -CN;

[0715] R2is acetyl:

[0716] R3is methoxy;

[0717] R4is hydrogen, and

[0718] Y is -NH- or -O-.

[0719] • a further preferred drug conjugate of formula [D-(X) b -(AA) w -(T) g -(L)-] n -Ab according to the application is a drug

[0720] conjugate of formula: wherein L, (AA) w , (X) b , and (T) g as defined above, wherein D is selected from:

[0721]

[0722] Or its pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer; wherein the wavy line indicates covalently linked to (X). b (If it exists), or (AA) w (If it exists), or to (T) g (If it exists), or a covalent connection point to L.

[0723] ●According to the formula [D-(X)] of the present invention b -(AA) w -(T) g -(L)-] n Another preferred drug conjugate for -Ab is the following drug conjugate, wherein L, (AA) w (X) b (T) g And D as defined above, and the portion Ab containing at least one antigen-binding site is an antigen-binding peptide.

[0724] ●According to the formula [D-(X)] of the present invention b -(AA) w -(T) g -(L)-] n Another preferred drug conjugate for -Ab is the following drug conjugate, wherein L, (AA) w (X) b (T) g And D as defined above, and a portion of Ab containing at least one antigen-binding site is an antibody, a single-domain antibody or its antigen-binding fragment.

[0725] ●According to the formula [D-(X)] of the present invention b -(AA) w -(T) g -(L)-] n Another preferred drug conjugate for -Ab is the following drug conjugate, wherein L, (AA) w (X) b (T) g And D, as defined above, includes at least one antigen-binding site in the portion of Ab that is a monoclonal, polyclonal, or bispecific antibody, wherein the antibody or its antigen-binding fragment is derived from any species, preferably human, mouse, or rabbit.

[0726] ●According to the formula [D-(X)] of the present invention b -(AA) w -(T) g -(L)-] n Another preferred drug conjugate for -Ab is the following drug conjugate, wherein L, (AA) w (X) b (T)g and D are as defined above, and the moiety Ab comprising at least one antigen binding site is an antibody or an antigen binding fragment thereof selected from the group consisting of a human antibody, an antigen binding fragment of a human antibody, a humanized antibody, an antigen binding fragment of a humanized antibody, a chimeric antibody, an antigen binding fragment of a chimeric antibody, a glycosylated antibody, and a glycosylated antigen binding fragment.

[0727] • another preferred drug conjugate of the formula [D-(X) b -(AA) w -(T) g -(L)-] n Ab according to the present application is a drug conjugate wherein L, (AA) w , (X) b , (T) g and D are as defined above, and the moiety Ab comprising at least one antigen binding site is an antibody or an antigen binding fragment thereof, wherein the antibody or antigen binding fragment thereof is an antigen binding fragment selected from the group consisting of a Fab fragment, a Fab' fragment, a F(ab')2 fragment, and a Fv fragment.

[0728] • another preferred drug conjugate of the formula [D-(X) b -(AA) w -(T) g -(L)-] n Ab according to the present application is a drug conjugate wherein L, (AA) w , (X) b , (T) g and D are as defined above, and the moiety Ab comprising at least one antigen binding site is an antibody or an antigen binding fragment thereof, wherein the antibody or antigen binding fragment thereof is a monoclonal antibody that immunospecifically binds to a cancer cell antigen, a viral antigen, an antigen of a cell producing an autoantibody associated with an autoimmune disease, a microbial antigen, and preferably a monoclonal antibody that immunospecifically binds to a cancer cell antigen.

[0729] • another preferred drug conjugate of the formula [D-(X) b -(AA) w -(T) g -(L)-] n Ab according to the present application is a drug conjugate wherein L, (AA) w , (X) b , (T) gand D is as defined herein, and the moiety Ab comprising at least one antigen binding site is an antibody selected from the group consisting of Alemtuzumab, Anetumab, Atezolizumab, Avelumab, Bevacizumab, Blinatomumab, Brentuximab, Catumaxomab, Cetuximab, Coltuximab, Daratumumab, Denintuzumab, Denosumab, Depatuxizumab, Dinutuximab, Durvalumab, Elotuzumab, Enfortumab, Glembatumumab, Gemtuzumab, Ibritumomab, Indatuximab, Indusatumab, Inotuzumab, Ipilimumab, Labetuzumab, Ladiratuzumab, Laprituximab, Lifastuzumab, Lorvotuzumab, Milatuzumab, Mirvetuximab, Naratuximab, Necitumumab, Nimotuzumab, Nivolumab, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Pertuzumab, Pinatuzumab, Polatuzumab, Ramucirumab, Rovalpituzumab, Sacituzumab, Siltuximab, Sirtratumab, Sofituzumab,Vadastuximab, Vorsetuzumab, an anti-HER2 antibody (e.g. Trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody and an anti-CD30 antibody or an antigen binding fragment or immunologically active portion thereof, wherein preferably the antibody is selected from the group consisting of Alemtuzumab, Anatumomab, Atacicept, Avonclix, Bevacizumab, Blontuvetumab, Brentuximab, Catumaxomab, Cetuximab, Daratumumab, Denintuzumab, Denosumab, Depatuxizumab, Dinutuximab, Durvalumab, Elotuzumab, Infliximab, Gavdorimab, Gemtuzumab, Ibritumomab, Indatuxizumab, Indusomab, Inozutumab, Ipilimumab, Labetuzumab, Labetuzumab, Labetuzumab, Milatuzumab, Nacolomab, Naxitamab, Nimotuzumab, Nivolumab, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Pertuzumab, Polatuzumab, Ramucirumab, Rilotumumab, Satumomab, Siltuximab, Sigumumab, Tremelimumab, Vadastuximab, Vorsetuzumab, an anti-HER2 antibody (e.g. Trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody and an anti-CD30 antibody or an antigen binding fragment or immunologically active portion thereof, and more preferably Alemtuzumab, Atacicept, Avonclix, Bevacizumab, Blontuvetumab, Brentuximab, Cetuximab, Daratumumab, Denosumab, Durvalumab, Elotuzumab, Gemtuzumab, Ibritumomab, Inozutumab, Ipilimumab, Labetuzumab, Nimotuzumab, Nivolumab, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Pertuzumab, Ramucirumab, Rilotumumab, Siltuximab, an anti-HER2 antibody (e.g. Trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody and an anti-CD30 antibody or an antigen binding fragment or immunologically active portion thereof. Especially preferred is Brentuximab, Gemtuzumab, Inozutumab, Rilotumumab, an anti-HER2 antibody (e.g. Trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody and an anti-CD30 antibody or an antigen binding fragment or immunologically active portion thereof; or the antibody is selected from the group consisting of an anti-HER2 antibody (e.g. Trastuzumab) and an anti-CD13 antibody or an antigen binding fragment or immunologically active portion thereof, especially Trastuzumab or an antigen binding fragment or immunologically active portion thereof.

[0730] • Formula [D-(X) b -(AA) w -(T) g -(L)-]n -Preferred drug conjugates of Ab include the following:

[0731] (a) A pharmaceutical conjugate according to the invention, wherein:

[0732] L can choose from the following groups:

[0733]

[0734] in:

[0735] The wavy line indicates a covalent connection to Ab (the wavy line on the right) and to (T). g (If it exists), or (AA) w (If it exists), or to (X) b (If it exists), or to point D (the wavy line on the left);

[0736] R 19 Selected from -C1-C 12 Alkylene-, -O-(C1-C 12 Alkylene, monocyclic or polycyclic, optionally substituted with one or more R groups. x Replacement -C6-C 12 Alpha-aryl, -C1-C 12 Alkylene-C6-C 12 arylene-(wherein the arylene group is monocyclic or polycyclic, and may be selectively replaced by one or more substituents R) x Replacement), -C6-C 12 Aspartic-C1-C 12 alkylene-(wherein the arylene group is monocyclic or polycyclic, and may be selectively substituted by one or more substituents R) x Replacement), -C5-C 12 Heterocyclic group - (wherein the heterocyclic group may be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, the group may be selectively substituented by one or more R groups) x Replacement), -C1-C 12 Alkylene-(C5-C 12 Heterocyclic group) - (wherein the heterocyclic group may be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, the group may be selectively substituented by one or more R groups. x Replace), -(C5-C 12 (heterocyclic group)-C1-C 12 Alkylene (wherein the heterocyclic group may be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, the group may be selectively substituented by one or more R groups) xsubstituted), -(OCH2CH2) r - and -CH2-(OCH2CH2) r - wherein each of the above alkylene substituents, whether alone or attached to another moiety, can optionally be substituted with one or more substituents R x ; and

[0737] R 30 is a -Ci-C6alkylene- group;

[0738] M is selected from the group consisting of -Ci-C6alkylene-, -Ci-C6alkylene-(C3-C8carbocyclyl)-, and phenylene optionally substituted with one or more substituents R x substituted phenylene;

[0739] r is an integer in the range of 1-6;

[0740] (AA) w having the formula (II):

[0741]

[0742] wherein the wavy line indicates covalent attachment to (X) b (if present), or to a drug moiety (left wavy line) and to (T) g (if present), or to a linker (right wavy line);

[0743] R 21 is selected from the group consisting of hydrogen, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, -CH2OH, -CH(OH)CH3, -CH2CH2SCH3, -CH2CONH2, -CH2COOH, -CH2CH2CONH2, -CH2CH2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)3NH2, -(CH2)3NHCOCH3, -(CH2)3NHCHO, -(CH2)4NHC(=NH)NH2, -(CH2)4NH2, -(CH2)4NHCOCH3, -(CH2)4NHCHO, -(CH2)3NHCONH2, -(CH2)4NHCONH2, -CH2CH2CH(OH)CH2NH2, 2-pyridylmethyl-, 3-pyridylmethyl-, 4-pyridylmethyl-, phenyl, cyclohexyl,

[0744]

[0745] w is an integer in the range of 0 to 12;

[0746] wherein X is an extension group selected from:

[0747] in case of conjugation of D via an amino group: -COO-(Ci-C6-alkylene)NH-, -COO-CH2- (optionally substituted with one or more substituents R x substituted phenylene)-NH-, -COO-(Ci-C6-alkylene)NH-COO-CH2- (optionally substituted with one or more substituents R x substituted phenylene)-NH-, -COCH2NH-COCH2-NH-, -COCH2-NH-, -COO-(Ci-C6-alkylene)S-, -COO-(Ci-C6-alkylene)NHCO(Ci-C6-alkylene)S-; or

[0748] in case of conjugation of D via a hydroxyl group: -CONH-(Ci-C6-alkylene)NH-, -COO-CH2- (optionally substituted with one or more substituents R x substituted phenylene)-NH-, -CONH-(Ci-C6-alkylene)NH-COO-CH2- (optionally substituted with one or more substituents R x substituted phenylene)-NH-, -COCH2NH-COCH2-NH-, -COCH2NH-, -CONH-(Ci-C6-alkylene)S-, and -CONH-(Ci-C6-alkylene)NHCO(Ci-C6-alkylene)S-;

[0749] b is 0 or 1, preferably 1 ;

[0750] wherein T is an extension group selected from -CO-(Ci-C6-alkylene)-NH-, -CO-(Ci-C6-alkylene)- [O-(C2-C6-alkylene)] j -NH-, and -COO-(Ci-C6-alkylene)-[O-(C2-C6-alkylene)] j -NH-, and -COO-(Ci-C6-alkylene)-[O-(C2-C6-alkylene)]

[0751] g is 0 or 1 ;

[0752] D is a drug moiety of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein:

[0753] R2is C(=O)R a , wherein R a is selected from the group consisting of hydrogen and substituted or unsubstituted Ci-C6-alkyl, wherein the optional substituents are one or more substituents R x ;

[0754] R3is hydrogen or -OR b , wherein R bis a substituted or unsubstituted C1-C6alkyl group, wherein the optional substituent is one or more substituents R x ;

[0755] R4is selected from the group consisting of hydrogen, -CH2OH, and -CH2NH2;

[0756] The moiety Ab comprising at least one antigen binding site is an antibody or an antigen binding fragment thereof, and is selected from the group consisting of a human antibody, an antigen binding fragment of a human antibody, a humanized antibody, an antigen binding fragment of a humanized antibody, a chimeric antibody, an antigen binding fragment of a chimeric antibody, a glycosylated antibody, and a glycosylated antigen binding fragment; and

[0757] n is a group [D-(X) b -(AA) w -(T) g -(L)-] in a ratio of 1 to 12 and in the range of 1 to 12.

[0758] (b) a drug conjugate according to the application selected from the group consisting of formulae (IV), (V) and (VI):

[0759]

[0760] wherein:

[0761] R 19 is selected from the group consisting of -C1-C8alkylene-, -O-(C1-C8alkylene), -C1-C8alkylene-C6-C 12 arylene- (wherein the arylene group is monocyclic or polycyclic, optionally substituted with one or more substituents R x ), and -C6-C 12 arylene-C1-C8alkylene- (wherein the arylene group is monocyclic or polycyclic, optionally substituted with one or more substituents R x ), wherein each alkylene substituent above, whether alone or connected to another moiety, is optionally substituted with one or more substituents R x ;

[0762] R 30 is a -C2-C4alkylene- group;

[0763] M is selected from the group consisting of -C1-C3alkylene- and -C1-C3alkylene-(C5-C7carbocyclyl)-;

[0764] (AA) w has the formula (II)

[0765]

[0766] wherein:

[0767] wavy line denotes covalent attachment to (X) b (if present), or to a drug moiety (left wavy line) and to (T) g (if present), or to a linker (right wavy line);

[0768] R 21 is selected from the group consisting of hydrogen, methyl, isopropyl, sec-butyl, benzyl, indolylmethyl, -(CH2)3NHCONH2, -(CH2)4NH2, -(CH2)3NHC(=NH)NH2, and -(CH2)4NHC(=NH)NH2 at each occurrence;

[0769] w is an integer from 0 to 6;

[0770] X is an extension group selected from the group consisting of:

[0771] in case of conjugation of D via an amino group: -COO-(C2-C4alkylene)NH-, -COO-CH2-phenylene-NH- (wherein the phenylene is optionally substituted by one to four substituents R x substituted from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, halogen atoms, nitro and cyano), -COO-(C2-C4alkylene)NH-COO-CH2- (optionally substituted by one to four substituents R x substituted from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, halogen atoms, nitro and cyano), -COO-(C2-C4alkylene)NH-COO-CH2- (optionally substituted by one to four substituents R

[0772] in case of conjugation of D via a hydroxyl group: -CONH-(C2-C4alkylene)NH-, -COO-CH2-phenylene-NH- (wherein the phenylene is optionally substituted by one to four substituents R x substituted from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, halogen atoms, nitro and cyano), -CONH-(C2-C4alkylene)NH-COO-CH2- (optionally substituted by one to four substituents R xSubstituted phenylene)-NH-, -COCH2NH-COCH2-NH-, -CONH-(C2-C4 alkylene)S- and -CONH-(C2-C4 alkylene)NHCO(C1-C3 alkylene)S-;

[0773] b is 0 or 1, preferably 1;

[0774] Where T is an extended group selected from the following: -CO-(C1-C4 alkylene)-NH-, -CO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH- and -COO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH-, where j is an integer from 1 to 10;

[0775] g is 0 or 1;

[0776] D is the pharmaceutical part of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer thereof, wherein:

[0777] R2 is an acetyl group;

[0778] R3 is hydrogen or methoxy, preferably R3 is methoxy;

[0779] R4 is hydrogen or -CH2OH, preferably R4 is hydrogen;

[0780] A portion of the Ab containing at least one antigen-binding site is an antibody or its antigen-binding fragment, wherein the antibody or antigen-binding fragment is a monoclonal antibody that specifically binds to cancer cell antigens, viral antigens, antigens of cells that produce autoimmune antibodies associated with autoimmune diseases, or microbial antigens, and is preferably a monoclonal antibody that specifically binds to cancer cell antigens; and

[0781] n represents the group [D-(X)] b -(AA) w -(T) g The ratio of -(L)-] (where L is as defined in formula (IV), (V) or (VI)) to a portion of Ab containing at least one antigen binding site and is in the range of 3 to 8.

[0782] (c) The pharmaceutical conjugates selected from formulas (IV), (V) and (VI) according to the present invention:

[0783]

[0784] in:

[0785] R 19selected from -C1-C6alkylene-, -phenylene-C1-C6alkylene- (wherein the phenylene is optionally substituted with one or more substituents R selected from the group consisting of alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, a halogen atom, a nitro group and a cyano group) x wherein each alkylene substituent above, whether alone or in conjunction with another moiety in the carbon chain, is optionally substituted with one or more substituents R selected from the group consisting of alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, aryl of 6 to 12 carbon atoms, a halogen atom, a nitro group and a cyano group x and R 19 is preferably a C1-C6alkylene group;

[0786] R 30 is a -C2-C4alkylene- group;

[0787] M is -C1-C3alkylene-(C5-C7carbocyclyl)-;

[0788] w is 0 or 2 and in the case where w is 2, (AA) w of formula (III):

[0789]

[0790] wherein the wavy line indicates covalent attachment to (X) b (if present), or to a drug moiety (wavy line on the left) and to (T) g (if present), or to a linker (wavy line on the right);

[0791] R 22 is selected from methyl, benzyl, isopropyl, sec-butyl and indolylmethyl;

[0792] R 23 is selected from methyl, -(CH2)4NH2, -(CH2)3NHCONH2 and -(CH2)3NHC(=NH)NH2;

[0793] X is an extension group selected from the group consisting of -COO-(C2-C4alkylene)NH-, -COO-CH2-phenylene-NH- (wherein the phenylene is optionally substituted with one to four substituents R selected from the group consisting of alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, a halogen atom, a nitro group and a cyano group) x -COO-CH2- (optionally substituted with one to four substituents R selected from the group consisting of alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, a halogen atom, a nitro group or a cyano group) xSubstituted phenylene)-NH-, -COCH2NH-COCH2-NH-, -COO-(C2-C4 alkylene)S- and -COO-(C2-C4 alkylene)NHCO(C1-C3 alkylene)S-;

[0794] b is 0 or 1, preferably 1;

[0795] Where T is an extended group selected from the following: -CO-(C1-C4 alkylene)-NH-, -CO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH- and -COO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH-, where j is an integer from 1 to 5;

[0796] g is 0 or 1;

[0797] D is the pharmaceutical part of formula (I) or (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer thereof, wherein:

[0798] R1 is CN;

[0799] R2 is an acetyl group:

[0800] R3 is a methoxy group;

[0801] R4 is hydrogen;

[0802] Y is either -NH- or -O-;

[0803] The moiety Ab comprising at least one antigen binding site is a monoclonal antibody selected from the group consisting of alemtuzumab, anatumomab mafenatox, atezolizumab, avelumab, bevacizumab, blinatumomab, brentuximab vedotin, catumaxomab, cetuximab, coltuximab, daratumumab, denintuzumab mafodotin, denosumab, depatuxizumab mafodotin, dinutuximab, durvalumab, elotuzumab, inotuzumab, gavdorimab, gemtuzumab, ibritumomab tiuxetan, indatuxizumab ivitux, inebilizumab, lintuzumab, lifastuzumab, lilotomab, mirikizumab, milatuzumab, naptumomab, necitumumab, nimotuzumab, nivolumab, obinutuzumab, ofatumumab, olaratumab, panitumumab, pembrolizumab, patritumab, polatuzumab, ramucirumab, rovalpituzumab, sacituzumab, secolizumab, situximab, sofituzumab, vadastuximab, vesatuximab, anti-HER2 antibodies (e.g. trastuzumab), anti-CD4 antibodies, anti-CD5 antibodies, anti-CD13 antibodies and anti-CD30 antibodies or an antigen binding fragment or immunologically active portion thereof, wherein the antibody is preferably selected from alemtuzumab, anatumomab mafenatox, atezolizumab, avelumab, bevacizumab, blinatumomab, brentuximab vedotin, catumaxomab, cetuximab, daratumumab, denintuzumab mafodotin, denosumab, depatuxizumab mafodotin, dinutuximab, durvalumab, elotuzumab, inotuzumab, gavdorimab, gemtuzumab, ibritumomab tiuxetan, indatuxizumab ivitux, inebilizumab, lintuzumab, lifastuzumab, lilotomab, mirikizumab, milatuzumab, naptumomab, necitumumab, nimotuzumab, nivolumab, obinutuzumab, ofatumumab, olaratumab, panitumumab, pembrolizumab, patritumab, polatuzumab, ramucirumab, rovalpituzumab, sacituzumab, secolizumab, situximab, sofituzumab, vadastuximab, vesatuximab, anti-HER2 antibodies (e.g. trastuzumab), anti-CD4 antibodies, anti-CD5 antibodies, anti-CD13 antibodies and anti-CD30 antibodies or an antigen binding fragment or immunologically active portion thereof,and more preferably is alemtuzumab, atezolizumab, avelumab, bevacizumab, blinatumomab, brentuximab vedotin, catumaxomab, cetuximab, daratumumab, denosumab, dinutuximab, durvalumab, elotuzumab, gemtuzumab, ibritumomab tiuxetan, inotuzumab ozagamicin, ipilimumab, labetuzumab, necitumumab, nimotuzumab, nivolumab, obinutuzumab, ofatumumab, olaratumab, panitumumab, pembrolizumab, pertuzumab, ramucirumab, rovalpituzumab, siltuximab, anti-HER2 antibodies (e.g. trastuzumab), anti-CD4 antibodies, anti-CD5 antibodies, anti-CD13 antibodies and anti-CD30 antibodies or antigen-binding fragments or immunologically active portions thereof. Among these, especially preferred are brentuximab vedotin, gemtuzumab ozogamicin, inotuzumab ozagamicin, rovalpituzumab, anti-HER2 antibodies (e.g. trastuzumab), anti-CD4 antibodies, anti-CD5 antibodies, anti-CD13 antibodies and anti-CD30 antibodies, or antigen-binding fragments or immunologically active portions thereof; or the antibody is selected from anti-HER2 antibodies (e.g. trastuzumab) and anti-CD13 antibodies or antigen-binding fragments or immunologically active portions thereof, especially trastuzumab or antigen-binding fragments or immunologically active portions thereof; and,

[0804] n is a group [D-(X) b -(AA) w -(T) g -(L)-] (wherein L is defined as in formula (IV), (V) or (VI)) to a moiety Ab comprising at least one antigen binding site in a ratio and in the range of 3 to 5.

[0805] (d) a drug conjugate according to the application selected from formulae (IV), (V) and (VI):

[0806]

[0807] wherein:

[0808] R 19 is -C2-C6alkylene-;

[0809] R 30 is -C2-C4alkylene-;

[0810] M is -C1-C3alkylene-(C5-C7carbocyclyl)-;

[0811] w is 0 or 2 and in case w is 2, (AA) w has formula (III):

[0812]

[0813] wherein R 22 is isopropyl, R23 is selected from the group consisting of methyl and -(CH2)3NHCONH2, wherein the wavy line indicates a covalent linkage to (X) b (if present), or to a drug moiety (wavy line on the left) and to (T) g (if present), or to a linker (wavy line on the right);

[0814] X is an extension group selected from the group consisting of -COO-(C2-C4alkylene)NH-, -COO-CH2-phenylene-NH- (wherein the phenylene is optionally substituted with one to four substituents R selected from the group consisting of alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, halogen atoms, nitro and cyano), -COO-(C2-C4alkylene)NH-COO-CH2- (optionally substituted phenylene)-NH-, -COCH2NH-COCH2-NH-, -COO-(C2-C4alkylene)S-, and -COO-(C2-C4alkylene)NHCO(C1-C3alkylene)S-; x substituted phenylene)-NH-, -COCH2NH-COCH2-NH-, -COO-(C2-C4alkylene)S-, and -COO-(C2-C4alkylene)NHCO(C1-C3alkylene)S-; x substituted phenylene)-NH-, -COCH2NH-COCH2-NH-, -COO-(C2-C4alkylene)S-, and -COO-(C2-C4alkylene)NHCO(C1-C3alkylene)S-;

[0815] b is 0 or 1, preferably 1; wherein T is an extension group selected from -CO-(C1-C4alkylene)-NH-, -CO-(C1-C4alkylene)-[O-(C2-C4alkylene) j -NH- and -COO-(C1-C4alkylene)-[O-(C2-C4alkylene) j -NH-, wherein j is an integer from 1 to 5;

[0816] g is 0 or 1;

[0817] D is a drug moiety selected from the group consisting of:

[0818]

[0819] or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof; wherein the wavy line indicates a covalent linkage to (X) b (if present), or (AA) w (if present), or to (T) g (if present), or to (L);

[0820] The moiety Ab comprising at least one antigen binding site is selected from brentuximab, gemtuzumab, inotuzumab, lorvotuzumab, an anti-HER2 antibody (e.g. trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody and an anti-CD30 antibody, or an antigen binding fragment or immunologically active portion thereof, and more preferably it is selected from an anti-HER2 antibody (e.g. trastuzumab) and an anti-CD13 antibody, or an antigen binding fragment or immunologically active portion thereof, in particular trastuzumab or an antigen binding fragment or immunologically active portion thereof; and

[0821] n is a group [D-(X) b -(AA) w -(T) g -(L)-] (wherein L is as defined in formula (IV), (V) or (VI)) to the moiety Ab comprising at least one antigen binding site is in the range of 3 to 5.

[0822] (e) a pharmaceutical conjugate according to the application selected from formulae (IV), (V) and (VI):

[0823]

[0824] wherein:

[0825] R 19 is -C2-C6alkylene-;

[0826] R 30 is -C2-C4alkylene-;

[0827] M is -C1-C3alkylene-(C5-C7carbocyclyl)-;

[0828] w is 0 or 2 and in case w is 2, (AA) w has the formula (III):

[0829]

[0830] wherein R 22 is isopropyl, R 23 is selected from methyl and -(CH2)3NHCONH2, and the wavy line indicates the point of covalent attachment to (X) b (if present), or to the drug moiety (wavy line on the left) and to (T)g (if present), or to the linker (wavy line on the right);

[0831] X is an extension group selected from the group consisting of -COO-(C2-C4alkylene)NH-, -COO-CH2-phenylene-NH- (wherein the phenylene is optionally substituted by one to four substituents R selected from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, halogen atoms, nitro and cyano x -COO-(C2-C4alkylene)NH-COO-CH2- (optionally substituted by one to four substituents R selected from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, halogen atoms, nitro and cyano x -COO-(C2-C4alkylene)NH-COO-CH2- (optionally substituted by one to four substituents R selected from the group consisting of alkyl having 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms, halogen atoms, nitro and cyano

[0832] b is 0 or 1, preferably 1 ;

[0833] wherein T is an extension group selected from -CO-(C1-C4alkylene)-NH-, -CO-(C1-C4alkylene)-[O-(C2-C4alkylene) j -NH- and -COO-(C1-C4alkylene)-[O-(C2-C4alkylene) j -NH-, wherein j is an integer from 1 to 5;

[0834] g is 0 or 1 ;

[0835] D is a drug moiety selected from

[0836]

[0837] or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof; wherein the wavy line indicates the point of covalent attachment to (X) b (if present), or (AA) w (if present), or to (T) g (if present), or to (L)

[0838] The moiety Ab comprising at least one antigen binding site is selected from brentuximab vedotin, gemtuzumab ozogamicin, inotuzumab ozogamicin, lorvotuzumab mertansine, an anti-HER2 antibody (e.g. trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody and an anti-CD30 antibody, or an antigen binding fragment or immunologically active portion thereof, and more preferably it is selected from an anti-HER2 antibody (e.g. trastuzumab) and an anti-CD13 antibody, or an antigen binding fragment or immunologically active portion thereof, in particular trastuzumab or an antigen binding fragment or immunologically active portion thereof; and

[0839] n is a group [D-(X) b -(AA) w -(T) g -(L)-] (where L is defined as in formula (IV), (V) or (VI)) to a moiety comprising at least one antigen binding site in a ratio and in the range of 3 to 5.

[0840] (f) a drug conjugate of formula (IV) according to the application:

[0841]

[0842] wherein:

[0843] R 19 is C2-C5alkylene-;

[0844] w is 0 or 2 and in case w is 2, (AA) w of formula (III):

[0845]

[0846] wherein R 22 is isopropyl, R 23 is selected from methyl and -(CH2)3NHCONH2, and the wavy line indicates the point of covalent attachment to (X) b (the left wavy line) and to (T) g (if present), or to a linker (the right wavy line); and

[0847] X is a -COOCH2-phenylene-NH- group;

[0848] b is 1 ;

[0849] T is a spacer group of formula -CO-(Ci-C4alkylene)-[O-(C2-C4alkylene)]4-NH-;

[0850] g is 0 or 1 ;

[0851] or of formula (V)

[0852]

[0853] wherein M is -methyl-cyclohexylidene-;

[0854] b is 1 ;

[0855] w is 0;

[0856] X is a spacer group selected from -(CH2)3S- and -(CH2)3NHCO(CH2)2S-

[0857] g is 0;

[0858] or having formula (VI)

[0859]

[0860] wherein R 19 is -C2-C5alkylene-;

[0861] R 30 is -C3alkylene-;

[0862] w is 0 or 2 and in case w is 2, (AA) w has formula (III):

[0863]

[0864] wherein R 22 is isopropyl, R 23 is selected from methyl and -(CH2)3NHCONH2, and the wavy line indicates the point of covalent attachment to (X) b (the left wavy line) and to (T) g (if present), or to a linker (the right wavy line); and

[0865] X is a -COOCH2-phenylene-NH- group;

[0866] b is 1 ;

[0867] T is an extension group of formula -CO-(Ci-C4alkylene)-[O-(C2-C4alkylene)]4-NH-;

[0868] g is 0 or 1 ;

[0869] D is a drug moiety selected from the group consisting of:

[0870]

[0871] or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof; wherein the wavy line indicates the point of covalent attachment to (X) b ;

[0872] The moiety Ab comprising at least one antigen binding site is brentuximab, gemtuzumab, inotuzumab, lorvotuzumab, an anti-HER2 antibody (e.g. trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody and an anti-CD30 antibody, or an antigen binding fragment or immunologically active portion thereof, and more preferably it is selected from an anti-HER2 antibody (e.g. trastuzumab) and an anti-CD13 antibody, or an antigen binding fragment or immunologically active portion thereof, in particular trastuzumab or an antigen binding fragment or immunologically active portion thereof; and

[0873] n is a group [D-(X) b -(AA) w -(T) g -(L)-] (wherein L is defined as in formula (IV)) to a moiety Ab comprising at least one antigen binding site in a ratio in the range of 3 to 5 and preferably 4.

[0874] g) an antibody drug conjugate according to the application selected from the group consisting of:

[0875] and

[0876]

[0877] wherein n is 2 to 6, more preferably 3, 4 or 5, and each is independently selected from brentuximab vedotin, gemtuzumab ozogamicin, inotuzumab ozagamicin, lorvotuzumab mertansine, an anti-HER2 antibody (e.g. trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody and an anti-CD30 antibody, or an antigen binding fragment or immunologically active moiety thereof, and more preferably it is selected from an anti-HER2 antibody (e.g. trastuzumab) and an anti-CD13 antibody, or an antigen binding fragment or immunologically active moiety thereof, in particular trastuzumab or an antigen binding fragment or immunologically active moiety thereof.

[0878] More preferably, the antibody drug conjugate is selected from the group consisting of:

[0879]

[0880] wherein n is 2 to 6, more preferably 3, 4 or 5, and is selected from an anti-HER2 antibody (e.g. trastuzumab) and an anti-CD13 antibody, or an antigen binding fragment or immunologically active moiety thereof, more preferably it is trastuzumab or an antigen binding fragment or immunologically active moiety thereof,

[0881]

[0882] wherein n is 2 to 6, more preferably 3, 4 or 5, and is selected from an anti-HER2 antibody (e.g. trastuzumab) and an anti-CD13 antibody, or an antigen binding fragment or immunologically active moiety thereof, more preferably it is trastuzumab or an antigen binding fragment or immunologically active moiety thereof,

[0883]

[0884] wherein n is 2 to 6, more preferably 3, 4 or 5, and selected from the group consisting of an anti-HER2 antibody (e.g. trastuzumab) and an anti-CD13 antibody or an antigen-binding fragment or immunologically active portion thereof, more preferably trastuzumab or an antigen-binding fragment or immunologically active portion thereof,

[0885]

[0886] wherein n is 2 to 6, more preferably 3, 4 or 5, and selected from the group consisting of an anti-HER2 antibody (e.g. trastuzumab) and an anti-CD13 antibody or an antigen-binding fragment or immunologically active portion thereof, more preferably trastuzumab or an antigen-binding fragment or immunologically active portion thereof.

[0887] It is especially preferred that the antibody drug conjugate according to the present application is in isolated or purified form.

[0888] the formula D-(X) according to the present application b -(AA) w -(T) g -L1or the formula D-(X) b -(AA) w -(T) g -H, wherein D, X, AA, T, L1,

[0889] • the formula D-(X) b -(AA) w -(T) g -L1or the formula D-(X) b -(AA) w -(T) g -H, wherein D, X, AA, T, L1,

[0890] b, g and w are each as defined herein; but further, if the compound is of the formula D-(X) b -(AA) w -(T) g -H, then b + w + g ≠ 0.

[0891] • the formula D-(X) according to the present application b -(AA) w -(T) g -L1or the formula D-(X) b -(AA) w -(T) g -H, wherein:

[0892] L1is a linker of the formula:

[0893]

[0894] wherein:

[0895] The wavy line indicates a covalent connection to (T). g (If it exists), or (AA) w (If it exists), or to (X) b (If it exists), or a point to D;

[0896] R 19 Selected from -C1-C 12 Alkylene-, -O-(C1-C 12 Alkylene, monocyclic or polycyclic, optionally substituted with one or more R groups. x Replacement -C6-C 12 Alpha-aryl, -C1-C 12 Alkylene-C6-C 12 arylene-(wherein the arylene group is monocyclic or polycyclic, and may be selectively replaced by one or more substituents R) x Replacement), -C6-C 12 Aspartic-C1-C 12 alkylene-(wherein the arylene group is monocyclic or polycyclic, and may be selectively substituted by one or more substituents R) x Replacement), -C5-C 12 Heterocyclic group - (wherein the heterocyclic group may be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, the group may be selectively substituented by one or more R groups) x Replacement), -C1-C 12 Alkylene-(C5-C 12 Heterocyclic group) - (wherein the heterocyclic group may be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, the group may be selectively substituented by one or more R groups. x Replace), -(C5-C 12 (heterocyclic group)-C1-C 12 Alkylene (wherein the heterocyclic group may be a saturated or unsaturated group having one or more rings and containing at least one oxygen, nitrogen, or sulfur atom in the one or more rings, the group may be selectively substituented by one or more R groups) x (substitute), -(OCH2CH2) r - and -CH2-(OCH2CH2) r - wherein, whether each of the above alkylene substituents is alone or attached to another part, the carbon chain can be selectively substituented by one or more R groups. x replace;

[0897] r is an integer in the range 1-6; and

[0898] D, R x, X, AA, T, b, g and w are each as defined in the application; but wherein if the compound is of formula D-(X) b -(AA) w -(T) g -H, then b + w + g ≠ 0.

[0899] • a compound of formula D-(X) b -(AA) w -(T) g -L1or formula D-(X) b -(AA) w -(T) g -H, wherein:

[0900] L1is a linker of formula:

[0901]

[0902] wherein:

[0903] the wavy line indicates covalent attachment to (T) g (if present), or (AA) w (if present), or to (X) b (if present), or to D;

[0904] R 19 is selected from -Ci-C8alkylene-, -0-(Ci-C8alkylene), -Ci-C8alkylene-C6-C 12 arylene- (wherein the arylene group is monocyclic or polycyclic, optionally substituted with one or more substituents R x ), and -C6-C 12 arylene-Ci-C8alkylene- (wherein the arylene group is monocyclic or polycyclic, optionally substituted with one or more substituents R x ), wherein each of the above alkylene substituents, whether alone or connected to another moiety, is optionally substituted with one or more substituents R x ;

[0905] (AA) w has formula (II):

[0906]

[0907] wherein the wavy line indicates covalent attachment to (X) b (if present), or to D (wavy line on the left) and to (T) g (if present), or L1or to a hydrogen atom (wavy line on the right);

[0908] wherein R 21Each time it appears, choose from the following groups: hydrogen, methyl, isopropyl, sec-butyl, benzyl, indolemethyl, -(CH2)3NHCONH2, -(CH2)4NH2, -(CH2)3NHC(=NH)NH2 and -(CH2)4NHC-(=NH)NH2, where w is an integer from 0 to 6;

[0909] X is an extension group selected from the group consisting of:

[0910] In the case of D via amino conjugation: -COO-(C2-C4 alkylene)NH-, -COO-CH2-phenylene-NH (wherein the phenylene may be selectively replaced by one to four substituents R selected from the group consisting of an alkyl group having 1 to 6 carbon atoms, an alkoxy group having 1 to 6 carbon atoms, a halogen atom, a nitro group, and a cyano group). x Substitution), -COO-(C2-C4 alkylene)NH-COO-CH2- (optionally substituents R selected from one to four groups consisting of an alkyl group having 1 to 6 carbon atoms, an alkoxy group having 1 to 6 carbon atoms, a halogen atom, a nitro group, and a cyano group). x Substituted phenylene)-NH-, -COCH2NH-COCH2-NH-, -COO-(C2-C4 alkylene)S- and -COO-(C2-C4 alkylene)-NHCO(C1-C3 alkylene)S- or

[0911] In the case of D via hydroxyl conjugation: -CONH-(C2-C4 alkylene)NH-, -COO-CH2-phenylene-NH- (wherein the phenylene may be selectively replaced by one to four substituents R selected from the group consisting of an alkyl group having 1 to 6 carbon atoms, an alkoxy group having 1 to 6 carbon atoms, a halogen atom, a nitro group, and a cyano group). x Substitution), -CONH-(C2-C4 alkylene)NH-COO-CH2- (optionally substituents R selected from one to four groups consisting of an alkyl group having 1 to 6 carbon atoms, an alkoxy group having 1 to 6 carbon atoms, a halogen atom, a nitro group, and a cyano group). x Substituted phenylene)-NH-, -COCH2NH-COCH2-NH-, -CONH-(C2-C4 alkylene)S- and -CONH-(C2-C4 alkylene)NHCO(C1-C3 alkylene)S-;

[0912] T is selected from the following extended groups: -CO-(C1-C4 alkylene)-NH-, -CO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH- and -COO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH-, where j is an integer from 1 to 10;

[0913] b is 0 or 1 ;

[0914] g is 0 or 1 ;

[0915] wherein if the compound is of formula D-(X) b -(AA) w -(T) g -H, then b + w + g ≠ 0; and

[0916] D is a drug moiety of formula (I); and is covalently linked via a hydroxyl or amino group; or

[0917] is a drug moiety of formula (IH), or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof, wherein:

[0918]

[0919] wherein the wavy line of (IH) indicates covalent linkage to (X) b (if present), or (AA) w (if present), or to (T) g (if present), or to the point of L1;

[0920] R1is -OH or -CN;

[0921] R2is a -C(=O)R a group, wherein R a is selected from hydrogen and substituted or unsubstituted C1-C6 alkyl, wherein the optional substituent is one or more substituents R x .

[0922] R3is hydrogen or -OR b group, wherein R b is substituted or unsubstituted C1-C6 alkyl, wherein the optional substituent is one or more substituents R x .

[0923] R4is selected from hydrogen, -CH2OH and -CH2NH2; and

[0924] Y is -NH- or -O-.

[0925] • a compound of formula D-(X) b -(AA) w -(T) g -L1or formula D-(X) b -(AA) w -(T) g -H, wherein:

[0926] L1is a group of the formula:

[0927]

[0928] wherein:

[0929] the wavy line denotes a covalent attachment to (T) g (if present), or (AA) w (if present), or to (X) b (if present), or to D;

[0930] R 19 is selected from -Ci-C6alkylene-, phenylene-Ci-C6alkylene- (wherein the phenylene group is optionally substituted by one or more substituents R x substituents R x substituents R 19 is preferably Ci-C6alkylene;

[0931] w is 0 or 2 and in case w is 2, (AA) w has the formula (III):

[0932]

[0933] wherein the wavy line denotes a covalent attachment to X (wavy line on the left) and to (T) g (if present), or L1or to a hydrogen atom (wavy line on the right);

[0934] R 22 is selected from methyl, benzyl, isopropyl, sec-butyl and indolylmethyl;

[0935] R 23 is selected from methyl, -(CH2)4NH2, -(CH2)3NHCONH2and -(CH2)3NHC(=NH)NH2;

[0936] X is an extension group selected from:

[0937] in case D is conjugated via an amino group: -COO-CH2-phenylene-NH-, -COO(CH2)3NHCOO-CH2-phenylene-NH, -COO-(CH2)3)NH-, -COO(CH2)3-S- and -COO-(CH2)3NHCO-(CH2)2S- or

[0938] In the case of D via hydroxyl conjugation: -COO-CH2-phenylene-NH-, -CONH(CH2)3NHCOOCH2-phenylene-NH-, -CONH(CH2)3NH-, -CONH(CH2)3-S- and -CONH(CH2)3NHCO(CH2)2S-.

[0939] Where T is an extended group selected from the following: -CO-(C1-C4 alkylene)-NH-, -CO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH- and -COO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH-, where j is an integer from 1 to 5;

[0940] b is 0 or 1;

[0941] g is 0 or 1;

[0942] Where the compound is of formula D-(X) b -(AA) w -(T) g For compounds with -H, b + w + g ≠ 0; and

[0943] D is the pharmaceutical part of formula (I); and is covalently linked via a hydroxyl or amino group; or

[0944] The pharmaceutical moiety of formula (IH), or its pharmaceutically acceptable salt, ester, solvate, tautomer, or stereoisomer:

[0945]

[0946] The wavy line (IH) represents a covalent connection point;

[0947] R1 is -CN or -OH;

[0948] R2 is an acetyl group;

[0949] R3 is hydrogen or methoxy, preferably methoxy;

[0950] R4 is hydrogen or -CH2OH, preferably hydrogen;

[0951] Y can be either -NH- or -O-.

[0952] ●According to formula D-(X) of the present invention b -(AA) w -(T) g -L1 or formula D-(X) b -(AA) w -(T)g a compound of formula (H), wherein:

[0953] L1is a linker of formula:

[0954]

[0955] wherein:

[0956] the wavy line indicates a covalent linkage to (T) g (AA), if present, or (X) w (AA), if present, or to (T) b (AA), if present, or to D;

[0957] R 19 is -C2-C6alkylene-;

[0958] w is 0 or 2 and in case w is 2, (AA) w of formula (III):

[0959]

[0960] R 22 is isopropyl, R 23 is selected from methyl and -(CH2)3NHCONH2, wherein the wavy line indicates a covalent linkage to X (wavy line on the left) and to (T) g (AA), if present, or L1or a point to a hydrogen atom (wavy line on the right);

[0961] X is an extension group selected from -COO-CH2-phenylene-NH-, -COO(CH2)3NHCOO-CH2-phenylene-NH, -COO-(CH2)3)NH-, -COO(CH2)3-S- and -COO-(CH2)3NHCO-(CH2)2S-;

[0962] wherein T is an extension group selected from -CO-(C1-C4alkylene)-NH-, -CO-(C1-C4alkylene)-[O-(C2-C4alkylene) j -NH- and -COO-(C1-C4alkylene)-[O-(C2-C4alkylene) j -NH-, wherein j is an integer from 1 to 5;

[0963] b is 0 or 1 ;

[0964] g is 0 or 1 ;

[0965] wherein if the compound is of formula D-(X) b (AA) w (T) gcompounds of formula (I) wherein:

[0966] D is a drug moiety selected from the group consisting of:

[0967]

[0968] or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof; wherein the wavy line indicates the point of covalent attachment.

[0969] • a compound of formula D-(X) b -(AA) w -(T) g -L1or a compound of formula D-(X) b -(AA) w -(T) g -H, wherein:

[0970] L1is a group of formula:

[0971]

[0972] wherein:

[0973] the wavy line indicates the point of covalent attachment to (T) g (if present), or (AA) w (if present), or to (X) b (if present), or to D;

[0974] R 19 is -C2-C5alkylene-;

[0975] w is 0 or 2 and in case w is 2, (AA) w has the formula (III):

[0976]

[0977] wherein R 22 is isopropyl, R 23 is selected from the group consisting of methyl and -(CH2)3NHCONH2, wherein the wavy line indicates the point of covalent attachment to X (wavy line on the left) and to (T) g (if present), or L1or to a hydrogen atom (wavy line on the right);

[0978] X is a -COO-CH2-phenylene-NH- group;

[0979] T is a -CO-(CH2)2-[O-(CH2)2]4-NH- group;

[0980] b is 0 or 1 ;

[0981] g is 0 or 1 ;

[0982] wherein if the compound is of formula D-(X) b -(AA) w -(T) g -H, then b + w + g ≠ 0; and

[0983] D is a drug moiety selected from the group consisting of:

[0984]

[0985] or a pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer thereof; wherein the wavy line indicates the point of covalent attachment.

[0986] • a compound of formula D-X-(AA) w -(T) g -L1:

[0987]

[0988] The term "pharmaceutically acceptable salt, ester, solvate, tautomer or stereoisomer" in the context of the drug conjugates of the present application means any pharmaceutically acceptable salt, ester, solvate, hydrate or stereoisomeric form or any other compound which, upon administration to a patient, is capable of providing, directly or indirectly, a compound as described herein. However, it will be appreciated that non-pharmaceutically acceptable salts are also within the scope of the present application, since such salts can be useful in the preparation of pharmaceutically acceptable salts. The preparation of salts, prodrugs and derivatives can be carried out by methods known in the art.

[0989] For example, pharmaceutically acceptable salts of the compounds provided herein are synthesized from the parent compound that contains a basic or acidic moiety by conventional chemical methods. Generally, such salts are prepared either by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two. Generally, non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Examples of the acid addition salts include mineral acid salts such as hydrochlorides, hydrobromides, hydroiodides, sulfates, nitrates, phosphates, and organic acid salts such as acetates, trifluoroacetates, maleates, fumarates, citrates, oxalates, succinates, tartrates, malates, mandelates, methanesulfonates, and p-toluenesulfonates. Examples of the base addition salts include inorganic salts such as sodium salts, potassium salts, calcium salts and ammonium salts, and salts of organic bases such as ethylenediamine, ethanolamine, N,N-dialkyl-ethanolamine, triethanolamine, and basic amino acid salts.

[0990] The pharmaceutical conjugates of the present application can be in crystalline form, either as free compounds or as solvates (e.g., hydrates), and it is intended that both forms be within the scope of the present application. Methods of solvation are generally known in the art.

[0991] Any compound that is a prodrug of a pharmaceutical conjugate of the present application is within the scope of the present application. The term "prodrug" is used in its broadest sense and encompasses those derivatives that are transformed in vivo after administration to the compound of the present application. Such derivatives would readily occur to those skilled in the art and include, for example, compounds in which a free hydroxy group is converted into an ester derivative. Many appropriate prodrugs will be familiar to those skilled in the art and are described, for example, in Burger's Medicinal Chemistry and Drug Discovery, 6thEdition (Donald J. Abraham, Ed., 2001, Wiley) and in "Design and Applications of Prodrugs" (H. Bundgaard, Ed., 1985, Harwood Academic Publishers), the contents of which are incorporated herein by reference.

[0992] The pharmaceutically acceptable esters associated with the compounds of the present application are not particularly limited and can be selected by one skilled in the art. In the case of such esters, it is preferred that such esters can be cleaved by a biological process such as hydrolysis in vivo. The group making up the ester (when the group is represented as R, the ester is represented as -COOR) can be, for example: C1-C4alkoxyC1-C4alkyl, such as methoxyethyl, 1-ethoxyethyl, 1-methyl-l-methoxyethyl, 1-(isopropoxy)ethyl, 2-methoxyethyl, 2-ethoxyethyl, 1,1-dimethyl-l-methoxyethyl, ethoxymethyl, propoxymethyl, isopropoxymethyl, butoxymethyl, or tert-butoxymethyl; C1-C4alkoxylatedC1-C4alkoxyC1-C4alkyl, such as 2-methoxyethoxymethyl; C6-C 10 aryloxyC1-C4alkyl, such as phenoxyethyl; halogenatedC1-C4alkoxyC1-C4alkyl, such as 2,2,2-trichloroethoxymethyl or bis(2-chloroethoxy)methyl; C1-C4alkoxycarbonylC1-C4alkyl, such as methoxycarbonylmethyl; cyanoC1-C4alkyl, such as cyanomethyl or 2-cyanoethyl; C1-C4alkylthiomethyl, such as methylthiomethyl or ethylthiomethyl; C6-C 10 aryloxyC1-C4alkyl, such as phenoxyethyl; halogenatedC1-C4alkoxyC1-C4alkyl, such as 2,2,2-trichloroethoxymethyl or bis(2-chloroethoxy)methyl; C1-C4alkoxycarbonylC1-C4alkyl, such as methoxycarbonylmethyl; cyanoC1-C4alkyl, such as cyanomethyl or 2-cyanoethyl; C1-C4alkylthiomethyl, such as methylthiomethyl or ethylthiomethyl; C6-C 10arylcarbonyl C1-C4alkyl, such as phenylcarbonylmethyl, naphthylcarbonylmethyl, 2- phenylcarbonyl ethyl, 3-phenylcarbonylpropyl, 4-phenylcarbonylbutyl, 5- phenylcarbonyl pentyl, 6-phenylcarbonylhexyl, 2-naphthylcarbonyl ethyl, 3- naphthylcarbonylpropyl, 4-naphthylcarbonylbutyl, 5-naphthylcarbonyl pentyl, or 6- naphthylcarbonylhexyl; arylsulfonyl C1-C4alkyl, such as 2-phenylsulfonyl ethyl or 2- tolylsulfonyl ethyl; C1-C7aliphatic acyloxy C1-C4alkyl, such as formacyloxy methyl, acetyloxy methyl, propionyloxy methyl, butyryloxy methyl, pivaloyloxy methyl, valeryloxy methyl, isovaleryloxy methyl, caproyloxy methyl, 1-formacyloxy ethyl, 1- acetyloxy ethyl, 1-propionyloxy ethyl, 1-butyryloxy ethyl, 1-pivaloyloxy ethyl, 1- valeryloxy ethyl, 1-isovaleryloxy ethyl, 1-caproyloxy ethyl, 2-formacyloxy ethyl, 2- acetyloxy ethyl, 2-propionyloxy ethyl, 2-butyryloxy ethyl, 2-pivaloyloxy ethyl, 2- valeryloxy ethyl, 2-isovaleryloxy ethyl, 2-caproyloxy ethyl, 1-formacyloxy propyl, 1- acetyloxy propyl, 1-propionyloxy propyl, 1-butyryloxy propyl, 1-pivaloyloxy propyl, 1-valeryloxy propyl, 1-isovaleryloxy propyl, 1-caproyloxy propyl, 1- acetyloxy butyl, 1-propionyloxy butyl, 1-butyryloxy butyl, 1-pivaloyloxy butyl, 1- acetyloxy pentyl, 1-propionyloxy pentyl, 1-butyryloxy pentyl, 1-pivaloyloxy pentyl, or 1-pivaloyloxy hexyl; C5-C6cycloalkylcarbonyloxy C1-C4alkyl, such as cyclopentylcarbonyloxymethyl, cyclohexylcarbonyloxymethyl, 1-cyclopentylcarbonyloxy ethyl, 1- cyclohexylcarbonyloxy ethyl, 1-cyclopentylcarbonyloxy propyl, 1-cyclohexylcarbonyloxy propyl, 1-cyclopentylcarbonyloxy butyl, or 1-cyclohexylcarbonyloxy butyl; C6-C 10C5-C6-cycloalkyl-C1-C4-alkyl, such as cyclopropylmethyl, 1-(cyclopropyl)- ethyl, 1-(cyclopropyl)propyl, 1-(cyclopropyl)butyl, 1-(cyclopropyl)pentyl, 1-(cyclopropyl)hexyl, cyclobutylmethyl, 1-(cyclobutyl)ethyl, 1-(cyclobutyl)propyl, 1-(cyclobutyl)butyl, 1-(cyclobutyl)pentyl, 1-(cyclobutyl)hexyl, cyclopentylmethyl, 1-(cyclopentyl)ethyl, 1-(cyclopentyl)propyl, 1-(cyclopentyl)butyl, 1-(cyclopentyl)pentyl, 1-(cyclopentyl)hexyl, cyclohexylmethyl, 1-(cyclohexyl)ethyl, 1-(cyclohexyl)propyl, 1-(cyclohexyl)butyl, 1-(cyclohexyl)pentyl or 1-(cyclohexyl)hexyl; C1-C4-alkyl substituted by one or two substituents independently selected from the group consisting of halogen, hydroxy, cyano, C1-C4-alkoxy, C1-C4-alkylthio, di(C1-C4-alkyl)amino, C1-C4-alkylsulfinyl, C1-C4-alkylsulfonyl, phenylsulfonyl, phenylsulfinyl, phenyl, phenyl-C1-C4-alkyl, phenyl-C1-C4-alkoxy, phenoxy, phenylthio, heteroarylthio, heteroaryl-C1-C4-alkyl, heteroaryl-C1-C4-alkoxy, heteroaryloxy, heteroaryl, heteroaryl-C1-C4-alkyl, heteroaryl-C1-C4-alkoxy, heteroaryloxy, heteroarylthio, heteroarylsulfonyl, heteroarylsulfinyl, aryloxy, arythio, arylsulfonyl, arylsulfinyl, aryloxy-C1-C4-alkyl, aryloxy-C1-C4-alkoxy, arylthio-C1-C4-alkyl, arylthio-C1-C4-alkoxy, arylsulfonyl-C1-C4-alkyl, arylsulfonyl-C1-C4-alkoxy, arylsulfinyl-C1-C4-alkyl, arylsulfinyl-C1-C4-alkoxy, aryl-C1-C4-alkyl, aryl-C1-C4-alkoxy, arylthio, arylsulfonyl, arylsulfinyl, aryl, aryl-C1-C4-alkyl, aryl-C1-C4-alkoxy, aryloxy, arylthio, arylsulfonyl, arylsulfinyl, heteroaryloxy-C1-C4-alkyl, heteroaryloxy-C1-C4-alkoxy, heteroarylthio-C1-C4-alkyl, heteroarylthio-C1-C4-alkoxy, heteroarylsulfonyl-C1-C4-alkyl, heteroarylsulfonyl-C1-C4-alkoxy, heteroarylsulfinyl-C1-C4-alkyl, heteroarylsulfinyl-C1-C4-alkoxy, heteroaryl-C1-C4-alkyl, heteroaryl-C1-C4-alkoxy, heteroaryloxy, heteroarylthio, heteroarylsulfonyl, heteroarylsulfinyl, heteroaryl, heteroaryl-C1-C4-alkyl, heteroaryl-C1-C4-alkoxy, heteroaryloxy, heteroarylthio, heteroarylsulfonyl, heteroarylsulfinyl,[5-(phenyl, which is optionally substituted by C1-C4alkyl, C1-C4alkoxy or a halogen atom)-2-oxo-1,3- dioxol-4-yl]methyl, such as (5-phenyl-2-oxo-1,3-dioxol-4-yl)methyl, [5-(4-methylphenyl)-2-oxo-1,3- dioxol-4-yl]methyl, [5-(4-methoxyphenyl)-2-oxo-1,3-dioxol-4-yl]methyl, [5-(4-fluorophenyl)-2-oxo- 1,3-dioxol-4-yl]methyl or [5-(4-chlorophenyl)-2-oxo-1,3-dioxol-4-yl]methyl; or phthalidyl, which is optionally substituted by C1-C4alkyl or one or more C1-C4alkoxy, such as phthalidyl, dimethylphthalidyl or dimethoxyphthalidyl; and preferably pivaloyloxymethyl, phthalidyl or (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl, and more preferably (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl.

[0993] Any compound referred to herein is intended to represent such specific compound, as well as certain variants or forms. In particular, the compounds referred to herein can have asymmetric centers and therefore exist in different enantiomeric forms. All optical isomers and stereoisomers of the compounds referred to herein, and mixtures thereof, are intended to be within the scope of the present application. Thus, any given compound referred to herein is intended to represent any one of a racemate, one or more enantiomeric forms, one or more diastereomeric forms, one or more atropisomeric forms, and mixtures thereof. In particular, the compounds of formula [D-(X) b -(AA) w -(T) g -(L)] n pharmaceutical conjugates of -Ab and compounds of formula DX-(AA) w -(T) g -L1or DX-(AA) w -(T) g The compounds of -H can include enantiomers (depending on their asymmetry) or diastereomers. Stereoisomerism with respect to double bonds is also possible, so in some cases the molecule can exist in the form of an (E)-isomer or a (Z)-isomer. If the molecule contains several double bonds, each double bond will have its own stereochemistry, which can be the same or different from the stereochemistry of the other double bonds of the molecule. Single isomers and mixtures of isomers are within the scope of the present application.

[0994] In addition, the compounds referred to herein can exist in geometric isomeric (i.e., cis and trans isomers), tautomeric, or atropisomeric forms. In particular, the term tautomer refers to one of two or more structural isomers of a compound that exist in equilibrium with one another and are readily convertible to one another. Common tautomeric pairs are amine-imine, amide-imide, keto-enol, lactam-lactim, and the like. In addition, any compound referred to herein is intended to represent all such forms unless otherwise indicated. In addition, the compounds referred to herein can exist in isotopically-labeled forms. All geometric isomers, tautomers, atropisomers, hydrates, solvates, polymorphs, and isotopically-labeled forms of the compounds referred to herein, and mixtures thereof, are considered to be within the scope of the present application.

[0995] Protected forms of the compounds disclosed herein are considered to be within the scope of the present application. Suitable protecting groups are well known to those skilled in the art. A review of protecting groups in organic chemistry is provided by Wuts, PGM and Greene TW in Protecting Groups in Organic Synthesis, 4thedition Wiley-Interscience and Kocienski PJ in Protecting Groups, 3rdedition Georg Thieme Verlag. These references provide chapters on the protection of OH, amino and SH groups. All of these references are incorporated by reference in their entirety.

[0996] Within the scope of the present application, an OH protecting group is defined as an O-bonded moiety resulting from the protection of OH via formation of a suitable protected OH group. Examples of such protected OH groups include ethers, silyl ethers, esters, sulfonate esters, sulfenic and sulfinic esters, carbonates, and carbamates. In the case of ethers, the OH protecting group can be selected from methyl, methoxymethyl, methylthiomethyl, (phenyldimethylsilyl)methoxymethyl, benzyl- oxymethyl, p-methoxybenzyloxymethyl, [(3,4-dimethoxybenzyl)oxy]methyl, p-nitrobenzyloxy- methyl, o-nitrobenzyloxymethyl, [(R)-1-(2-nitrophenyl)ethoxy]methyl, (4-methoxyphenoxy)- methyl, guaithyiomethyl, [(p-phenylphenyl)oxy]methyl, t-butoxymethyl, 4-pentenyloxymethyl, silyloxymethyl, 2-methoxyethoxymethyl, 2-cyanoethoxymethyl, bis(2-chloroethoxy)methyl, 2,2,2-trichloroethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, menthoxy- methyl, O-bis(2-acetoxy-ethoxy)methyl, tetrahydropyranyl, fluorotetrahydropyranyl, 3-bromo- tetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl, 4- methoxy-tetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4- methyl)-phenyl]-4-methoxypiperidin-4-yl, 1-(2-fluorophenyl)-4-methoxypiperidin-4-yl, 1-(4- chlorophenyl)-4-methoxypiperidin-4-yl, 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2- chloroethoxy)ethyl, 2-hydroxyethyl, 2-bromoethyl, 1-[2-(trimethylsilyl)ethoxy]ethyl, 1-methyl- 1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 1-methyl-1- phenoxyethyl, 2,2,2-trichloroethyl, 1,1-bis-anisyl-2,2,2-trichloroethyl, 1,1,1,3,3,3-hexafluoro-2- phenylisopropyl, 1-(2-cyanoethoxy)ethyl, 2-trimethylsilylethyl, 2-(benzylthio)ethyl, 2-(phenyl- seleno)ethyl, t-butyl, cyclohexyl, 1-methyl-1'-cyclopropylmethyl, allyl, prenyl, cinnamyl, 2- phenallyl, propargyl, p-chlorophenyl, p-methoxyphenyl, p-nitrophenyl, 2,4-dinitrophenyl, 2,3,5,6- tetrafluoro-4-(trifluoromethyl)phenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, 2,6- dimethoxybenzyl, o-nitobenzyl, p-nitobenzyl, piperonyl nitrobenzyl, piperonyl nitropiperonyl, halobenzyl, 2,6-dichlorobenzyl, 3,4-dichlorobenzyl, 3,4-dimethoxybenzyl, 4-4-dichlorobenzyl, 2,6-difluorobenzyl, p-cyanobenzyl, fluorobenzyl, 4-fluoroalkoxybenzyl, trimethylsilylxylyl, p-phenylbenzyl, 2-phenyl-2-propyl, p-acylaminobenzyl, p-azidobenzyl, 4-azido-3-chlorobenzyl, 2-trifluoromethylbenzyl, 4-trifluoromethylbenzyl, p-(methylsulfinyl)benzyl, p-silacyclobutylbenzyl, 4-acetoxybenzyl, 4-(2-trimethylsilyl)ethoxymethoxybenzyl, 2-naphthylmethyl, 2-pyridylmethyl, 4-pyridylmethyl, 3-methyl-2-pyridylmethyl N-oxide, 2-quinolylmethyl, 6-methoxy-2-(4-methylphenyl)-4-quinolylmethyl, 1-pyrenylmethyl, diphenylmethyl, 4-methoxydiphenylmethyl, 4-phenyldiphenylmethyl, p,p'-dinitrodiphenylmethyl, 5-dibenzo-cycloheptyl, triphenylmethyl, tri(4-tert-butylphenyl)methyl, α-naphthyl-diphenylmethyl, p-methoxyphenyl-diphenylmethyl, di(p-methoxyphenyl)phenyl-methyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromobenzoylmethyloxy)diphenylmethyl, 4,4',4"-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-tris(acetylpropyloxypenyl)methyl, 4,4',4"-tris(benzoyloxypenyl)methyl, 4,4'-dimethoxy-3"-[N-(imidazolylmethyl)]trityl, 4,4'-dimethoxy-3"-[N-(imidazolylethyl)carbamoyl]trityl, bis(4-methoxyphenyl)-1'-pyrenylmethyl, 4-(17-tetraphenyl[a,c,g,i]fluorenylmethyl)-4,4"-dimethoxytrityl, 9-anthryl, 9-(9-phenyl)diphenanthryl, 9-phenylthioxanthyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiol-2-yl, 4,5-bis(ethoxycarbonyl)-[1,3]-dioxol-2-yl, benzoisothiazolyl S,S-dioxide. In the case of silyl ethers, the OH protecting group can be selected from trimethylsilyl, triethylsilyl, triisopropylsilyl, dimethylisopropylsilyl, diethylisopropylsilyl, dimethylhexylsilyl, 2-norbornyl-dimethylsilyl, t-butyl-dimethylsilyl, t-butyl-diphenylsilyl, triphenylsilyl, tri-p-cymene-silyl, triphenylsilyl, diphenylmethylsilyl, di-t-butylmethylsilyl, bis(t-butyl)-1-pyrenylmethoxysilyl, tris(trimethylsilyl)silyl, (2-hydroxystyryl)dimethylsilyl, (2-hydroxystyryl)diisopropylsilyl, t-butylmethoxyphenylsilyl, t-butoxydiphenylsilyl, 1,1,3,3-tetraisopropyl-3-[2-(triphenylmethoxy)ethoxy]disiloxan-1-yl, and fluorosilyl. In the case of esters, the OH protecting group can be selected fromOH protecting group together with the oxygen atom of the unprotected OH to which it is attached forms an ester which can be selected from formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trichloroacetamide, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, phenylacetate, diphenylacetate, 3-phenylpropionate, bis-fluorosulfonylpropionyl, 4-pentenoate, 4-oxovalerate, 4,4-(ethylenedithio)valerate, 5[3-bis(4-methoxyphenyl)hydroxymethylphenoxy]pent-2-enoate, pivaloate, 1-adamantaneacetate, crotonoate, 4-methoxy-crotonoate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate, 4-bromobenzoate, 2,5-difluorobenzoate, p-nitrobenzoate, pyridinecarboxylate, nicotinate, 2-(azidomethyl)benzoate, 4-azido-butyrate, (2-azidomethyl)phenylacetate, 2-{[(triphenylsulfanyl)oxy]methyl}benzoate, 2-{[(4-methoxytriphenylsulfanyl)oxy]methyl}benzoate, 2-{[methyl(triphenylsulfanyl)amino]methyl}benzoate, 2-{{[(4-methoxytriphenyl)sulfanyl]methylamino}methyl}benzoate, 2-(allyloxy)phenylacetate, 2-(isoprenyloxy methyl)benzoate, 6-(acryloyloxy methyl)-3-methoxy-2-nitrobenzoate, 6-(acryloyloxymethyl)-3-methoxy-4-nitrobenzoate, 4-benzyl-oxobutanoate, 4-trialkylsilyloxy-butanoate, 4-acetoxy-2,2-dimethylbutanoate, 2,2-dimethyl-4-pentenoate, 2-iodobenzoate, 4-nitro-4-methylvalerate, m-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 4-(methylsulfanyl-methoxy)butanoate, 2-(methylsulfanylmethoxymethyl)benzoate, 2-(chloroacetyloxymethyl)benzoate, 2-[(2-chloroacetyloxy)ethyl]benzoate, 2-[2-(benzyloxy)ethyl]benzoate, 2-[2-(4-methoxybenzyl-oxy)ethyl]benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenyl-acetate, isobutyrate, monosuccinate, (E)-2-methyl-2-butenoate, m-(methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate and 2-chlorobenzoate. In the case of sulfonates, sulfinates and sulfinic esters,The OH protecting group together with the oxygen atom of the unprotected OH to which it is attached forms a sulfonate, a sulphenate and a sulfinic acid ester which can be selected from the group consisting of sulfate, allyl sulfonate, methanesulfonate, benzenesulfonate, toluenesulfonate, 2-[(4-nitrophenyl)ethyl]sulfonate, 2-trifluoromethylbenzenesulfonate, 4-monomethoxytritylsulfenyl, 2,4-dinitrophenylsulfenyl alkyl carbonate, 2,2,5,5-tetramethylpyrrolidin-3-one-1-sulfinyl and dimethylphosphinylsulfinyl. In case of a carbonate, the OH protecting group together with the oxygen atom of the unprotected OH to which it is attached forms a carbonate which can be selected from the group consisting of methyl carbonate, methoxymethyl carbonate, 9-fluorenylmethyl carbonate, ethyl carbonate, bromoethyl carbonate, 2-(methylthiomethoxy)ethyl carbonate, 2,2,2-trichloroethyl carbonate, 1,1-dimethyl-2,2,2-trichloroethyl carbonate, 2-(trimethylsilyl)ethyl carbonate, 2-[dimethyl(2-naphthylmethyl)silyl]ethyl carbonate, 2-(phenylsulfonyl)ethyl carbonate, 2-(triphenylphosphonio)ethyl carbonate, cis-[4-[[(methoxytrityl)sulfenyl]oxy]tetrahydrofuran-3-yl]oxycarbonate, isobutyl carbonate, tert-butyl carbonate, vinyl carbonate, allyl carbonate, cinnamyl carbonate, propargyl carbonate, p-chlorophenyl carbonate, p-nitrophenyl carbonate, 4-ethoxy-1-naphthyl carbonate, 6-bromo-7-hydroxycoumarin-4-ylmethyl carbonate, benzyl carbonate, o-nitrobenzyl carbonate, p-nitrobenzyl carbonate, p-methoxybenzyl carbonate, 3,4-dimethoxybenzyl carbonate, anthraquinone-2-ylmethyl carbonate, 2-dansyl ethyl carbonate, 2-(4-nitrophenyl)ethyl carbonate, 2-(2,4-dinitrophenyl)ethyl carbonate, 2-(2-nitrophenyl)propyl carbonate, 2-(3,4-methylenedioxy-6-nitrophenyl)propyl carbonate, 2-cyano-1-phenylethyl carbonate, 2-(2-pyridyl)amino-1-phenylethyl carbonate, 2-[N-methyl-N-(2-pyridyl)]amino-1-phenylethyl carbonate, benzoylmethyl carbonate, 3',5'-dimethoxybenzoin carbonate, methyl dithio carbonate and S-benzyl thio carbonate.

[0997] Within the scope of the present application, an amino protecting group is defined as the N-bonded moiety resulting from the protection of an amino group via the formation of a suitable protected amino group. Examples of protected amino groups include carbamates, ureas, amides, heterocyclic systems, N-alkyl amines, N-alkenyl amines, N-alkynyl amines, N-aryl amines, imines, enamines, N-metal derivatives, N-N derivatives, N-P derivatives, N-Si derivatives, and N-S derivatives. In the case of carbamates, the amino protecting group together with the amino group to which it is attached forms a carbamate which can be selected from the group consisting of methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate, 2,6-di-tert-butyl-9-fluorenylmethyl carbamate, 2,7-bis(trimethylsilyl)fluorenylmethyl carbamate, 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluorenylmethyl carbamate, 17-tetraphenyl[a,c,g,i]fluorenylmethyl carbamate, 2-chloro-3-indenylmethyl carbamate, benzo[f]inden-3-ylmethyl carbamate, 1,1-dioxobenzo[b]-thiophene-2-ylmethyl carbamate, 2-methylsulfonyl-3-phenyl-1-prop-2-enyl carbamate, 2,7-di-tert-butyl-[9,(10,10-dioxo-10,10,10,10-tetrahydrothioxanthenyl)]methyl carbamate, 2,2,2-trichloroethyl carbamate, 2-trimethylsilylethyl carbamate, (2-phenyl-2-trimethylsilanyl)ethyl carbamate, 2-phenylethyl carbamate, 2-chloroethyl carbamate, 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate, 1,1-dimethyl-2,2,2-trichloroethyl carbamate, 2-(2'-pyridinyl)ethyl carbamate, 2-(4'-pyridinyl)ethyl carbamate, 2,2-bis(4'-nitrophenyl)ethyl carbamate, 2-[(2-nitrophenyl)dithio]-1-phenylethyl carbamate, 2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate, fluoro BOC carbamate, 1-adamantyl carbamate, 2-adamantyl carbamate, 1-(1-adamantyl)-1-methylethyl carbamate, 1-methyl-1-(4-biphenylyl)ethyl carbamate, 1-(3,5-di-tert-butylphenyl)-1-methylethyl carbamate, triisopropylsilylethoxycarbamate, vinyl carbamate, allyl carbamate, isoprenyl carbamate, 1-isopropylallyl carbamate, cinnamyl carbamate, 4-nitrocinnamyl carbamate, 3-(3'-pyridinyl)prop-2-enyl carbamate, hexynyl carbamate, propargyl carbamate, bis carbamate 1,4-but-2-yne, 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, dithioalkyl carbamate, benzyl carbamate, 3,5-di-tert-butylbenzyl, p-methoxyphenyl carbamate, p-nitrophenyl carbamate, p-bromophenyl carbamate, p-chlorophenyl carbamate, 2,4-dichlorophenyl carbamate, 4-methylsulfinylphenyl carbamate, 4-trifluoromethylphenyl carbamate, fluorophenyl carbamate, 2-naphthylmethyl carbamate, 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 4-phenylacetoxyphenyl carbamate, 4-azidophenyl carbamate, 4-azido-methoxyphenyl carbamate, m-chloro-p-acetoxyphenyl carbamate, p-(dihydroxyboryl)-phenyl carbamate, 5-benzoisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonyl ethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, 2-(4-nitrophenylsulfonyl)ethyl carbamate, 2-(2,4-dinitrophenylsulfonyl)ethyl carbamate, 2-(4-trifluoromethylphenylsulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methyl carbamate, 2-phosphoethyl carbamate, 2-[phenyl(methyl)dithio]ethyl carbamate, 1-methyl-1-(triphenylphosphonio)ethyl carbamate, 1,1-dimethyl-2-cyanoethyl carbamate, 2-dansyl ethyl carbamate, 2-(4-nitrophenyl)ethyl carbamate, 4-methylthiophenyl carbamate, 2,4-dimethylthiophenyl carbamate, m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, α-methyl nitrophenyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, 2-nitrophenylethyl carbamate, 6-nitroveratryl carbamate, 4-methoxybenzoylmethyl carbamate, 3',5'-dimethoxybenzoin carbamate, 9-dibenzosuberyl carbamate, N-methyl-N-(o-nitrophenyl) carbamate, t-amyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, cyclobutyl carbamate, cyclopentyl carbamate, cyclohexyl carbamate, isobutyl carbamate, isobornyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate, diisopropylmethyl carbamate, 2,2-dimethoxy-carbonylmethyl carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate, butynyl carbamate, 1,1-dimethylpropyne, 2-iodoethyl carbamate, 1-methyl-1-(4'-pyridyl)ethyl carbamate, 1-methyl-1-(p-phenylazo-phenyl)ethyl carbamate, p-(p'-methoxyphenylazo)benzyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-trimethylbenzyl carbamate, isoxazolidin-4-yl carbamate, 4-(trimethylammonio)benzyl carbamate, p-cyanobenzyl carbamate, bis(2-pyridyl)methyl carbamate, 2-furylmethyl carbamate, phenyl carbamate, 2,4,6-tri-tert-butylphenyl carbamate, 1-methyl-1-phenylethyl carbamate, and S-phenylmethylthiocarbamate. In the case of ureas, the amino protecting group can be selected from the group consisting of phenothiazinyl-(10)-carbonyl, N'-p-toluenesulfonylaminocarbonyl, N'-phenylaminothiocarbonyl, 4-hydroxyphenylaminocarbonyl, 3-hydroxytryptaminocarbonyl, and N'-phenylaminothiocarbonyl. In the case of amides, the amino protecting group together with the amino group to which it is attached forms an amide which can be selected from the group consisting of formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropionamide, pent-4-enamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalaninamide, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, 2,2-dimethyl-2-(o-nitrophenyl)acetamide, o-nitrophenoxyacetamide, 3-(o-nitrophenyl)propionamide, 2-methyl-2-(o-nitrophenoxy)propionamide, 3-methyl-3-nitrobutyramide, o-nitrocinnamide, o-nitrobenzamide, 3-(4-tert-butyl-2,6-dinitrophenyl)-2,2-dimethylpropionamide, o-(benzyloxy- methyl)benzamide, 2-(acetyloxymethyl)benzamide, 2-[(tert-butyldiphenylsilyl)oxy- methyl]benzamide, 3-(3',6'-dioxo-2',4',5'-trimethylcyclohexa-1',4'-dien)-3,3-dimethyl- propionamide, o-hydroxy-trans-cinnamide, 2-methyl-2-(p-phenylazo-phenyloxy)propionamide, 4-chlorobutyramide, aceto-acetamide, 3-(p-hydroxyphenyl)propionamide, (N'-dithiobenzyloxy- carbonylamino)acetamide, and N-acetylmethioninamide. In the case of heterocyclic systems, the amino protecting group together with the amino group to which it is attached forms a heterocyclic system which can be selected from the group consisting of 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dichlorophthalimide, N-tetrachlorophthalimide, N-4-nitrophthalimide, N-thiodiglycoloyl, N-dithiasuccinimide, N-2,3-diphenylmaleimide, N-2,3-dimethylmaleimide, N-2,5-dimethylpyrrole, N-2,5-bis(triisopropylsilyloxy)pyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct, N-1,1,3,3-tetramethyl-1,3-disilaisoindoline, N-diphenylsilyl diethylene, N-5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, N-5-substituted 1,3-benzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridinone and 1,3,5-dioxane. In the case of N-alkyl, N-alkenyl, N-alkynyl or N-aryl amines, the amino protecting group can be selected from N-methyl, N-tert-butyl, N-allyl, N-isoprenyl, N-cinnamyl, N-phenylallyl, N-propargyl, N-methoxymethyl, N-[2-(trimethylsilyl)ethoxy]methyl, N-3-acetoxypropyl, N-cyanomethyl, N-2-azonorbornene, N-benzyl, N-4-methoxybenzyl, N-2,4-dimethoxybenzyl, N-2-hydroxybenzyl, N-ferrocenylmethyl, N-2,4-dinitrophenyl, o-methoxyphenyl, p-methoxyphenyl, N-9-phenylfluorenyl, N-fluorenyl, N-2-pyridylmethyl amine N'-oxide, N-7-methoxycoumarin-4-ylmethyl, N-diphenylmethyl, N-bis(4-methoxyphenyl)methyl, N-5-dibenzo-cycloheptyl, N-triphenylmethyl, N-(4-methylphenyl)diphenylmethyl and N-(4-methoxyphenyl)diphenylmethyl. In the case of imines, the amino protecting group can be selected from N-1,1-dimethylthiomethylene, N-benzylidene, N-p-methoxybenzylidene, N-diphenylmethylene, N-[2-pyridyl)mesityl]methylene, N-(N',N'-dimethylaminomethylene), N-(N',N'-diphenylmethylaminomethylene), N-(N'-tert-butylaminomethylene), N,N'-isopropylidene, N-p-nitrobenzylidene, N-salicylidene, N-5-chlorosalicylidene, N-(5-chloro-2-hydroxyphenyl)phenylmethylene, N-cyclohexylidene and N-tert-butylidene. In the case of enamines, the amino protecting group can be selected from N-(5,5-dimethyl-3-oxo-1-cyclohexenyl), N-2,7-dichloro-9-fluorenylidene, N-1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl, N-(1,3-dimethyl-2,4,6-(1H,3H,5H)-trioxo-pyrimidin-5-yl)-methyl, N-4,4,4-trifluoro-3-oxo-1-butenyl and N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl). In the case of N-metal derivatives, the amino protecting group can be selected from N-borane, N-diphenylboronate, N-diethylboronate, N-9-borabicyclononane, N-difluoroboronate and 3,5-bis(trifluoromethyl)phenylboronic acid; and N-P derivatives in which the amino protecting group together with the amino group to which it is attached forms an N-P derivative selected from the group consisting of diphenylphosphinamide, dimethylsulfanylphosphinamide, diphenylsulfanylphosphinamide, dialkylamino phosphorodiamidate, dibenzylamino phosphorodiamidate, diaminophosphorodiamidate, and imino triphenylphosphorane. In the case of N-Si derivatives, the NH2protecting group can be selected from the group consisting of tert-butyldiphenylsilyl and triphenylsilyl. In the case of N-S derivatives, the protected amino group can be selected from the group consisting of N-sulphenyl or N-sulfonyl derivatives. The N-sulphenyl derivatives can be selected from the group consisting of benzenesulphenamide, 2-nitrobenzenesulphenamide, 2,4-dinitrobenzenesulphenamide, pentachlorobenzenesulphenamide, 2-nitro-4-methoxybenzenesulphenamide, triphenylmethylbenzenesulphenamide, 1-(2,2,2-trifluoro-1,1-diphenyl)ethylbenzenesulphenamide, and N-3-nitro-2-pyridylbenzenesulphenamide. The N-sulfonyl derivatives can be selected from the group consisting of methanesulfonyl amide, trifluoromethanesulfonyl amide, tert-butylsulfonyl amide, benzylsulfonyl amide, 2-(trimethylsilyl)ethanesulfonyl amide, p-toluenesulfonyl amide, benzenesulfonyl amide, o-anisidinesulfonyl amide, 2-nitrobenzenesulfonyl amide, 4-nitrobenzenesulfonyl amide, 2,4-dinitrobenzenesulfonyl amide, 2-naphthalenesulfonyl amide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonyl amide, 2-(4-methylphenyl)-6-methoxy-4-methylsulfonyl amide, 9-anthracenesulfonyl amide, pyridine-2-sulfonyl amide, benzothiazole-2-sulfonyl amide, benzoylmethylsulfonyl amide, 2,3,6-trimethyl-4-methoxybenzenesulfonyl amide, 2,4,6-trimethoxybenzenesulfonyl amide, 2,6-dimethyl-4-methoxybenzenesulfonyl amide, pentamethylbenzenesulfonyl amide, 2,3,5,6-tetramethyl-4-methoxybenzenesulfonyl amide, 4-methoxybenzenesulfonyl amide, 2,4,6-trimethylbenzenesulfonyl amide, 2,6-dimethoxy-4-methylbenzenesulfonyl amide, 3-methoxy-4-tert-butylbenzenesulfonyl amide, and 2,2,5,7,8-pentamethylchromane-6-sulfonyl amide.

[0998] Within the context of the present application, an SH protecting group is defined as an S-bond moiety resulting from the protection of an SH group via the formation of a suitable protected SH group. Examples of such protected SH groups include thioethers, disulfides, silylthioethers, thioesters, thiocarbonates, and thiocarbamates. In the case of thioethers, the SH protecting group can be selected from S-alkyl, S-benzyl, S-p-methoxybenzyl, S-o-hydroxybenzyl, S-p-hydroxybenzyl, S-o-acetoxybenzyl, S-p-acetoxybenzyl, S-p-nitrobenzyl, S-o-nitrobenzyl, S-2,4,6-trimethylbenzyl, S-2,4,6-trimethoxybenzyl, S-4-picolyl, S-2-picolyl-N-oxide, S-2-quinolylmethyl, S-9-anthrylmethyl, S-9-fluorenylmethyl, S-dibenzofulvenyl, S-diferrocenylmethyl, S-diphenylmethyl, S-bis(4-methoxyphenyl)methyl, S-5-dibenzocycloheptyl, S-triphenylmethyl, 4-methoxytriphenylmethyl, S-diphenyl-4-pyridylmethyl, S-phenyl, S-2,4-dinitrophenyl, S-2-quinolyl, S-t-butyl, S-1-adamantyl, S-methoxymethyl, S-isobutoxymethyl, S-benzyloxymethyl, S-1-ethoxyethyl, S-2-tetrahydropyranyl, S-benzylthiomethyl, S-phenylthiomethyl, S-acetamidomethyl (Acm), S-trimethylacetamidomethyl, S-benzamidomethyl, S-allyloxycarbonylamidomethyl, S-N-[2,3,5,6-tetrafluoro-4-(N'-piperidinyl)-phenyl-N-allyloxycarbonylamidomethyl, S-phthalimidomethyl, S-phenylacetamidomethyl, S-acetyl methyl, S-carboxymethyl, S-cyanomethyl, S-(2-nitro-l-phenyl)ethyl, S-2-(2,4-dinitrophenyl)ethyl, S-2-(4'-pyridyl)ethyl, S-2-cyanoethyl, S-2-(trimethylsilyl)ethyl, S-2,2-bis(ethoxycarbonyl)ethyl, S-(l-m-nitrophenyl-2-benzoyl)ethyl, S-2-phenylsulfonyl ethyl, S-l-(4-methylphenylsulfonyl)-2-methylpropan-2-yl, and S-p-hydroxybenzoylmethyl. In the case of disulfides, the protected SH group can be selected from S-ethyldisulfide, S-t-butyldisulfide, S-2-nitrophenyldisulfide, S-2,4-dinitrophenyldisulfide, S-2-benzenediazinyldisulfide, S-2-carboxyphenyldisulfide, and S-3-nitro-2-pyridyldisulfide. In the case of silylthioethers, the SH protecting group can be selected from those listed above for silyl ether protection of OH.In the case of a thioester, the SH protecting group can be selected from S-acetyl, S-benzoyl, S-2-methoxyisobutyryl, S-trifluoroacetyl, S-N-[[p-biphenyl)-isopropoxy]carbonyl]-N-methyl-γ-aminobutyric acid tert-butyl ester and S-N-(tert-butoxycarbonyl)-N-methyl-γ-aminobutyric acid tert-butyl ester. In the case of a thiocarbonic acid ester, the SH protecting group can be selected from S-2,2,2-trichloroethoxycarbonyl, S-tert-butoxycarbonyl, S-benzyloxycarbonyl, S-p-methoxybenzyloxycarbonyl and S-fluorenylmethylcarbonyl. In the case of a thiocarbamic acid ester, the protected SH group can be selected from S-(N-ethylcarbamic acid ester) and S-(N-methoxymethylcarbamic acid ester).

[0999] Reference to these groups should not be construed as limiting the scope of the application, as they are mentioned purely as examples of protecting groups for OH, amino and SH groups, but other groups known to the person skilled in the art having the said functions are also contemplated and it is understood that the application also encompasses them.

[1000] To provide a more concise description, some quantitative expressions given herein are not qualified by the term "about". It is understood that every quantity given herein is intended to encompass both the actual value and also the approximated value of the given value, whether or not expressly using the term "about", based on reasonable inferences of the person skilled in the art, including equivalent and approximate values obtained due to experimental and / or measuring conditions of the given value.

[1001] "Antibody-drug conjugates (ADC)" represent a targeted strategy for delivering cytotoxic molecules to cancer cells (see, for example, International Patent Applications WO-A-2004 / 010957, WO-A-2006 / 060533 and WO-A-2007 / 024536). Such compounds are typically referred to as "conjugates" of drugs, toxins and radionuclides. Tumor cells are killed when the drug conjugate binds to the tumor cell and releases and / or activates the cytotoxic activity of the drug moiety. The selectivity offered by the drug conjugate minimizes toxicity to normal cells, thereby enhancing drug tolerability in patients. Three examples of drug antibody conjugates of this type that have received marketing approval are: Gemtuzumab ozogamicin for acute myeloid leukemia, Brentuximab vedotin for relapsed and refractory Hodgkin lymphoma and anaplastic large cell lymphoma, and ado-Trastuzumab emtansine for breast cancer, especially HER2+.

[1002] The efficacy of drugs for cancer chemotherapy often relies on differences in growth rates, biochemical pathways, and physiological characteristics between cancer and normal tissues. As a result, most standard chemotherapeutic agents are relatively non-specific and exhibit dose-limiting toxicities that result in suboptimal therapeutic outcomes. One approach to selectively target malignant cells and unhealthy tissues is to use specific monoclonal antibodies (mAbs) that recognize tumor-associated antigens expressed on the surface of tumor cells [Meyer, DL and Senter, PD (2003) Recent advances in antibody drug conjugates for cancer therapy. Annu. Rep. Med. Chem., 38, 229-237; Chari, R.V. (2008) Targeted cancer therapy: conferring specificity to cytotoxic drugs. Acc. Chem. Res. 41, 98-107]. Over 30 IgG-type immunoglobulins (IgGs) and related agents have been approved over the past 25 years, primarily for cancer and inflammatory diseases.

[1003] An alternative strategy is to conjugate small anti-neoplastic molecules to mAbs, which serve as carriers (increasing half-life) and targeting agents (selectivity). Because of the high selectivity of monoclonal antibodies (mAbs) for tumor-associated antigens, good pharmacokinetics, and relatively low intrinsic toxicity, much work has been done toward targeted delivery of drugs using monoclonal antibodies (mAbs). mAb-drug conjugates (ADCs) are formed by covalently linking an anti-cancer drug (usually via a conditionally stable linker system) to a mAb. Once bound to a cell surface antigen, the mAb for most ADCs is actively transported to lysosomes or other intracellular compartments, where enzymes, low pH, or reducing agents facilitate drug release. However, there are very limited ADCs currently in development.

[1004] Antigens must have a high degree of tumor cell selectivity to limit toxicity and off-target effects. Numerous tumor-associated antigens have been studied in preclinical models and in clinical trials, including antigens overexpressed in B cells (e.g., CD20, CD22, CD40, CD79), T cells (CD25, CD30), cancer cells (HER2, EGFR, EpCAM, EphB2, PSMA), endothelium (endoglin), or stromal cells (fibroblast activation protein) [Teicher BA. Antibody-drug conjugate targets. Curr Cancer Drug Targets 9(8):982-1004, 2009]. An important property of ADC targets is their internalization ability; this can be an intrinsic characteristic of the antibody itself, or it can be induced by the antibody binding to its antigen. Indeed, internalization of the ADC is critical to reduce toxicity associated with extracellular delivery of the drug payload.

[1005] A limited number of cytotoxic drug families are currently actively investigated in clinical trials as payloads in ADCs in relation to conjugated small molecules and in comparison to a variety of putative antigen targets: calicheamycin (Pfizer), duocarmycin (Synthon), pyrrolobenzo-diazepine (Spirogen), irinotecan (Immunomedics), maytansinoid (DM1 and DM4; ImmunoGen + Genentech / Roche, Sanofi-Aventis, Biogen Idee, Centocor / Johnson & Johnson, Millennium / Takeda), and auristatin (MMAE and MMAF; Seattle Genetics + Genentech / Roche, Medimmune / AstraZeneca, Bayer-Schering, Celldex, Progenics, Genmab). Calicheamycin, duocarmycin, and pyrrolobenzo-diazepine are DNA minor groove binders, irinotecan is a topoisomerase I inhibitor, and maytansinoid and auristatin are microtubulin depolymerizing agents.

[1006] Interestingly, three of these ADCs that have acquired cytotoxicity have entered late-stage clinical trials. Trastuzumab-mertansine (T-DM1), trastuzumab linked to a semisynthetic maytansinoid via a stable linker (FDA approved on February 22, 2013 for advanced HER2-positive breast cancer); Inotuzumab ozogamicin (CMC-544), a humanized anti-CD22 mAb (G5 / 44, IgG4) conjugated with calicheamicin via an acid-labile linker (acetylphenoxy-butyric acid) (B-cell non-Hodgkin's lymphoma); Brentuximab vedotin, a humanized anti-CD30 mAb linked to monomethyl auristatin E (MMAE) via a maleimide caproyl-valyl-citrulline-based-p-aminobenzylcarbamate linker (FDA approved on August 19, 2011 for anaplastic large-cell lymphoma and Hodgkin's lymphoma).

[1007] Linkers represent a critical component of the ADC structure. Several classes of second-generation linkers have been investigated, including acid-labile hydrazone linkers (lysosomal) (e.g., gemtuzumab ozogamicin and inotuzumab ozogamicin); disulfide-based linkers (reducing intracellular environment); noncleavable thioether linkers (catabolic degradation in lysosomes) (e.g., trastuzumab-mertansine); peptide linkers (e.g., citrulline-valine) (lysosomal proteases, such as cathepsin B) (e.g., brentuximab vedotin): see, e.g., WO-A-2004 / 010957, WO-A-2006 / 060533, and WO-A-2007 / 024536. Purification of antibody-drug conjugates by size exclusion chromatography (SEC) has also been described [see, e.g., Liu et al., Proc. Natl. Acad. Sci. USA, 93:8618-8623 (1996) and Chari et al., Cancer Research, 52: 127-131 (1992)].

[1008] Trastuzumab (Herceptin) is a monoclonal antibody that interferes with the HER2 / neu receptor. Its primary use is in the treatment of certain breast cancers. HER receptors are proteins embedded in cell membranes that carry molecular signals from outside the cell to inside the cell and turn genes on and off. HER proteins stimulate cell proliferation. In some cancers, especially certain types of breast cancer, HER2 is overexpressed, and this causes the cancer cells to replicate uncontrollably.

[1009] In 20-30% of early stage breast cancers, the HER2 gene is amplified, which causes it to overexpress epidermal growth factor (EGF) receptors in the cell membrane. In some types of cancer, HER2 can send signals that make an impact on the cell without growth factors reaching and binding to the receptor; however, trastuzumab is ineffective in this case.

[1010] When the HER2 pathway functions normally, it promotes cell growth and division; however, when it is overexpressed, cell growth accelerates beyond its normal limits. In some types of cancer, the pathway is used to promote rapid cell growth and proliferation, thus forming tumors. In cancer cells, the HER2 protein can be overexpressed up to 100 times more than in normal cells (two million versus 20,000 per cell). This overexpression causes strong and constant proliferation signaling, thus forming tumors. Overexpression of HER2 also causes checkpoint inactivation, allowing proliferation to increase even more.

[1011] In the compounds of the present application, Ab is a moiety comprising at least one antigen binding site. In an alternative embodiment, Ab can be any suitable agent capable of binding to a target cell, preferably an animal cell and more preferably a human cell. Examples of such agents include lymphokines, hormones, growth factors, and nutrient delivery molecules (e.g., transferrin). In another example, Ab can be an aptamer, and can include a nucleic acid or a peptide aptamer.

[1012] Where Ab is a moiety comprising at least one antigen binding site, the moiety is preferably an antigen binding peptide or polypeptide. In a preferred embodiment, the moiety is an antibody or an antigen binding fragment thereof.

[1013] The term "antibody" in the drug conjugates of the present application refers to any immunoglobulin, preferably a full-length immunoglobulin. Preferably, the term encompasses monoclonal antibodies, polyclonal antibodies, multispecific antibodies such as bispecific antibodies, and antibody fragments thereof, as long as they exhibit the desired biological activity. Antibodies can be derived from any species, but are preferably of rodent, e.g., rat or mouse, human, or rabbit origin. Alternatively, the antibodies, preferably monoclonal antibodies, can be humanized antibodies, chimeric antibodies, or antibody fragments thereof. The term "chimeric antibody" can also include "primatized" antibodies comprising variable domain antigen binding sequences derived from a non-human primate (e.g., Old World Monkey, Ape, etc.) and human constant region sequences. The immunoglobulin molecule can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgGl, IgG2, IgG3, IgG4, IgAl and IgA2) or subclass of immunoglobulin molecule.

[1014] The term "monoclonal antibody" refers to a population of substantially homogeneous antibody molecules (i.e., each antibody molecule in the population is identical except for possible naturally occurring mutations that can be present in minor amounts) produced by a single clone of B cell line (often a hybridoma). Importantly, each monoclonal antibody has the same antigenic specificity, i.e., it is directed against a single determinant (epitope) on an antigen.

[1015] Monoclonal antibodies can be produced by methods known in the art. However, for example, monoclonal antibodies can be produced by the hybridoma method (Kohler et al. (1975) Nature 256:495), the human B cell hybridoma technique (Kozbor et al., 1983, Immunology Today 4:72), or the EBV-hybridoma technique (Cole et al., 1985, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96). Alternatively, recombinant DNA methods (see US 4816567) can be used to produce monoclonal antibodies, or the techniques described in Clackson et al. (1991) Nature, 352:624-628; Marks et al. (1991) J. Mol. Biol., 222:581-597 can be used to isolate from phage antibody libraries.

[1016] Polyclonal antibodies are antibodies directed against different determinants (epitopes) of the same antigen. This population of different antibodies can be obtained from the sera of animals immunized with the antigen, using methods well known in the art.

[1017] The term "bispecific antibody" refers to an artificial antibody composed of two different monoclonal antibodies. It can be designed to bind to either one to two adjacent epitopes on a single antigen, thereby increasing avidity and specificity, or to two different antigens for various applications, but particularly for recruiting cytotoxic T cells and natural killer (NK) cells, or to re-target toxins, radionuclides or cytotoxic drugs for cancer therapy (Holliger and Hudson, Nature Biotechnology, 2005, 23(9), 1126-1136). Bispecific antibodies can have a hybrid immunoglobulin heavy chain with a first binding specificity in one arm, and a hybrid immunoglobulin heavy chain-light chain pair (providing a second binding specificity) in the other arm. This asymmetric structure helps to separate the desired bispecific compound from unwanted immunoglobulin chain combinations, as only one half of the bispecific molecule contains an immunoglobulin light chain, which provides a convenient handle for separation (WO94 / 04690; Suresh et al., Methods in Enzymology, 1986, 121 :210; Rodrigues et al., 1993, J. of Immunology 151 :6954-6961; Carter et al., 1992, Bio / Technology 10:163-167; Carter et al., 1995, J. of Hematotherapy 4:463-470; Merchant et al., 1998, Nature Biotechnology 16:677-681).

[1018] Methods of making hybrid or bispecific antibodies are known in the art. In one method, bispecific antibodies can be produced by chemical cross-linking of two different Fab or scFv modules or by genetic fusion of two fusion tumors into a single "quadroma" (Holliger and Hudson, Nature Biotechnology, 2005, 23(9), 1126-1136).

[1019] The term "chimeric" antibody refers to an antibody wherein different portions are derived from different animal species. For example, a chimeric antibody can have variable regions from a mouse and constant regions from a human. In contrast, a "humanized antibody" is derived primarily from a human, even though it can contain non-human portions. Specifically, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or non-human primate having the desired specificity, affinity, and capacity. In some instances, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies can comprise residues that are not found in the recipient antibody or the donor antibody. These modifications are made to further optimize antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally also can comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.

[1020] Recombinant antibodies, such as chimeric and humanized monoclonal antibodies, can be produced by recombinant DNA techniques known in the art. Fully human antibodies can be produced using transgenic mice that are incapable of expressing endogenous immunoglobulin heavy and light chains genes but can express human heavy and light chain genes. The transgenic mice are immunized in the normal fashion with a selected antigen. Monoclonal antibodies to the antigen can be obtained using conventional hybridoma technology. The human immunoglobulin transgenes that have been introduced into the transgenic mice rearrange and subsequently undergo class switching and somatic mutation to generate high affinity human antibodies. Consequently, the mice are capable of generating a repertoire of human antibodies in response to antigen challenge. In particular, the mice can generate human antibodies specific for antigens that are normally immunogenic in humans but not in the mouse. For an overview of this technology for producing human antibodies, see Lonberg and Huszar (1995, Int. Rev. Immunol. 13:65-93). For a detailed discussion of this technology for producing human antibodies and human monoclonal antibodies and for producing such antibodies from transgenic mice see e.g. U.S. Patents 5625126, 5633425, 5569825, 5661016, 5545806; each incorporated herein by reference in its entirety. Other human antibodies can be purchased from e.g. Abgenix, Inc. (Freemont, CA) and Genpharm (San Jose, CA).

[1021] The term "antigen binding fragment" in the context of the pharmaceutical conjugates of the present invention refers to a portion of a full-length antibody wherein such antigen binding fragment of an antibody retains the antigen binding function of the full-length antibody. An antigen binding fragment can comprise a portion of an antibody variable region that comprises at least one, two, preferably three CDRs selected from CDR1, CDR2, and CDR3. An antigen binding fragment can also comprise a portion of an immunoglobulin light chain and heavy chain. Examples of antibody fragments include Fab, Fab', F(ab')2, scFv, di-scFv, sdAb, and BiTE (Bi-specific T-cell Engaging molecule), Fv fragments (including nanobodies, diabodies, diabody-Fc fusions, triabodies, and tetrabodies); minibodies; linear antibodies; fragments produced by a Fab expression library, anti-idiotypic (anti-Id) antibodies, CDRs (complementarity determining regions), and antigenic determinant binding fragments of any of the above that immunospecifically bind to a target antigen (such as a cancer cell antigen, a viral antigen, or a microbial antigen); single chain or single domain antibody molecules, including heavy chain only antibodies, e.g., camelid VHH domains and shark V-NAR; and multi-specific antibodies formed from antibody fragments. In contrast, a full-length antibody, which is referred to as an "antibody," is an antibody comprising VL and VH domains, as well as complete light and heavy chain constant domains.

[1022] Antibodies can also have one or more effector functions, which refer to biological activities attributable to the Fc region of the antibody (a native sequence Fc region or an amino acid sequence variant Fc region engineered according to methods in the art to alter receptor binding). Examples of antibody effector functions include CIq binding; complement dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors (such as B cell receptor; BCR), etc.

[1023] Antibodies can also be functionally active fragments (also referred to herein as immunologically active portions), antibody derivatives or analogs that immunospecifically bind to a target antigen (such as a cancer cell antigen, viral antigen or microbial antigen), or other antibodies that bind to tumor cells. In this regard, functionally active means that the fragment, derivative or analog is capable of eliciting an anti-idiotypic antibody that recognizes the same antigen as the antibody from which the fragment, derivative or analog is derived. In particular, in one exemplary embodiment, the antigenicity of the idiotype of an immunoglobulin molecule can be enhanced by deleting the C-terminal of the framework and CDR sequences that are specific for the antigen-recognizing CDR sequences. To determine which CDR sequences bind to an antigen, synthetic peptides containing the CDR sequences can be used in binding assays with the antigen by any of the binding assay methods known in the art (e.g., BIAcore®assay), see, e.g., Kabat et al., 1991, Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md; Kabat E et al., 1980, J. of Immunology 125(3):961-969).

[1024] The term "antibody" can also include fusion proteins of an antibody or functionally active fragment thereof, e.g., where the antibody is fused at the N- or C-terminus via a covalent linkage (e.g., a peptide bond) to an amino acid sequence of another protein (or a portion thereof, such as a 10, 20 or 50 amino acid portion of the protein) that is not the antibody. The antibody or fragment thereof can be covalently linked at the N-terminus of a constant domain to the other protein.

[1025] In addition, antibodies or antigen-binding fragments of the application can include analogs and derivatives of antibodies or antigen-binding fragments thereof that are modified, such as by covalent attachment of any type of molecule, so long as such covalent attachment allows the antibody to retain its antigen-binding immunospecificity. Examples of modifications include glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting groups / blocking groups, proteolytic cleavage, attachment to a cell antibody unit or other protein, etc. Any of numerous chemical modifications can be introduced into an antibody by known techniques, including but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis in the presence of tunicamycin, etc. Additionally, an analog or derivative can contain one or more unnatural amino acids.

[1026] The antibodies or antigen-binding fragments of the application can also have modifications (e.g., substitutions, deletions, or additions) in the Fc domain of the antibody. In particular, the modifications can be in the Fc-hinge region and result in increased binding to the FcRn receptor (WO 97 / 34631).

[1027] In one embodiment, the antibody in the pharmaceutical conjugate of the application can be any antibody or antigen-binding fragment thereof, preferably a monoclonal antibody that can be used in the treatment of a disease, preferably a cancer, more preferably a cancer selected from the group consisting of lung cancer (including NSCLC), gastric cancer, colorectal cancer, breast cancer, pancreatic cancer, endometrial cancer, bladder cancer, cervical cancer, esophageal cancer, gall bladder cancer, uterine cancer, salivary ductal carcinoma, ovarian cancer, renal cancer, leukemia, multiple myeloma, and lymphoma, wherein the cancer is preferably a HER2-positive cancer, wherein the HER2-positive cancer includes HER2-positive lung cancer (including HER2-positive NSCLC), HER2-positive gastric cancer, HER2-positive colorectal cancer, HER2-positive breast cancer, HER2-positive pancreatic cancer, HER2-positive endometrial cancer, HER2-positive bladder cancer, HER2-positive cervical cancer, HER2-positive esophageal cancer, HER2-positive gall bladder cancer, HER2-positive uterine cancer, HER2-positive salivary ductal carcinoma, and HER2-positive ovarian cancer, more preferably HER2-positive breast cancer, HER2-positive ovarian cancer, and HER2-positive gastric cancer, most preferably HER2-positive breast cancer.

[1028] Antibodies that can be useful in treating cancer include, but are not limited to, antibodies to CA125 (ovarian), CA15-3 (carcinoma), CA19-9 (carcinoma), L6 (carcinoma), Lewis Y (carcinoma), Lewis X (carcinoma), alpha fetoprotein (carcinoma), CA 242 (colorectal), placental alkaline phosphatase (carcinoma), prostate specific antigen (prostate), prostatic acid phosphatase (prostate), epidermal growth factor (carcinoma) (e.g., EGF receptor 2 protein (breast cancer)), MAGE-I (carcinoma), MAGE-2 (carcinoma), MAGE-3 (carcinoma), MAGE-4 (carcinoma), anti-transferrin receptor (carcinoma), p97 (melanoma), MUC1-KLH (breast cancer), CEA (colorectal), gplOO (melanoma), MARTl (melanoma), PSA (prostate), IL-2 receptor (T cell leukemia and lymphoma), CD20 (non-Hodgkin's lymphoma), CD52 (leukemia), CD33 (leukemia), CD22 (lymphoma), human chorionic gonadotropin (carcinoma), CD38 (multiple myeloma), CD40 (lymphoma), mucin (carcinoma), P21 (carcinoma), MPG (melanoma), and Neu oncogene product (carcinoma). Some specific antibodies that are useful include, but are not limited to, BR96 mAb (Trail, P. A. et al., Science (1993) 261, 212-215), BR64 (Trail, P. A. et al., Cancer Research (1997) 57, 100-105); mAbs to CD40 antigen, such as S2C6 mAb (Francisco, J. A. et al., Cancer Res. (2000) 60:3225-3231); mAbs to CD70 antigen, such as 1F6 mAb; and mAbs to CD30 antigen, such as AC10 (Bowen, M. A. et al., (1993) J. Immunol., 151:5896-5906; Wahl et al., 2002 Cancer Res. 62(13):3736- 3742). Many other internalizing antibodies that bind to tumor-associated antigens can be used and have been reviewed (Franke, A. E. et al., Cancer Biother Radiopharm. (2000) 15:459-476; Murray, J. L., (2000) Semin Oncol, 27:64-70; Breitling, F. and Dubel, S., Recombinant Antibodies, John Wiley, and Sons, New York, 1998).

[1029] Other tumor-associated antigens include, but are not limited to, BMPR1B, E16, STEAP1, STEAP2, 0772P, MPF, Napi3b, Sema5b, PSCA hlg, ETBR, MSG783, TrpM4, CRIPTO, CD21, CD79b, FcRH2, HER2, NCA, MDP, IL20R alpha, Brevican, EphB2R, ASLG659, PSCA, GEDA, BAFF-R, CD79A, CXCR5, HLA-DOB, P2X5, CD72, LY64, FCRH1, IRTA2, and TENB2.

[1030] In another embodiment, the antibody or antigen-binding fragment binds to an epitope present on a cell, such as a tumor cell. Preferably, where the cell is a tumor cell, the tumor cell epitope is not present on a non-tumor cell, or is present at a lower concentration or in a different spatial configuration than in the tumor cell.

[1031] In one embodiment, the antibody or antigen-binding fragment binds to an epitope present on one of the following antigens: CA125, CA15-3, CA19-9 L6, Lewis Y, Lewis X, alpha fetoprotein, CA 242, placental alkaline phosphatase, prostate specific antigen, prostatic acid phosphatase, epidermal growth factor (e.g., EGF receptor 2 protein), MAGE-I, MAGE-2, MAGE-3, MAGE-4, anti-transferrin receptor, p97, MUC1-KLH, CEA, gp100, MARTl, PSA, IL-2 receptor, CD20, CD52, CD33, CD22, human chorionic gonadotropin, CD38, CD40, Mucin, P21, MPG, Neu oncogene product, BMPR1B, E16, STEAP1, STEAP2, 0772P, MPF, Napi3b, Sema5b, PSCA hlg, ETBR, MSG783, TrpM4, CRIPTO, CD21, CD79b, FcRH2, HER2, NCA, MDP, IL20R alpha, Brevican, EphB2R, ASLG659, PSCA, GEDA, BAFF-R, CD79A, CXCR5, HLA-DOB, P2X5, CD72, LY64, FCRH1, IRTA2, TENB2.

[1032] In one embodiment, where the antigen is ErBB2 (also known as ErBB2, CD340, or HER2; such terms are used interchangeably), the antibody or antigen-binding fragment can bind to one or more of the following epitopes: ARHCL (SEQ ID NO: 1), QNGS (SEQ ID NO: 2), and PPFCVARC PSG (SEQ ID NO: 3). These epitopes correspond to positions 557-561, 570-573, and 593-603, respectively, of the human HER2 polypeptide sequence (Accession No: NM_004448, Version: NM_004448.3).

[1033] An antibody that binds a molecular target or antigen of interest, such as an ErbB2 antigen, is an antibody that is capable of binding said antigen with sufficient affinity so that the antibody is useful in targeting cells expressing the antigen. Where the antibody is one that binds ErbB2, it generally preferentially binds ErbB2 as opposed to other ErbB receptors, and can be one that does not significantly cross-react with other proteins such as EGFR, ErbB 3, or ErbB4. In such embodiments, the extent of binding of the antibody to these non-ErbB2 proteins, e.g., cell surface binding to endogenous receptors, will be less than 10% as determined by fluorescence activated cell sorting (FACS) analysis or radioimmunoprecipitation (RIA). At times, the anti-ErbB2 antibody does not significantly cross-react with the rat neu protein, e.g., as described in Schecter et al., Nature 312:513-516 (1984) and Drebin et al., Nature 312:545-548 (1984).

[1034] In another embodiment, the antibody or target of the pharmaceutical conjugate of the present application can be selected from the antibodies or targets in the following table. Such antibodies are immunospecific for the target antigen and are either commercially available or produced by any method known in the art, such as recombinant expression techniques.

[1035] Table 1: Therapeutic monoclonal antibodies

[1036]

[1037]

[1038]

[1039]

[1040]

[1041]

[1042]

[1043]

[1044]

[1045]

[1046]

[1047]

[1048]

[1049]

[1050]

[1051]

[1052]

[1053] In addition to the above, the antibody of the pharmaceutical antibody conjugate of the present application can be Vitaxin, which is a humanized antibody for the treatment of sarcoma; Smart IDIO, which is a humanized anti-HLA-DR antibody for the treatment of non-Hodgkin's lymphoma; Oncolym, which is a radiolabeled murine anti-HLA-Drlo antibody for the treatment of non-Hodgkin's lymphoma; and Allomune, which is a humanized anti-CD2 mAb for the treatment of Hodgkin's disease or non-Hodgkin's lymphoma.

[1054] The antibody of the pharmaceutical conjugate of the present application can also be any antibody fragment known for the treatment of any disease, preferably cancer. Again, such antibody fragments are immunospecific for the target antigen and are either commercially available or produced by any method known in the art, such as recombinant expression techniques. Examples of such antibodies that can be utilized include any from the following table.

[1055] Table 2: Therapeutic monoclonal antibody fragments

[1056]

[1057]

[1058]

[1059]

[1060]

[1061] Fab antigen binding fragment (one arm)

[1062] F(ab')2 antigen binding fragment, including hinge region (two arms)

[1063] Fab' antigen binding fragment, including hinge region (one arm)

[1064] scFv single chain variable fragment

[1065] di-scFv dimeric single chain variable fragment

[1066] (Holliger and Hudson, Nature Biotechnology, 2005, 23(9), 1126-1136).

[1067] In a preferred embodiment, the antibody in the pharmaceutical conjugate of the application targets a cell surface antigen.

[1068] In a preferred embodiment, the antibody in the pharmaceutical conjugate of the application can bind to a receptor encoded by an ErbB gene. The antibody can specifically bind to an ErbB receptor selected from the group consisting of EGFR, HER2, HER3 and HER4. Preferably, the antibody in the pharmaceutical conjugate can specifically bind to the extracellular domain of the HER2 receptor and inhibit the growth of tumor cells overexpressing said HER2 receptor. The antibody of the pharmaceutical conjugate can be a monoclonal antibody, such as a murine monoclonal antibody, a chimeric antibody or a humanized antibody. Preferably, the humanized antibody can be huMAb 4D5-1, huMAb 4D5-2, huMAb 4D5-3, huMAb 4D5-4, huMAb 4D5-5, huMAb 4D5-6, huMAb 4D5-7 or huMAb 4D5-8 (trastuzumab), especially preferably trastuzumab. The antibody can also be an antibody fragment, such as a Fab fragment.

[1069] Other preferred antibodies include:

[1070] (i) an anti-CD4 antibody. The antibody of the pharmaceutical conjugate can be a monoclonal antibody, such as a murine monoclonal antibody, a chimeric antibody or a humanized antibody;

[1071] (ii) an anti-CD5 antibody. The antibody of the pharmaceutical conjugate can be a monoclonal antibody, such as a murine monoclonal antibody, a chimeric antibody or a humanized antibody;

[1072] (iii) an anti-CD13 antibody. The antibody of the pharmaceutical conjugate can be a monoclonal antibody, such as a murine monoclonal antibody, a chimeric antibody or a humanized antibody;

[1073] (iv) Anti-CD20 antibodies. The antibody of the drug conjugate can be a monoclonal antibody, such as a murine monoclonal antibody, a chimeric antibody, or a humanized antibody. Preferably, the humanized antibody is rituximab or an antibody fragment thereof, such as a Fab fragment; and

[1074] (v) Anti-CD30 antibodies. The antibody of the drug conjugate can be a monoclonal antibody, such as a murine monoclonal antibody, a chimeric antibody, or a humanized antibody. Preferably, the humanized antibody is brentuximab vedotin or an antibody fragment thereof.

[1075] In one embodiment of the application, the drug antibody conjugate can exhibit one or more of the following: (i) increased cytotoxicity (or decreased cell survival), (ii) increased cytostatic activity (cytostasis), (iii) increased affinity for binding to the target antigen or antigenic determinant, (iv) increased internalization of the conjugate, (v) reduced side effects in patients, and / or (vi) improved toxicity profile. Such increases can be relative to known drug antibody conjugates that bind to the same or different antigenic determinant or antigen in the art.

[1076] Methods for preparing drug antibody conjugates

[1077] The drug antibody conjugates of the present application can be prepared according to techniques well known in the art. Methods for conjugating moieties comprising at least one antigen binding site, such as antibodies, to a large number of different drugs using different methods have been described previously and are exemplified, for example, in WO-A-2004 / 010957, WO-A-2006 / 060533, and WO-A-2007 / 024536, the contents of which are incorporated herein by reference. These include the use of linker groups that derivatize the drug, toxin, or radionuclide in such a way that the drug, toxin, or radionuclide can then be attached to the moiety, such as an antibody. Typically, attachment to the moiety, such as an antibody, is via one of three routes: via the free thiol group in a cysteine after reduction of a disulfide moiety in the antibody; via the free amino group in a lysine in the antibody; and via the free hydroxyl group in a serine and / or threonine in the antibody. The method of attachment varies depending on the site of attachment on the moiety, such as an antibody. Purification of antibody-drug conjugates by size exclusion chromatography (SEC) has also been described [see, e.g., Liu et al., Proc. Natl. Acad. Set (USA), 93:8618-8623 (1996) and Chari et al., Cancer Research, 52: 127-131 (1992)].

[1078] As indicated earlier, a process for preparing a drug conjugate according to the application is provided, which comprises conjugating a moiety Ab comprising at least one antigen binding site to a drug D of formula (I) or (IH), Ab and D being as defined herein.

[1079] An example of a process for preparing a drug conjugate of the application comprises preparing a drug antibody conjugate of the application of formula (G) or (G') below:

[1080]

[1081] The process comprises the following steps:

[1082] (i) reacting a drug (D-H) of formula (IH)-H:

[1083]

[1084] wherein the substituents in the definition of (IH)-H are as defined above for formula (IH), with a compound of formula (D') or (E):

[1085]

[1086] to give a compound of formula (F) or (F'), respectively:

[1087]

[1088] (ii) partially reducing one or more disulfide bonds in the antibody to be conjugated, to give a partially reduced antibody with free thiol groups Ab-SH:

[1089] and

[1090] (iii) reacting the partially reduced antibody with free thiol groups Ab-SH with the compound of formula (F) or (F') produced in step (i), to give the desired drug antibody conjugate of formula (G) or (G'), respectively:

[1091]

[1092] In another preferred embodiment of this method, the antibody is selected from brentuximab, gemtuzumab, inotuzumab, lorvotuzumab, an anti-HER2 antibody (e.g., trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody, and an anti-CD30 antibody, or an antigen-binding fragment or immunologically active portion thereof, or is selected from an anti-HER2 antibody (e.g., trastuzumab) and an anti-CD13 antibody, or an antigen-binding fragment or immunologically active portion thereof, and is most preferably trastuzumab or an antigen-binding fragment or immunologically active portion thereof. Furthermore, the partial reduction of this monoclonal antibody is performed using tris[2-carboxyethyl]phosphine hydrochloride (TCEP).

[1093] Another embodiment of the method for preparing the drug antibody conjugate of the present application includes preparing a drug antibody conjugate of the following formula (W) or (W'):

[1094]

[1095]

[1096] The method comprises the steps of:

[1097] (i) reacting the antibody with 2-iminothiolane hydrochloride (Traut's reagent) to obtain a thiol-activated antibody:

[1098]

[1099] (ii) reacting the thiol-activated antibody with the compound of formula (F) or (F'), respectively, to obtain the desired drug antibody conjugate of formula (W) or (W').

[1100]

[1101] In another preferred embodiment of this method, the antibody is selected from brentuximab, gemtuzumab, inotuzumab, lorvotuzumab, an anti-HER2 antibody (e.g., trastuzumab), an anti-CD4 antibody, an anti-CD5 antibody, an anti-CD13 antibody, and an anti-CD30 antibody, or an antigen-binding fragment or immunologically active portion thereof, or is selected from an anti-HER2 antibody (e.g., trastuzumab) and an anti-CD13 antibody, or an antigen-binding fragment or immunologically active portion thereof, and is most preferably trastuzumab or an antigen-binding fragment or immunologically active portion thereof.

[1102] Another example of the method for preparing the drug antibody conjugate of the present application includes preparing a drug antibody conjugate of the following formula (O) or (P):

[1103]

[1104] The method comprises the steps of:

[1105] (i):

[1106] (a) Drugs (DH) that make up (IH)-H:

[1107]

[1108] The substituents in (IH)-H are as defined above. When reacted with a compound of formula X2-C(O)-X1, where X1 and X2 are leaving groups, the compound of formula (B) is obtained:

[1109]

[1110] Furthermore, the connection point of the -(C=O)X1 portion is the free -NH2 group of the compound of formula DH, or

[1111] (b) Reaction of a drug (DH) of formula (IH)-H as defined above with 4-nitrophenylchloroformate yields a compound of formula (J):

[1112]

[1113] Furthermore, the connection point of the (4-nitrophenyl)-O-CO- group is the same as the connection point of the X1(CO) part in (a) above;

[1114] (ii):

[1115] (c) Reconstitute the compound of formula (B) produced in step (i) with formula HO-(CH2). 1-6 NHProt NH The reaction of hydroxyl compounds and the removal of self-coupled compounds from Prot NH Groups, yielding compound of formula (C):

[1116]

[1117] Then, the compound of formula (C) is reacted with the compound of formula Me-SS-(CH2). 1-3 The reaction of the -CO2H compound yields the compound of formula (K):

[1118] or

[1119] (d) Make the compound (J) produced in step (i) react with the formula HO-(CH2). 1-3 SProt SH The compound reacts and the self-coupled compound removes Prot SH Groups were used to obtain compounds of formula (L):

[1120]

[1121] (iii) reacting (K) or (L) produced in step (ii) with dithiothreitol under disulfide reducing conditions to produce compounds of formula (M) and (N), respectively:

[1122]

[1123] (iv) reacting the antibody to be conjugated with succinimidyl-4-(N- maleimidomethyl)cyclohexane-1-carboxylate to derivatize the antibody with succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate at one or more lysine residues:

[1124]

[1125] (v) reacting the derivatized antibody produced in step (iv) with (M) or (N) produced in step (iii) to produce the desired drug antibody conjugate of formula (O) or (P):

[1126]

[1127] The compound of formula X2-C(O)-X1is preferably 1,1'- carbonyldiimidazole. Similarly, the hydroxyl compound that is reacted with the compound of formula (B) is preferably HO-(CH2) 2-4 -NHProt NH , and more preferably HO-(CH2)3-NHProt NH .

[1128] In a preferred embodiment of the application, the compound that is reacted with the compound of formula (C) to produce the compound of formula (K) is 3- (methyl dithiasilyl)propionic acid.

[1129] In another preferred embodiment, the compound that is reacted with the compound of formula (J) to produce the compound of formula (L) is HO-(CH2) 1-3 SProt SH HO-(CH2)3SProt SH .

[1130] In case the linkage to the drug attachment moiety is via the free thiol group in a cysteine after partial reduction of the disulfide bond moiety in a moiety comprising at least one antigen binding site, such as a monoclonal antibody, said partial reduction is typically performed by first dilution to a suitable concentration and buffer solution, followed by partial reduction of the disulfide bond by means of addition of a suitable reducing agent, such as tris[2-carboxyethyl]phosphine hydrochloride (TCEP) or dithiothreitol (DTT). By choosing the appropriate ratio of the moiety to be reduced, such as a monoclonal antibody, to the reducing agent, the reaction conditions and time of reduction, the desired ratio of free thiol groups to moiety can be obtained, e.g. four free thiol groups per monoclonal antibody.

[1131] The partially reduced moiety, such as a partially reduced monoclonal antibody with free thiol groups, prepared as described above is then reacted with a drug- linker compound of formula D-(X) b -(AA) w -(T) g L1of the present application, wherein the group L1in such a compound is a maleimido group which freely reacts with a thiol group. The resulting drug antibody conjugate is purified by any suitable means known in the art, e.g. by size exclusion chromatography (SEC) [see e.g. Liu et al., Proc. Natl. Acad. Sci. USA, 93: 8618-8623 (1996) and Chari et al., Cancer Research, 52: 127-131 (1992)].

[1132] In a preferred embodiment of the present application, the partially reduced monoclonal antibody is an anti-HER2 antibody, such as trastuzumab, or an anti-CD13 antibody or an antigen binding fragment or immunologically active portion thereof, preferably trastuzumab or an antigen binding fragment or immunologically active portion thereof; or preferably an anti-CD13 antibody or an antigen binding fragment or immunologically active portion thereof.

[1133] In an alternative embodiment of the present application, the lysines in a moiety comprising at least one antigen binding site, such as a monoclonal antibody, can first be reacted with succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate. The free amino groups on the antibody can react with the N-hydroxysuccinimide ester to give a maleimide-activated antibody:

[1134]

[1135] The maleimide-activated antibody can then be reacted with a compound of formula D-(X) b -(AA) w -(T) g -H having an active thiol moiety.

[1136] In an alternative embodiment of the application, lysines in a moiety comprising at least one antigen binding site, such as a monoclonal antibody, can first be reacted with 2-iminothiolane (Traut's reagent). The free amines on the antibody can be reacted with the iminothiolactone to yield a thiol-activated antibody.

[1137]

[1138] Figure 1 shows conjugation via free thiol in cysteine after partial reduction of disulfide bond groups in an antibody for the preparation of the pharmaceutical antibody conjugate of formula [D-(X) b -(AA)w-(T) g -(L)-] n A particular example of a method for the preparation of a pharmaceutical antibody conjugate of formula [D-(X)

[1139] Figure 2 shows conjugation with free amine in lysine after reaction of an antibody with Traut's reagent for the preparation of the pharmaceutical antibody conjugate of formula [D-(X) b -(AA) w -(T) g -(L)-] n Another particular example of a method for the preparation of a pharmaceutical antibody conjugate of formula [D-(X)

[1140] Compositions comprising the pharmaceutical antibody conjugate of the application and uses thereof

[1141] Also provided is a pharmaceutical composition comprising the pharmaceutical conjugate according to the application and a pharmaceutically acceptable carrier. The pharmaceutical conjugate of formula [D-(X) b -(AA) w -(T) g -(L)-] n Examples of administration forms of the pharmaceutical conjugate of formula [D-(X)

[1142] The pharmaceutically acceptable carrier or vehicle can be particulate, such that the composition is, for example, in tablet or powder form. One or more carriers can be liquid, wherein the composition is, for example, an oral syrup or injectable liquid. Additionally, one or more carriers can be gaseous, so as to provide an aerosol composition useful, for example, in inhalable administration. The term "carrier" refers to a diluent, adjuvant, or vehicle with which a pharmaceutical antibody conjugate of the present application is administered. Such a pharmaceutical carrier can be a liquid such as water and oil, including those of either vegetable or animal origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. The carrier can be saline, gum acacia, gelatin, starch paste, talc, keratin, colloidal silica, urea and the like. Additionally, auxiliary, stabilizing, thickening, lubricating and coloring agents can be used. In one embodiment, the pharmaceutical antibody conjugate or composition of the present application, and the pharmaceutically acceptable carrier, when administered to an animal, are sterile. Water is a preferred carrier when the pharmaceutical antibody conjugate of the present application is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical carriers also include excipients such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol and the like. The composition of the application, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents.

[1143] When oral administration is intended, the pharmaceutical composition is preferably in either solid or liquid form, with semi-solid, semi-liquid, suspension and gel forms included within the forms considered herein as either solid or liquid.

[1144] As a solid composition for oral administration, the composition can be formulated as a powder, granule, compressed tablet, pill, capsule, chewing gum, wafer or the like. Such a solid composition typically contains one or more inert diluents. In addition, one or more of the following can be present: binders such as carboxymethylcellulose, ethyl cellulose, microcrystalline cellulose, or gelatin; excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, corn starch and the like; lubricants such as magnesium stearate; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; flavoring agents such as peppermint, methyl salicylate or citrus flavoring; and coloring agents.

[1145] When the composition is in the form of a capsule (for example, a gelatin capsule), it can contain, in addition to the above types of materials, a liquid carrier such as polyethylene glycol or oil.

[1146] The composition can be in liquid form, such as an elixir, syrup, solution, emulsion, or suspension. The liquid can be for oral administration or for delivery by injection. When intended for oral administration, the composition can include one or more of a sweetening agent, preservatives, dye / colorant, and flavoring agent. In a composition intended to be administered by injection, one or more of a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer, and isotonic agent can also be included.

[1147] The preferred route of administration is non-oral, including, but not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, intranasal, intracerebral, intraventricular, intrathecal, intravaginal, or transdermal. The preferred mode of administration is at the discretion of the medical practitioner and can be dependent, in part, on the location of the medical condition, such as the location of the cancer. In a more preferred embodiment, the pharmaceutical antibody conjugate of the present application is administered intravenously.

[1148] The liquid compositions of the present application, whether solutions, suspensions or other like form, can also include one or more of the following: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono- or diglycerides, polyethylene glycols, glycerin, or other solvents; antibacterial agents such as benzyl alcohol or methyl parabens; and agents for adjusting tonicity such as sodium chloride or dextrose. The non-oral compositions can be enclosed in ampuls, disposable syringes or multiple dose vials made of glass, plastic or other material. Physiological saline is a preferred adjuvant.

[1149] The amount of the pharmaceutical conjugate of the present application effective to treat a particular disorder or condition will depend on the nature of the disorder or condition, and can be determined by standard clinical techniques. In addition, in vitro or in vivo assays can optionally be employed to help identify the optimal dosage ranges. The precise dose to be employed in the compositions will also depend on the route of administration and the seriousness of the disease or disorder, and should be decided according to the judgment of the medical practitioner and each patient's circumstances.

[1150] The composition comprises an effective amount of the pharmaceutical conjugate of the present application in order to achieve the appropriate dosage. The proper dosage of the compound will vary according to the particular formulation, mode of application, and the particular site, host and disease, e.g., cancer and what type of tumor, if cancer, being treated. Other factors like age, body weight, sex, diet, time of administration, rate of excretion, host condition, drug combinations, reaction sensitivities and severity of the disease shall be taken into account. Administration can be carried out continuously or periodically throughout the course of treatment.

[1151] The pharmaceutical conjugate or composition of the present application can be administered by any appropriate route, such as by infusion or rapid injection, by absorption through epithelial or mucocutaneous linings.

[1152] In certain embodiments, it can be desirable to administer one or more of the pharmaceutical conjugates or compositions of the application locally to an area in need of treatment. In one embodiment, administration can be by direct injection at the site of a cancer, tumor or tumor tissue or pre-tumor tissue (or formation site). In another embodiment, administration can be by direct injection at the site of manifestation (or formation site) of an autoimmune disease.

[1153] Pulmonary administration can also be employed, e.g., through the use of an inhaler or nebulizer, and formulation with an aerosol or via a perfusion liquid in fluorocarbons or synthetic lung surfactants. In certain embodiments, the pharmaceutical antibody conjugates or compositions of the application can be formulated as suppositories with traditional binders and carriers such as triglycerides.

[1154] The compositions of the application can take the form of solutions, suspensions, emulsions, tablets, pills, pellets, capsules, capsules containing liquids, powders, sustained release formulations, suppositories, emulsions, aerosols, sprays, suspensions, or any other form suitable for use. Additional examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E. W. Martin.

[1155] The pharmaceutical compositions can be prepared by methods well known in the art of pharmacy. For example, compositions intended to be administered by injection can be prepared by combining a pharmaceutical conjugate of the application with water and rendering the resulting solution sterile. A surfactant can be added to facilitate uniform solution or suspension.

[1156] It has been found that the pharmaceutical conjugates and compositions of the application are particularly effective in the treatment of cancer.

[1157] Thus, as previously mentioned, the present application provides a method of treating a patient, particularly a human, in need thereof, suffering from cancer, comprising administering to said diseased individual a therapeutically effective amount of a pharmaceutical conjugate or composition of the present application. The present application provides a pharmaceutical conjugate according to the present application for use in the treatment of cancer, more preferably a cancer selected from the group consisting of lung cancer (including NSCLC), gastric cancer, colorectal cancer, breast cancer, pancreatic cancer, endometrial cancer, bladder cancer, cervical cancer, esophageal cancer, gall bladder cancer, uterine cancer, salivary ductal carcinoma, ovarian cancer, renal cancer, leukemia, multiple myeloma and lymphoma. The most preferred cancer is breast cancer. A preferred cancer is a HER2 positive cancer, wherein a HER2 positive cancer includes a HER2 positive lung cancer (including a HER2 positive NSCLC), a HER2 positive gastric cancer, a HER2 positive colorectal cancer, a HER2 positive breast cancer, a HER2 positive pancreatic cancer, a HER2 positive endometrial cancer, a HER2 positive bladder cancer, a HER2 positive cervical cancer, a HER2 positive esophageal cancer, a HER2 positive gall bladder cancer, a HER2 positive uterine cancer, a HER2 positive salivary ductal carcinoma, and a HER2 positive ovarian cancer, more preferably a HER2 positive breast cancer, a HER2 positive ovarian cancer and a HER2 positive gastric cancer, most preferably a HER2 positive breast cancer.

[1158] The pharmaceutical conjugates and compositions of the present application can be used to inhibit the proliferation of tumor or cancer cells, or to treat cancer in an animal. The pharmaceutical conjugates and compositions of the present application can thus be used in a variety of settings for the treatment of cancer in an animal. The conjugates of the present application comprising a drug-linker- moiety comprising at least one antigen binding site can be used to deliver a drug or drug unit to a tumor or cancer cell. Without being bound by theory, in one embodiment, the moiety comprising at least one antigen binding site of the pharmaceutical conjugates of the present application binds or associates with a cancer or tumor cell associated antigen, and the pharmaceutical conjugates of the present application can be taken up within the tumor or cancer cell via receptor-mediated endocytosis. The antigen can be attached to the tumor or cancer cell or can be an extracellular matrix protein associated with the tumor or cancer cell. Once inside the cell, one or more specific sequences within the linker unit are hydrolytically cleaved by one or more tumor or cancer cell associated proteases or hydrolases, thereby releasing the drug or drug-linker compound. Subsequently, the released drug or drug-linker compound is free to migrate within the cell and elicit cytotoxic activity. In an alternative embodiment, the drug or drug unit is cleaved from the pharmaceutical conjugates of the present application outside the tumor or cancer cell, and the drug or drug-linker compound subsequently penetrates the cell.

[1159] In one embodiment, the moiety comprising at least one antigen binding site binds to a tumor cell or cancer cell. In another embodiment, the moiety comprising at least one antigen binding site binds to a tumor cell or cancer cell antigen on the surface of a tumor cell or cancer cell. In another embodiment, the moiety comprising at least one antigen binding site binds to a tumor cell or cancer cell antigen that is an extracellular matrix protein associated with a tumor cell or cancer cell.

[1160] The specificity of the moiety comprising at least one antigen binding site for a particular tumor cell or cancer cell is important in determining those tumors or cancers that are most effectively treated. For example, the drug conjugates of the present application having a unit of trastuzumab can be used to treat antigen-positive cancers, including leukemias, lung cancer, colon cancer, lymphomas (e.g., Hodgkin's disease, non-Hodgkin's lymphoma), solid tumors such as sarcomas and carcinomas, multiple myeloma, renal cancer, and melanoma. Preferably, the cancer can be lung cancer, colorectal cancer, breast cancer, pancreatic cancer, renal cancer, leukemia, multiple myeloma, lymphoma, or ovarian cancer. For example, the drug conjugates of the present application having a unit of rituximab can be used to treat tumors expressing CD-20, such as hematological cancers, including leukemias and lymphomas. For example, the drug conjugates of the present application having a unit of an anti-CD4 antibody can be used to treat tumors expressing CD-4, such as hematological cancers, including lymphomas. For example, the drug conjugates of the present application having a unit of an anti-CD5 antibody can be used to treat tumors expressing CD-5, such as hematological cancers, including leukemias and lymphomas. For example, the drug conjugates of the present application having a unit of an anti-CD13 antibody can be used to treat tumors expressing CD-13, such as hematological cancers, including leukemias and lymphomas.

[1161] Other specific types of cancers that can be treated with the drug conjugates of the present application include, but are not limited to: blood-borne cancers, including all forms of leukemia; lymphomas, such as Hodgkin's disease, non-Hodgkin's lymphoma, and multiple myeloma.

[1162] In particular, the drug conjugates and compositions of the present application exhibit excellent activity in the treatment of breast cancer.

[1163] The drug conjugates and compositions of the present application provide tumor or cancer targeting specific to the conjugate, thus reducing the overall toxicity of these conjugates. The linker unit stabilizes the drug antibody conjugate in the blood, but can be cleaved by tumor-specific proteases and hydrolases within the cell, releasing the drug.

[1164] The drug conjugates and compositions of the present application can be administered to animals that have also undergone surgery as a treatment for cancer. In one embodiment of the present application, the additional method of treatment is radiation therapy.

[1165] In a particular embodiment of the application, the pharmaceutical conjugate or composition of the application can be administered with radiotherapy. The radiotherapy can be administered simultaneously with, prior to or after the treatment with the pharmaceutical conjugate or composition of the application. In one embodiment, the pharmaceutical conjugate or composition of the application is administered simultaneously with the radiotherapy. In another particular embodiment, the administration of the pharmaceutical conjugate or composition of the application is preceded or followed by, preferably preceded by, at least one hour, five hours, 12 hours, one day, one week, one month, more preferably several months (e.g. up to three months) of administration of the radiotherapy.

[1166] As to the radiation, any radiotherapy regimen can be used depending on the type of cancer to be treated. For example, but not by way of limitation, x-ray radiation can be administered; in particular, high-energy megavoltage (radiation above 1 MeV energy) can be used for deep tumors, and electron beam and orthovoltage radiation can be used for skin cancers. Radioactive isotopes emitting gamma rays, such as radium, cobalt and other elements, can also be administered.

[1167] In the present application, a kit comprising a therapeutically effective amount of a pharmaceutical conjugate according to the application and a pharmaceutically acceptable carrier is provided. In one embodiment, a kit comprising a composition according to the application and optionally instructions for use in the treatment of a cancer, and more preferably a cancer selected from the group consisting of lung cancer (including NSCLC), gastric cancer, colorectal cancer, breast cancer, pancreatic cancer, endometrial cancer, bladder cancer, cervical cancer, esophageal cancer, gallbladder cancer, uterine cancer, salivary ductal carcinoma, ovarian cancer, renal cancer, leukemia, multiple myeloma and lymphoma is provided.

[1168] In one embodiment, the kit according to this aspect is used in the treatment of a cancer, and more preferably a cancer selected from the group consisting of lung cancer (including NSCLC), gastric cancer, colorectal cancer, breast cancer, pancreatic cancer, endometrial cancer, bladder cancer, cervical cancer, esophageal cancer, gallbladder cancer, uterine cancer, salivary ductal carcinoma, ovarian cancer, renal cancer, leukemia, multiple myeloma and lymphoma. Most preferably, the kit is used in the treatment of breast cancer. BRIEF DESCRIPTION OF DRAWINGS

[1169] The application is schematically illustrated by way of example in the accompanying drawings in which:

[1170] Figure 1 is a schematic representation of a method according to the application wherein the conjugation to the antibody is via a free thiol group;

[1171] Figure 2 is a schematic representation of a method according to the application wherein the conjugation to the antibody is via a free amino group.

[1172] EXAMPLE

[1173] The present application is further illustrated by the following non-limiting examples. In the examples, the following abbreviations are used:

[1174] CDI, 1,1'-carbonyldiimidazole

[1175] DIPEA, N,N-diisopropylethylamine

[1176] Hex, hexane

[1177] EtOAc, ethyl acetate

[1178] DCM, dichloromethane

[1179] NMP, N-methyl-2-pyrrolidone

[1180] DMF, dimethylformamide

[1181] EDC, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride

[1182] EDTA, ethylenediaminetetraacetic acid

[1183] MeOH, methanol

[1184] DTT, dithiothreitol

[1185] Py, pyridine

[1186] THF, tetrahydrofuran

[1187] TCEP, tris[2-carboxyethyl]phosphine hydrochloride

[1188] MC, 6-maleimidocaproyl

[1189] Fmoc, 9-fluorenylmethyloxycarbonyl

[1190] Cit, citrulline

[1191] Val, valine

[1192] DMSO, dimethyl sulfoxide

[1193] Trt, trityl

[1194] HOBt, 1-hydroxybenzotriazole

[1195] DIPCDI, N,N'-diisopropylcarbodiimide

[1196] TFA, trifluoroacetic acid PABOH, 4-aminobenzyl alcohol bis-PNP, bis(4-nitrophenyl) carbonate NAC, N-acetylcysteine SEC, size exclusion chromatography

[1197] HPLC, high performance liquid chromatography

[1198] ADC, antibody drug conjugate ATCC, American Type Culture Collection DMEM, Dulbecco's Modified Eagle Medium RPMI, Roswell Park Memorial institute medium ITS, Insulin- transferrin-selenite supplement FCS, fetal calf serum

[1199] SRB, Sulforhodamine B

[1200] PBS, phosphate buffered saline

[1201] DR, dose response

[1202] UV, ultraviolet light

[1203] SMCC, succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate LAR, linker to antibody ratio

[1204] Synthesis of drug

[1205] Compounds 1 and 2 were obtained following the procedure described in WO2003066638 (Examples 69 and 65, respectively, pages 112-116).

[1206] Compound 4 was obtained following the procedure described in WO2003066638 (Example 12, pages 61-62).

[1207] Compounds 8-S and 8-R were obtained by the procedure described in WO2018197663 (Example 8, pages 97-98).

[1208] Compound 16-S was obtained by the procedure described in WO2018197663 (Example 19, page 117).

[1209] Example 1-1

[1210] A)

[1211]

[1212] To a solution of 4 (35 mg, 0.054 mmol) in acetic acid (0.7 mL, 0.08 M) was added L-tryptophan (36 mg, 0.189 mmol, Sigma-Aldrich). The reaction mixture was stirred at 50 °C for 3 h, then the acetic acid was evaporated. Saturated aqueous NaHC03was added, and the mixture was extracted with CH2CI2. The combined organic layers were dried over Na2S04. Flash column chromatography (hexanes:EtOAc, 1 : 1) gave the pure compound 5-S (28 mg, 63%).

[1213] R f = 0.25 (hexanes:EtOAc, 1 : 1).

[1214] 1 H NMR (400 MHz, CDC13): δ 7.72 (s, 1H), 7.35 (d, J = 7.9 Hz, 1H), 7.26 (d, J = 7.9 Hz, 1H), 7.12 (t, J = 7.9 Hz, 1H), 7.02 (t, J = 8.0 Hz, 1H), 6.62 (s, 1H), 6.25 (s, 1H), 6.03 (s, 1H), 5.91-5.80 (m, 1H), 5.75 (s, 1H), 5.17-5.04 (m, 3H), 4.60 (s, 1H), 4.41 (s, 1H), 4.36 (d, J = 11.5 Hz, 1H), 4.29 (dd, J = 11.7, 2.1 Hz, 1H), 4.22 (d, J = 2.7 Hz, 1H), 3.81 (s, 3H), 3.59-3.44 (m, 3H), 3.35 (dd, J = 11.1, 9.0 Hz, 1H), 2.97-2.64 (m, 5H), 2.61 (dd, J = 15.3, 4.6 Hz, 1H), 2.43-2.29 (m, 1H), 2.37 (s, 3H), 2.28 (s, 3H), 2.05 (s, 3H).

[1215] ESI-MS m / z: C 44 H 45 Calcd for N5O9S: 819.3. Found: 820.3 (M + 1) + .

[1216] B)

[1217]

[1218] To a solution of 5-S (26 mg, 0.032 mmol) in CH2Cl2was added PdCl2(PPh3)2(13 mg, 0.02 mmol), acetic acid (0.069 mL, 1.2 mmol), and HSniBu3(0.17 mL, 0.64 mmol). The reaction mixture was stirred at 23 °C for 2 h. The crude product was concentrated in vacuo. Flash column chromatography (hexanes:EtOAc, 1 :9 to 9:1) afforded pure compound 6-S (17 mg, 68%).

[1219] R f = 0.15 (hexanes:EtOAc, 1 :1).

[1220] 1 H NMR (400 MHz, CDC13): δ 7.74 (s, 1H), 7.38 (d, J = 7.9 Hz, 1H), 7.26 (d, J = 7.9 Hz, 1H), 7.12 (t, J = 8.0 Hz, 1H), 7.02 (t, J = 7.9 Hz, 1H), 6.63 (s, 1H), 6.26 (s 1H), 6.03 (s, 1H), 5.82 (s, 1H), 5.15 (d, J = 11.6 Hz, 1H), 4.59 (s, 1H), 4.50 (d, J = 5.1 Hz, 1H), 4.41 (s, 1H), 4.29 (dd, J = 11.7, 2.1 Hz, 1H), 4.22 (d, J = 2.7 Hz, 1H), 3.90 - 3.81 (m, 1H), 3.80 (s, 3H), 3.67 - 3.49 (m, 1H), 3.49 (d, J = 5.2 Hz, 1H), 3.36 (dd, J = 11.0, 9.1 Hz, 1H), 3.11 - 2.85 (m, 3H), 2.60 (dd, J = 15.3, 4.5 Hz, 1H), 2.42 (d, J = 15.3 Hz, 1H), 2.38 - 2.22 (m, 2H), 2.35 (s, 3H), 2.25 (s, 3H), 2.06 (s, 3H).

[1221] ESI-MS m / z: C 41 H 41 Calcd for N5O9S: 779.3. Found: 780.2 (M + 1) + .

[1222] C)

[1223]

[1224] To a solution of 6-S (14 mg, 0.018 mmol) in CH3CN:H2O (1.39:1, 1.3 mL, 0.015 M) was added AgNO3(61 mg, 0.36 mmol). After 17 h at 23 °C, the reaction was quenched with a 1:1 mixture of brine and saturated aqueous NaHCO3, stirred for 15 min, diluted with CH2Cl2, stirred for 5 min, and extracted with CH2Cl2. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The resulting residue was purified by flash column chromatography (CH2Cl2:CH3OH, 99:1 to 85:15) to give pure 7-S (3 mg, 22%).

[1225] R f = 0.15 (CH2Cl2:CH3OH, 9:1).

[1226] 1 H NMR (500 MHz, CD3OD): δ 7.70 (s, 1H), 7.35 (d, J = 7.9 Hz, 1H), 7.26 (d, J = 7.9 Hz, 1H), 7.11 (t, J = 8.2 Hz, 1H), 7.02 (t, J = 8.2 Hz, 1H), 6.63 (s, 1H), 6.23 (s, 1H), 6.01 (s, 1H), 5.76 (s, 1H), 5.26 (d, J = 11.5 Hz, 1H), 4.92 (s, 1H), 4.54 (s, 1H), 4.48 (s, 2H), 4.37 (d, J = 5.3 Hz, 1H), 4.21 (d, J = 10.2 Hz, 1H), 3.80 (s, 3H), 3.67-3.50 (m, 4H), 3.36 (t, J = 10.2 Hz, 1H), 3.04-2.82 (m, 3H), 2.61 (dd, J = 15.2, 5.8 Hz, 1H), 2.42-2.28 (m, 2H), 2.36 (s, 3H), 2.27 (s, 3H), 2.02 (s, 3H).

[1227] ESI-MS m / z: C 40 H 42 N4O 10 S. Calcd: 770.3. Found: 753.2 (M-H2O+1) + .

[1228] Example 1-2

[1229] A)

[1230]

[1231] To a solution of 4 (400 mg, 0.62 mmol) in acetic acid (8 ml, 0.08 M) was added 8-S (468 mg, 2.13 mmol). The reaction mixture was stirred at 52 °C for 17 h, then acetic acid was evaporated. Saturated aqueous NaHC03was added, and the mixture was extracted with CH2Cl2. The combined organic layers were dried over Na2S04, filtered, and concentrated in vacuo. Flash column chromatography (hexanes:EtOAc, 1 : 1) gave pure compound 9-S (325 mg, 62%).

[1232] R f = 0.30 (hexanes:EtOAc, 1 : 1).

[1233] 1 H NMR (400 MHz, CDC13): δ 7.65 (s, 1H), 7.16 (d, J = 8.6 Hz, 1H), 6.78 (s, 1H), 6.77 (m, 1H), 6.62 (s, 2H), 6.23 (d, J = 1.3 Hz, 2H), 6.02 (d, J = 1.3 Hz, 2H), 5.85 (dddd, J = 17.1, 10.2, 6.8, 5.8 Hz, 1H), 5.75 (s, 1H), 5.15-5.00 (m, 3H), 4.59 (s, 1H), 4.43-4.22 (m, 4H), 3.80 (s, 3H), 3.78 (s, 3H), 3.53 (d, J = 12.9 Hz, 2H), 3.46 (d, J = 5.0 Hz, 1H), 3.38 (s, 1H), 2.93 (s, 1H), 2.86 (d, J = 4.4 Hz, 1H), 2.85-2.70 (m, 2H), 2.58 (dd, J = 15.2, 4.6 Hz, 1H), 2.42-2.30 (m, 2H), 2.37 (s, 3H), 2.26 (s, 3H), 2.04 (s, 3H).

[1234] ESI-MS m / z: C 45 H 47 N5O 10 Calcd for S: 849.9. Found: 850.3 (M + 1) + .

[1235] B)

[1236]

[1237] To a solution of 9-S (325 mg, 0.38 mmol) in CH2Cl2was added PdCl2(PPh3)2(160 mg, 0.23 mmol), acetic acid (0.82 mL, 14.2 mmol) and HSniBu3(1.7 mL, 6.27 mmol). The reaction mixture was stirred at 23 °C for 1.5 h. The crude product was concentrated in vacuo. Flash column chromatography (hexanes:EtOAc, 1:9 to 9:1) afforded pure compound 10-S (180 mg, 59%).

[1238] R f = 0.15 (hexanes:EtOAc, 1:1).

[1239] 1 H NMR (400 MHz, CDC13): δ 7.19 (s, 1H), 6.79 (m, 2H), 6.65 (s, 1H), 6.26 (s, 1H), 6.03 (d, J = 1.4 Hz, 2H), 5.77 (d, J = 11.5 Hz, 1H), 5.10 (s, 1H), 4.59 (s, 1H), 4.48 (d, J = 4.9 Hz, 1H), 4.39-4.29 (m, 3H), 3.79 (s, 3H), 3.79 (s, 3H), 3.64-3.33 (m, 4H), 3.03-2.90 (m, 4H), 2.59 (d, J = 14.6 Hz, 2H), 2.44-2.32 (m, 2H), 2.37 (s, 3H), 2.26 (s, 3H), 2.04 (s, 3H).

[1240] ESI-MS m / z: C 42 H 43 N5O 10 Calcd for S: 809.3. Found: 810.3 (M + 1) + .

[1241] C)

[1242]

[1243] To a solution of 10-S (180 mg, 0.22 mmol) in CH3CN:H2O (1.39:1, 16 ml, 0.015 μm) was added AgNO3(756 mg, 4.40 mmol). After 18 h at 23 °C, the reaction was quenched with a 1:1 mixture of brine and saturated aqueous NaHCO3, stirred for 15 min, diluted with CH2Cl2, stirred for 5 min, and extracted with CH2Cl2. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The resulting residue was purified by flash column chromatography (CH2Cl2:CH3OH, 99:1 to 85:15) to give pure 11-S (100 mg, 56%).

[1244] R f = 0.35 (CH2Cl2:CH3OH, 9:1).

[1245] 1 H NMR (500 MHz, CD3OD): δ 7.15 (dd, J = 8.8, 0.6 Hz, 1H), 6.82 (dd, J = 2.5, 0.6 Hz, 1H), 6.68 (dd, J = 8.9, 2.5 Hz, 1H), 6.56 (s, 1H), 6.27 (d, J = 1.3 Hz, 1H), 6.08 (d, J = 1.3 Hz, 1H), 5.31 (d, J = 11.5 Hz, 1H), 4.62-4.55 (m, 1H), 4.44 (ddtd, J = 4.9, 1.5, 1.0, 0.5 Hz, 2H) 4.38-4.27 (m, 1H), 4.25-4.18 (m, 1H), 3.75 (s, 3H), 3.74 (s, 3H), 3.64 (d, J = 4.8 Hz, 1H), 3.61-3.42 (m, 3H), 3.13-2.95 (m, 3H), 2.80 (dd, J = 10.4, 5.4 Hz, 2H), 2.68 (dd, J = 15.1, 4.2 Hz, 2H), 2.55 (d, J = 15.4 Hz, 1H), 2.51-2.36 (m, 3H), 2.34 (s, 3H), 2.29 (s, 3H), 2.00 (s, 3H).

[1246] 13C NMR (126 MHz, CD3OD): d 172.6, 169.2, 155.1, 148.0, 147.2, 144.7, 142.4, 142.1, 133.1, 132.6, 132.2, 131.1, 128.2, 125.5, 122.2, 122.0, 116.3, 112.9, 112.8, 111.4, 109.0, 103.5, 100.9, 91.0, 66.6, 65.0, 61.8, 60.3, 59.2, 57.1, 56.1, 51.7, 47.2, 45.5, 43.8, 39.0, 28.2, 25.4, 20.6, 16.3, 9.5.

[1247] ESI-MS m / z: C 41 H 44 N4O 11 S. Found: 783.4 (M-H2O+1) + .

[1248] (+)-HR-ESI-TOF-MS m / z 800.2796 [M+H] + (C 41 H 44 N4O 11 S. Found: 783.4 (M-H2O+1)

[1249] Example 1-3

[1250] A)

[1251]

[1252] To a solution of 4 (400 mg, 0.62 mmol) in acetic acid (8 ml, 0.08 m) was added 8-R (468 mg, 2.13 mmol). The reaction mixture was stirred at 52 °C for 17 h, then acetic acid was evaporated. Saturated aqueous NaHC03was added, and the mixture was extracted with CH2CI2. The combined organic layers were dried over Na2S04, filtered, and concentrated in vacuo. Flash column chromatography (hexanes:EtOAc, 1:1) gave the pure compound 9-R (390 mg, 77%).

[1253] R f = 0.30 (hexanes:EtOAc, 1:1).

[1254] 1H NMR (400 MHz, CDC13): δ 7.64 (s, 1H), 7.14 (d, J = 8.8 Hz, 1H), 6.81 (d, J = 2.6 Hz, 1H), 6.74 (dd, J = 8.8, 2.4 Hz, 1H), 6.59 (s, 1H), 6.18 (d, J = 1.4 Hz, 1H), 5.97 (d, J = 1.4 Hz, 1H), 5.91 - 5.80 (m, 1H), 5.79 (s, 1H), 5.15 - 4.92 (m, 3H), 4.62 (s, 1H), 4.42 - 4.23 (m, 2H), 4.23 - 4.03 (m, 3H), 3.79 (s, 3H), 3.78 (s, 3H), 3.68 - 3.48 (m, 2H), 3.43 (d, J = 5.1 Hz, 2H), 3.01 - 2.68 (m, 3H), 2.57 - 2.41 (m, 3H), 2.39 (s, 3H), 2.25 (s, 3H), 2.22 - 2.20 (m, 1H), 2.07 (s, 3H).

[1255] ESI-MS m / z: C 45 H 47 N5O 10 S. Calcd: 849.9. Found: 850.4 (M + 1) + .

[1256] B)

[1257]

[1258] To a solution of 9-R (390 mg, 0.46 mmol) in CH2Cl2was added PdCl2(PPh3)2(193 mg, 0.28 mmol), acetic acid (1.0 mL, 17.2 mmol), and HS1Bu3(2.04 mL, 7.60 mmol). The reaction mixture was stirred at 23 °C for 1.5 h. The crude product was concentrated in vacuo. Flash column chromatography (hexanes:EtOAc, 1:9 to 9:1) afforded pure compound 10-R (210 mg, 57%).

[1259] R f = 0.15 (hexanes:EtOAc, 1:1).

[1260] 1H NMR (400 MHz, CDC13): δ 7.16 (d, J = 8.8 Hz, 1H), 6.82 (d, J = 2.4 Hz, 1H), 6.76 (dd, J = 8.8, 2.5 Hz, 1H), 6.62 (s, 1H), 6.22 (d, J = 1.5 Hz, 1H), 6.00 (d, J = 1.5 Hz, 1H), 5.03 (d, J = 11.5 Hz, 1H), 4.62 (s, 1H), 4.51 (d, J = 5.0 Hz, 1H), 4.36 (s, 1H), 4.23 - 4.11 (m, 3H), 3.85 (dd, J = 8.1, 4.1 Hz, 1H), 3.80 (s, 3H), 3.78 (s, 3H), 3.72 - 3.57 (m, 1H), 3.45 (d, J = 5.2 Hz, 2H), 3.12 (d, J = 17.6 Hz, 1H), 2.99 (dd, J = 17.9, 9.6 Hz, 1H), 2.54 (s, 2H), 2.46 (d, J = 14.7 Hz, 2H), 2.38 (s, 3H), 2.33 - 2.19 (m, 1H), 2.26 (s, 3H), 2.08 (s, 3H).

[1261] ESI-MS m / z: C 42 H 43 N5O 10 S. Calcd: 809.3. Found: 810.5 (M + 1) + .

[1262] C)

[1263]

[1264] To a solution of 10-R (210 mg, 0.26 mmol) in CH3CN:H2O (1.39:1, 18 mL, 0.015 μm) was added AgNO3(883 mg, 5.20 mmol). After 18 h at 23 °C, the reaction was quenched with a 1:1 mixture of brine and saturated aqueous NaHCO3, stirred for 15 min, diluted with CH2Cl2, stirred for 5 min, and extracted with CH2Cl2. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The resulting residue was purified by flash column chromatography (CH2Cl2:CH3OH, 99:1 to 85:15) to give pure 11-R (140 mg, 66%).

[1265] R f = 0.30 (CH2Cl2:CH3OH, 9:1).

[1266] 1H NMR (500 MHz, CD3OD): δ 7.13 (dd, J = 8.8, 0.6 Hz, 1H), 6.82 (d, J = 8.8 Hz, 1H), 6.67 (dd, J = 8.8, 2.5 Hz, 1H), 6.61 (s, 1H), 6.25 (d, J = 1.3 Hz, 1H), 6.07 (d, J = 1.4 Hz, 1H), 5.21 (d, J = 11.3 Hz, 1H), 4.80 - 4.72 (m, 2H), 4.58 (s, 1H), 4.45 (d, J = 5.4 Hz, 1H), 4.21 (d, J = 2.7 Hz, 1H), 4.16 - 4.06 (m, 1H), 3.75 (s, 3H), 3.72 (s, 3H), 3.64 (d, J = 7.7 Hz, 2H), 3.55 - 3.48 (m, 3H), 3.16 (d, J = 17.6 Hz, 1H), 3.03 (dd, J = 17.7, 9.8 Hz, 1H), 2.76 - 2.63 (m, 2H), 2.32 (s, 3H), 2.29 (s, 3H), 2.25 - 2.11 (m, 2H), 2.04 (s, 3H).

[1267] 13 C NMR (126 MHz, CD3OD): δ 171.6, 153.6, 146.6, 145.9, 143.4, 141.3, 140.9, 132.1, 131.1, 130.8, 129.7, 126.4, 121.2, 120.7, 114.8, 112.1, 111.5, 110.0, 108.8, 107.6, 107.6, 102.1, 99.5, 89.6, 65.4, 63.1, 60.1, 59.0, 57.8, 55.9, 54.7, 52.7, 45.9, 26.6, 25.1, 24.2, 19.4, 19.1, 14.8, 13.0, 8.2.

[1268] ESI-MS m / z: C 41 H 44 N4O 11 S of calculated: 800.3. Experimental: 783.3 (M-H2O+1) + .

[1269] (+)-HR-ESI-TOF-MS m / z 800.2781 [M+H] + (C 41 H 44 N4O 11 S of calculated: 800.2727).

[1270] Example 1-4

[1271] A)

[1272]

[1273] To a solution of 4 (350 mg, 0.54 mmol) in acetic acid (7 ml, 0.08 m) was added 2-benzofuran-3-yl-ethylamine hydrochloride (12) (1.52 g, 7.70 mmol, Sigma Aldrich). The reaction mixture was stirred at 52 °C for 72 h, then acetic acid was evaporated. Saturated aqueous NaHC03solution was added, the mixture was extracted with CH2Cl2. The combined organic layers were dried over anhydrous Na2S04, filtered, and concentrated in vacuo. Flash column chromatography (hexane:EtOAc, 1 : 1) gave pure 13 (180 mg, 42%).

[1274] R f = 0.5 (hexane:EtOAc, 1 : 1).

[1275] 1 H NMR (400 MHz, CDC13): δ 7.39 - 7.29 (m, 2H), 7.23 - 7.07 (m, 2H), 6.64 (s, 1H), 6.19 (d, J = 1.3 Hz, 1H), 6.04 (d, J = 1.3 Hz, 1H), 5.97 - 5.80 (m, 1H), 5.78 (s, 1H), 5.19 - 4.97 (m, 3H), 4.54 (s, 1H), 4.36 (dd, J = 4.8, 1.6 Hz, 1H), 4.31 (s, 1H), 4.20 (dd, J = 11.4, 1.9 Hz, 2H), 3.80 (s, 3H), 3.59 - 3.49 (m, 1H), 3.47 (dd, J = 7.0, 2.9 Hz, 1H), 3.25 (ddd, J = 11.4, 8.1, 5.0 Hz, 1H), 3.04 (d, J = 18.0 Hz, 1H), 2.98 - 2.72 (m, 5H), 2.59 - 2.49 (m, 2H), 2.37 (s, 3H), 2.27 (s, 3H), 2.23 - 2.12 (m, 1H), 2.07 (s, 3H).

[1276] ESI-MS m / z: C 43 H 42 Calculated for N4O9S: 790.9. Found: 791.5 (M + 1) + .

[1277] B)

[1278]

[1279] To a solution of 13 (320 mg, 0.40 mmol) in CH2Cl2was added PdCl2(PPh3)2(170 mg, 0.24 mmol), acetic acid (0.86 mL, 15 mmol), and SnBu3H (1.78 mL, 6.60 mmol). The reaction mixture was stirred at 23 °C for 1.5 h. The crude product was concentrated in vacuo. Flash column chromatography (hexanes:EtOAc, 1:9 to 9:1) afforded pure 14 (130 mg, 43%).

[1280] R f = 0.15 (hexanes:EtOAc, 1:1).

[1281] 1 H NMR (400 MHz, CDC13): δ 7.40-7.31 (m, 2H), 7.19-7.10 (m, 2H), 6.65 (s, 1H), 6.19 (d, J = 1.5 Hz, 1H), 6.04 (d, J = 1.5 Hz, 1H), 5.05 (d, J = 11.5 Hz, 1H), 4.43 (s, 1H), 4.51-4.47 (m, 1H), 4.30 (s, 1H), 4.21 (d, J = 2.3 Hz, 2H), 3.86-3.76 (m, 2H), 3.79 (d, J = 1.9 Hz, 2H), 3.46 (d, J = 4.7 Hz, 1H), 3.29-3.22 (m, 1H), 3.19 (d, J = 17.9 Hz, 1H), 2.99 (dd, J = 17.9, 9.4 Hz, 1H), 2.83 (s, 1H), 2.53 (dt, J = 7.9, 4.8 Hz, 2H), 2.35 (s, 3H), 2.33-2.23 (m, 1H), 2.27 (s, 3H), 2.20-2.14 (m, 1H), 2.07 (m, 3H).

[1282] ESI-MS m / z: C 40 H 38 Calcd for N4O9S: 750.8. Found: 751.9 (M + 1) + .

[1283] C)

[1284]

[1285] To a solution of 14 (130 mg, 0.17 mmol) in CH3CN:H2O (1.39:1, 12 mL, 0.015 M) was added AgN03(578 mg, 3.40 mmol). After 3 h at 23 °C, a 1 : 1 mixture of saturated aqueous NaHC03and brine was added, stirred for 15 min, diluted with CH2CI2, stirred for 5 min, and extracted with CH2CI2. The combined organic layers were dried over anhydrous Na2S04, filtered, and concentrated in vacuo. The resulting residue was purified by flash column chromatography (CH2CI2:CH3OH, 99: 1 to 85: 15) to give pure 15 (80 mg, 64%).

[1286] R f = 0.25 (CH2CI2:CH3OH, 9: 1).

[1287] 1 H NMR (500 MHz, CD3OD): δ 7.45-7.34 (m, 2H), 7.26-7.09 (m, 2H), 6.60 (s, 1H), 6.06 (d, J = 1.1 Hz, 1H), 6.24 (d, J = 1.1 Hz, 1H), 5.24 (d, J = 11.5 Hz, 1H), 4.74 (s, 1H), 4.52 (s, 1H), 4.47 (d, J = 4.9 Hz, 1H), 4.19-4.09 (m, 2H), 3.74 (s, 3H), 3.64 (d, J = 9.2 Hz, 1H), 3.57 (d, J = 4.9 Hz, 1H), 3.43-3.37 (m, 1H), 3.20-3.09 (m, 1H), 3.04 (dd, J = 17.8, 9.5 Hz, 1H), 2.96-2.90 (m, 1H), 2.83 (d, J = 15.4 Hz, 1H), 2.59-2.56 (m, 2H), 2.34 (s, 3H), 2.30 (s, 3H), 2.10-2.02 (m, 1H), 2.05 (s, 3H).

[1288] 13 C NMR (126 MHz, CD3OD): δ 171.9, 170.7, 156.0, 150.5, 148.7, 147.0, 144.8, 142.4, 142.1, 132.6, 131.2, 128.6, 125.5, 124.7, 123.8, 122.3, 121.2, 120.2, 116.8, 114.9, 114.0, 112.3, 103.5, 91.4, 90.7, 63.7, 62.3, 60.4, 58.7, 57.1, 47.2, 43.5, 40.8, 39.3, 28.2, 21.5, 20.6, 16.2, 9.6.

[1289] ESI-MS m / z: C 39 H 39 N3O 10 Calcd for S: 741.8. Found: 724.9 (M-H2O+1) + .

[1290] (+)-HR-ESI-TOF-MS m / z: 741.2416 [M+H] + (C 39 H 39 N3O 10 Calcd for S: 741.8. Found: 724.9 (M-H2O+1)

[1291] Example 1-5

[1292] A)

[1293]

[1294] To a solution of 4 (150 mg, 0.24 mmol) in CH3CN (15 mL, 0.016 M) was added 16-S (230 mg, 1.20 mmol) and cyanuric chloride (TCT) (45 mg, 30%). The reaction mixture was stirred at 85 °C for 24 h, then saturated aqueous NaHCO3 was added and the mixture was extracted with CH2Cl2. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Flash column chromatography (hexanes:EtOAc, 9:1 to 1:9) afforded the pure compound 17-S (145 mg, 73% yield).

[1295] 1H NMR (400 MHz, CDC13): δ 7.35 (dt, J = 8.2, 0.9 Hz, 1H), 7.31 (ddd, J = 7.6, 1.5, 0.7 Hz, 1H), 7.20 (ddd, J = 8.4, 7.2, 1.5 Hz, 1H), 7.13 (td, J = 7.4, 1.1 Hz, 1H), 6.62 (s, 1H), 6.20 (d, J = 1.5 Hz, 1H), 6.05 (d, J = 1.4 Hz, 1H), 5.85 (m, 1H), 5.74 (s, 1H), 5.16 - 5.08 (m, 3H), 4.58 (s, 1H), 4.40 - 4.32 (m, 2H), 4.28 (dd, J = 11.5, 2.2 Hz, 1H), 4.19 (d, J = 2.9 Hz, 1H), 3.80 (s, 3H), 3.58 - 3.53 (m, 1H), 3.50 (dd, J = 11.3, 4.1 Hz, 2H), 3.42 - 3.30 (m, 1H), 2.96 (s, 1H), 2.90 - 2.73 (m, 4H), 2.58 (dd, J = 15.7, 4.9 Hz, 1H), 2.52 (d, J = 15.0 Hz, 1H), 2.37 (s, 3H), 2.36 - 2.26 (m, 2H), 2.28 (s, 3H), 2.04 (s, 3H).

[1296] ESI-MS m / z: 821.3 (M+H) + .

[1297] B)

[1298]

[1299] To a solution of 17-S (140 mg, 0.17 mmol) in CH2Cl2was added PdCl2(PPh3)2(19 mg, 0.027 mmol), acetic acid (0.097 mL, 1.70 mmol), and SnBu3H (1.65 mL, 6.12 mmol). The reaction mixture was stirred at 23 °C for 3 h. The crude product was concentrated in vacuo. Flash column chromatography (hexanes:EtOAc, 1:9 to 9:1) afforded pure compound 18-S (94 mg, 71% yield).

[1300] 1H NMR (400 MHz, CDC13): δ 7.35 (dt, J = 8.2, 0.9 Hz, 1H), 7.31 (dt, J = 7.6, 1.0 Hz, 1H), 7.20 (ddd, J = 8.3, 7.2, 1.5 Hz, 1H), 7.13 (td, J = 7.4, 1.1 Hz, 1H), 6.62 (s, 1H), 6.20 (d, J = 1.4 Hz, 1H), 6.05 (d, J = 1.4 Hz, 1H), 5.09 (dd, J = 11.5, 1.1 Hz, 1H), 4.58 (s, 1H), 4.52 (d, J = 5.0 Hz, 1H), 4.39 - 4.35 (m, 1H), 4.27 (dd, J = 11.5, 2.1 Hz, 1H), 4.20 (d, J = 2.6 Hz, 1H), 3.85 (d, J = 18.3 Hz, 1H), 3.79 (s, 3H), 3.54 - 3.44 (m, 2H), 3.34 (dd, J = 11.2, 9.2 Hz, 1H), 3.04 - 2.97 (m, 2H), 2.92 (tt, J = 8.6, 4.3 Hz, 1H), 2.60 - 2.47 (m, 2H), 2.35 (s, 3H), 2.34 - 2.28 (m, 2H), 2.28 (s, 3H), 2.05 (s, 3H).

[1301] ESI-MS m / z: 781.3 (M + H) + .

[1302] C)

[1303]

[1304] To a solution of 18-S (90 mg, 0.11 mmol) in CH3CN:H2O (1.39:1, 8 mL, 0.015 M) was added AgNO3(580 mg, 3.45 mmol). After 18 h at 23 °C, a mixture of 1:1 brine and saturated aqueous NaHCO3was added, stirred for 15 min, diluted with CH2Cl2, stirred for 5 min, and extracted with CH2Cl2. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The resulting residue was purified by flash column chromatography (CH2Cl2:CH3OH, 99:1 to 85:15) to give pure 19-S (60 mg, 68% yield).

[1305] 1H NMR (400 MHz, CDC13): δ 7.35 (d, J = 8.1 Hz, 1H), 7.32-7.28 (m, 1H), 7.19 (td, J = 8.3, 7.8, 1.4 Hz, 1H), 7.16-7.09 (m, 1H), 6.58 (s, 1H), 6.18 (d, J = 1.5 Hz, 1H), 6.04 (d, J = 4.6 Hz, 1H), 5.18 (d, J = 11.3 Hz, 1H), 4.91 (s, 1H), 4.63 (s, 1H), 4.60-4.46 (m, 2H), 4.18 (d, J = 10.8 Hz, 2H), 3.83-3.71 (m, 2H), 3.78 (s, 1H), 3.69 (s, 1H), 3.56-3.44 (m, 2H), 3.32 (t, J = 10.3 Hz, 1H), 3.08-2.86 (m, 2H), 2.54 (dd, J = 15.6, 5.0 Hz, 2H), 2.37-2.23 (m, 2H), 2.32 (s, 3H), 2.27 (s, 3H), 2.04 (s, 3H).

[1306] ESI-MS m / z: 754.3 (M-H20+H) + .

[1307] Synthesis of Linker

[1308] Preparation of LIN 1 : MC-Val-Cit-PABC-PNP

[1309] Reaction Mechanism

[1310]

[1311] (a) Preparation of LIN 1-1 : MC-Val-Cit-OH

[1312] LIN 1-1

[1313]

[1314] Cl-Trt Cl-resin (20 g, 1.49 mmol / g) (Iris Biotech, Ref: BR-1065, 2-Chlorotrityl chloride resin (200-400 mesh, 1% DVB, 1.0-1.6 mmol / g), CAS 42074-68-0) was placed in a filter plate. 100 mL of DCM was added to the resin and the mixture was stirred for 1 h. The solvent was removed by filtration under vacuum. A solution of Fmoc-Cit-OH (11.83 g, 29.78 mmol) and DIPEA (17.15 mL, 98.45 mmol) in DCM (80 mL) was added and the mixture was stirred for 10 min. After this, DIPEA (34.82 mmol, 199.98 mmol) was added and the mixture was stirred for 1 h. The reaction was terminated by the addition of MeOH (30 mL) after 15 min of stirring. The thus produced Fmoc-Cit-O-Trt Cl-resin was subjected to the following washings / treatments: DCM (5 x 50 mL x 0.5 min), DMF (5 x 50 mL x 0.5 min), piperidine:DMF (1:4, 1 x 1 min, 2 x 10 min), DMF (5 x 50 mL x 0.5 min), DCM (5 x 50 mL x 0.5 min). The final piperidine wash resulted in NH2-Cit-O-Trt Cl-resin. Calculated loading: 1.15 mmol / g.

[1315] The above produced NH2-Cit-O-Trt Cl-resin was washed with DMF (5 x 50 mL x 0.5 min) and a solution of Fmoc-Val-OH (31.22 g, 91.98 mmol), HOBt (11.23 g, 91.98 mmol) in DMF (100 mL) was added to the NH2-Cit-O-Trt Cl-resin, stirred and DIPCDI (14.24 mL, 91.98 mmol) was added and the mixture was stirred for 1.5 h. The reaction was terminated by washing with DMF (5 x 50 mL x 0.5 min). The thus produced Fmoc-Val-Cit-O-Trt Cl-resin was treated with piperidine:DMF (1:4, 1 x 1 min, 2 x 10 min) and washed with DMF (5 x 50 mL x 0.5 min). The final piperidine wash resulted in NH2-Val-Cit-O-Trt Cl-resin.

[1316] A solution of 6-maleimidocaproic acid (MC-OH) (9.7 g, 45.92 mmol), HOBt (6.21 g, 45.92 mmol) in DMF (100 mL) was added to the above produced NH2-Val-Cit-O-Trt Cl-resin, stirred and DIPCDI (7.12 mL, 45.92 mmol) was added and the mixture was stirred for 1.5 h. The reaction was terminated by washing with DMF (5 x 50 mL x 0.5 min) and DCM (5 x 50 mL x 0.5 min).

[1317] The peptide was cleaved from the resin by treatment with TFA:DCM (1 :99, 5 x 100 mL). The resin was washed with DCM (7 x 50 mL x 0.5 min). The combined filtrates were evaporated to dryness under reduced pressure and the obtained solid was triturated with Et20 and filtered to obtain LIN 1-1 as a white solid (7.60 g, 71%).

[1318] 1 H NMR (500 MHz, DMSO-d6): δ 12.47 (s, 1H), 8.13 (d, J = 7.3 Hz, 1H), 7.74 (d, J = 9.0 Hz, 1H), 6.99 (s, 2H), 5.93 (s, 1H), 5.35 (s, 2H), 4.20 (dd, J = 9.0, 6.8 Hz, 1H), 4.15-4.07 (m, 1H), 3.36 (t, J = 7.0 Hz, 2H), 3.00-2.88 (m, 2H), 2.21-2.12 (m, 1H), 2.11-2.03 (m, 1H), 1.98-1.86 (m, 1H), 1.74-1.62 (m, 1H), 1.61-1.50 (m, 1H), 1.50-1.31 (m, 6H), 1.21-1.11 (m, 2H), 0.84 (d, J = 6.8 Hz, 3H), 0.80 (d, J = 6.8 Hz, 3H).

[1319] ESI-MS m / z: C 21 H 33 Calculated for N5O7: 467.2. Found: 468.3 (M+H) + .

[1320] (b) Preparation of LIN 1-2: MC-Val-Cit-PABOH

[1321] LIN 1-2

[1322]

[1323] To a solution of LIN 1-1 (1.6 g, 3.42 mmol) and 4-aminobenzyl alcohol (PABOH) (0.84 g, 6.84 mmol) in DCM (60 mL) was added a solution of HOBt (0.92 g, 6.84 mmol) in DMF (5 mL). DIPCDI (1.05 mL, 6.84 mmol) was added, the reaction mixture was stirred at 23 °C for 2 h, Et2O (150 mL) was added, and the resulting solid was filtered on a filter plate under vacuum to give LIN 1-2 (1.31 g, 67%).

[1324] 1 H NMR (500 MHz, DMSO-d6): δ 9.88 (s, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.77 (dd, J = 12.2, 8.5 Hz, 1H), 7.53 (d, J = 8.2 Hz, 2H), 7.21 (d, J = 8.2 Hz, 2H), 6.99 (s, 3H), 6.01 - 5.92 (m, 1H), 5.39 (s, 2H), 5.07 (s, 1H), 4.41 (s, 2H), 4.39 - 4.31 (m, 1H), 4.23 - 4.12 (m, 1H), 3.36 (t, J = 7.0 Hz, 2H), 3.06 - 2.97 (m, 1H), 2.96 - 2.90 (m, 1H), 2.22 - 2.03 (m, 2H), 2.01 - 1.88 (m, 1H), 1.76 - 1.62 (m, 1H), 1.63 - 1.28 (m, 6H), 1.25 - 1.11 (m, 2H), 0.84 (d, J = 6.9 Hz, 3H), 0.81 (d, J = 6.8 Hz, 3H).

[1325] ESI-MS m / z: C 28 H 40 Calculated for N6O7: 572.3. Found: 573.3 (M+H) + .

[1326] (c) Preparation of LIN 1: MC-Val-Cit-PAB-PNP

[1327] LIN 1

[1328]

[1329] To a solution of LIN 1-2 (500 mg, 0.87 mmol) and bis-PNP (2.64 g, 8.72 mmol) in DCM:DMF (8:2, 25 mL) was added DIPEA (0.45 mL, 2.61 mmol). The reaction mixture was stirred at 23 °C for 20 h and poured onto a silica gel column (DCM:CH3OH, 50:1 to 10:1) to give the pure target LIN 1 (364 mg, 57%).

[1330] R f = 0.40 (CH2Cl2:CH3OH, 9:1).

[1331] 1 H NMR (400 MHz, CDC13 / CD30D): δ 9.45 (s, 1H), 8.23 (d, J = 8.3 Hz, 2H), 7.59 (d, J = 8.5 Hz, 2H), 7.35 (d, J = 8.3 Hz, 2H), 7.34 (d, J = 8.5 Hz, 2H), 6.65 (s, 2H), 5.20 (s, 2H), 4.56 (dt, J = 10.5, 5.4 Hz, 1H), 4.15 (d, J = 7.2 Hz, 1H), 3.46 (dd, J = 8.0, 6.4 Hz, 2H), 3.16 - 2.89 (m, 2H), 2.21 (dd, J = 8.3, 6.6 Hz, 2H), 2.06 - 1.97 (m, 1H), 1.90 - 1.83 (m, 1H), 1.73 - 1.46 (m, 7H), 1.34 - 1.20 (m, 2H), 0.91 (d, J = 6.7 Hz, 3H), 0.90 (d, J = 6.7 Hz, 3H).

[1332] 13 C NMR (125 MHz, CDC13 / CD30D) δ 174.4, 172.4, 171.1, 170.6, 160.5, 155.5, 152.5, 145.3, 138.7, 134.1, 129.9, 129.5, 125.2, 121.8, 120.0, 70.6, 59.0, 53.2, 37.5, 35.8, 30.6, 29.6, 29.3, 28.1, 26.2, 26.2, 25.1, 19.1, 18.1.

[1333] ESI-MS m / z: C 35 H 43 N7O 11 Calculated: 737.3. Found: 738.3 (M+H) + .

[1334] Preparation of LIN-2: MC2-PEG4-Val-Cit-PABC-PNP

[1335] Reaction mechanism

[1336]

[1337] a) Preparation of LIN 2-1 : MC2-PEG4-Val-Cit-OH

[1338] LIN 2-1

[1339]

[1340] Cl-TrtCl-resin (5 g, 1.49 mmol / g) was placed in a filter plate. CH2Cl2(25 mL) was added to the resin and the mixture was stirred at 23 °C for 1 h. The solvent was removed by filtration under vacuum. A solution of Fmoc-Cit-OH (2.95 g, 7.44 mmol) and DIPEA (4.29 mL, 24.61 mmol) in CH2Cl2(20 mL) was added and the mixture was stirred at 23 °C for 10 min. Additional DIPEA (8.70 mL, 49.99 mmol) was added and the mixture was stirred at 23 °C for 1 h. The reaction was stopped by addition of MeOH (10 mL) and stirred at 23 °C for 15 min. Fmoc-Cit-O-TrtCl-resin was subjected to the following washings / treatments: CH2Cl2(5 x 15 mL x 0.5 min), DMF (5 x 15 mL x 0.5 min), piperidine:DMF (1 :4, 15 mL, 1 x 1 min, 2 x 10 min), DMF (5 x 15 mL x 0.5 min), CH2Cl2(5 x 15 mL x 0.5 min). The calculated loading was 1.17 mmol / g.

[1341] NH2-Cit-O-TrtCl-resin was washed with DMF (5 x 15 mL x 0.5 min) and a solution of Fmoc-Val-OH (7.80 g, 22.99 mmol) and HOBt (2.80 g, 24.5 mmol) in DMF (25 mL) was added to the NH2-Cit-O-TrtCl-resin followed by DIPCDI (3.56 mL, 24.5 mmol) at 23 °C. The reaction mixture was stirred at 23 °C for 1.5 h. The reaction was stopped by washing with DMF (5 x 15 mL x 0.5 min). Fmoc-Val-Cit-O-TrtCl-resin was treated with piperidine:DMF (1 :4, 15 mL, 1 x 1 min, 2 x 10 min) and washed with DMF (5 x 15 mL x 0.5 min).

[1342] A solution of 15-(9-fluorenylmethoxycarbonyl)amino-4,7,10,13- tetraoxa-pentadecanoic acid (Fmoc-NH-PEG4-OH) (4.27 g, 8.75 mmol) and HOBt (1.18 g, 8.72 mmol) in DMF (30 mL) was added to NH2-Val-Cit-O-Trt Cl-resin followed by the addition of DIPCDI (1.35 mL, 8.72 mmol) at 23 °C. The reaction mixture was stirred at 23 °C for 24 h. The reaction was stopped by washing with DMF (5 x 15 mL x 0.5 min). The Fmoc-NH-PEG4-Val-Cit-O-Trt Cl-resin was treated with piperidine:DMF (1:4, 15 mL, 1 x 1 min, 2 x 10 min) and washed with DMF (5 x 15 mL x 0.5 min).

[1343] A solution of 3-(maleimidopropionic acid) (MC2-OH) (3.95 g, 23.35 mmol) and HOBt (3.16 g, 23.37 mmol) in DMF (30 mL) was added to NH2-PEG4-Val-Cit-O-Trt Cl-resin followed by the addition of DIPCDI (3.62 mL, 23.37 mmol) at 23 °C. The reaction mixture was stirred at 23 °C for 2 h. The reaction was stopped by washing with DMF (5 x 15 mL x 0.5 min) and CH2Cl2(5 x 15 mL x 0.5 min).

[1344] The peptide was cleaved from the resin by treatment with TFA:CH2Cl2(1:99, 5 x 50 mL). The resin was washed with CH2Cl2(7 x 50 mL x 0.5 min). The combined filtrates were evaporated to dryness under reduced pressure, the obtained solid was triturated with Et2O and filtered to obtain LIN 2-1 as a white solid (4.59 g, 87% yield).

[1345] 1H NMR (300 MHz, CDC13): δ 7.67-7.57 (m, 1H), 7.44 (d, J = 8.3 Hz, 1H), 7.11 (t, J = 5.4 Hz, 1H), 6.73 (s, 2H), 4.49 (d, J = 7.2 Hz, 1H), 4.35 (t, J = 7.7 Hz, 1H), 3.82 (t, J = 7.0 Hz, 2H), 3.74 (t, J = 6.2 Hz, 2H), 3.68-3.56 (m, 13H), 3.56-3.45 (m, 2H), 3.39 (q, J = 5.4 Hz, 2H), 3.17 (s, 2H), 2.55 (q, J = 7.0, 6.0 Hz, 4H), 2.16-1.99 (m, 1H), 1.91 (s, 1H), 1.75 (s, 1H), 1.43 (s, 2H), 0.94 (d, = 9.7 Hz, 3H), 0.93 (d, = 9.7 Hz, 3H).

[1346] ESI-MS m / z: 673.3 (M + H) + .

[1347] (b) Preparation of LIN 2-2: MC2-PEG4-Val-Cit-PABOH

[1348] LIN 2-2

[1349]

[1350] To a solution of LIN 2-1 (1.5 g, 2.22 mmol) and 4-aminobenzyl alcohol (PABOH) (0.55 g, 4.45 mmol) in CH2Cl2(60 mL) at 23 °C was added a solution of HOBt (0.60 g, 4.45 mmol) in DMF (5 mL) followed by DIPCDI (0.69 mL, 4.45 mmol). The reaction mixture was stirred at 23 °C for 5 h, Et2O (150 mL) was added, and the resulting solid was vacuum filtered to give crude LIN 2-2 (2.37 g, >100% yield) which was used in the next step without further purification.

[1351] 1H NMR (500 MHz, DMSO-d6): δ 7.57 (d, J = 8.6 Hz, 2H), 7.30 (d, J = 8.6 Hz, 2H), 6.81 (s, 2H), 4.58 (s, 1H), 4.56 (s, 2H), 4.50 (dd, J = 9.1, 5.1 Hz, 1H), 4.21 (d, J = 7.0 Hz, 1H), 3.80 - 3.68 (m, 4H), 3.65 - 3.59 (m, 12H), 3.55 - 3.47 (m, 1H), 3.20 (dd, J = 13.6, 6.9 Hz, 1H), 3.12 (dt, J = 13.5, 6.7 Hz, 1H), 2.55 (td, J = 6.1, 2.1 Hz, 2H), 2.46 (t, J = 6.9 Hz, 2H), 2.15 - 2.07 (m, 1H), 1.95 - 1.88 (m, 1H), 1.79 - 1.70 (m, 1H), 1.67 - 1.50 (m, 2H), 0.99 (d, J = 7.0 Hz, 3H), 0.98 (d, J = 7.0 Hz, 3H).

[1352] ESI-MS m / z: 778.4 (M+H) + .

[1353] (c) Preparation of LIN 2: MC2-PEG4-Val-Cit-PABC-PNP

[1354]

[1355] To a solution of LIN 2-2 (1.73 g, 2.22 mmol) and bis-PNP (3.38 g, 11.12 mmol) in DCM:DMF (8:2, 75 mL) was added DIPEA (1.16 mL, 6.07 mmol) at 23 °C. The reaction mixture was stirred at 23 °C for 19 h and poured onto a silica gel column (CH2Cl2:CH3OH, 50:1 to 10:1) to give pure LIN 2 (945 mg, 45% yield).

[1356] 1H NMR (500 MHz, CD3OD): δ 8.22 (d, J = 9.2 Hz, 2H), 7.61 (d, J = 8.6 Hz, 2H), 7.34 (d, J = 9.2 Hz, 2H), 7.33 (d, J = 8.6 Hz, 2H), 6.67 (s, 2H), 4.57-4.47 (m, 1H), 4.23-4.12 (m, 1H), 3.78-3.76 (m, 12H), 3.63-3.50 (m, 16H), 3.49-3.41 (m, 2H), 3.34-3.25 (m, 2H), 3.18-3.03 (m, 2H), 2.51 (t, J = 5.9 Hz, 2H), 2.45 (t, J = 7.2 Hz, 2H), 2.13-1.99 (m, 1H), 1.92-1.84 (m, 1H), 1.73-1.62 (m, 1H), 1.55-1.45 (m, 2H), 0.92 (d, J = 6.8 Hz, 3H), 0.90 (d, J = 6.8 Hz, 3H).

[1357] 13 C NMR (75 MHz, CDC13 / CD3OD): δ 174.4, 172.9, 172.4, 172.4, 171.6, 170.9, 170.8, 170.7, 163.7, 155.8, 155.7, 152.5, 145.4, 138.8, 134.1, 131.3, 130.4, 129.2, 128.7, 125.7, 124.9, 121.8, 119.8 (x 2), 115.1, 70.2 (x 2), 70.1 (x 2), 70.0, 69.9, 69.8, 69.0, 66.9, 59.2, 53.5, 39.0, 36.0, 34.4, 34.1, 30.4, 29.0, 18.5, 17.5.

[1358] ESI-MS m / z: 943.4 (M + H) + .

[1359] R f = 0.20 (CH2Cl2: CH3OH, 9: 1).

[1360] Preparation of LIN 3: MC2-PEG4-Val-Ala-PABC-PNP

[1361] Reaction mechanism

[1362]

[1363] (a) Preparation of LIN 3-1: MC2-PEG4-Val-Ala-OH

[1364] LIN 3-1

[1365]

[1366] Cl-TrtCl-resin (5 g, 1.49 mmol / g) was placed in a filter plate. CH2Cl2(25 mL) was added to the resin and the mixture was stirred at 23 °C for 1 h. The solvent was removed by filtration under vacuum. A solution of Fmoc-Ala-OH (2.31 g, 7.41 mmol) and DIPEA (4.28 mL, 24.61 mmol) in CH2Cl2(20 mL) was added and the mixture was stirred at 23 °C for 10 min. Additional DIPEA (8.60 mL, 49.37 mmol) was added and the reaction mixture was stirred at 23 °C for 1 h. The reaction was stopped by the addition of MeOH (10 mL) and stirred at 23 °C for 15 min. Fmoc-Ala-O-TrtCl-resin was subjected to the following wash / treatment: CH2Cl2(5 x 15 mL x 0.5 min), DMF (5 x 15 mL x 0.5 min), piperidine:DMF (1:4, 15 mL, 1 x 1 min, 2 x 10 min), DMF (5 x 15 mL x 0.5 min), CH2Cl2(5 x 15 mL x 0.5 min). The loading was calculated: 1.34 mmol / g.

[1367] NH2-Ala-O-TrtCl-resin was washed with DMF (5 x 15 mL x 0.5 min) and a solution of Fmoc-Val-OH (9.09 g, 26.79 mmol) and HOBt (3.62 g, 26.79 mmol) in DMF (25 mL) was added to the NH2-Ala-O-TrtCl-resin followed by the addition of DIPCDI (4.14 mL, 26.79 mmol) at 23 °C. The mixture was stirred at 23 °C for 1.5 h. The reaction was stopped by washing with DMF (5 x 15 mL x 0.5 min). Fmoc-Val-Ala-O-TrtCl-resin was treated with piperidine:DMF (1:4, 15 mL, 1 x 1 min, 2 x 10 min) and washed with DMF (5 x 15 mL x 0.5 min).

[1368] A solution of 15-(9-fluorenylmethoxycarbonyl)amino-4,7,10,13- tetraoxa-pentadecanoic acid (Fmoc-NH-PEG4-OH) (4.90 g, 8.75 mmol) and HOBt (1.35 g, 9.98 mmol) in DMF (30 mL) was added to NH2-Val-Ala-O-Trt Cl-resin followed by the addition of DIPCDI (1.55 mL, 10.0 mmol) at 23 °C. The reaction mixture was stirred at 23 °C for 22 h. The reaction was stopped by washing with DMF (5 x 15 mL x 0.5 min). The Fmoc-NH-PEG4-Val-Ala-O-Trt Cl-resin was treated with piperidine:DMF (1:4, 15 mL, 1 x 1 min, 2 x 10 min) and washed with DMF (5 x 15 mL x 0.5 min).

[1369] A solution of 3-(maleimidopropionic acid) (MC2-OH) (4.53 g, 26.78 mmol) and HOBt (3.62 g, 26.77 mmol) in DMF (30 mL) was added to NH2-PEG4-Val-Ala-O-Trt Cl-resin followed by the addition of DIPCDI (4.15 mL, 26.80 mmol) at 23 °C. The reaction mixture was stirred at 23 °C for 2 h. The reaction was stopped by washing with DMF (5 x 15 mL x 0.5 min) and CH2Cl2(5 x 15 mL x 0.5 min).

[1370] The peptide was cleaved from the resin by treatment with TFA:CH2Cl2(1:99, 5 x 50 mL). The resin was washed with CH2Cl2(7 x 50 mL x 0.5 min). The combined filtrates were evaporated to dryness under reduced pressure, the obtained solid was triturated with Et2O and filtered to obtain L 3-1 as a white solid (4.73 g, 87% yield).

[1371] 1H NMR (500 MHz, CDC13): δ 7.67 (bs, 1 H), 7.31 (d, J = 8.9 Hz, 1 H), 7.17 (d, J = 7.0 Hz, 1 H), 6.85 (t, J = 5.6 Hz, 1 H), 6.72 (s, 2 H), 4.51 (q, J = 7.1 Hz, 1 H), 4.38 (dd, J = 8.9, 6.9 Hz, 1 H), 3.84 (t, J = 7.1 Hz, 2 H), 3.75 (t, J = 5.9 Hz, 2 H), 3.69 - 3.59 (m, 12 H), 3.55 (t, J = 5.1 Hz, 2 H), 3.41 (qd, J = 5.0, 1.7 Hz, 2 H), 2.62 - 2.49 (m, 4 H), 2.19 - 2.01 (m, 1 H), 1.44 (d, J = 7.2 Hz, 3 H), 0.95 (d, J = 11.9 Hz, 1 H), 0.94 (d, J = 11.9 Hz, 1 H).

[1372] (b) Preparation of LIN 3-2: MC2-PEG4-Val-Ala-PABOH

[1373] LIN 3-2

[1374]

[1375] To a solution of LIN 3-1 (1.84 g, 3.13 mmol) and 4-aminobenzyl alcohol (PABOH) (0.77 g, 6.27 mmol) in CH2Cl2(70 mL) at 23 °C was added a solution of HOBt (0.84 g, 6.27 mmol) in DMF (5 mL) followed by DIPCDI (0.97 mL, 6.27 mmol). The reaction mixture was stirred at 23 °C for 5 h, Et20 (150 mL) was added, and the resulting solid was vacuum filtered to give crude LIN 3-2 (1.74 g, 81% yield) which was used in the next step without further purification.

[1376] 1H NMR (500 MHz, DMSO-d6): δ 7.58 (d, J = 8.5 Hz, 2H), 7.30 (d, J = 8.5 Hz, 2H), 6.81 (s, 2H), 4.56 (s, 2H), 4.52-4.41 (m, 1H), 4.21 (d, J = 6.7 Hz, 1H). 3.91 (p, J = 6.5 Hz, 1H), 3.81-3.67 (m, 4H), 3.65-3.54 (m, 12H), 3.49 (t, J = 5.5 Hz, 2H), 2.56 (dd, J = 6.6, 5.5 Hz, 2H), 2.46 (t, J = 6.9 Hz, 2H), 2.12 (h, J = 6.8 Hz, 1H), 1.45 (d, J = 7.2 Hz, 3H), 1.00 (d, J = 12.1 Hz, 3H), 0.98 (d, J = 12.1 Hz, 3H).

[1377] (c) Preparation of LIN 3: MC2-PEG4-Val-Ala-PABC-PNP

[1378] LIN 3

[1379]

[1380] To a solution of LIN 3-2 (1.74 g, 2.51 mmol) and bis-PNP (3.82 g, 12.57 mmol) in CH2Cl2: DMF (8: 1, 70 mL) was added DIPEA (1.31 mL, 7.54 mmol) at 23 °C. The reaction mixture was stirred at 23 °C for 20 h and poured onto a silica gel column (CH2Cl2: CH3OH, 50: 1 to 10: 1) to give pure LIN 3 (1.26 g, 59% yield).

[1381] 1H NMR (500 MHz, CDC13): δ 8.82 (s, 1H), 8.27 (d, J = 9.2 Hz, 2H), 7.73 (d, J = 8.6 Hz, 2H), 7.38 (d, J = 9.1 Hz, 4H), 7.15 (dd, J = 21.8, 7.2 Hz, 2H), 6.69 (s, 2H), 6.62 (t, J = 5.7 Hz, 1H), 5.24 (s, 2H), 4.67 (p, J = 7.2 Hz, 1H), 4.24 (dd, J = 6.8, 5.7 Hz, 1H), 3.91 - 3.76 (m, 2H), 3.71 (ddd, J = 10.1, 6.1, 4.3 Hz, 1H), 3.66 - 3.54 (m, 14H), 3.53 (t, J = 5.1 Hz, 1H), 3.46 - 3.33 (m, 2H), 2.76 - 2.57 (m, 1H), 2.57 - 2.42 (m, 2H), 2.33 - 2.19 (m, 1H), 1.46 (d, J = 7.1 Hz, 3H), 1.01 (d, J = 12.1 Hz, 3H), 1.00 (d, J = 12.1 Hz, 3H).

[1382] 13 C NMR (75 MHz, CD3OD): δ 173.0, 172.1, 171.6 (x2), 170.7, 163.8, 155.7, 152.5, 145.4, 140.3, 138.9, 134.1, 130.4, 129.1, 125.6, 124.8, 121.9, 119.7, 115.1, 70.2, 70.1 (x3), 70.0, 69.9, 69.8, 69.0, 66.9, 59.1, 53.4, 49.7, 39.0, 36.0, 34.3, 34.1, 30.4, 18.3, 17.3, 16.6.

[1383] ESI-MS m / z: 857.3 (M+H) + .

[1384] R f = 0.45 (CH2Cl2: CH3OH, 9: 1).

[1385] Example 3: Synthesis of formula D-X-(AA) w -(T) g -L1

[1386] Preparation of compound DL-1

[1387]

[1388] To a solution of 1 (15 mg, 0.019 mmol) and L1 (14 mg, 0.019 mmol) in 1 -methyl-2-pyrrolidinone (NMP) (1 mL, 0.019 M) was added DIPEA (3 μL, 0.019 mmol) at 23 °C. After 72 h, EtOAc was added and the reaction mixture was washed with water, the organic layer was dried over anhydrous Na2S04, filtered, and concentrated in vacuo. The resulting residue was purified by HPLC prep to give pure DL1 (7.5 mg, 29% yield).

[1389] 1H NMR (500 MHz, CD3OD): δ 7.58 (d, J = 8.6 Hz, 1H), 7.50 (d, J = 8.6 Hz, 1H), 7.32 (d, J = 8.6 Hz, 1H), 7.21 (d, J = 8.6 Hz, 1H), 7.11 (dd, J = 8.7, 1.8 Hz, 1H), 6.82 (t, J = 2.0 Hz, 1H), 6.77 (s, 2H), 6.67 (ddd, J = 8.9, 2.5, 1.3 Hz, 1H), 6.58 (s, 1H), 6.23 (dd, J = 3.0, 1.3 Hz, 1H), 6.07 (t, J = 1.4 Hz, 1H), 5.64 (ddd, J = 12.4, 4.9, 1.8 Hz, 1H), 5.21 (dd, J = 22.0, 11.1 Hz, 1H), 5.19 - 5.11 (m, 1H), 5.14 - 5.04 (m, 1H), 5.04 - 4.96 (m, 1H), 4.75 (s, 1H), 4.70 (s, 1H), 4.58 (s, 1H), 4.50 (ddd, J = 8.7, 5.1, 3.3 Hz, 1H), 4.30 (d, J = 3.1 Hz, 1H), 4.22 - 4.11 (m, 3H), 3.75 (s, 3H), 3.74 (s, 3H), 3.58 - 3.53 (m, 1H), 3.50 - 3.44 (m, 2H), 3.35 (s, 3H), 3.24 - 3.17 (m, 2H), 3.11 (ddd, J = 13.7, 10.6, 6.6 Hz, 1H), 3.02 (dd, J = 17.5, 9.8 Hz, 1H), 2.90 - 2.84 (m, 2H), 2.76 (dd, J = 15.3, 2.4 Hz, 1H), 2.59 (dd, J = 7.0, 4.9 Hz, 2H), 2.36 - 2.24 (m, 6H), 2.14 - 2.07 (m, 1H), 2.10 - 1.97 (m, 4H), 2.04 (s, 3H), 1.93 - 1.86 (m, 1H), 1.79 - 1.71 (m, 1H), 1.66 - 1.60 (m, 2H), 1.59 - 1.53 (m, 4H), 1.35 - 1.25 (m, 4H), 0.97 (m, 6H).

[1390] ESI-MS m / z: 1352.2 (M-H20+H) +

[1391] Preparation of compound DL-2

[1392]

[1393] To a solution of 2 (21 mg, 0.027 mmol) in dimethylformamide (DMF) (2 mL, 0.013 M) at 23 °C was added L1 (22 mg, 0.029 mmol), 1-hydroxybenzotriazole (HOBT, 3.9 mg, 0.029 mmol) and DIPEA (26 μL, 0.15 mmol). After 72 h, EtOAc was added and the reaction mixture was washed with water, the organic layer was dried over anhydrous Na2S04, filtered, and concentrated in vacuo. The residue obtained was purified by HPLC prep to give pure DL2 (3.5 mg, 9% yield).

[1394] 1 H NMR (400 MHz, CDC13): δ 7.74 (d, J = 7.8 Hz, 1H), 7.47 (dd, J = 21.6, 8.1 Hz, 2H), 7.23 (d, J = 7.8 Hz, 1H), 7.12 (d, J = 8.2 Hz, 1H), 7.07 (d, J = 8.2 Hz, 1H), 6.77 (s, 2H), 6.64 (s, 2H), 6.54 (s, 1H), 6.16 (s, 1H), 5.97 (s, 1H), 5.63 (d, J = 17.2 Hz, 1H), 5.11 (d, J = 12.5 Hz, 1H), 5.01 (s, 1H), 4.90 (d, J = 12.2 Hz, 1H), 4.66 (s, 1H), 4.50 (s, 1H), 4.29 - 4.19 (m, 2H), 4.13 - 4.08 (m, 1H), 3.74 (s, 3H), 3.70 (s, 3H), 3.68 (s, 3H), 3.43 (t, J = 7.1 Hz, 2H), 3.34 (t, J = 1.9 Hz, 1H), 3.33 (s, 2H), 3.08 (s, 2H), 2.98 - 2.72 (m, 5H), 2.50 (d, J = 16.0 Hz, 1H), 2.33 (s, 3H), 2.26 (s, 3H), 2.22 - 2.14 (m, 3H), 1.99 (s, 3H), 1.81 (s, 1H), 1.63 - 1.50 (t, J = 7.4 Hz, 4H), 1.48 - 1.39 (m, 4H), 1.28 - 1.19 (m, 3H), 0.90 - 0.86 (m, 6H).

[1395] ESI-MS m / z: 1379.5 (M+H) + .

[1396] Example 4: Preparation of antibody-drug conjugate (ADC)

[1397] In this example, the synthesis of antibody-drug conjugates of the application is described. It should be noted that these syntheses are exemplary and that the methods described can be applied to all the compounds and antibodies described herein.

[1398] Preparation of anti-CD13 monoclonal antibodies

[1399] Anti-CD13 monoclonal antibodies were obtained according to well-known methods commonly used in the art. Briefly, BALB / c mice were immunized with human endothelial cells isolated from umbilical cord. To this end, 1.5E7 cells were injected intraperitoneally to mice on day -45 and day -30, and intravenously to mice on day -3. On day 0, the spleens were removed from these animals and the splenocytes were fused with SP2 mouse myeloma cells in a 4:1 ratio according to standard techniques to generate hybridomas and distributed on 96-well tissue culture plates (Costar, Cambridge, MA). After 2 weeks, the hybridoma culture supernatants were harvested and tested for reactivity against the cell line used in the immunization step by flow cytometry. Positive supernatants were analyzed by immunofluorescence staining of the corresponding cells used as antigens. Fused tumors exhibiting specific staining, immunoprecipitation pattern and cell distribution were selected and cloned and subcloned by limiting dilution.

[1400] Once the clones were selected, the cells were cultured in RPMI-1640 medium supplemented with 10% (v / v) fetal bovine serum, 2 mM glutamine, 100 U / mL penicillin and 100 μg / mL streptomycin at 37°C for 3-4 days until the medium turned light yellow. At that time, two-thirds of the medium volume was removed, centrifuged at 1,000 x g for 10 min to pellet the cells, and the supernatant was centrifuged again at 3,000 x g for 10 min to further wash, or filtered through a 22 μm pore size membrane. The clarified supernatant was precipitated with 55% saturated ammonium sulfate, and the resulting pellet was resuspended in 100 mM Tris-HCl pH 7.8 (1 mL per 100 mL of the original clarified supernatant) and dialyzed against 5 L of 100 mM Tris-HCl pH 7.8 with 150 mM NaCl for 16-24 h at 4°C, changing the dialysis solution at least three times. The dialyzed material was finally loaded onto a protein A-sepharose gel column and the corresponding monoclonal antibody was eluted with 100 mM sodium citrate pH 3.0 or, alternatively, with 1 M glycine pH 3.0. Those eluted fractions containing the antibody were neutralized with 2 M Tris-HCl pH 9.0 and finally dialyzed against PBS, and stored at -80°C until their use.

[1401] Preparation of antibody-drug conjugates ADC1 with trastuzumab and DL1

[1402] (a) Preparation of trastuzumab

[1403] Trastuzumab (trastuzumab was purchased from Roche as a white lyophilized powder for the preparation of concentrated solutions for infusion) was dissolved in 5 mL of phosphate buffer (50...

Claims

1. A drug conjugate comprising a drug moiety D covalently linked to the remainder of the drug conjugate, the drug conjugate having the formula: [D-(X b -(AA) w -(T) g -(L)-] n -Ab, wherein: D is a drug moiety having the following formula (IH), or a pharmaceutically acceptable salt thereof, wherein: a wavy line indicates that D is covalently linked to (X) b , if (X) b is present; or (AA) w , if (AA) w is present; or to a point (T) g , if (T) g is present; or to (L). Y is -NH- or -O-; R1is -OH or -CN; R2is -C(=O)R a group; R3is hydrogen or -OR b group; R4is selected from hydrogen and -CH2OH; R a is unsubstituted C1-C6alkyl; R b is unsubstituted C1-C6alkyl; X is an extension group selected from: -COO-(Ci-C6alkylene)NH-; -COO-CH2-(unsubstituted phenylene)-NH-; -COO-(Ci-C6alkylene)NH-COO-CH2-(unsubstituted phenylene)-NH-; -COCH2NH-COCH2-NH-; -COCH2NH-; -COO-(Ci-C6alkylene)S-; -COO-(Ci-C6alkylene)NHCO(Ci-C6alkylene)S-; T is an extension group selected from the group consisting of: -CO-(Ci-C6alkylene)-NH-; -CO-(Ci-C6alkylene)-[O-(C2-C6alkylene)] j -NH-; -COO-(Ci-C6alkylene)-[O-(C2-C6alkylene)] j -NH-; wherein j is an integer from 1 to 25; (AA) w selected from formula (II): where the left wavy line denotes covalent attachment to (X) b , if (X) b is present; or to a point of a drug moiety D, and the right wavy line denotes covalent attachment to (T) g , if (T) g is present; or to a point of a linker L; and R 21 is selected for each occurrence from the group consisting of hydrogen, methyl, isopropyl, isobutyl, sec-butyl, benzyl, -CH2CH2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)3NH2, -(CH2)4NHC(=NH)NH2, -(CH2)4NH2, -(CH2)3NHCONH2, -(CH2)4NHCONH2, phenyl, cyclohexyl, and w is an integer ranging from 0 to 6; L is a linker group selected from the group consisting of: wherein: the wavy line on the right represents the point of covalent attachment to Ab, and the wavy line on the left represents the point of covalent attachment to (T) g , if (T) g is present; or (AA) w ; or to (X) b , if (X) b is present, or to a point of a drug moiety D; R 19 unsubstituted -C1-C 12 alkylene-; R 30 -C1-C6alkylene; M is selected from the group consisting of -Ci-C6alkylene- and -Ci-C6alkylene-(C3-C8carbocyclyl)-; b is an integer of 0 or 1 ; g is an integer of 0 or 1 ; Ab is an anti-HER2 antibody or antigen binding fragment thereof; and n is the ratio of the group [D-(X) b -(AA) w -(T) g -(L)-] to the Ab moiety and is in the range of 1 to 20.

2. The drug conjugate of claim 1, wherein R4is hydrogen.

3. The drug conjugate of claim 1, wherein Y is -NH-.

4. The drug conjugate of claim 1, wherein Y is -O-.

5. The drug conjugate of claim 1, wherein R1is -OH.

6. The drug conjugate of claim 1, wherein R1is -CN.

7. The drug conjugate of claim 1, wherein R2 is -C(=0)R a group, wherein R a is selected from unsubstituted methyl, unsubstituted ethyl, unsubstituted n-propyl, unsubstituted isopropyl, unsubstituted n-butyl, unsubstituted isobutyl, unsubstituted sec-butyl, and unsubstituted t-butyl.

8. The drug conjugate of claim 7, wherein R2is acetyl.

9. The drug conjugate of claim 1, wherein R3 is hydrogen or -OR b wherein R b is selected from unsubstituted methyl, unsubstituted ethyl, unsubstituted n-propyl, unsubstituted isopropyl, unsubstituted n-butyl, unsubstituted isobutyl, unsubstituted sec-butyl, and unsubstituted t-butyl.

10. The drug conjugate of claim 9, wherein R3is hydrogen.

11. The drug conjugate of claim 9, wherein R3 is -OR b wherein R b is selected from unsubstituted methyl, unsubstituted ethyl, unsubstituted n-propyl, unsubstituted isopropyl, unsubstituted n-butyl, unsubstituted isobutyl, unsubstituted sec-butyl, and unsubstituted t-butyl.

12. The drug conjugate of claim 11, wherein R3is methoxy.

13. The drug conjugate of claim 1, wherein D is a compound of the following formula: or a pharmaceutically acceptable salt thereof.

14. The drug conjugate of claim 13, wherein D is a compound of the following formula: or a pharmaceutically acceptable salt thereof.

15. The drug conjugate of claim 13, wherein D is a compound of the following formula: or a pharmaceutically acceptable salt thereof.

16. The drug conjugate of claim 1, wherein the salt is selected from the group consisting of hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate, acetate, trifluoroacetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulfonate, p-toluenesulfonate, sodium salt, potassium salt, calcium salt, ammonium salt, ethylenediamine salt, ethanolamine salt, N,N-dialkylenethanolamine salt, triethanolamine salt, and basic amino acid salt.

17. The drug conjugate of claim 1, selected from the group consisting of formulas (IV), (V), and (VI): wherein: X, T, AA, w, b, g, D, Ab, and n are as defined in claim 1; R 19 is unsubstituted -Ci-C8alkylene-; R 30 is a -C2-C4alkylene- group; and M is unsubstituted -Ci-C3alkylene-(C5-C7carbocyclyl)-.

18. The drug conjugate of claim 1, selected from the group consisting of formulas (IV), (V), and (VI): wherein: X, T, AA, w, b, g, D, Ab, and n are as defined in claim 1; M is -C1-C3 alkylene-(C5-C7 carbocyclyl)-.

19. The drug conjugate of claim 1, wherein: w is an integer ranging from 0 to 6. wherein: R 19 -C1-C6alkylene-; R 30 is -C2-C4alkylene-; and 21. The drug conjugate of claim 1, wherein X is an extension group selected from the group consisting of: -COO-(C2-C4 alkylene)NH-; R 21 is selected, at each occurrence, from the group consisting of: hydrogen, methyl, isopropyl, sec-butyl, benzyl, -(CH2)3NHCONH2, -(CH2)4NH2, -(CH2)3NHC(=NH)NH2, and -(CH2)4NHC(=NH)NH2; and -COO-CH2-unsubstituted phenylene-NH-; 20. The drug conjugate of claim 1, wherein w is 0 or 2 and when w is 2, (AA) w also has the formula (III): -COO-(C2-C4 alkylene)NH-COO-CH2-(unsubstituted phenylene)-NH-; the left wavy line denotes covalent attachment to (X) b , if (X) b is present; or to a point of a drug moiety D, and the right wavy line denotes covalent attachment to (T) g , if (T) g is present; or to a point of a linker L; R 22 selected from the group consisting of methyl, benzyl, isopropyl, sec-butyl and and R 23 is selected from the group consisting of methyl, -(CH2)4NH2, -(CH2)3NHCONH2, and -(CH2)3NHC(=NH)NH2. -COCH2NH-COCH2-NH-; -COO-(C2-C4 alkylene)S-; -COO-(C2-C4 alkylene)NHCO(C1-C3 alkylene)S-; and b is 0 or 1.

22. The drug conjugate of claim 21, wherein b is 1.

23. The drug conjugate of claim 21, wherein X is an extension group selected from the group consisting of: -COO-CH2-unsubstituted phenylene-NH-; -COO(CH2)3NHCOOCH2-unsubstituted phenylene-NH-; -COO(CH2)3NH-; -COO(CH2)3-S-; -COO(CH2)3NHCO(CH2)2S-; and b is 0 or 1.

24. The drug conjugate of claim 23, wherein b is 1.

27. The drug conjugate of claim 1, wherein D is a drug moiety of formula (IH), or a pharmaceutically acceptable salt thereof, wherein R1is CN or -OH; R2is acetyl; R3is hydrogen or methoxy; R4is hydrogen or -CH2OH; and Y is -NH- or -O-.

28. The drug conjugate of claim 27, wherein R3is methoxy.

29. The drug conjugate of claim 1, wherein D is a drug moiety of formula (IH), or a pharmaceutically acceptable salt thereof, wherein: R1is CN; R2is acetyl; R3is methoxy; R4is hydrogen; and Y is -NH- or -O-.

30. The drug conjugate of claim 1, wherein the Ab moiety is trastuzumab or an antigen binding fragment thereof.

25. The pharmaceutical conjugate according to claim 1, wherein T is an extension group selected from the group consisting of: -CO-(C1-C4 alkylene)NH-; -CO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH-;-COO-(C1-C4 alkylene)-[O-(C2-C4 alkylene)] j -NH-; where j is an integer from 1 to 10; and g is 0 or 1.

26. The drug conjugate of claim 25, wherein T is an extensional group selected from the group consisting of -CO-(Ci-C4alkylene)NH-; -CO-(Ci-C4alkylene)-[O-(C2-C4alkylene)] -NH-; -COO-(Ci-C4alkylene)-[O-(C2-C4alkylene)] -NH-; wherein j is an integer from 1 to 5; and g is 0 or 1. j -NH-; -COO-(Ci-C4alkylene)-[O-(C2-C4alkylene)] -NH-; wherein j is an integer from 1 to 5; and g is 0 or 1. j -NH-; -COO-(Ci-C4alkylene)-[O-(C2-C4alkylene)] -NH-; wherein j is an integer from 1 to 5; and g is 0 or 1.

31. The drug conjugate of claim 1, which is an antibody drug conjugate selected from the group consisting of:

32. The drug conjugate of claim 31, wherein n is 3, 4, or 5.

34. The drug conjugate of claim 1, wherein the antibody drug conjugate is in isolated form.

35. The drug conjugate of claim 1, wherein the antibody drug conjugate is in purified form.

36. The drug conjugate of claim 1, wherein b+g+w is not 0.

37. The drug conjugate of claim 1, wherein b+w is not 0. b is 1 when w is not 0. b is 1 when w is 0. ​ ​ ​ ​ ​ ​ ​ and wherein n is 2 to 6, and each is an anti-HER2 antibody or antigen-binding fragment thereof. ​ 33. The drug conjugate of claim 31, wherein each is selected from trastuzumab or an antigen-binding fragment thereof. ​ ​ ​ ​ 38. The drug conjugate of claim 1, wherein, ​ 39. The drug conjugate of claim 1, wherein, ​ 40. The drug conjugate of claim 1, wherein, n is an integer in the range of 1-12.

41. The drug conjugate of claim 40, wherein, n is an integer in the range of 1-8.

42. The drug conjugate of claim 41, wherein, n is 1, 2, 3, 4, 5, or 6.

43. The drug conjugate of claim 41, wherein n is an integer in the range of 3-8.

44. The drug conjugate of claim 42, wherein n is 3, 4, or 5.

45. The drug conjugate of claim 44, wherein n is 4.

46. A compound of formula D-(X) b - (AA) w - (T) g - L1or formula D-(X) b - (AA) w - (T) g - H. L1is a linker selected from the group consisting of the following formulas: wherein the wavy line represents a covalent attachment to (T) g , if (T) g is present; or to (AA) w , if (AA) w is present; or to (X) b , if (X) b is present; or to a point of D. R 19 -C1-C 12 alkylene-; b is an integer of 0 or 1; g is an integer of 0 or 1; w is an integer in the range of 0 to 6; wherein, for the compound of formula D-(X) b -(AA) w -(T) g -H, b + w + g ≠ 0; wherein each of D, X, T, and AA is as described in claim 1.

47. A compound of formula D-X according to claim 46 b - (AA) w - (T) g a compound of -L1, selected from: and 48. A compound of formula D-(X) b -(AA) w -(T) g A compound of formula -L1, wherein each of D, X, AA, T, L1, b, g, and w is as described in claim 46.

49. Equation D-(X) b -(AA) w -(T) g Compounds of -H, wherein each of D, X, AA, T, b, g, and w is as described in claim 1; but further, wherein b + w + g ≠ 0.

50. The compound of claim 46, wherein b+g+w is not 0.

51. The compound of claim 46, wherein b+w is not 0.

52. The compound of claim 46, wherein, b is 1 when w is not 0.

53. The compound of claim 46, wherein, b is 1 when w is 0.

54. A pharmaceutical composition comprising the drug conjugate according to claim 1 and a pharmaceutically acceptable carrier.

55. Use of a drug conjugate according to any one of claims 1 to 45 for the manufacture of a medicament for the treatment of a cancer, wherein the cancer is selected from the group consisting of HER2-positive lung cancer, HER2-positive gastric cancer, HER2-positive colorectal cancer, HER2-positive breast cancer, HER2-positive pancreatic cancer, and HER2-positive ovarian cancer.

56. The use of claim 55, wherein the cancer is selected from the group consisting of HER2-positive NSCLC, HER2-positive breast cancer, HER2-positive ovarian cancer, and HER2-positive gastric cancer.

57. The use of claim 56, wherein the cancer is HER2-positive breast cancer.

58. A kit comprising a therapeutically effective amount of a drug conjugate according to any one of claims 1 to 45 and a pharmaceutically acceptable carrier.

59. Use of the kit of claim 58 for the manufacture of a medicament for the treatment of a cancer, wherein the cancer is selected from the group consisting of HER2-positive lung cancer, HER2-positive gastric cancer, HER2-positive colorectal cancer, HER2-positive breast cancer, HER2-positive pancreatic cancer, and HER2-positive ovarian cancer.

60. The use of claim 59, wherein the cancer is selected from the group consisting of HER2-positive NSCLC, HER2-positive breast cancer, HER2-positive ovarian cancer, and HER2-positive gastric cancer.

61. The use of claim 60, wherein the cancer is HER2-positive breast cancer.

62. A method of making a drug conjugate according to any one of claims 1 to 45, comprising conjugating an Ab moiety and a drug D, Ab and D being as defined in any one of claims 1 to 45.

63. The method of claim 62 for making a drug conjugate of formula (G) or (G’): wherein R 22 selected from the group consisting of methyl, benzyl, isopropyl, sec-butyl and R 23 is selected from the group consisting of methyl, -(CH2)4NH2, -(CH2)3NHCONH2, and -(CH2)3NHC(=NH)NH2; and n ranges from 1 to 20; the method comprising the steps of: (i) reacting a drug D-H of formula (IH)-H: wherein the substituents in the definition of (IH)-H are as defined in any one of claims 1-45, with a compound of formula (D’) or (E): wherein R 22 and R 23 as defined above in formula (G) and (G'), respectively. to give a compound of formula (F) or (F’), respectively: (ii) partially reducing one or more disulfide bonds in the antibody Ab to be conjugated, resulting in a reduced antibody Ab-SH having free thiol groups: and (iii) reacting the partially reduced antibody Ab-SH having free thiol groups with the compound of formula (F) or (F') produced in step (i), resulting in a drug conjugate of formula (G) or (G'), respectively:

64. The method of claim 63, wherein the antibody is an anti-HER2 antibody, or antigen binding fragment thereof, that is partially reduced in step (ii) using tris[2- carboxyethyl]phosphine hydrochloride.

65. The method of claim 64, wherein the antibody is trastuzumab, or antigen binding fragment thereof, that is partially reduced in step (ii) using tris[2-carboxyethyl]phosphine hydrochloride.

66. The method of claim 62 for preparing a drug conjugate of formula (W) or (W'): wherein R 22 selected from the group consisting of methyl, benzyl, isopropyl, sec-butyl and R 23 is selected from the group consisting of methyl, -(CH2)4NH2, -(CH2)3NHCONH2, and -(CH2)3NHC(=NH)NH2; and n ranges from 1 to 20; the method comprising the steps of: (i) reacting an antibody to be conjugated with 2-iminothiolane hydrochloride, resulting in a thiol-activated antibody: (ii) reacting the thiol-activated antibody with the compound of formula (F) or (F'), resulting in a drug conjugate of formula (W) or (W'), respectively 67. The method of claim 66, wherein the antibody is an anti-HER2 antibody, or antigen binding fragment thereof.

68. The method of claim 67, wherein the antibody is trastuzumab, or antigen binding fragment thereof.

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