N-Substituted Indole Derivatives and Conjugates for the Treatment of Cancer

Abstract

The present invention relates to N-substituted derivatives of indoles of formula (I): JPEG2025521735000104.jpg53170, and their use in the treatment of cancer. The present invention further provides protein-drug conjugates, more specifically antibody-drug conjugates, from the compounds of formula (I).

Description

Technical Field

[0001] The present invention relates to the pharmaceutical field, and more specifically to N-substituted derivatives of indole and their use as therapeutic compounds. In particular, such derivatives are used in the treatment of cancer. Some of them are also used in the production of conjugates with proteins or antibodies.

Background Art

[0002] Kinesin is a superfamily of motor proteins with ATPase activity. They are involved in the normal biological activities of various cells, including mitosis, meiosis, and intracellular vesicle transport. Kinesin family member 20A (KIF20A, also known as MKlp2) is located on chromosome 5q31.2 and plays an important role in the appearance and development of tumors. Recently, several studies have demonstrated that KIF20A can play an important role in the development and progression of many different types of cancer, such as melanoma, breast cancer, nasopharyngeal cancer, pancreatic cancer, hepatocellular carcinoma, lung cancer, and colorectal cancer.

[0003] Over the past few years, a class of indole derivatives as compounds targeting MKlp2 (KIF20A), such as the compounds described in international patent application WO2014 / 086964, have been developed. These compounds are selective inhibitors for MKlp2 (KIF20A) with nanomolar efficiency, despite their poor solubility in water at both neutral and acidic pH.

[0004] It is well known that solubility is one of the most important parameters to achieve the desired concentration of a drug for a pharmacological response in systemic circulation, regardless of the administration used. In fact, low aqueous solubility is a major problem encountered during the development of pharmacologically active chemical entities and the formulations containing them.

[0005] Therefore, the aqueous solubility of this class of indole derivatives needs to be enhanced to ensure the solubility of such derivatives at the absorption site.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Patent Document 6

Patent Document 7

Non-Patent Documents

[0007]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

[0008] In this context, the inventors have provided highly water - soluble prodrugs derived from indole derivatives, particularly those reported in WO2014 / 086964. More specifically, the nitrogen atom of indole is linked to either a phosphonic acid containing promoieties or a cleavable group, such as a metabolic liable group. The cleavable group can also include a conjugate protein linker in order to, for example, provide a protein conjugate with albumin to increase the circulation half - life of the drug, and to promote the intracellular and / or extracellular release of these drugs into tumors by, for example, increasing the internalization of these drugs within cancer cells by an antibody conjugate.

[0009] Therefore, the present invention relates to a compound of formula (I):

[0010] [Chemical formula]

[0011] [In the formula, - X represents a nitrogen atom, a C-CN unit or an N+-O- unit, - R1 and R1' independently represent hydrogen, halogen, (C1-C6) alkoxy, or a -SO2-CH3 group, provided that when R1 or R1' is (C1-C6) alkoxy, R2 is not halogen, - R2 is · (C1-C6) alkoxy, (C3-C6) cycloalkoxy, (C3-C6) heterocycloalkoxy, aryloxy, heteroaryloxy, (C1-C6) alkylaryloxy, (C1-C6) alkylheteroaryloxy groups, which are optionally substituted by at least one halogen, said groups, or thio (C1-C6) alkyl, thioaryl, thioheteroaryl, thio (C1-C6) alkylaryl or thio (C1-C6) -alkylheteroaryl groups, which are optionally substituted by at least one halogen or by a (C1-C6) alkoxy group, said groups, · -NR4R5 unit, O-(C1-C6) alkyl-NR4R5 unit or S-(C1-C6) alkyl-NR4R5 unit (wherein R4 and R5 represent H, (C1-C6) alkyl groups, or R4 and R5 together form a 3- to 7-membered ring optionally interrupted by one or several heteroatoms, provided that at least one of R4 and R5 is not H), · NHCOR6 unit (wherein R6 represents a (C1-C6) alkyl group), · aryl or heteroaryl groups optionally substituted by at least one halogen, trifluoromethyl group or (C1-C3) alkoxy group, or · halogen represents, - R3 represents hydrogen, a (C1-C3) alkyl group, a (C1-C3) alkoxy group or halogen, - Ra is · Formula (A):

[0012]

Chem.

[0013] or formula (A'):

[0014]

Chem.

[0015] group (wherein n is an integer included between 1 and 12), and · formula (B):

[0016]

Chem.

[0017] group (wherein · L1 is a cleavable group selected from among pH-sensitive groups, photoinducible cleavable groups, in vivo reductive cleavable groups, and enzymatically cleavable groups, · L2 is a tert-butoxycarbonyl group or a conjugated protein conjugate, · m is an integer equal to 0 or 1) is a group selected from the group consisting of] or a pharmaceutically acceptable salt thereof.

[0018] In certain embodiments, R2 is · a (C1-C6) alkoxy group, preferably a methoxy group, · a (C3-C6) heterocycloalkoxy group, preferably an oxetanoxy group, or · a heteroaryl group, preferably a furanyl or triazolyl group represents.

[0019] In a preferred embodiment, the compound of formula (I) is - X represents a C-CN unit, - R1 represents a halogen, preferably bromine, - R1' represents hydrogen, - R2 represents a (C1-C6) alkoxy group, preferably a methoxy group, - R3 represents hydrogen and is of the following type.

[0020] In a further preferred embodiment, the compound of formula (I) - X represents a C-CN unit, - R1 represents hydrogen, - R1' represents a (C1-C6) alkoxy group, preferably a methoxy group, - R2 represents a (C1-C6) alkoxy group, preferably a methoxy group, - R3 represents hydrogen and is of the following type.

[0021] In certain embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof is such that Ra is of formula (A):

[0022]

Chemical formula

[0023] or formula (A'):

[0024]

Chemical formula

[0025] and is a group [wherein n is an integer included between 1 and 12]. Preferably, n is an integer included between 2 and 10. More preferably, n is an integer included between 2 and 8, advantageously between 2 and 6. Even more preferably, n is 2, 3, 4, 5, or 6.

[0026] In certain embodiments, the pharmaceutically acceptable salts of the compounds of formula (I) are selected from sodium salts, disodium salts, lysine salts, dilysine salts, arginine salts or diarginine salts, preferably from sodium salts or disodium salts, more preferably sodium salts.

[0027] In preferred embodiments, the compounds of formula (I) are - Example 1: (Z)-5-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonic acid, - Example 2: (Z)-3-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid, - Example 3: (Z)-7-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-7-oxoheptylphosphonic acid, - Example 4: (Z)-5-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-5-oxopentylphosphonic acid, - Example 5: (Z)-7-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-7-oxoheptylphosphonic acid, - Example 6: (Z)-Hydrogen 5-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonic acid sodium, - Example 7: (Z)-Hydrogen 3-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid sodium, - Example 8: (Z)-Hydrogen 4-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonic acid sodium, - Example 9: Sodium (Z)-6-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxohexylphosphonate, - Example 10: Sodium (Z)-7-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-7-oxoheptylphosphonate, - Example 11: Sodium (Z)-5-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-5-oxopentylphosphonate, - Example 12: Sodium (Z)-7-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-7-oxoheptylphosphonate, - Example 13: Disodium (Z)-4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonate, - Example 14: Disodium (Z)-5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate, - Example 15: (S)-2,6-Diaminohexanoic acid compound and (Z)-3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid (2:1), - Example 16: (S)-2,6-Diaminohexanoic acid compound and (Z)-4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonic acid (2:1), - Example 17: 3-[Hydroxy(dioxo)-lambda6-phosphanyl]propyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate, and - Example 18: 4-[Hydroxy(dioxo)-lambda6-phosphanyl]butyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate is selected from the group consisting of

[0028] In another embodiment of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof is such that Ra is of formula (B):

[0029] [Chemical formula]

[0030] group [wherein · L1 is a cleavable group selected from among pH-sensitive groups, photoinducible cleavable groups, in vivo reducible cleavable groups, and enzymatically cleavable groups, · L2 is a tert-butoxycarbonyl group or a conjugated protein linker, · m is an integer equal to 0 or 1] is such that

[0031] In a particular embodiment, L1 is an enzymatically cleavable group cleaved by protease, peptidase, esterase, beta-glucuronidase, glycosidase, phosphodiesterase, phosphatase, pyrophosphatase, tubulin tyrosine ligase or lipase.

[0032] In a preferred embodiment, L1 is p-aminobenzyloxycarbonyl-AA1 w -AA2 x -AA3 y -AA4 za base, where AA1, AA2, AA3, and AA4 are independently selected from the group consisting of alanine, valine, citrulline, phenylalanine, lysine, glycine, aspartic acid, asparagine, glutamic acid, and derivatives thereof, preferably citrulline, valine, cysteic acid, glycine, and glutamic acid, more preferably citrulline, valine, and cysteic acid, and w, x, y, and z are independently integers equal to 0 or 1.

[0033] According to a particular embodiment, m is 1 and L2 is of the formula:

[0034] [Chemical formula]

[0035] [wherein r is an integer between 1 and 36, preferably between 1 and 24, more preferably an integer equal to 2, 4, 8, 12, 16, 20, or 24] a conjugated protein conjugate having the formula.

[0036] In a preferred embodiment, the compound of formula (I) is - Example 19: (2S,3S,4S,5R,6S)-6-[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenoxy]-3,4,5-trihydroxytetrahydropyran-2-carboxylic acid, - Example 20: [4-[[(2S)-2-[[(2S)-2-(tert-butoxycarbonylamino)-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate, - Example 21: [4-[[(2S)-2-[[(2S)-2-Aminopropanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate hydrochloride, - Example 22: [4-[[(2S)-2-[[(2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy](peg4)propanoylamino]-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate, - Example 23: [4-[[(2S)-2-[[(2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamido](peg24)propanoylamino]-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate, - Example 24: 3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-(tert-butoxycarbonylamino)-3-oxopropane-1-sulfonic acid, - Example 25: 2-Amino-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-3-oxopropane-1-sulfonic acid, and - Example 26: (2R)-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy]propanoylamino]-3-oxopropane-1-sulfonic acid is selected from the group consisting of.

[0037] A further object of the present invention is a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable excipient.

[0038] In particular, such pharmaceutical compositions are for use in the treatment of cancer. Preferably, the cancer is selected from leukemia, acute myeloid leukemia, lymphoma, breast cancer, pancreatic cancer, lung cancer or colon cancer.

[0039] A further object of the present invention is a conjugate of formula (II):

[0040] [Chemical formula]

[0041] [wherein, - X, R1, R 1' , R2, R3, and L1 are as defined herein, - L2 is a binding protein conjugate, - P is a peptide or protein capable of binding to a target of interest, preferably an antibody, antibody fragment, or antigen-binding fragment, - v is an integer from 1 to 10] is.

Brief Description of the Drawings

[0042]

Figure 1

Mode for Carrying Out the Invention

[0043] Definition According to the present invention, the following terms have the following meanings.

[0044] Terms referred to in this specification with prefixes such as C1 - C3, C1 - C6 or C2 - C6 can also be used with a smaller number of carbon atoms such as C1 - C2, C1 - C5, or C2 - C5. For example, when the term C1 - C3 is used, it means that the corresponding hydrocarbon chain can contain from 1 to 3 carbon atoms, especially 1, 2 or 3 carbon atoms. For example, when the term C1 - C6 is used, it means that the corresponding hydrocarbon chain can contain from 1 to 6 carbon atoms, especially 1, 2, 3, 4, 5 or 6 carbon atoms. For example, when the term C2 - C6 is used, it means that the corresponding hydrocarbon chain can contain from 2 to 6 carbon atoms, especially 2, 3, 4, 5 or 6 carbon atoms.

[0045] The term "alkyl" refers to a saturated, straight-chain or branched-chain aliphatic group. The term "(C1 - C3)alkyl" more specifically means methyl, ethyl, propyl, or isopropyl. The term "(C1 - C6)alkyl" more specifically means methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl or hexyl. In a preferred embodiment, "alkyl" is methyl, ethyl, propyl, isopropyl, or tert-butyl, more preferably methyl.

[0046] The term "alkoxy" or "alkyloxy" corresponds to an alkyl group as defined above that is bonded to the molecule by an -O- (ether) linkage. (C1-C3) alkoxy includes methoxy, ethoxy, propyloxy, and isopropyloxy. (C1-C6) alkoxy includes methoxy, ethoxy, propyloxy, isopropyloxy, butyloxy, isobutyloxy, tert-butyloxy, pentyloxy, and hexyloxy. In a preferred embodiment, "alkoxy" or "alkyloxy" is methoxy.

[0047] The term "thio" corresponds to an alkyl group as defined above that is bonded to the molecule by an -S- (thioether) linkage. Thio (C1-C6) alkyl groups include thiomethyl, thioethyl, thiopropyl, thiobutyl, thiopentyl, and thiohexyl.

[0048] The term "halogen" corresponds to a fluorine, chlorine, bromine, or iodine atom, preferably fluorine, chlorine, or bromine, more preferably chlorine or bromine.

[0049] The term "aryl" corresponds to a monocyclic or bicyclic aromatic hydrocarbon having from 6 to 12 carbon atoms. For example, the term "aryl" includes phenyl, biphenyl, or naphthyl. In a preferred embodiment, aryl is phenyl.

[0050] As used herein, the term "heteroaryl" corresponds to an aromatic monocyclic or polycyclic group containing between 5 and 14 atoms and containing at least one heteroatom such as a nitrogen, oxygen or sulfur atom. Examples of such monocyclic or polycyclic heteroaryl groups are pyridinyl, thiazolyl, thiophenyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, benzofuranyl, thianaphthalenyl, indolyl, indolinyl, quinolinyl, isoquinolinyl, benzimidazolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, triazinyl, thianthrenyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxanthinyl, isothiazolyl, isoxazolyl, pyrazinyl, pyridazinyl, indolizinyl, isoindolyl, indazolyl, purinyl, quinolidinyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl, chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, indolinyl, isoindolinyl, oxazolidinyl, benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolinyl, benzothienyl, benzothiazolyl, isatinyl, dihydropyridyl, pyrimidinyl, s-triazinyl, oxazolyl, or thiophenyl. In preferred embodiments, the heteroaryl group is pyridinyl, furanyl, pyrazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, and isoxazolyl. Preferably, heteroaryl is pyridyl, thiazolyl, furanyl, pyranyl, pyrrolyl, imidazolyl, tetrazolyl, benzofuranyl, pyrrolinyl, triazinyl, pyrazinyl, pyridazinyl, triazolyl or tetrazolyl. More preferably, heteroaryl is furanyl or triazolyl.

[0051] The term "cycloalkyl" corresponds to a saturated or unsaturated monocyclic, bicyclic, or tricyclic alkyl group containing from 3 to 20 carbon atoms. It also includes cycloalkyl groups that are fused, bridged, or spiro-linked. The term "cycloalkyl" includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In a preferred embodiment, "cycloalkyl" is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0052] The term "heterocycloalkyl" corresponds to a saturated or unsaturated cycloalkyl group as defined above, further containing at least one heteroatom such as a nitrogen, oxygen, or sulfur atom. It also includes heterocycloalkyl groups that are fused, bridged, or spiro-linked. Representative heterocycloalkyl groups include, but are not limited to, 3-dioxolane, benzo[1,3]dioxolyl, azetidinyl, oxetanyl, pyrazolinyl, pyranyl, thiomorpholinyl, pyrazolidinyl, piperidyl, piperazinyl, 1,4-dioxanyl, imidazolinyl, pyrrolinyl, pyrrolidinyl, piperidinyl, imidazolidinyl, morpholinyl, 1,4-dithianyl, pyrrolidinyl, oxazolinyl, oxazolidinyl, isoxazolinyl, isoxazolidinyl, thiazolinyl, thiazolidinyl, isothiazolinyl, isothiazolidinyl, dihydropyranyl, tetrahydropyranyl, tetrahydrofuranyl, and tetrahydrothiophenyl.

[0053] The terms "cycloalkoxy", "heterocycloalkoxy", "aryloxy", and "heteroaryloxy" correspond to "cycloalkyl", "heterocycloalkyl", "aryl", and "heteroaryl" as defined above, which are bonded to the molecule by an -O- (ether) bond. For example, "cycloalkoxy", "heterocycloalkoxy", "aryloxy", and "heteroaryloxy" correspond to -O-cycloalkyl, -O-heterocycloalkyl, -O-aryl, and -O-heteroaryl. A preferred embodiment of "heterocycloalkoxy" is oxetanyloxy or oxetanoxy (-O-oxetanyl).

[0054] The term "Boc" corresponds to the following tert-butoxycarbonyl group "-C(=O)-O-C(CH3)3".

[0055] The expression "substituted by at least ~" means that the group is substituted by one or several of the groups in the list.

[0056] "Pharmaceutically acceptable salts" include inorganic and organic acid salts. Representative examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, etc. Representative examples of suitable organic acids include formic acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, propionic acid, benzoic acid, cinnamic acid, citric acid, fumaric acid, maleic acid, methanesulfonic acid, etc. Further examples of pharmaceutically acceptable inorganic or organic acid addition salts include the pharmaceutically acceptable salts described in J. Pharm. Sci. 1977, 66, 2, as well as Handbook of Pharmaceutical Salts: Properties, Selection, and Use, edited by P. Heinrich Stahl and Camille G. Wermuth, 2002. In a preferred embodiment, the salt is selected from the group consisting of maleate, hydrochloride, hydrobromide, and methanesulfonate. "Pharmaceutically acceptable salts" also include inorganic and organic base salts. Representative examples of suitable inorganic bases include alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as calcium or magnesium salts, or ammonium salts. Representative examples of suitable salts with organic bases include, for example, salts with methylamine, dimethylamine, trimethylamine, piperidine, morpholine, tris-(2-hydroxyethyl)amine. "Pharmaceutically acceptable salts" further include salts with amino acids such as aspartic acid, lysine, and arginine. In a preferred embodiment, the salt is selected from the group consisting of sodium, lysine, and arginine salts. It also includes disodium, diarginine, and dilysine salts.

[0057] The present invention provides highly water-soluble indole derivatives (i.e., N-substituted indole derivatives) substituted on the nitrogen of the indole core.

[0058] As used herein, derivatives or compounds not substituted on the nitrogen of the indole core are also referred to, without limitation, as "indole derivatives", "payloads", "active ingredients", "active compounds", "pharmaceutically active ingredients", "biologically active ingredients", "biologically active forms", "drugs", or "molecules". Such "indole derivatives" etc. have the following formula:

[0059] [Chemical formula]

[0060] [wherein X, R1, R 1' , R2, and R3 are as defined in the present disclosure] have.

[0061] As used herein, N-substituted indole derivatives are also referred to as "prodrugs".

[0062] Generally, "prodrugs", as used herein, are "N-substituted indole derivatives" of formula (I) which may themselves be less active or even inactive, but upon administration are converted to the corresponding biologically active form under physiological conditions, e.g., by metabolism, solvolysis, or otherwise.

[0063] In particular, the prodrugs of indole derivatives as defined herein may be compounds of formula (I) obtained by subjecting the amino group of an indole derivative to acylation, alkylation or phosphorylation (for example, compounds of formula (I) obtained by subjecting the amino group of an indole derivative to eicosanoylation, alanylation, pentylaminocarbonylation, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylation, tetrahydrofuranylation, pyrrolidylmethylation, pivaloyloxymethylation and tert-butylation).

[0064] In certain embodiments, prodrugs of indole derivatives as defined herein include, but are not limited to, any prodrug suitable for a drug containing an amino group such as an indole derivative drug. For example, it may include amides, carbamates and thiocarbamates (O-thiocarbamate or S-thiocarbamate), N-acyl-oxyalkyl derivatives, N-acyl-oxyacyloxycarbonyl derivatives, β-aminoketones, (oxodioxolenyl)methyl derivatives, N-Mannich bases, imines (Schiff bases), enamines and enamino ketones, azo compounds, lactonization systems, tetrahydro-(2H)-1,3,5-thiadiazine-2-thione (THTT), redox systems, and prodrugs such as PEG, as disclosed by Simplicio et al. (Molecules 2008, 13, 519-547).

[0065] In one embodiment, the prodrug of the indole derivative as defined herein is of the amide prodrug type. More specifically, the nitrogen atom of the indole core is substituted by a -CO-R' group, where R' can represent H, halogen which may or may not be substituted, cycloalkyl, aryl or heteroaryl optionally substituted (C1-C6) alkyl. In a further embodiment, the prodrug of the indole derivative as defined herein is of the carbamate prodrug type. More specifically, the nitrogen atom of the indole core is substituted by a -CO2-R' group, where R' can represent H, halogen which may or may not be substituted, cycloalkyl, aryl or heteroaryl optionally substituted (C1-C6) alkyl. In a further embodiment, the prodrug of the indole derivative as defined herein is an N-acyl-oxyalkylamine. More specifically, the nitrogen atom of the indole core is substituted by a -CH(R'')-CO2-R' group, where R' can represent H, halogen which may or may not be substituted, cycloalkyl, aryl or heteroaryl optionally substituted (C1-C6) alkyl, and R'' can represent H, (C1-C6) alkyl optionally substituted by halogen. The amide, carbamate, and N-acyl-oxyalkylamine prodrug types can release the drug by amidase, peptidase, or esterase.

[0066] In one embodiment, the prodrug of the indole derivative as defined herein is of the sulfenamide prodrug type. More specifically, the nitrogen atom of the indole core is substituted by a -S-R' group, where R' can represent H, halogen which may or may not be substituted, cycloalkyl, aryl or heteroaryl optionally substituted (C1-C6) alkyl.

[0067] In certain embodiments, the prodrug of the indole derivative as defined herein is of the phosphate prodrug type. More specifically, the nitrogen atom of the indole core is (C1-C6) alkyl-O-P(O)(OH)(O - ) group, -NH-P(O)-(OH)2 group, -N-P(O)-(OH) group, -N-P-(OH)2 group or -NO-P(O)-(OH)2 substituted. The phosphate prodrug type can release the drug in the presence of phosphatases such as alkaline phosphatase.

[0068] In certain embodiments, the prodrug of the indole derivative as defined herein is of the N-Mannich base prodrug type. More specifically, the nitrogen atom of the indole core is substituted by a (C1-C6) alkyl-NH-CO-R' group, where R' can represent H, halogen which may or may not be substituted, cycloalkyl, aryl or heteroaryl optionally substituted by (C1-C6) alkyl.

[0069] In certain embodiments, the prodrug of the indole derivative as defined herein is

[0070] [Chemical formula]

[0071] [wherein R 1 may be H, alkyl or especially C1-C8 alkyl, R 2 may be alkyl, cycloalkyl, aryl or heteroaryl, which may or may not be substituted, or haloalkyl, R 3 may be H, metal, R2 or a substituted or unsubstituted primary, secondary or tertiary amine, R 4 may be H, metal, ammonium salt, or alkyl, R5 may be a substituted or unsubstituted natural amino acid, R a or R b may be H, alkyl or aryl, or NR a or NR b may be an amino acid, X may be C or O, k may be 1 or 2, m may be from 2 to 22, or (CH k ) m may be a saturated, unsaturated or conjugated hydrocarbon, n may be from 0 to 2, the metal may be Na, K, Li, Ca, Mg, Ag or Zn is selected from the group consisting of.

[0072] In a further specific embodiment, the prodrug of the indole derivative as defined herein is of the phosphothiolate prodrug type. More specifically, the nitrogen atom of the indole core is substituted by (C1-C6)alkyl-NH-P(O)(OR')-C(V)-X-S-R'' or by -P(O)(OR')-C(V)-X-S-R'', where V represents (C1-C8)alkyl, X represents CR3(R4), R' independently represents an optionally substituted aliphatic or aromatic residue, optionally, R' is (C1-C8)alkoxy n [where n is 1, 2, 3, 4, 5 or 6], F, CI, Br, I, -NO2, N(C1-C8)alkyl, -NH2, -N((C1-C8)alkyl)2, =O, C3-C8-cycloalkyl, -S-S-(C1-C8)alkyl, hydroxy-((C1-C8)alkoxy) n[Here, n is 1, 2, 3, 4, 5, or 6], represents (C1-C8) alkyl optionally substituted with at least one of C2-C8-alkynyl or optionally substituted phenyl, or optionally, R' is (C1-C8) alkyl, (C1-C8) alkoxy n [Here, n is 1, 2, 3, 4, 5, or 6], represents phenyl optionally independently substituted with at least one of F, Cl, I, Br, NO2, N(C1-C8) alkyl, NH2, or -N((C1-C8) alkyl)2, or optionally, R' represents a 5- or 6-membered heteroaromatic system R3 represents H or (C1-C8) alkyl R4 represents H or (C1-C8) alkyl

[0073] R'' represents optionally substituted (C1-C8) alkyl, optionally substituted phenyl, optionally substituted aromatic 5- or 6-membered heterocyclic system, amino acid, peptide, protein, antibody, sugar, polysaccharide, nucleotide, oligonucleotide, or polymer

[0074] The phosphorothioate prodrug type was developed by Tubulis and is more specifically disclosed in U.S. Patent No. 11,161,873B2 and U.S. Patent Application No. 2020 / 0390901A1

[0075] In a further specific embodiment, the prodrug of the indole derivative as defined herein is of the carbohydrate prodrug type. The carbohydrate prodrug type can release the drug in the presence of glycosidase enzymes. In particular, the carbohydrate prodrug type includes the prodrugs disclosed by H. Martin et al. (Chem. Soc. Rev., 2022, 51, 9694) or in U.S. Patent Application No. 2009 / 0227617. In a specific embodiment, it is of the following formula:

[0076]

Chemical formula

[0077] [In the formula, X, R1, R 1' , R2, and R3 are defined in the present disclosure, Y is -O- or -NH-CO-O-, R is H, -OH, -CH2OH, -CH3, -COOR', where R' is H, -CH3, -CH2-CH=CH2, -(CH2) x

[0078] [Chemical formula]

[0079] CH, -CH2-CH2-OH, -CH2-CH2-OCH3, or a residue having the formula

[0080] [Chemical formula]

[0081] and is, n is an integer from 1 to 40, preferably from 1 to 10, more preferably from 2 to 6, x is an integer from 1 to 5, preferably from 1 to 3, more preferably 1 or 2, p is an integer from 1 to 5, preferably from 1 to 3, more preferably 1 or 2, alkyl is a linear or branched alkyl residue having 1 to 10 carbon atoms, preferably 1 to 3 carbon atoms] and includes a glycoconjugate having

[0082] In a further specific embodiment, it is the following formula:

[0083] [Chemical formula]

[0084] [In the formula, X, R1, R 1' , R2, and R3 are as defined in the present disclosure, Y is -O- or -NH-CO-O-, R is H, -OH, -CH2OH, -CH3, -COOR', where R' is H, -CH3, -CH2-CH=CH2, -(CH2) x

[0085]

Chemical formula

[0086] CH, -CH2-CH2-OH, -CH2-CH2-OCH3, or a residue having the formula

[0087]

Chemical formula

[0088] and is a residue having n is an integer from 1 to 40, preferably from 1 to 10, more preferably from 2 to 6, x is an integer from 1 to 5, preferably from 1 to 3, more preferably 1 or 2, p is an integer from 1 to 5, preferably from 1 to 3, more preferably 1 or 2, alkyl is a linear or branched alkyl residue having 1 to 10 carbon atoms, preferably 1 to 3 carbon atoms] and includes a glycoconjugate having

[0089] In a further embodiment according to the present invention, the term "N-substituted indole derivative" corresponds to an indole derivative in which the nitrogen atom of the indole core is substituted by a substituent Ra.

[0090] Therefore, the present invention relates to one of the compounds of formula (I) or a pharmaceutically acceptable salt thereof:

[0091]

Chemical formula

[0092] [wherein, - X represents a nitrogen atom, a C-CN unit or an N+-O- unit, - R1 and R1' independently represent hydrogen, halogen, (C1-C6) alkoxy, or a -SO2-CH3 group, provided that when R1 or R1' is (C1-C6) alkoxy, R2 is not halogen, - R2 is ·(C1-C6) alkoxy, (C3-C6) cycloalkoxy, (C3-C6) heterocycloalkoxy, aryloxy, heteroaryloxy, (C1-C6) alkylaryloxy, (C1-C6) alkylheteroaryloxy groups which are optionally substituted by at least one halogen, said groups, or thio (C1-C6) alkyl, thioaryl, thioheteroaryl, thio (C1-C6) alkylaryl or thio (C1-C6) -alkylheteroaryl groups which are optionally substituted by at least one halogen or by a (C1-C6) alkoxy group, said groups, ·-NR4R5 unit, O-(C1-C6) alkyl-NR4R5 unit or S-(C1-C6) alkyl-NR4R5 unit (wherein R4 and R5 represent H, a (C1-C6) alkyl group, or R4 and R5 together form a 3- to 7-membered ring optionally interrupted by one or several heteroatoms, provided that at least one of R4 and R5 is not H), ·NHCOR6 unit (wherein R6 represents a (C1-C6) alkyl group), ·aryl or heteroaryl groups optionally substituted by at least one halogen, trifluoromethyl group or (C1-C3) alkoxy group, or ·halogen represents, - R3 represents hydrogen, a (C1-C3) alkyl group, a (C1-C3) alkoxy group or halogen, - Ra is ·Formula (A):

[0093] [Chemistry]

[0094] or formula (A'):

[0095] [Chemistry]

[0096] group (wherein n is an integer included between 1 and 12), and · formula (B):

[0097] [Chemistry]

[0098] group (wherein · L1 is a cleavable group selected from among pH-sensitive groups, photoinducible cleavable groups, in vivo reductive cleavable groups, and enzymatically cleavable groups, · L2 is a tert-butoxycarbonyl group or a conjugate protein linker, · m is an integer equal to 0 or 1) is a group selected from the group consisting of or a pharmaceutically acceptable salt thereof is provided.

[0099] Indole derivatives, etc. ("payload") In certain embodiments, the indole derivative and the compound of formula (I) are the (Z)-isomer. In further certain embodiments, the indole derivative and the compound of formula (I) are the (E)-isomer.

[0100] According to the present invention, R1 and R1' independently represent hydrogen, halogen, (C1-C6) alkoxy, or -SO2-CH3 group, provided that when R1 or R1' is (C1-C6) alkoxy, R2 is not halogen. In certain embodiments, R1 and R1' are such that one is H and the other represents halogen, (C1-C6) alkoxy, or -SO2-CH3 group, provided that when R1 or R1' is (C1-C6) alkoxy, R2 is not halogen. Preferably, R1 and R1' are such that one is H and the other represents halogen, typically bromine, chlorine or fluorine, advantageously bromine or chlorine, more specifically bromine.

[0101] According to the present invention, R2 is · (C1-C6) alkoxy, (C3-C6) cycloalkoxy, (C3-C6) heterocycloalkoxy, aryloxy, heteroaryloxy, (C1-C6) alkylaryloxy, (C1-C6) alkylheteroaryloxy group, which is optionally substituted by at least one halogen, said group, or thio (C1-C6) alkyl, thioaryl, thioheteroaryl, thio (C1-C6) alkylaryl or thio (C1-C6) -alkylheteroaryl group, which is optionally substituted by at least one halogen or by (C1-C6) alkoxy group, said group, · -NR4R5 unit, O-(C1-C6) alkyl-NR4R5 unit or S-(C1-C6) alkyl-NR4R5 unit (wherein R4 and R5 represent H, (C1-C6) alkyl group, or R4 and R5 together form a 3- to 7-membered ring optionally interrupted by one or several heteroatoms, provided that at least one of R4 and R5 is not H), · NHCOR6 unit (wherein R6 represents (C1-C6) alkyl group), · aryl or heteroaryl group optionally substituted by at least one halogen, trifluoromethyl group or (C1-C3) alkoxy group, or · halogen represents.

[0102] In particular, R2 is · a (C1-C6) alkoxy, phenoxy or oxetanoxy group, optionally substituted by at least one halogen, preferably bromine, chlorine or fluorine, more preferably fluorine such as trifluoromethyl, etc., of said group, · a halogen, preferably bromine, chlorine or fluorine, more preferably bromine or chlorine, · an R4-N-R5 unit or an S-(C1-C6)alkyl-NR4R5 unit (wherein R4 and R5 represent H or a (C1-C6)alkyl group, provided that at least one of R4 and R5 is not H), · an NHCOR6 unit (wherein R6 represents a (C1-C6)alkyl group, preferably methyl, ethyl or tert-butyl), or · an aryl group optionally substituted by at least one halogen or trifluoromethyl group, or · a heteroaryl group, preferably furan, triazole, pyridine, thiazole, pyran, pyrrole, imidazole, tetrazole, benzofuran, triazinyl, pyrazinyl, pyridazinyl, or tetrazole represents.

[0103] Preferably, R2 is · a (C1-C6) alkoxy group selected from the group consisting of a methoxy group, an ethoxy group, an isopropoxy group, optionally substituted by fluorine such as trifluoromethyl or oxetanoxy, preferably a methoxy group and an isopropoxy group, or a phenoxy group, · a halogen selected from the group consisting of fluorine and chlorine, · an R4-N-R5 unit or an S-(C1-C6)alkyl-NR4R5 unit (wherein R4 and R5 represent a methyl or ethyl group), · a group selected from the group consisting of a thiomethyl group, a thioethyl group, a thiobenzyl group, a thiopyridinyl group and a thiophenyl group, optionally substituted by at least one fluorine or trifluoromethyl group, · A phenyl group optionally substituted by at least one bromine or trifluoromethyl group, or · A heteroaryl group selected from the group consisting of furan or triazole represents.

[0104] More preferably, R2 is · A (C1-C6) alkoxy group, preferably a methoxy group, · A (C3-C6) heterocycloalkoxy group, preferably an oxetanoxy group, or · A heteroaryl group, preferably a furanyl or triazolyl group represents.

[0105] According to the present invention, R3 represents hydrogen, a (C1-C3) alkyl group, preferably methyl, ethyl or isopropyl, a (C1-C3) alkoxy group, preferably methoxy, ethoxy or isopropoxy, or halogen, preferably fluorine. Preferably, R3 is hydrogen, methoxy or fluorine. More preferably, R3 is hydrogen.

[0106] In a preferred embodiment: - X represents a C-CN unit, - R1 represents halogen, preferably bromine, - R1' represents hydrogen, - R2 represents a (C1-C6) alkoxy group, preferably a methoxy group, - R3 represents hydrogen.

[0107] In a further preferred embodiment: - X represents a C-CN unit, - R1 represents hydrogen, - R1' represents a (C1-C6) alkoxy group, preferably a methoxy group, - R2 represents a (C1-C6) alkoxy group, preferably a methoxy group, - R3 represents hydrogen.

[0108] In certain embodiments, the indole derivatives correspond to the indole derivatives disclosed in WO2014 / 086964, and more specifically to the indole derivatives within the scope of the claims of EP2928864B1.

[0109] In preferred embodiments, indole derivatives and the like are - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-chloropyridin-3-yl)acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-chloropyridin-3-yl)acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-methoxypyridin-3-yl)acrylonitrile; - (E)-2-(5-Bromo-1H-indol-3-yl)-3-(4-methoxypyridin-3-yl)acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-(dimethylamino)pyridin-3-yl)acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-dimethylamino)pyridin-3-yl)acrylonitrile, hydrochloride; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(dimethylamino)pyridin-3-yl)acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-methoxypyridin-3-yl)acrylonitrile; - (E)-2-(5-Chloro-1H-indol-3-yl)-3-(4-methoxypyridin-3-yl)acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-phenoxypyridin-3-yl)acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-phenoxypyridin-3-yl)acrylonitrile; - (Z)-2-(5-Methoxy-1H-indol-3-yl)-3-(4-methoxypyridin-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-ethoxypyridin-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-isopropoxypyridin-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(methylthio)pyridin-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(ethylthio)pyridin-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(3-bromophenyl)pyridin-3-yl)-acrylonitrile; - (Z)-3-(4-(3-Bromophenyl)pyridin-3-yl)-2-(5-chloro-1H-indol-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(phenylthio)pyridin-3-yl)-acrylonitrile; - (Z)-3-(4-(Benzylthio)pyridin-3-yl)-2-(5-bromo-1H-indol-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(3,4-dimethoxy)thio)pyridin-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(4-fluorophenoxy)pyridin-3-yl)-acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-(4-fluorophenoxy)pyridin-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(diethylamino)pyridin-3-yl)-acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(4-(trifluoromethyl)phenyl)pyridin-3-yl)acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-(4-(trifluoromethyl)phenyl)pyridin-3-yl)acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-((4-fluorophenyl)thio)pyridin-3-yl)acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-((4-fluorophenyl)thio)pyridin-3-yl)acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-(furan-3-yl)pyridin-3-yl)acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-(pyridin-2-ylthio)pyridin-3-yl)acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(pyridin-2-ylthio)pyridin-3-yl)acrylonitrile; - (Z)-3-(4-(1H-1,2,4-Triazol-1-yl)pyridin-3-yl)-2-(5-bromo-1H-indol-3-yl)acrylonitrile; - (Z)-3-(4-(1H-1,2,4-Triazol-1-yl)pyridin-3-yl)-2-(5-chloro-1H-indol-3-yl)acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(furan-3-yl)pyridin-3-yl)acrylonitrile; - (E)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(furan-3-yl)pyridin-3-yl)acrylonitrile; - (Z)-2-(5-Chloro-1H-indol-3-yl)-3-(4-methoxypyridin-3-yl)acrylonitrile; - (Z)-2-(5-Bromo-1H-indol-3-yl)-3-(4-(2-(dimethylamino)ethylthio)pyridin-3-yl)acrylonitrile; - (Z)-3-(2-(5-Bromo-1H-indol-3-yl)-2-cyanovinyl)-4-(4-fluorophenoxy)benzonitrile; - (Z)-3-(2-(5-Bromo-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile; - (E)-3-(2-(5-Bromo-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile; - (Z)-3-(2-(5-Bromo-1H-indol-3-yl)-2-cyanovinyl)-4-(dimethylamino)benzonitrile; - (Z)-3-(2-(5-Chloro-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile; - (Z)-3-(2-(5-Chloro-1H-indol-3-yl)-2-cyanovinyl)-4-(dimethylamino)benzonitrile; - (Z)-3-(2-(5-Chloro-1H-indol-3-yl)-2-cyanovinyl)-4-(ethylthio)benzonitrile; - (Z)-2-(6-Bromo-1H-indol-3-yl)-3-(4-methoxypyridin-3-yl)acrylonitrile; - (Z)-2-(6-Fluoro-1H-indol-3-yl)-3-(4-methoxypyridin-3-yl)acrylonitrile; - (Z)-2-(6-Chloro-1H-indol-3-yl)-3-(4-methoxypyridin-3-yl)acrylonitrile; - (Z)-3-(2-(5-Bromo-1H-indol-3-yl)-2-cyanovinyl)-4-(furan-3-yl)pyridine 1-oxide; - (Z)-3-(2-(5-Chloro-1H-indol-3-yl)-2-cyanovinyl)-4-methoxypyridine 1-oxide; - (Z)-3-(2-(5-Chloro-1H-indol-3-yl)-2-cyanovinyl)-4-(trifluoromethoxy)benzonitrile; - (Z)-3-(2-(5-Bromo-1H-indol-3-yl)-2-cyanovinyl)-4-(trifluoromethoxy)benzonitrile; and is selected from the group consisting of those pharmaceutically acceptable salts.

[0110] In a more preferred embodiment, the indole derivative is Example 38 of WO2014 / 086964: the following formula:

[0111]

Chemical formula

[0112] (Z)-3-(2-(5-Bromo-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile having

[0113] Phosphonic acid series The present invention provides the compound of formula (I) as a "phosphonic acid prodrug" of the indole derivative as defined above.

[0114] Therefore, in a particular embodiment, Ra is of formula (A):

[0115]

Chemical formula

[0116] or of formula (A'):

[0117]

Chemical formula

[0118] group of [wherein, n is an integer included between 1 and 12]. In a preferred embodiment, n is an integer included between 2 and 10, more preferably between 2 and 8, and even more preferably between 2 and 6. Advantageously, n is 2, 3, 4, 5, or 6.

[0119] In certain embodiments, the "phosphonic acid prodrug" includes their pharmaceutically acceptable salts. Preferably, the pharmaceutically acceptable salt is selected from sodium salt, disodium salt, lysine salt, dilysine salt, arginine salt or diarginine salt. More preferably, the pharmaceutically acceptable salt is selected from sodium salt or disodium salt, and even more preferably sodium salt.

[0120] In a preferred embodiment, the compound of formula (I) is - Example 1: (Z)-5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonic acid, - Example 2: (Z)-3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid, - Example 3: (Z)-7-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-7-oxoheptylphosphonic acid, - Example 4: (Z)-5-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-5-oxopentylphosphonic acid, - Example 5: (Z)-7-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-7-oxoheptylphosphonic acid, - Example 6: (Z)-sodium 5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate, - Example 7: Sodium (Z)-3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonate, - Example 8: Sodium (Z)-4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonate, - Example 9: Sodium (Z)-6-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxohexylphosphonate, - Example 10: Sodium (Z)-7-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-7-oxoheptylphosphonate, - Example 11: Sodium (Z)-5-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-5-oxopentylphosphonate, - Example 12: Sodium (Z)-7-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-7-oxoheptylphosphonate, - Example 13: Disodium (Z)-4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonate, - Example 14: Disodium (Z)-5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate, - Example 15: (S)-2,6-Diaminohexanoic acid compound and (Z)-3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid (2:1), - Example 16: (S)-2,6-diaminohexanoic acid compound and (Z)-4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonic acid (2:1), - Example 17: 3-[hydroxy(dioxo)-λ6-phosphanyl]propyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate, and - Example 18: 4-[hydroxy(dioxo)-λ6-phosphanyl]butyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate selected from the group consisting of.

[0121] L1 series The present invention further provides a compound of formula (I) as an "L1 prodrug" of an indole derivative as defined above. As used herein, an "L1 prodrug" corresponds to a prodrug containing a cleavable group L1.

[0122] Thus, in certain embodiments, Ra is of formula (B):

[0123]

Chemical formula

[0124] group [wherein, · L1 is a cleavable group selected from among a pH-sensitive group, a photoinducible cleavable group, a bioreductive cleavable group, and an enzymatically cleavable group, · L2 is a tert-butoxycarbonyl group or a conjugate protein linker, · m is an integer equal to 0 or 1] is.

[0125] According to the present invention, L1 is a cleavable group selected from among a pH-sensitive group, a photoinducibly cleavable group, a reductively cleavable group, and an enzymatically cleavable group.

[0126] In certain embodiments, L1 is a pH-sensitive group. As used herein, the term "pH-sensitive group" is highly sensitive to hydrolysis and can be cleaved at a specific pH value, particularly under acidic conditions in the pH range of 4 to 6. Thus, such a pH-sensitive group may be an acid-inducibly cleavable group or an acid-labile group, which is relatively stable in neutral blood (pH 7.3 - 7.5) and can be hydrolyzed in weakly acidic endosomes (pH 5.0 - 6.5) and lysosomes (pH 4.5 - 5.0). Examples of hydrolyzable pH-sensitive groups include, but are not limited to, hydrazine, hydrazone, semicarbazone, thiosemicarbazone, cis-aconitamide, orthoester, carbonate, acetal, ketal, boronic acid ester, and the like. In a preferred embodiment, the pH-sensitive group as defined above is an acid-inducibly cleavable group.

[0127] In a further specific embodiment, L1 is a photoinducibly cleavable group.

[0128] In a further specific embodiment, L1 is a biodegradable reducible group. As used herein, a "biodegradable reducible" group is cleavable under biological reducing conditions. For example, the group is cleavable in the presence of a reducing agent such as glutathione or dithiothreitol. Thus, a glutathione-sensitive group can be cleaved by a disulfide exchange reaction with intracellular glutathione species. Preferably, L1 is a glutathione-sensitive group or a group having a disulfide or a group having a sulfonamide. For example, various disulfide groups are known in the art, including those that can be formed using SATA (N-succinimidyl-5-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), and SMPT (N-succinimidyl oxycarbonyl alpha methyl alpha(2-pyridyldithio)toluene). As used herein, a sulfonamide group is a sulfonyl group linked to an amine group, where the sulfur-nitrogen bond can be cleaved.

[0129] In a further specific embodiment, L1 is an enzymatically cleavable group. As used herein, an "enzymatically cleavable" group is a group that is cleaved by an enzyme such as protease, peptidase, esterase, beta-glucuronidase, glycosidase, phosphodiesterase, phosphatase, pyrophosphatase, tubulin tyrosine ligase, or lipase.

[0130] In one embodiment, the enzymatically cleavable group L1 is cleaved by a protease or peptidase.

[0131] Examples of proteases include, but are not limited to, cathepsin B, VAGP tetrapeptide, etc. Preferably, the enzymatically cleavable group L1 is cleaved by a peptidase. In this embodiment, the enzymatically cleavable group L1 contains a peptide that is the cleavage site of the linker, thereby facilitating the release of the drug upon exposure to intracellular proteases such as lysosomal enzymes. The peptide can be designed and optimized for enzymatic cleavage by a specific enzyme, such as a tumor-related protease, cathepsin B, C, and D, or a plasmin protease. Examples of peptides having two amino acids include, but are not limited to, alanine-alanine (ala-ala), valine-alanine (val-ala), valine-citrulline (vc or val-cit), valine-arginine (val-arg), valine-lysine (val-lys), alanine-phenylalanine (af or ala-phe); phenylalanine-lysine (fk or phe-lys); phenylalanine-homolys; and N-methyl-valine-citrulline (Me-val-cit). Examples of peptides having three amino acids include, but are not limited to, glycine-valine-citrulline (gly-val-cit), aspartic acid-valine-citrulline (asp-val-cit), alanine-alanine-asparagine (ala-ala-asn), alanine-phenylalanine-lysine (ala-phe-lys), glutamic acid-cysteine-glycine (glu-cys-gly), glycine-glycine-phenylalanine (gly-gly-phe), and glycine-glycine-glycine (gly-gly-gly). Examples of peptides having four amino acids include, but are not limited to, glycine-glycine-valine-citrulline (gly-gly-val-cit), glycine-glycine-phenylalanine-glycine (gly-gly-phe-gly), and glutamic acid-glycine-glutamic acid-glutamic acid (glu-gly-glu-glu). The above amino acid combinations can also exist in the reverse order (i.e., cit-val).

[0132] Examples of peptides are provided in particular in Table A (Table 1).

[0133]

Table 1

[0134] The peptides of the present disclosure can include L- or D-isomers of amino acid residues. The term "naturally occurring amino acid" refers to Ala, Asp, Asx, Cit, Cys, Glu, Phe, Glx, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, and Tyr. In contrast to the configuration in naturally occurring ("L-") amino acids, "D-" designates an amino acid having the "D" (dextrorotary) configuration. The amino acids described herein can be purchased commercially or synthesized using methods known in the art.

[0135] Preferably, L1 is a p-aminobenzyloxycarbonyl-AA1 w -AA2 x -AA3 y -AA4 z group, where AA1, AA2, AA3, and AA4 independently represent amino acids as defined above, and w, x, y, and z are independently integers equal to 0 or 1. As used herein, the "p-aminobenzyloxycarbonyl" group is also referred to as PABC. More preferably, L1 is a p-aminobenzyloxycarbonyl-AA1 w -AA2 x -AA3 y -AA4 za base, where AA1, AA2, AA3, and AA4 are independently selected from the group consisting of alanine, arginine, valine, citrulline, phenylalanine, lysine, glycine, aspartic acid, asparagine, glutamic acid, and derivatives thereof, preferably citrulline, valine, cysteic acid, glycine, and glutamic acid, more preferably citrulline, valine, and cysteic acid, and w, x, y, and z are independently integers equal to 0 or 1. Even more preferably, L1 is a group selected from the group consisting of -PABC-Cit-Val and -PABC-Cit-Val-cysteic acid.

[0136] In certain embodiments, the enzymatically cleavable group L1 is cleaved by pyrophosphatase. In this embodiment, L1 contains a pyrophosphate group.

[0137] In certain embodiments, the enzymatically cleavable group L1 is cleaved by beta-glucuronidase. In preferred embodiments, L1 is a -PABC-glucuronide group.

[0138] In certain aspects, an "L1 prodrug" is one in which L1 is as defined above and L2 is absent. In this aspect, the compound of formula (I) is such that Ra is of formula (B):

[0139]

Chemical formula

[0140] a group [wherein L1 is as defined above and m is an integer equal to 0] and is such that.

[0141] According to this aspect, preferred compounds are - Example 19: (2S,3S,4S,5R,6S)-6-[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenoxy]-3,4,5-trihydroxytetrahydropyran-2-carboxylic acid, - Example 21: [4-[[(2S)-2-[[(2S)-2-aminopropanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl 5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate hydrochloride, and - Example 25: 2-amino-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-3-oxopropane-1-sulfonic acid is a compound selected from the group consisting of.

[0142] In one embodiment, the "L1 prodrug" is such that L1 is as defined above, m is an integer equal to 1, and L2 is a tert-butoxycarbonyl (Boc) group. In this embodiment, the compound of formula (I) is such that Ra is of formula (B):

[0143]

Chemical formula

[0144] group [wherein, L1 is as defined above, and m is an integer equal to 0] is such a one.

[0145] According to this embodiment, preferred compounds are - Example 20: [4-[[(2S)-2-[[(2S)-2-(tert-Butoxycarbonylamino)-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate, and - Example 24: 3-[[(1S)-1-[[(1S)-1-[[4-[[5-Bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-(tert-butoxycarbonylamino)-3-oxopropane-1-sulfonic acid is a compound selected from the group consisting of.

[0146] In one embodiment, the "L1 prodrug" is such that L1 is defined as above, m is an integer equal to 1, and L2 is a binding protein conjugate.

[0147] As used herein, a "binding protein conjugate" is a group that can react or bind or conjugate with a peptide, protein or antibody. More specifically, a "binding protein conjugate" includes any chemical moiety or group that can bind a peptide, protein or antibody via a stable and covalent bond. For example, a "binding protein conjugate" has the functionality to react with free cysteine present in a peptide, protein or antibody.

[0148] Examples of "binding protein conjugates" include, but are not limited to, SMCC (succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate) groups, succinimidyl thioether groups. Further examples of "binding protein conjugates" are the following formula:

[0149] [Chemical formula]

[0150] [In the formula, r is an integer included between 1 and 36, preferably between 1 and 24, more preferably an integer equal to 2, 4, 8, 12, 16, 20, or 24] comprises a group having

[0151] According to this embodiment, preferred compounds are - Example 22: [4-[[(2S)-2-[[(2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy](peg4)propanoylamino]-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate, and - Example 26: (2R)-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy]propanoylamino]-3-oxopropane-1-sulfonic acid is a compound selected from the group consisting of

[0152] Conjugate The present invention relates to a conjugate of formula (II):

[0153]

Chemical formula

[0154] [In the formula, - X, R1, R 1' , R2, R3, and L1 are as defined herein, - L2 is a binding protein conjugate, - P is a peptide or protein, preferably an antibody, antibody fragment, or antigen-binding fragment, capable of binding to a target of interest, - v is an integer from 1 to 10] is also provided.

[0155] According to the present invention, the conjugate of formula (II) is - X, R1, R 1' , R2, R3 are defined in the above "indole derivative" or "payload" section including all specific and preferred embodiments, - L1 is a cleavable group defined in the above "L1 series" section including all specific and preferred embodiments, - L2 is a binding protein linker defined in the above "L1 series" section including all specific and preferred embodiments of such a kind.

[0156] According to the present invention, the peptide or protein "P" may be conjugated to more than one compound of formula (I), wherein m is 1 and L2 is a binding protein linker. For example, the peptide or protein "P" may be conjugated to 1 to 10, 1 to 9, 1 to 8, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 compounds of formula (I), wherein m is 1 and L2 is a binding protein linker. In a preferred embodiment, v is an integer from 1 to 8, from 2 to 8. In a particularly preferred embodiment, v is an integer equal to 1, 2, 3, 4, 5, 6, 7, or 8.

[0157] According to the present invention, P is a peptide or protein capable of binding to a target of interest. As used herein, "a peptide or protein capable of binding to a target of interest" refers to any peptide and / or protein that recognizes and binds to a cell surface molecule, particularly a cell surface marker, antigen, epitope, or receptor. In certain embodiments, protein P binds to another protein not limited to the polypeptide moiety. Protein P that targets specific cells, tissues, or locations may also have certain therapeutic effects such as antiproliferative (cell growth inhibitory and / or cytotoxic) activity against the target cells or pathways. In certain embodiments, protein P can contain or be genetically engineered to contain at least one chemically reactive group such as a carboxylic acid, amine, thiol, or an amino acid moiety or side chain with chemical reactivity. In certain embodiments, protein P can contain a targeting moiety that binds or forms a complex with a cell surface molecule such as a cell surface receptor or antigen for a given target cell population. Following specific binding or complex formation with the receptor, the cell allows uptake of the targeted conjugate, which is then internalized by the cell.

[0158] In certain embodiments, P is a protein, preferably albumin. These albumin conjugates of formula (II) increase the circulating half-life of the payload and facilitate intracellular delivery to tumors.

[0159] In a preferred embodiment, P is an antibody, antibody fragment, or antigen-binding fragment.

[0160] An antibody is a protein produced by the immune system that can recognize and bind to a specific antigen. The target antigen generally has multiple binding sites, also called epitopes or antigenic determinants, which are recognized by the CDRs of multiple antibodies. Each antibody that specifically binds to a different epitope has a different structure. Thus, one antigen can have more than one corresponding antibody. The term "antibody" is used herein in the broadest sense and specifically encompasses monoclonal antibodies, single-domain antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments as long as they exhibit the desired biological activity. Antibodies may be murine, human, humanized, chimeric, or derived from other species.

[0161] A monoclonal antibody is a homogeneous population of antibodies against a specific antigenic determinant (e.g., a cancer cell antigen, a viral antigen, a microbial antigen, a protein, a peptide, a carbohydrate, a chemical substance, a nucleic acid, or a fragment thereof). Monoclonal antibodies (mAbs) against the antigen of interest can be prepared by using any technique known in the art that provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, human B cell hybridoma technology and EBV-hybridoma technology. Such antibodies may be of any immunoglobulin class, including IgG, IgM, IgE, IgA, and IgD, as well as any of their subclasses. The hybridomas that produce the mAbs used in the present disclosure may be cultured in vitro or in vivo.

[0162] Useful monoclonal antibodies include, but are not limited to, human monoclonal antibodies, humanized monoclonal antibodies, antibody fragments, or chimeric human-mouse (or other species) monoclonal antibodies. Human monoclonal antibodies may be made by any of a number of techniques known in the art.

[0163] In certain embodiments, the antibody can be a bispecific antibody (BsAb), i.e., a protein, typically an engineered protein, that can simultaneously bind two different types of antigens or two different epitopes on the same antigen. The BsAb can be a bispecific monoclonal antibody (BsMAb). Methods for making bispecific antibodies are known in the art. Traditional generation of full-length bispecific antibodies is based on the co-expression of two immunoglobulin heavy chain-light chain pairs, where the two chains have different specificities. The main types of production methods are quadromas, chemical conjugation, and genetic recombination, each method yielding a unique format. BsAbs are often produced by the quadroma or hybridoma method.

[0164] In the quadroma procedure, due to the random pairing of immunoglobulin heavy and light chains, a potential mixture of 10 different antibody molecules is generated, only one of which has the correct bispecific structure. The purification of the correct molecule, usually carried out using an affinity chromatography step, is quite cumbersome and the product yield is low.

[0165] According to a different approach, antibody variable domains (antibody-antigen combining sites) with the desired binding specificities are fused to immunoglobulin constant domain sequences. The fusion can be with an immunoglobulin heavy chain constant domain that includes at least a portion of the hinge, C H 2, and C H 3 regions. The first heavy chain constant region (C H(1) may contain a site necessary for light chain binding present in at least one of the fusions. Nucleic acids encoding immunoglobulin heavy chain fusions and, if desired, immunoglobulin light chains are inserted into separate expression vectors and co-transfected into a suitable host organism. This provides excellent flexibility in adjusting the mutual ratio of the three polypeptide fragments in a manner where the unequal ratios of the three polypeptide chains used in the construction provide an optimal yield. However, if expression at an equal ratio of at least two polypeptide chains results in a high yield, or if the ratio is not particularly significant, it is possible to insert the coding sequences for two or all three polypeptide chains in one expression vector.

[0166] Bispecific antibodies may 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 facilitates the separation of the desired bispecific compound from unwanted immunoglobulin chain combinations, since the presence of the immunoglobulin light chain only in half of the bispecific molecule provides an easy separation technique. Using such techniques, bispecific antibodies can be prepared for the treatment or prevention of diseases as defined herein.

[0167] Hybrid or bifunctional antibodies may be derived either biologically, i.e., by cell fusion techniques, or chemically, in particular using cross-linking agents or disulfide bridge-forming reagents, and may include whole antibodies or fragments thereof.

[0168] The antibody can be a functionally active fragment, derivative or analog of an antibody that immunospecifically binds to a cancer cell antigen, a viral antigen, or a microbial antigen, or another antibody that is bound to a tumor cell or matrix. In this regard, "functionally active" means that an anti-idiotype antibody that recognizes the same antigen as the antibody from which the fragment, derivative or analog is derived can be induced by the fragment, derivative or analog. Specifically, in an exemplary embodiment, the antigenicity of the idiotype of an immunoglobulin molecule can be enhanced by a framework at the C-terminus of the CDR sequence that specifically recognizes the antigen and by deletion of the CDR sequence. To determine which CDR sequence binds to the antigen, synthetic peptides containing the CDR sequence can be used in a binding assay with the antigen by any binding assay method known in the art.

[0169] Other useful antibodies include antibody fragments such as F(ab')2 fragments containing the variable region, the light chain constant region and the C domain of the heavy chain, which can be produced by pepsin digestion of the antibody molecule, and Fab fragments that can be produced by reducing the disulfide bridges of the F(ab')2 fragments, but are not limited thereto. Other useful antibodies include heavy and light chain dimers of the antibody, or any minimal fragment thereof such as Fvs or single chain antibodies (SCAs), or any other molecule having the same specificity as the antibody. As used herein, an antibody fragment is a fragment of an antibody as defined herein. H

[0170] ​In addition, recombinant antibodies such as chimeric and humanized monoclonal antibodies, which contain both human and non-human parts that can be produced using standard recombinant DNA techniques, are useful antibodies. Chimeric antibodies are molecules in which different parts are derived from different animal species, such as those having variable regions derived from murine monoclonal antibodies and human immunoglobulin constant regions. Humanized antibodies are antibody molecules derived from non-human species that have one or more complementarity-determining regions (CDRs) derived from non-human species and framework regions derived from human immunoglobulin molecules. Such chimeric and humanized monoclonal antibodies can be generated by recombinant DNA techniques known in the art.

[0171] Fully human antibodies can be generated using transgenic mice that are unable to express endogenous immunoglobulin heavy and light chain genes but can express human heavy and light chain genes. The transgenic mice are immunized in the usual way with a selected antigen, such as all or part of the polypeptides of the present disclosure. Monoclonal antibodies directed against the antigen can be obtained using conventional hybridoma technology. The human immunoglobulin transgenes carried by the transgenic mice are rearranged during B cell differentiation and then undergo class switching and somatic hypermutation. Thus, it is possible to generate therapeutically useful IgG, IgA, IgM, and IgE antibodies using such techniques. For an overview of this technology for generating human antibodies, see Lonberg and Huszar (1995, Int. Rev. Immunol. 13:65-93). Other human antibodies are commercially available, for example, from Abgenix, Inc. (Freemont, Calif.) and Genpharm (San Jose, Calif.).

[0172] Fully human antibodies that recognize a selected epitope can be produced using a technique called "guided selection." In this approach, a selected non-human monoclonal antibody, such as a mouse antibody, is used to guide the selection of a fully human antibody that recognizes the same epitope. Human antibodies can also be generated using various techniques known in the art, including phage display libraries.

[0173] The antibody can be a fusion protein in which the antibody, or a functionally active fragment thereof, and the amino acid sequence of another protein that is not an antibody (or a portion thereof, such as at least 10, 20, or 50 amino acid moieties of the protein) are fused via a covalent bond (e.g., a peptide bond), for example, at either the N-terminus or C-terminus of the amino acid sequence. For example, the antibody or a fragment thereof may be covalently linked to another protein at the N-terminus of the constant domain.

[0174] The antibody includes analogs and derivatives, where either is modified by a covalent bond of any type of molecule, provided that such a covalent bond allows the antibody to retain its antigen-binding immunospecificity. For example, without limitation, derivatives and analogs of the antibody may be further modified, for example, by glycosylation, acetylation, PEGylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, conjugation with cell antibody units or other proteins, etc. Any of a number of chemical modifications can be performed by known techniques, including but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis in the presence of tunicamycin, etc. In addition, the analog or derivative can contain one or more non-natural amino acids.

[0175] The antibody conjugate can include an antibody having a modification (e.g., substitution, deletion, or addition) at an amino acid residue that interacts with an Fc receptor. In particular, the antibody includes an antibody having a modification at an amino acid residue identified as being involved in the interaction between the anti-Fc domain and the FcRn receptor. An antibody immunospecific for a cancer cell antigen can be obtained commercially, for example, from Genentech (San Francisco, Calif.), or can be generated by any method known to those of skill in the art, such as, for example, chemical synthesis or recombinant expression techniques. A nucleotide sequence encoding an antibody immunospecific for a cancer cell antigen can be obtained, for example, from the GenBank database or a similar database, a published literature, or by routine cloning and sequencing.

[0176] In certain embodiments, the antibody conjugate can be a monoclonal antibody, such as a murine monoclonal antibody, a chimeric antibody, or a humanized antibody. In some embodiments, the antibody can be an antibody fragment, such as a Fab fragment.

[0177] Known antibodies that can be conjugated for the treatment or prevention of cancer are described herein. Antibodies immunospecific for cancer cell antigens can be obtained commercially or generated by any method known to those skilled in the art, such as recombinant expression techniques. Nucleotide sequences encoding antibodies immunospecific for cancer cell antigens can be obtained, for example, from the GenBank database or similar databases, published literature, or by routine cloning and sequencing. Examples of antibodies available for the treatment of cancer include humanized anti-HER2 monoclonal antibodies for the treatment of patients with metastatic breast cancer; the chimeric anti-CD20 monoclonal antibody RITUXAN® (rituximab; Genentech) for the treatment of patients with non-Hodgkin lymphoma; the murine antibody OvaRex (oregovomab; AltaRex Corporation, MA) for the treatment of ovarian cancer; the murine IgG2a antibody Panorex (edrecolomab, Glaxo Wellcome, NC) for the treatment of colorectal cancer; the anti-EGFR IgG chimeric antibody Cetuximab Erbitux® (cetuximab, Imclone Systems Inc., NY) for the treatment of epithelial growth factor-positive cancers such as head and neck cancer; the humanized antibody Vitaxin (etaracizumab, MedImmune, Inc., MD) for the treatment of sarcoma; the humanized IgG1 antibody Campath I / H (alemtuzumab, Leukosite, MA) for the treatment of chronic lymphocytic leukemia (CLL); the humanized anti-CD33 IgG antibody Smart MI95 (Protein Design Labs, Inc., CA) for the treatment of acute myeloid leukemia (AML); the humanized anti-CD22 IgG antibody Lymphocide (epratuzumab, Immunomedics, Inc., NJ) for the treatment of non-Hodgkin lymphoma; the humanized anti-HLA-DR antibody Smart ID 10 (Protein Design Labs, Inc., CA) for the treatment of non-Hodgkin lymphoma; the radiolabeled murine anti-HLA-Dr10 antibody OncoLim (Techniclone, Inc.alloMune (BioTransplant, CA), a humanized anti-CD2 mAb for the treatment of Hodgkin's disease or non-Hodgkin's lymphoma; Avastin (bevacizumab, Genentech, Inc., CA), an anti-VEGF humanized antibody for the treatment of lung and colorectal cancer; epratuzumab (Epratuzamab) (Immunomedics, Inc., NJ and Amgen, CA), an anti-CD22 antibody for the treatment of non-Hodgkin's lymphoma; and CEA-Side (Immunomedics, NJ), a humanized anti-CEA antibody for the treatment of colorectal cancer, among others but not limited to these.

[0178] Other useful antibodies include but are not limited to brentuximab (anti-CD30), polatuzumab (anti-CD79b), enfortumab (anti-nectin 4), tisotumab (tisotumad) (anti-tissue factor), loncastuximab (anti-CD19), rituximab (anti-CD20), trastuzumab (anti-HER2), inotuzumab (anti-CD22), gemtuzumab (anti-CD33), pertuzumab (anti-HER2), obinutuzumab (anti-CD20), ofatumumab (anti-CD20), orlatumumab (anti-PDGFR-α), isatuximab (anti-CD38), sacituzumab (anti-TROP2), U3-1784 (anti-FGFR4), daratumumab (anti-CD38), STI-6129 (anti-CD38), lintuzumab (anti-CD33), huMy9-6 (anti-CD33), belantamab (anti-BCMA), indatuximab (anti-CD138), cetuximab (anti-EGFR), dinutuximab (anti-GD2), anti-CD38 A2 antibody, HuAT13 / 5 antibody, alemtuzumab (anti-CD52), ibritumomab (anti-CD20), tositumomab (anti-CD20 conjugated to I231), bevacizumab (anti-VEGF), panitumumab (anti-EGFR), tremelimumab (previously ticilimumab - anti-CTA-4), catumaxomab (anti-CD3 and EpCAM), oregovomab (anti-CA125), h1959 antibody (anti-Gal-3BP), and bertuzumab (anti-CD20).

[0179] Other useful antibodies include, but are not limited to, antibodies against the following antigens: CA125 (ovary), CA15-3 (cancer), CA19-9 (cancer), L6 (cancer), Lewis Y (cancer), Lewis X (cancer), alpha-fetoprotein (cancer), CA 242 (colorectal), placental alkaline phosphatase (cancer), prostate specific antigen (prostate), prostate acid phosphatase (prostate), epidermal growth factor (cancer), MAGE-1 (cancer), MAGE-2 (cancer), MAGE-3 (cancer), MAGE-4 (cancer), anti-transferrin receptor (cancer), p97 (melanoma), MUC1-KLH (breast cancer), CEA (colorectal), gp100 (melanoma), MART1 (melanoma), PSA (prostate), IL-2 receptor (T cell leukemia and lymphoma), CD20 (non-Hodgkin lymphoma), CD52 (leukemia), CD33 (leukemia), CD22 (lymphoma), human chorionic gonadotropin (cancer), CD38 (multiple myeloma), CD40 (lymphoma), mucin (cancer), P21 (cancer), MPG (melanoma), and Neu cancer gene product (cancer). Some specific useful antibodies 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 against the CD40 antigen such as S2C6 mAb (Francisco, J. A. et al., Cancer Res. (2000) 60:3225-3231), mAbs against the CD70 antigen such as 1F6 mAb, and mAbs against the CD30 antigen such as AC10. Many other internalizing antibodies that bind to tumor-associated antigens can be used and are encompassed.

[0180] Other antigens to which the conjugate of the present invention can bind are 5T4, ACE, ADRB3, AKAP-4, ALK, androgen receptor, AOC3, APP, Axinl, AXL, B7H3, B7-H4, BCL2, BCMA, bcr-abl, BORIS, BST2, C242, C4.4a, CA125, CA6, CA9, CAIX, CCL11, CCR5, CD123, CD133, CD138, CD142, CD15, CD15-3, CD171, CD179a, CD18, CD19, CD19-9, CD2, CD20, CD22, CD23, CD24, CD25, CD27L, CD28, CD3, CD30, CD31, CD300LF, CD33, CD352, CD37, CD38, CD4, CD40, CD41, CD44, CD44v6, CD5, CD51, CD52, CD54, CD56, CD62E, CD62P, CD62L, CD70, CD71, CD72, CD74, CD79a, CD79b, CD80, CD90, CD97, CD125, CD138, CD141, CD147, CD152, CD154, CD326, CEA, CEACAM5, CFTR, clamping factor, cKit, claudin 3, claudin 18.2, CLDN6, CLEC12A, CLL-1, cll3, c-MET, Cripto protein, CS1, CTLA-4, CXCR2, CXORF61, cyclin B1, CYP1B1, cadherin-3, cadherin-6, DLL3, E7, EDNRB, EFNA4, EGFR, EGFRvIII, ELF2M, EMR2, ENPP3, EPCAM, EphA2, ephrin A4, ephrin B2, EPHB4, ERBB2 (Her2 / neu), ErbB3, ERG (TMPRSS2 ETS fusion gene), ETBR, ETV6-AML, FAP, FCAR, FCRL5, FGFR1, FGFR2, FGFR3, FGFR4, FLT3, folate receptor alpha, folate receptor beta, FOLR1, Fos-related antigen 1, fucosyl Gal-3BP, GM1, GCC, GD2, GD3, GloboH, GM3, GPC1, GPC2, GPC3, gplOO, GPNMB, GPR20, GPRC5D, GUCY2C, HAVCR1, HER2, HER3, HGF, HMF24, HMWMAA, HPVE6, hTERT, human telomerase reverse transcriptase, ICAM, ICOS-L, IFN-α, IFN-γ, IGF-I receptor, IGLL1, IL-2 receptor, IL-4 receptor, IL-13Ra2, IL-1 IRa, IL-1, IL-12, IL-23, IL-13, IL-22, IL-4, IL-5, IL-6, interferon receptor, integrin (including α4, αvβ3, αvβ5, αvβ6, α1β4, α4β1, α4β7, α5β1, α6β4, αIIbβ3 integrin), integrin alpha V, intestinal carboxylesterase, KIT, LAGE-la, LAIRl, LAMP-1, LCK, legumain, Lewis Y, LFA-l (CD11a), L-selectin (CD62L), LILRA2, LIV-1, LMP2, LRRC15, LY6E, LY6K, LY75, MAD-CT-1, MAD-CT-2, MAGE Al, Melan A / MART1, mesothelin, ML-IAP, MSLN, mucin, MUC1, MUC16, mut hsp70-2, MYCN, myostatin, NA17, NaPi2b, NCA-90, NCAM, nectin-4, NGF, NOTCH1, NOTCH2, NOTCH3, NOTCH4, NY-BR-1, NY-ESO-1, o-acetyl-GD2, OR51E2, OY-TES1, p53, p53 variant, PANX3, PAP, PAX3, PAX5, p-CAD, PCTA-1 / galectin 8, PD-L1, PD-L2, PDGFR, PDGFR-beta, phosphatidylserine, PIK3CA, PLAC1, polysialic acid, Prostase, prostate cancer cells, prostain, Pseudomonas aeruginosa, rabies, survivin and telomerase, PRSS21, PSCA, PSMA, PTK7, RAGE-1, RANKL, Ras variant, respiratory syncytial virus, Rh factor, RhoC, RON, ROR1, ROR2, RU1, RU2, sarcoma translocation breakpoint, SART3, SLAMF7, SLC44A4, sLe, SLITRK6, sperm protein 17, sphingosine-1-phosphate, SSEA-4, SSX2, STEAP1, TAG72, TARP, TCRb, TEM1 / CD248, TEM7R, tenascin C, TF, TGF-1, TGF-b2, TNF-a, TGS5, Tie2. It includes, but is not limited to, TIM-1, Tn Ag, TRAC, TRAIL-R1, TRAIL-R2, TROP-2, TRP-2, TRPV1, TSHR, tumor antigen CTAA16.88, tyrosinase, UPK2, VEGF, VEGFR1, VEGFR2, vimentin, WT1, and / or XAGE1.

[0181] Antibodies that bind to antigens related to antigen-presenting cells such as CD40, OX40L, endoglin, DEC-205, 4-1BBL, CD36, CD36, CD204, MARCO, DC-SIGN, CLEC9A, CLEC5A, dectin-2, CLEC10A, CD206, CD64, CD32A, CD1A, HVEM, CD32B, PD-L1, BDCA-2, XCR-1, and CCR2 can also be conjugated.

[0182] The antibody conjugate can bind to both receptors or receptor complexes expressed on activated lymphocytes. The receptor or receptor complex can include members of the immunoglobulin gene superfamily, TNF receptor superfamily, integrin, cytokine receptor, chemokine receptor, major histocompatibility protein, lectin, or complement regulatory protein. Non-limiting examples of suitable members of the immunoglobulin superfamily are CD2, CD3, CD4, CD8, CD19, CD22, CD28, CD79, CD90, CD152 / CTLA-4, PD-1, and ICOS. Non-limiting examples of suitable members of the TNF receptor superfamily are CD27, CD40, CD95 / Fas, CD134 / OX40, CD137 / 4-1BB, TNF-R1, TNFR-2, RANK, TACI, BCMA, osteoprotegerin, Apo2 / TRAIL-R1, TRAIL-R2, TRAIL-R3, TRAIL-R4, and APO-3. Non-limiting examples of suitable integrins are CD11a, CD11b, CD11c, CD18, CD29, CD41, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD103, and CD104. Non-limiting examples of suitable lectins are C-type, S-type, and I-type lectins.

[0183] Preferably, the antibody is 3F8, 8H9, Avagovomab, Abciximab (REOPRO®), Bevacizumab, Abrezekimab, Abirilumab, Actoxumab, Adalimumab (HUMIRA®), Adecatumumab, Aducanumab, Afasevikumab, Aferimomab, Afutuzumab, Alacizumab, ALD518, Alemtuzumab (CAMPATH®), Alirocumab (PRALUENT®), Altumomab, Amacizumab, Anatumomab, Andecaliximab, Anetumab, Anifrolumab, Anrukinzumab, Anti-TnC-A1-SIP (F16), Apolizumab, Aptusumab, Arcitumomab (CEA-SCAN®), Ascrinvacumab, Acerizumab, Atidortoxumab, Atolizumab (Tocilizumab, ACTEMRA®, ROACTEMRA®), Atezolizumab (TECENTRIQ®), Atinumumab, Atorlimumab, Avelumab (Bavencio), Azintuzumab, Balantamab, Bapineuzumab, Basiliximab (SIMULECT®), Babiximab, BCD-100, Bectumomab (LYMPHOSCAN®), Begelomab, Belantamab, Belimumab (BENLYSTA®), Bemarituzumab, Benralizumab (FASENRA®), Bemnacizumab, Bensumtumumab, Bepotastimab (SCINITIMUN®), Bevacizumab (AVASTIN®), Bezlotoxumab (ZINPLAVA®), Biciromab (FIBRISCINT®), Bimagrumab, Bimekizumab, Biltratumab, Bivatuzumab, Brecelumab, Blinatumomab, Blontuvetmab, Brodalumab (SILIQ™), Brodalumab (BEOVU®), Bronchituximab, Brosimumab (CRYSVITA®), Cabiralizumab, Caplacizumab (CABLIVI®), Camidanlumab, Camrelizumab,Canakinumab (ILARIS®), Cantuzumab, Capromab, Carlumab, Carotuximab, Catumaxomab (REMOVAB®), cBR96, CC49, Cemiplimab (LIBTAYO®), Celguntuzumab, Setrelizumab (certrelimab), Certolizumab, Cetuximab (ERBITUX®), Sibrotuzumab, Siltuximab, Cituzumab, Cixutumumab, Clazakizumab, Crenoliximab, Cribakizumab, Codrituzumab, Cofetuzumab, Cortuximab, Conatumumab, Consitumumab, Cosfiriviximab, CR6261, Crenezumab, Crisantaspase (ADAKVEO®), Clotetumomab, Cusatuzumab, Dacetuzumab, Daclizumab (ZINBRYTA®), Darotuzumab, Dapirilizumab, Daratumumab (DARZALEX®), Declotrexumab, Demcizumab, Denintuzumab, Denosumab (PROLIA®), Depatuxizumab, Derlotuximab, Detumomab, Dezamizumab, Dinutuximab (UNITUXIN®), Diridavumab, Dmagrozumab, Dostarlimab, Dorlimomab, Dorlixizumab, Dorzigumab, DS-8201, Duligotuzumab, Dupilumab (DUPIXENT®), Durvalumab (IMFINZI®), Ducesigumab, Eculizumab, Eculizumab (SOLIRIS®), Edobacomab, Edrecolomab (PANOREX®), Efalizumab (RAPTIVA®), Efungumab (MYCOGRAB®), Eledumab, Elesclomol, Elgemtuzumab, Elotuzumab (EMPLICITI®), Elsirimumab, Emactuzumab, Emapalumab (GAMIFANT®), Emibetuzumab, Emicizumab (HEMLIBRA®), Enapotamab, Enavatuzumab, Enfortumab (PADCEV®), Enlimomab, Enoblituzumab, Enokizumab, Enoticumab, Ensituximab,Epitumomab, Eptinezumab (VYEPTI®), Epratuzumab, Erenumab (AIMOVIG®), Erlizumab, Ertumaxomab (REXOMUN®), Etaracizumab (ABEGRIN®), Etigilimab, Etrolizumab, Ebinecumab, Evolocumab (REPATHA®), Exbibulumab, Fanolesomab (NEUTROSPEC®), Faralimomab, Faricimab, Farettuzumab, Fasinumab, FBTA05, Felvizumab, Fezakinumab, Fibatuzumab, Ficlatuzumab, Figitumumab, Philimab, Flanvotumab, Fretikumab, Flotetuzumab, Fontolizumab (HUZAF®), Foralumab, Forabilumab, Fremanezumab (AJOVY®), Fresolimumab, Flovobosumab, Fmnevetmab, Fluranumab, Futsuximab, Galcanezumab (EMGALITY®), Galiximab, Gantacumab, Ganitumab, Gantenerumab, Gavrilimomab, Gezibumab, Gemtuzumab, Geboxizumab, Gilberitumab, Gimsilumab, Gilenuximab, Glembatumumab, Golimumab (SIMPONI®), Gomiliximab, Guselkumab (TREMFYA®), huMy9-6, OR000213, Ianalmab, Ibalizumab (TROGARZO®), IBI308, Ibritumomab, Icrucumab, Idarucizumab (PRAXBFND®), Ifabotuzumab, Igovomab (INDIMACIS-125), Iladatuzumab, IMAB362, Imalumab, Imaprelimab, Imciromab (MYOSCINT®), Imugatuzumab, Inclacumab, Indatuximab, Indusatumab, Inebilizumab, Infliximab (REMICADE®), Intertumumab, Inolimomab, Inotuzumab, iomab-B, Ipilimumab, Irratumumab, Isatuximab (SARCLISA®), Iscalimab, Istiratumab, Itolizumab,Ixekizumab (TALTZ®), Certolizumab, Labetuzumab (CEA-CIDE™), Lacutamab, Radretumab, Lampalizumab, Lanadelumab (TAKHZYRO®), Landogrozumab, Laprituximab, Larcabuximab, Lebrikizumab, Lemalesomab, Lendalizumab, Lenvervimab, Lenzilumab, Lerdelimumab, Leronlimab, Lesofavumab, Retuzumab, Rexatumumab, Rivibizumab, Rifatumumab, Regerizumab, Relotoma, Linuzumab, Lirilumab, Rodocytumab, Locibetumab, Loncastuximab, Lorvotuzumab, Losatuxizumab, Lucatumumab, Lulizumab, Lumiliximab, Lumretuzumab, Lupartumab, Lutikizumab, Mapatumumab, Margetuximab, Marstacimab, Maslimomab, Matsuzumab, Mabrilimumab, Mepolizumab (NUCALA®), Mepelimomab, Miratumumab, Minretomomab, Mirikizumab, Milbeximab, Mitsumomab, Modotuximab, Molalizumab, Mogamulizumab (POTELIGEO®), Morolimumab, Mosunetuzumab, Motavizumab (NUMAX®), Moxetumomab (LUMOXITI®), Muromonab-CD3 (ORTHOCLONE OKT3®), Nacolomab, Namilumab, Napumomab, Natalizumab (TYSABRI®), Navicixizumab, Navivumab, Nakitamab, Nebacumab, Necitumumab (PORTRAZZA®), Nemolizumab, NEODOOl, Nerelemomab, Nesvacumab, Netakimab, Nimotuzumab (THERACIM®), Nirsevimab, Nivolumab, Nofetumomab, Obiltoxaximab (ANTHIM®), Obinutuzumab, Ocaratuximab, Ocrelizumab (OCREVUS®), Odulimomab, Ofatumumab (ARZERRA®),Lartruvo (registered trademark), Ocrelizumab, Olaratumab, Orozumab, Omalizumab (Xolair (registered trademark)), Onartuzumab, OMS721, Ofatumumab, Ontecizumab, Ontuxizumab, Onbasilimab, Opcizumab, Oportuzumab, Oregovomab (OVAREX), Ortucumab, Otelixizumab, Otilimab, Otrelizumab, Oxelumab, Ozanezumab, Ozogamicin, Ozoralizumab, Pagibaximab, Palivizumab (Synagis (registered trademark)), Pembrolizumab, Panitumumab (Vectibix (registered trademark)), Pancumumab, Panobacumab, Parsatuzumab, Pascorizumab, Pasotuxizumab, Patritumab, PDR001, Pembrolizumab, Pemtumomab (Theragyn (registered trademark)), Perakizumab, Pertuzumab (Omnitarg (registered trademark)), Pexelizumab, Pidilizumab, Pinatuzumab, Pintumomab, Prazumab, Polatuzumab (Polivy), Prezalumab, Prozalizumab, Pogalizumab, Ponezumab, Porgabiximab, Placinezumab, Prezalumab, Priliximab, Pritoxaximab, Prutumab, PRO140, Crizanlizumab, Lacotumomab, Radretumab, Rafivirumab, Larapansizumab, Ramucirumab, Ranevetmab, Ranibizumab (Lucentis (registered trademark)), Rabagalizumab, Ravulizumab (Ultomiris (registered trademark)), Laxibacumab, Refametinib, Regabilimab, REGN-EB3, Renatlimab, Remtulizumab, Reslizumab (Cinqair (registered trademark)), Rilotumumab, Linukumab, Risankizumab (Skyrizi (registered trademark)), Rituximab (Rituxan (registered trademark)), Ribavizumab, rmab, Robatumumab, Lorelumab, Romilkimab, Romosozumab (Evenity (registered trademark)), Rontalizumab, Rossmantuzumab, Robalpituzumab, Robelizumab (Leukarrest (registered trademark)),Rozanolixizumab, Rupulizumab (ANTOVA), SA237, Sacituzumab, Samalizumab, Samrotamab, Sarilumab (KEVZARA®), Satralizumab, Satumumab Pentide, Secukinumab (COSENTYX®), Celrelizumab, Seribantumab, Setoxaximab, Setmsumab, Sevimmab, SGN-CD19A, SHP647, Sibrotuzumab, Sipatrilimab, Silukiximab, Simtuzumab, Cipilizumab, Sirtratumab, Silkumab, Sofituzumab, Solanezumab, Solitomab, Sonepcizumab, Sonetuzumab, Spartalizumab, Stamulumab, STI-6129, Sulesomab (LEUKOSCAN®), Sputabumab, Stimlimab, Subizumab, Subratoxumab, Tabalumab, Takatsuzumab (AFP-CIDE®), Tadocizumab, Talacotuzumab, Talisizumab, Tamtuvetmab, Tanezumab, Taplitumomab Paptox, Tarextumab, Tabolumab, Tefibazumab (AUREXIS®), Telimomab, Telisotuzumab, Tesidolumab, Tetraxetan, Tetulomab, Tenatumomab, Tenelixizumab, Teplizumab (TEPEZZA®), Teplizumab, Tezepelumab, TGN1412, Chibrizumab, Tislelizumab (TREMELIMUMAB®), Tigatuzumab, Timigutuzumab, Timolumab, Tiragolumab, Tiragotumab, Chisrelizumab, Chisotuzumab, Chiuxetan, Tildrakizumab (ILUMYA®), TNX-650, Tocilizumab (Actemra®, Atolizumab), Tomzotuximab, Tralizumab, Tosatoxumab, Tositumomab (BEXXAR®), Tovetumab, Tralokinumab, Trastuzumab (HERCEPTIN®), TRBS07, Tregalizumab,Tremelimumab, Trevoglumab, Tsukotsuzumab, Tubulizumab, Ultuxizumab, Ustekinumab (STELERA®), Ublituximab, Urocuplumab, Urelumab, Utomilumab, Badastuximab, Vanalimab, Vandortuzumab, Bantiktuzumab, Vanucizumab, Vapaliximab, Barisacumab, Barilumab, Vatelizumab, Vedolizumab, Bertuzumab, including Zumab, Vepalimomab, Besencumab, Vizilimumab (NUVION®), Bobalirazumab, Borosikizumab (HUMASPECT®), Bonrerolizumab, Bopratelimab, Bolcesetuzumab, Bortezomab, Bunakizumab, Xencizumab, XMAB-5574, Zalutumumab (HuMEX-EGFr), Zanomlimab (HuMAX-CD4), Zatuximab, Zenocizumab, Ziralimumab, Solbecizumab or Zolimomab.

[0184] An antibody that "binds" to a target such as a molecular target or an antigen of interest is one that can bind to that antigen with sufficient affinity, and thus the antibody is useful when targeting cells expressing the antigen.

[0185] In a further specific embodiment, P is a peptide that preferably has the ability to specifically target protein receptors overexpressed in tumor tissue. These peptides often have a precedent in the literature of having strong binding affinity for target sites within the nanomolar scale, measured by different techniques including, for example, surface plasmon resonance (SPR), biolayer interferometry (BLI), and isothermal titration calorimetry (ITC). More specifically, the peptide is a tripeptide RGD (arg - gly - asp) that targets integrins (α5β1, α8β1, and αIIbβ3) or GnRH (gonadotropin - releasing hormone) that targets GnRH - R (hormone receptor version) or SST (somatostatin) that targets SSTR1 - 5 (somatostatin receptors) or EGF (epidermal growth factor) that targets EGFR:HER1, HER2, HER3, HER4 or angiopep - 2 that targets LRP - 1 (low - density lipoprotein receptor - related protein - 1) or bradykinin potentiating peptide (BPP) that targets ACE (angiotensin - converting enzyme receptor).

[0186] The conjugate of formula (II) as defined above can also be represented by the following formula III:

[0187]

Chemical formula

[0188] It can also be represented by.

[0189] In this embodiment, X, R1, R 1' , R2, and R3 are as defined in the above "payload" section including all specific and preferred embodiments. P and v are as defined above.

[0190] In this embodiment, L3 represents a linker that enables the peptide or protein P to bind to a "payload" (a molecule or an indole derivative). As used herein, the term "linker" refers to any chemical moiety that can link a peptide or protein "P" to a "molecule".

[0191] In certain embodiments, the linker L3 can contain a heterobifunctional group. As used herein, the term "heterobifunctional group" refers to a chemical moiety that links the linker, in part, to a peptide or protein P. A heterobifunctional group is characterized by having different reactive groups at both ends of the chemical moiety. The binding to "P" can be accomplished through chemical or enzymatic conjugation, or a combination of both. Chemical conjugation involves a controlled reaction of accessible amino acid residues on the surface of protein P with the reaction handles in the heterobifunctional group. Examples of chemical conjugation include, but are not limited to, lysine amide coupling, cysteine coupling, and coupling via non-natural amino acids incorporated by genetic engineering, where non-natural amino acid residues with the desired reaction handles are installed in "P". In enzymatic conjugation, the enzyme mediates the coupling of the linker to accessible amino residues in the peptide or protein P. Examples of enzymatic conjugation include, but are not limited to, peptide transfer using sortase, peptide transfer using microbial transglutaminase, and N-glycan engineering. Chemical conjugation and enzymatic conjugation can be used sequentially. For example, enzymatic conjugation can be used to install unique reaction handles in "P" for use in subsequent chemical conjugation. In particular, the heterobifunctional group is

[0192]

Chemical formula

[0193] [wherein,

[0194] [Chemical formula]

[0195] is the binding point with the remaining part of linker L3,

[0196] [Chemical formula]

[0197] is the binding point with "P". is selected from.

[0198] In certain embodiments, linker L3 is a group that can be cleaved enzymatically by proteases such as cathepsin B, VGAP tetrapeptide, etc. In particular, linker L3 is

[0199] [Chemical formula]

[0200] [wherein q is an integer from 2 to 10, and Z 1 , Z 2 , Z 3 , and Z 4 are each independently either absent or a naturally occurring amino acid residue in the L- or D-configuration, provided that at least two of Z 1 , Z 2 , Z 3 , and Z 4 are amino acid residues,

[0201] [Chemical formula]

[0202] is the binding point with the molecule,

[0203] [Chemical formula]

[0204] is a protease-cleavable linker selected from the junction points with "P". Preferably, Z 1 Z 2 Z 3 and Z 4 are, independently, absent or selected from the group consisting of L-valine, D-valine, L-citrulline, D-citrulline, L-alanine, D-alanine, L-glutamine, D-glutamine, L-glutamic acid, D-glutamic acid, L-aspartic acid, D-aspartic acid, L-asparagine, D-asparagine, L-phenylalanine, D-phenylalanine, L-lysine, D-lysine, and glycine, provided that Z 1 Z 2 Z 3 and Z 4 at least two of are amino acid residues. More preferably, Z 1 is absent or glycine, and Z 2 is absent or selected from L-glutamine, D-glutamine, L-glutamic acid, D-glutamic acid, L-aspartic acid, D-aspartic acid, L-alanine, D-alanine, and glycine, Z 3 is selected from L-valine, D-valine, L-alanine, D-alanine, L-phenylalanine, D-phenylalanine, and glycine, Z 4 is selected from L-alanine, D-alanine, L-citrulline, D-citrulline, L-asparagine, D-asparagine, L-lysine, D-lysine, L-phenylalanine (phenylalamine), D-phenylalanine, and glycine. Even more preferably, L3 is

[0205]

Chemical formula

[0206] [wherein q is 5,

[0207]

Chemical formula

[0208] is a bonding point with the molecule,

[0209] [Chemical formula]

[0210] is a bonding point with "P" is.

[0211] In certain embodiments, linker L3 is a pyrophosphatase-cleavable linker, preferably of the following formula:

[0212] [Chemical formula]

[0213] [wherein q is an integer from 2 to 10,

[0214] [Chemical formula]

[0215] is a bonding point with the molecule,

[0216] [Chemical formula]

[0217] is a bonding point with "P" is a pyrophosphatase-cleavable linker having.

[0218] In certain embodiments, linker L3 is preferably

[0219] [Chemical formula]

[0220] [wherein, q is an integer from 2 to 10, ---- does not exist or is a bond,

[0221]

Chem.

[0222] is a bonding point with the molecule,

[0223]

Chem.

[0224] is a bonding point with "P"]] is a beta-glucuronidase-cleavable linker selected from

[0225] In certain embodiments, linker L3 is in vivo reducible. In vivo reducible linkers take advantage of the difference in reduction potential between intracellular compartments and plasma. Reduced glutathione, which is presented in the cytoplasm of tumor cells, is up to 1000-fold higher than that present in the cytoplasm of normal cells, and tumor cells also contain enzymes that can contribute to reduction in intracellular compartments. The linker keeps the conjugate intact during systemic circulation and is selectively cleaved by the high intracellular concentration of glutathione to release the active drug from the non-toxic prodrug at the tumor site. Preferably, L3 is

[0226]

Chem.

[0227] [wherein, q is an integer from 2 to 10, and R, R', R", and R'" are each independently selected from hydrogen, C1-C6 alkoxy C1-C6 alkyl, (C1-C6)2NC1-C6 alkyl, and C1-C6 alkyl, or two geminal R groups can together with the carbon atom to which they are attached form a cyclobutyl or cyclopropyl ring,

[0228] [Chem.]

[0229] is a binding point with the molecule,

[0230] [Chem.]

[0231] is a binding point with "P" and is a bioreducible linker selected from

[0232] In certain embodiments, linker L3 is acid-cleavable. Preferably, L3 is

[0233] [Chem.]

[0234] [wherein q is an integer from 2 to 10,

[0235] [Chem.]

[0236] is a binding point with the molecule,

[0237] [Chem.]

[0238] is a binding point with "P" and is an acid-cleavable linker selected from

[0239] In certain embodiments, L3 is a click-to-release linker, where release of the molecule is chemically induced by tetrazine or related compounds. Preferably, L3 is

[0240] [Chemical]

[0241] [wherein, q is an integer from 2 to 10,

[0242] [Chemical]

[0243] is a bonding point with the molecule,

[0244] [Chemical]

[0245] is a bonding point with "P"]] is a click reaction-releasing linker selected from

[0246] In certain embodiments, L3 is linker L1-L2, where L1 is a cleavable group as defined in the above "L1 series" section, including all specific and preferred embodiments, and L2 is a binding protein conjugate as defined in the above "L1 series" section, including all specific and preferred embodiments.

[0247] The present invention further provides a "payload" or "drug" as defined above conjugated to nanoparticles, particularly silica nanoparticles, and a compound of formula (I) as defined herein. In certain embodiments, the "payload" can be substituted with a linker to form a linker-payload conjugate, which binds to the silica nanoparticles, thereby forming a nanoparticle-drug conjugate. Examples of such nanoparticle-drug conjugates (NDCs) are described, for example, in International Patent Applications WO2022 / 093800 and WO2022 / 093794.

[0248] Preferred embodiments are (a) Nanoparticles comprising a silica-based core and a silica shell surrounding at least a portion of the core, polyethylene glycol (PEG) covalently bound to the surface of the nanoparticles, and optionally a fluorescent compound covalently encapsulated within the core of the nanoparticles, (b) A targeting ligand that binds to the folate receptor, wherein the targeting ligand is selected from the group consisting of folic acid, dihydrofolic acid, tetrahydrofolic acid, and any folate receptor binding derivatives of the foregoing, and the targeting ligand is bound to the nanoparticles either directly or indirectly via a spacer group, (c) Ra is a group of formula (B)

[0249] [Chemical formula]

[0250] [wherein, L1 is a cleavable group selected from among pH-sensitive groups, photoinducible cleavable groups, bioreductive cleavable groups, and enzymatically cleavable groups, and m is equal to 0] A compound of formula (I) as defined herein, wherein the compound of formula (I) is bound to the nanoparticles either directly or indirectly via a spacer group A nanoparticle-drug conjugate comprising.

[0251] Therapeutic use The present invention relates to - A pharmaceutical composition comprising any compound having formula (I), (II), (III) as defined above, including any one of the disclosed embodiments, and / or - A pharmaceutical composition comprising any compound having formula (I), (II), or (III) as defined above, including any one of the disclosed embodiments, and a pharmaceutically acceptable excipient, and / or - (a) Any compound having or comprising any one of the disclosed embodiments of formula (I), (II), or (III) as defined above, and (b) an additional active ingredient, preferably an additional anti-tumor agent, a pharmaceutical composition, and / or - A pharmaceutical composition as defined above or any compound having formula (I), (II), or (III) as defined above, comprising any one of the disclosed embodiments for use as a drug, and / or - A pharmaceutical composition as defined above or any compound having formula (I), (II), or (III) as defined above for use in the treatment of cancer, and / or - A product or kit containing, as a combined preparation, (a) any compound of formula (I), (II), or (III) as disclosed above, comprising any one of the disclosed embodiments, and (b) an additional active ingredient, preferably an additional anti-tumor agent, for simultaneous, separate, or sequential use, particularly in the treatment of cancer, and / or - A combined preparation comprising (a) any compound of formula (I), (II), or (III) as disclosed above, comprising any one of the disclosed embodiments, and (b) an additional active ingredient, preferably an additional anti-tumor agent, for simultaneous, separate, or sequential use, particularly in the treatment of cancer, and / or - A pharmaceutical composition as defined above or any compound having formula (I), (II), or (III) as defined above for use in combination with radiotherapy, surgery (e.g., tumor resection), hyperthermia, and / or other anti-tumor therapies, or before, simultaneously with, or after surgery (e.g., tumor resection) in the treatment of cancer, and / or - Use of a pharmaceutical composition as defined above or any compound having formula (I), (II), or (III) as defined above, comprising any one of the disclosed embodiments, for the manufacture of a medicament for the treatment of cancer, and / or - Use of a pharmaceutical composition as defined above, or any compound having formula (I), (II) or (III) as defined, comprising any one of the disclosed embodiments, for the manufacture of a medicament for the treatment of cancer, and (b) an additional active ingredient, preferably an additional anti-tumor agent, and / or - A method for treating cancer in a subject in need thereof, comprising administering an effective amount of a pharmaceutical composition as defined above, or any compound having formula (I), (II) or (III) as defined, comprising any one of the disclosed embodiments, and / or - A method for treating cancer in a subject in need thereof, comprising administering an effective amount of a pharmaceutical composition as defined above, or any compound having formula (I), (II) or (III) as defined, and (b) an additional active ingredient, preferably an additional anti-tumor agent, comprising any one of the disclosed embodiments, and / or - A method for treating cancer in a subject in need thereof, comprising administering an effective amount of a pharmaceutical composition as defined above, or any compound having formula (I), (II) or (III) as defined, in combination with radiotherapy, surgery (e.g., tumor resection), hyperthermia and / or other anti-tumor therapies, comprising any one of the disclosed embodiments relates to.

[0252] As used herein, the term "cancer" refers to the presence of cells having characteristics typical of cancer-causing cells, such as uncontrolled growth, immortality, the potential for metastasis, rapid growth and proliferation rates, and certain characteristic morphological features. Cancer can be a solid tumor or a hematopoietic tumor. Examples of cancer include, for example, leukemia, lymphoma, blastoma, carcinoma, and sarcoma. More specific examples of such cancers include chronic myeloid leukemia, acute lymphoblastic leukemia, Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL), squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, glioma, gastrointestinal cancer, kidney cancer, ovarian cancer, liver cancer, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, neuroblastoma, osteosarcoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, stomach cancer, bladder cancer, hepatocellular carcinoma, breast cancer, esophageal cancer, colon cancer, and head and neck cancer, gastric cancer, germ cell tumor, pediatric sarcoma, sinonasal natural killer, multiple myeloma, acute myeloid leukemia (AML), chronic lymphocytic leukemia, mastocytosis, and any symptoms associated with mastocytosis. Preferably, the cancer is leukemia, acute myeloid leukemia, lymphoma, breast cancer, pancreatic cancer, lung cancer, or colon cancer. More preferably, the cancer is acute myeloid leukemia or pancreatic cancer. Optionally, the cancer is associated with abnormal regulation of MKlp2 or its pathway. In particular, the cancer is associated with overexpression of MKlp2.

[0253] As used herein, the terms "treatment", "treating", or "treatment" refer to any act intended to relieve the health condition of a patient, such as therapy, prevention, prophylaxis, and retardation of a disease. In certain embodiments, such terms refer to remission or eradication of a disease or symptoms associated with the disease. In other embodiments, the term refers to minimizing the spread or worsening of a disease caused by administration of one or more therapeutic agents to a subject having such a disease.

[0254] "Effective amount" means the amount of the pharmaceutical composition of the present invention that prevents, eliminates or reduces the adverse effects of the disease to be treated in mammals including humans. It is understood that the dosage administered can be adapted by those skilled in the art according to the patient, pathology, mode of administration, etc. For example, the compounds of the present invention may be used at a dosage of 0.01 to 500 mg / kg body weight per day. In certain embodiments, the pharmaceutical composition according to the present invention comprises from 0.01 to 500 mg / kg of the compound of the present invention. It is understood that the dosage administered can be adapted by those skilled in the art according to the patient, pathology, mode of administration, etc.

[0255] The route of administration can be topical, transdermal, oral, rectal, sublingual, intranasal, intrathecal, intratumoral or parenteral (including subcutaneous, intramuscular, intravenous and / or intradermal). Preferably, the route of administration is parenteral, oral or topical. The pharmaceutical composition is adapted to one or several of the routes mentioned above. The pharmaceutical composition, kit, product or combination preparation is preferably administered by injection or by intravenous infusion or by a suitable sterile liquid preparation, or via the gastrointestinal tract in the form of a liquid or solid dosage.

[0256] The pharmaceutical composition can be formulated as a solution in a pharmaceutically compatible solvent, or as an emulsion, suspension or dispersion in a suitable pharmaceutical solvent or vehicle, or as a pill, tablet or capsule containing a solid vehicle by methods known in the art. The formulations of the present invention suitable for oral administration are in the form of discrete units such as capsules, sachets, tablets or lozenges, each containing a predetermined amount of the active ingredient; in the form of powders or granules; in the form of solutions or suspensions in aqueous or non-aqueous liquids; or in the form of oil-in-water emulsions or water-in-oil emulsions. The formulations for rectal administration may be in the form of suppositories incorporating the active ingredient and a carrier such as cocoa butter, or in the form of enemas. The formulations suitable for parenteral administration conveniently include a sterile oily or aqueous preparation of the active ingredient which is preferably isotonic with the blood of the recipient. All such formulations may also contain other pharmaceutically compatible non-toxic adjuvants such as stabilizers, antioxidants, binders, dyes, emulsifiers or flavoring substances. The formulations of the present invention thus comprise the active ingredient in association with a pharmaceutically acceptable carrier and optionally other therapeutic ingredients. The carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to its recipient. The pharmaceutical composition is advantageously applied by injection or intravenous infusion of a suitable sterile liquid preparation, or by oral administration via the gastrointestinal tract. Methods for the safe and effective administration of most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in standard literature.

[0257] Additional anti-tumor agents can be selected from a non-exhaustive list of anti-tumor agents consisting of inhibitors of topoisomerase I or II, anti-mitotic agents, DNA alkylating agents, anti-metabolites, targeted agents such as kinase inhibitors, therapeutic antibodies designed to mediate cytotoxicity to cancer cells or to modulate one of their major biological functions, and / or antibody-drug conjugates.

[0258] Antimitotic agents include, but are not limited to, paclitaxel, docetaxel, and analogs such as larotaxel (also known as XRP9881; Sanofi-Aventis), XRP6258 (Sanofi-Aventis), BMS-184476 (Bristol-Meyer-Squibb), BMS-188797 (Bristol-Meyer-Squibb), BMS-275183 (Bristol-Meyer-Squibb), ortataxel (also known as IDN 5109, BAY 59-8862 or SB-T-101131; Bristol-Meyer-Squibb), RPR 109881A (Bristol-Meyer-Squibb), RPR 116258 (Bristol-Meyer-Squibb), NBT-287 (TAPESTRY), PG-paclitaxel (also known as CT-2103, PPX, paclitaxel poliglumex, polyglutamate paclitaxel or Xyotax™), ABRAXANE® (also known as nab-paclitaxel; ABRAXIS BIOSCIENCE), tesetaxel (also known as DJ-927), IDN 5390 (INDENA), taxao-prexin (also known as docosahexanoic acid-paclitaxel; PROTARGA), DHA-paclitaxel (also known as Taxoprexin®), and MAC-321 (WYETH).

[0259] Inhibitors of topoisomerase I and / or II include, but are not limited to, etoposide, topotecan, camptothecin, irinotecan, amsacrine, intoplicine, anthracyclines such as doxorubicin, epirubicin, daunorubicin, idarubicin, and mitoxantrone. Inhibitors of topoisomerase I and II include, but are not limited to, intoplicine.

[0260] DNA alkylating agents include, but are not limited to, cisplatin, carboplatin and oxaliplatin. In a preferred embodiment, the DNA alkylating agent is cisplatin.

[0261] Antimetabolites block the enzymes responsible for nucleic acid synthesis or are incorporated into DNA, thereby generating inaccurate genetic codes and inducing apoptosis. Its non-exhaustive examples include, but are not limited to, folic acid antagonists, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors. More specifically, they include methotrexate, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatin, 5-fluorouracil, gemcitabine, and capecitabine.

[0262] Antitumor agents can be alkylating agents, including, but not limited to, nitrogen mustards, ethyleneimine derivatives, alkyl sulfonates, nitrosoureas, metal salts, and triazenes. Its non-exhaustive examples include uracil mustard, chloromethine, cyclophosphamide (CYTOXAN®), ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, cisplatin, carboplatin, oxaliplatin, thiotepa, streptozocin, dacarbazine, and temozolomide.

[0263] The anti-tumor agent can also be a targeting agent, particularly a kinase inhibitor. The kinase may be selected from the group consisting of intracellular tyrosine or serine / threonine kinases, receptor tyrosine or serine / threonine kinases. For example, the agent may have the ability to inhibit angiogenesis based on its inhibitory activity against VEGFR and PDGFR kinases. In particular, the targeting agent can be selected from a plurality of already approved kinase inhibitor drugs: Gleevec that inhibits Abl, as well as Iressa and Tarceva that both inhibit EGFR, Sorafenib (Nexavar, BAY 43-9006) that inhibits Raf, Dasatinib (BMS-354825) and Nilotinib (AMN-107, Tasigna) that also inhibit Abl, Lapatinib that also inhibits EGFR, Temsirolimus (Torisel, CCI-779) that targets the mTOR pathway, Sunitinib (Sutent, SU11248) that inhibits several targets including VEGFR, and specific antibodies that inactivate kinase receptors: Herceptin and Avastin.

[0264] Antibody-drug conjugates include, but are not limited to, Gemtuzumab Ozogamicin, Brentuximab Vedotin, Trastuzumab Emtansine, Inotuzumab Ozogamicin, Polatuzumab Vedotin, Enfortumab Vedotin, Trastuzumab Deruxtecan (Enhertu®), Sacituzumab Govitecan (Trodelvy®), Belantamab Mafodotin (Blenrep®), Trastuzumab Duocarmycin, BAT8001, Mirvetuximab Soravtansine, SAR408701, Loncastuximab Tesirine, Camidanlumab Tesirine, Badastuximab Talirine, Lobapituizumab Tesirine or Depatuxizumab Mafodotin.

[0265] As used herein, the term "therapy" refers to any type of treatment of cancer (i.e., anti-tumor therapy), including adjuvant therapy and neoadjuvant therapy. Therapy includes radiotherapy, as well as therapies such as hormonal therapy, chemotherapy, immunotherapy and monoclonal antibody therapy, preferably systemic therapy.

[0266] As used herein, the term "adjuvant therapy" refers to any type of treatment for cancer that is usually given as an additional treatment after surgical removal of the primary tumor in patients at risk of metastasis and / or likely to recur. The aim of such adjuvant treatment is to improve the prognosis. Adjuvant therapy includes radiation therapy, as well as therapies such as hormone therapy, chemotherapy, immunotherapy and monoclonal antibody therapy, preferably systemic therapy.

[0267] The term "hormone therapy" or "therapy with hormones" refers to cancer treatment aimed at blocking, adding or removing hormones. For example, in breast cancer, the female hormones estrogen and progesterone can promote the growth of some breast cancer cells. Therefore, in these patients, hormone therapy is given to block estrogen, and a non-exhaustive list of commonly used drugs includes tamoxifen, toremifene, anastrozole, exemestane, letrozole, goserelin / leuprolide, megestrol acetate, and fluoxymesterone.

[0268] As used herein, the term "chemotherapy treatment" or "chemotherapy" refers to a cancer treatment using chemical or biological substances, in particular one or more antineoplastic agents.

[0269] The term "radiation treatment" or "radiation therapy" is a term commonly used in the art to refer to multiple types of radiation therapy, including internal and external radiation therapy or radioimmunotherapy, as well as the use of various types of radiation, including X-rays, gamma rays, alpha particles, beta particles, photons, electrons, neutrons, radioisotopes, and other forms of ionizing radiation.

[0270] Further aspects and advantages of the present invention are described in the following examples, which should be regarded as illustrative rather than limiting.

Examples

[0271] I. "Phosphonic Acid Prodrug" General Synthetic Scheme I:

[0272]

Chem.

[0273] A phosphonoester is obtained by the condensation of dibenzyl phosphite with bromoalkanoate, and thus a phosphonic acid is obtained after basic hydrolysis. Subsequently, the acid is coupled with the NH - indole of the drug via a peptide coupling agent to obtain a benzylphosphonate prodrug, which is hydrolyzed to the prodrug phosphonic acid in an acidic medium. Treatment with a base yields the phosphonic acid prodrug as a salt.

[0274] Procedure for the Preparation of Phosphonic Acid Prodrug Phosphonoalkanoic acid was synthesized according to C. Zingle et al., Bioorganic & Medicinal Chemistry letters, 22(21), 6563 - 6567 (2012).

[0275] A. Phosphonoalkanoic Acid Ester (Step 1) 1. Ethyl 5 - (Bis(benzyloxy)phosphoryl)pentanoate A suspension of sodium hydride (1.141 g, 28.5 mmol) in DMF (40 mL) was slowly added with dibenzyl phosphite (7 mL, 28.5 mmol) in DMF (20 mL) at 0 °C under an argon atmosphere. The reaction mixture was stirred at room temperature for 15 minutes, cooled again to 0 °C, and ethyl 5-bromovalerate (4.51 mL, 28.5 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour, poured into 150 mL of water, and extracted with 150 mL of EtOAc. The organic layer was washed twice with water (75 mL). The combined aqueous layers were extracted again with 75 mL of EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give ethyl 5-(bis(benzyloxy)phosphoryl)pentanoate as a colorless oil (11.41 g). Yield = 87% LC-MS: [M+H] = 391 1 1H NMR (300 MHz, DMSO) δ ppm: 7.45 - 7.20 (m, 10H), 5.10 - 4.86 (m, 4H), 4.02 (q, J = 7.3 Hz, 2H), 2.29 (dt, J = 14.3, 7.4 Hz, 2H), 1.81 (ddd, J = 10.7, 7.9, 5.9 Hz, 2H), 1.70 - 1.35 (m, 4H), 1.15 (t, J = 7.1 Hz, 3H)

[0276] The following compounds were prepared using the same procedure as above: 2. Ethyl 3-(bis(benzyloxy)phosphoryl)propanoate Yield = 25% LC-MS: [M+H] = 363 1H NMR (300 MHz, DMSO) δ ppm: 7.37 (s, 10H), 4.99 (qd, J = 12.1, 8.1 Hz, 4H), 4.03 (q, J = 7.1 Hz, 2H), 2.37 (t, J = 7.3 Hz, 2H), 2.25 - 2.05 (m, 2H), 1.15 (t, J = 7.1 Hz, 3H) 3. Ethyl 7-(bis(benzyloxy)phosphoryl)heptanoate Yield = 99% LC-MS: [M+H] = 419 1H NMR (300 MHz, DMSO) δ ppm: 7.43 - 7.24 (m, 10H), 4.97 (qd, J = 12.1, 8.3 Hz, 4H), 4.03 (q, J = 7.1, 2H), 2.24 (dt, J = 12.2, 7.3 Hz, 2H), 1.88 - 1.67 (m, 2H), 1.57 - 1.31 (m, 6H), 1.34 - 1.18 (m, 2H), 1.16 (t, J = 7.1, 3H)

[0277] B. Phosphonoalkanoate acids (Step 2) 1. 5-(bis(benzyloxy)phosphoryl)pentanoic acid To a solution of ethyl 5-(bis(benzyloxy)phosphoryl)pentanoate (11.41 g, 29.2 mmol) in ethanol (100 mL) was added 1N sodium hydroxide (58.5 mL, 58.5 mmol). The reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted with 200 mL of water and washed with ethyl acetate (3 × 100 mL). The combined organic layers were washed with water. The aqueous layer was acidified to pH = 3 - 4 with 1N HCl (60 mL) and then extracted three times with ethyl acetate (100 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated to dryness to give 5-(bis(benzyloxy)phosphoryl)pentanoic acid as a colorless oil (7.93 g). Yield = 74% LC-MS: [M+H] = 363 1 1H NMR (300 MHz, DMSO) δ ppm: 7.46 - 7.23 (m, 10H), 5.10 - 4.85 (m, 4H), 2.18 (t, J = 7.0 Hz, 2H), 1.88 - 1.71 (m, 2H), 1.61 - 1.38 (m, 4H)

[0278] The following compounds were prepared using the same procedure: 2. 3-(Bis(benzyloxy)phosphoryl)propanoic acid Yield = 87% LC-MS: [M+H] = 335 1 H NMR (300 MHz, DMSO) δ ppm: 12.35 (s, 1H), 7.43 - 7.27 (m, 10H), 5.11 - 4.88 (m, 4H), 2.45 - 2.34 (m, 2H), 2.13 - 1.97 (m, 2H) 3. 7-(Bis(benzyloxy)phosphoryl)heptanoic acid Yield = 83% LC-MS: [M+H] = 391 1 H NMR (300 MHz, DMSO) δ ppm: 7.45 - 7.24 (m, 10H), 4.96 (qd, J = 12.1, 8.2 Hz, 4H), 2.17 (dt, J = 10.8, 7.3 Hz, 2H), 1.86 - 1.67 (m, 2H), 1.53 - 1.33 (m, 4H), 1.23 (ddt, J = 32.4, 13.4, 6.6 Hz, 4H)

[0279] C. Prodrug moiety coupling (Step 3) 1. (Z)-Dibenzyl-5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate A solution of (Z)-3-(2-(5-bromo-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile (2.10 g, 5.55 mmol), 5-(bis(benzyloxy)phosphoryl)pentanoic acid (2.40 g, 6.62 mmol) and PyBOP (4.31 g, 8.28 mmol) in DMF (30 mL) was added with TEA (1.539 mL, 11.04 mmol). The reaction mixture was stirred at room temperature overnight and then poured into 200 mL of water. The resulting suspension was stirred at room temperature for 30 minutes, filtered, washed twice with water (50 mL) and dried under vacuum to give (Z)-dibenzyl 5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate (4.10 g) as a pale yellow solid. Yield = 51% LC-MS: [M+H] = 722 1 H NMR (300 MHz, DMSO) δ ppm: 8.39 (d, J = 8.5 Hz, 2H), 8.29 (d, J = 1.7 Hz, 1H), 8.16 (d, J = 1.8 Hz, 1H), 8.01 (dd, J = 8.7, 2.1 Hz, 1H), 7.94 (s, 1H), 7.62 (dd, J = 8.9, 1.9 Hz, 1H), 7.40 - 7.30 (m, 11H), 5.08 - 4.90 (m, 4H), 3.97 (s, 3H), 3.15 (t, J = 7.1 Hz, 2H), 2.01 - 1.85 (m, 2H), 1.75 (dd, J = 14.2, 7.0 Hz, 2H), 1.62 (m, 2H)

[0280] The following compounds were prepared using the same procedure: 2. (Z)-dibenzyl-3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonate Yield = 74% LC-MS: [M+H] = 694 1H NMR (300 MHz, DMSO) δ ppm: 8.35 (d, J = 7.6 Hz, 2H), 8.30 (d, J = 1.8 Hz, 1H), 8.15 (d, J = 1.8 Hz, 1H), 8.01 (dd, J = 8.7, 2.1 Hz, 1H), 7.97 (s, 1H), 7.62 (dd, J = 8.9, 1.9 Hz, 1H), 7.43 - 7.23 (m, 11H), 5.14 - 4.94 (m, 4H), 3.98 (s, 3H), 3.43 - 3.35 (m, 2H), 2.29 (dt, J = 8.3, 7.3 Hz, 2H) 3. (Z)-Dibenzyl 7-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-7-oxoheptylphosphonate Yield = 52% LC-MS: [M+H] = 750 1H NMR (300 MHz, CDCl3) δ ppm: 8.38 (d, J = 8.9 Hz, 1H), 8.21 (d, J = 1.8 Hz, 1H), 7.93 (d, J = 1.8 Hz, 1H), 7.79 (s, 1H), 7.72 (s, 1H), 7.66 (dd, J = 8.7, 2.0 Hz, 1H), 7.47 (dd, J = 8.9, 1.9 Hz, 1H), 7.26 (s, 10H), 6.97 (d, J = 8.7 Hz, 1H), 4.93 (ddd, J = 30.1, 11.8, 8.5 Hz, 4H), 3.93 (s, 3H), 2.83 (t, J = 7.3 Hz, 2H), 1.86 - 1.34 (m, 10H) 4. (Z)-Dibenzyl 5-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-5-oxopentylphosphonate Yield = 52% LC-MS: [M+H] = 674 11H NMR (300 MHz, DMSO) δ ppm: 8.28 (d, J = 1.8 Hz, 1H), 8.15 (s, 1H), 8.05 (d, J = 2.4 Hz, 1H), 8.02 - 7.97 (m, 1H), 7.93 (s, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.44 - 7.25 (m, 11H), 7.06 (dd, J = 8.8, 2.4 Hz, 1H), 4.99 (p, J = 12.1 Hz, 4H), 3.97 (s, 3H), 3.83 (s, 3H), 3.13 (t, J = 6.9 Hz, 2H), 2.02 - 1.85 (m, 2H), 1.75 (dd, J = 13.9, 7.0 Hz, 2H), 1.69 - 1.53 (m, 2H) 5. (Z)-Dibenzyl 7-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-7-oxoheptylphosphonate Yield = 46% LC-MS: [M+H] = 702 1H NMR (300 MHz, DMSO) δ ppm: 8.27 (d, J = 1.9 Hz, 1H), 8.14 (s, 1H), 8.06 (d, J = 2.4 Hz, 1H), 7.99 (dd, J = 8.7, 2.1 Hz, 1H), 7.94 (s, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.40 - 7.30 (m, 11H), 7.06 (dd, J = 8.8, 2.4 Hz, 1H), 4.98 (qd, J = 12.1, 8.2 Hz, 4H), 3.98 (d, J = 5.1 Hz, 3H), 3.09 (t, J = 7.1 Hz, 2H), 1.88 - 1.75 (m, 2H), 1.71 - 1.59 (m, 2H), 1.47 (d, J = 7.7 Hz, 2H), 1.41 - 1.32 (m, 4H)

[0281] D. Phosphonic acid prodrug (Step 4)

[0282] (Example 1) (Z)-5-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonic acid

[0283] [Chemical formula]

[0284] (Z)-Dibenzyl 5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate (1.531 g, 2.119 mmol) was dissolved in 4 M HCl (47.7 mL, 190.8 mmol) in 1,4-dioxane to obtain a pale yellow solution. The reaction mixture was heated at 90 °C for 2 hours. After cooling, the precipitate was filtered and washed successively twice with dioxane (3 mL) and diisopropyl ether (20 mL) to give (Z)-5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonic acid as a light grey solid (1.003 g). Yield = 86% LC-MS: [M+H] = 541.9 1 H NMR (300 MHz, DMSO) δ ppm: 8.48 - 8.35 (m, 2H), 8.30 (d, J = 1.9 Hz, 1H), 8.16 (d, J = 1.8 Hz, 1H), 8.02 (dd, J = 8.7, 2.1 Hz, 1H), 7.96 (s, 1H), 7.62 (dd, J = 8.9, 1.9 Hz, 1H), 7.38 (d, J = 8.8 Hz, 1H), 3.99 (s, 3H), 3.18 (t, J = 7.1 Hz, 2H), 1.69 (dd, J = 49.3, 8.9 Hz, 6H)

[0285] Using the same procedure, the following compounds were prepared:

[0286] (Example 2) (Z)-3-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid

[0287]

Chem.

[0288] Yield = 96% LC-MS: [M+H] = 513.9 1 H NMR (300 MHz, DMSO) δ ppm: 8.42 - 8.31 (m, 2H), 8.28 (d, J = 1.7 Hz, 1H), 8.17 (d, J = 1.8 Hz, 1H), 8.04 - 7.94 (m, 2H), 7.63 (dd, J = 8.9, 1.9 Hz, 1H), 7.37 (d, J = 8.8 Hz, 1H), 3.99 (s, 3H), 3.33 (d, J = 6.8 Hz, 2H), 2.04 - 1.90 (m, 2H)

[0289] (Example 3) (Z)-7-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-7-oxoheptylphosphonic acid

[0290]

Chem.

[0291] Yield = 98% LC-MS: [M+H] = 570 11H NMR (300 MHz, DMSO) δ ppm: 8.40 (d, J = 7.7 Hz, 2H), 8.28 (s, 1H), 8.14 (s, 1H), 8.00 (d, J = 8.6 Hz, 1H), 7.94 (s, 1H), 7.60 (d, J = 8.9 Hz, 1H), 7.37 (d, J = 8.8 Hz, 1H), 3.97 (s, 3H), 3.15 (t, J = 7.1 Hz, 2H), 1.69 (s, 2H), 1.44 (d, J = 25.9 Hz, 8H)

[0292] (Example 4) (Z)-5-(3-(1-Cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-5-oxopentylphosphonic acid

[0293] [Chemical formula]

[0294] Yield = 69% LC-MS: [M+H] = 494 1 1H NMR (300 MHz, DMSO) δ ppm: 8.25 (s, 1H), 8.16 (s, 1H), 8.04 (s, 1H), 7.98 (d, 1H), 7.91 (s, 1H), 7.86 (d, 1H), 7.35 (d, 1H), 7.03 (dd, 1H), 3.97 (s, 3H), 3.83 (s, 3H), 3.15 (t, J = 7 Hz, 2H), 1.77 (t, J = 7 Hz, 2H), 1.61 (m, 4H)

[0295] (Example 5) (Z)-7-(3-(1-Cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-7-oxoheptylphosphonic acid

[0296] [Chemical]

[0297] Yield = 73% LC-MS: [M+H] = 522

[0298] E. Sodium phosphonate prodrug (Step 5)

[0299] (Example 6) (Z)-Sodium 5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate

[0300] [Chemical]

[0301] (Z)-5-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonic acid (700 mg, 1.291 mmol) was suspended in water (25 mL), and then 1N sodium hydroxide (1.291 mL, 1.291 mmol) was added at +5 °C. The reaction mixture was stirred at room temperature for 0.5 h to obtain a clear solution. The solution was lyophilized to give (Z)-sodium 5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate as a pale yellow solid (0.72 g). Yield = 98% LC-MS: [M+H] = 562 1 H NMR (300 MHz, DMSO) δ ppm: 12.12 (brs, 1H), 8.5 -7.0 (m, 8H), 3.96 (brs, 3H), 3.53 (m, 6H), 1.69 (m, 2H)

[0302] Using the same protocol, the following compounds were obtained:

[0303] (Example 7) (Z)-Sodium hydrogen 3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonate

[0304] [Chemical formula]

[0305] Yield = 97% LC-MS: [M-H] = 511.9 1 1H NMR (300 MHz, DMSO) δ ppm: 12.04 (brs, 1H), 8.21 (s, 1H), 8.08 (s, 1H), 7.90 (m, 2H), 7.70 (s, 1H), 7.48 (d, 1H), 7.35 (m, 2H), 3.97 (s, 3H), 3.41 (brs, 4H)

[0306] (Example 8) (Z)-Sodium hydrogen 4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonate

[0307] [Chemical formula]

[0308] LC-MS: [M+H] = 528

[0309] (Example 9) (Z)-Sodium hydrogen 6-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxohexylphosphonate

[0310] [Chemical formula]

[0311] LC-MS: [M+H] = 556

[0312] (Example 10) (Z)-Sodium 7-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-7-oxoheptylphosphonate

[0313]

Chemical formula

[0314] Yield = 98% LC-MS: [M-H] = 568 1 H NMR (300 MHz, DMSO) δ ppm: 12.11 (brs, 1H), 8.25 - 7.35 (m, 8H), 3.97 (s, 3H), 3.59 (brs, 8H), 1.37 (brs, 4H)

[0315] (Example 11) (Z)-Sodium 5-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-5-oxopentylphosphonate

[0316]

Chemical formula

[0317] Yield quantitative LC-MS: [M+H] = 494 1 H NMR (300 MHz, D2O) δ ppm: 7.5-6.5 (m, 8H), 3.59 (s, 6H), 3.25 (brs, 2H), 1.57 (m, 6H)

[0318] (Example 12) (Z)-Sodium 7-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-7-oxoheptylphosphonate

[0319]

Chemical formula

[0320] Yield = 96% LC-MS: [M+H] = 522

[0321] (Example 13) (Z)-Disodium 4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonate

[0322]

Chemical formula

[0323] (Z)-4-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonic acid (74.2 mg, 0.14 mmol) and sodium bicarbonate (23.6 mg, 0.281 mmol) were suspended in water (10 mL) and heated at 40 °C for 10 minutes to obtain a clear solution. The solution was lyophilized to obtain (Z)-disodium 4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonate as a pale yellow solid (0.72 g). LC-MS: [M+H] = 528 1 1H NMR (500 MHz, D2O) δ ppm: 7.9-6.7 (m, 10H), 3.78 (s, 3H), 2.9 (brs, 2H), 1.9 (d, 2H), 1.5 (d, 2H)

[0324] (Example 14) (Z)-5-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonic acid disodium

[0325] [Chemical formula]

[0326] (Z)-5-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonic acid (229 mg, 0.422 mmol) was suspended in water (10 mL), and then 1N sodium hydroxide (0.845 mL, 0.845 mmol) was added. Acetone (50 mL) was slowly added, and the precipitate was filtered and washed twice with acetone (5 mL) to obtain (Z)-5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate disodium salt as a pale yellow solid (0.214 g). LC-MS: [M+H] = 542

[0327] F. Lysine salt of phosphonic acid prodrug (Step 5)

[0328] (Example 15) (S)-2,6-Diaminohexanoic acid compound and (Z)-3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid (2:1)

[0329] [Chemical formula]

[0330] (Z)-Hydrogen 3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid (0.399 g, 0.776 mmol) was suspended in water (10 mL). L-Lysine (0.227 g, 1.552 mmol) was added, and the reaction mixture was stirred at room temperature for 10 minutes until completely solubilized. Then, acetone (50 mL) was slowly added, and the precipitate was filtered, washed with acetone (2 × 5 mL), and dried to obtain a yellow solid (0.52 g). Yield = 78% LC-MS: [M-H] = 511.9

[0331] (Example 16) (S)-2,6-Diaminohexanoic acid compound and (Z)-4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonic acid (2:1)

[0332]

Chemical formula

[0333] LC-MS: [M-H] = 525.8

[0334] General synthetic scheme II

[0335]

Chemical formula

[0336] Condensation of dibenzyl phosphite with THP-protected bromoalcohol gives phosphono-THP-protected alcohol, which upon acid hydrolysis gives phosphonoalcohol. Then, the NH-indole of the drug is treated with triphosgene to give the corresponding chloroformate, which is reacted with phosphonoalcohol to give the benzyl phosphonate prodrug. These latter compounds are hydrolyzed to the prodrug phosphonic acid in an acidic medium.

[0337] (Example 17) 3-[Hydroxy(dioxo)-lambda6-phosphanyl]propyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate

[0338] [Chemical formula]

[0339] A) Dibenzyl (3-hydroxypropyl)phosphonate Dibenzyl phosphite (4.4 mL, 20 mmol) was added to a stirred suspension of HNa (1 g, 25 mmol) in dry DMF at 0 °C under nitrogen. After 15 minutes at room temperature, the solution was cooled to 0 °C, then 2-(3-bromopropoxy)tetrahydro-2H-pyran (3.4 mL, 20 mmol) was added and the reaction mixture was stirred at room temperature for 5 hours. The solution was diluted with water (100 mL) and extracted with diethyl ether (2 × 150 mL). The combined extracts were washed with aqueous NH4Cl (4 × 30 mL), brine (50 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by flash chromatography on SiO2 (eluent: 0 to 100% EtOAc in petroleum ether) to give 5.41 g of a colorless liquid. Yield = 67%

[0340] PPTS (0.025 g, 0.1 mmol) was added to a stirred solution of the previous compound (0.404 g, 1 mmol) in EtOH (10 mL) and the solution was heated at 55 °C for 3 hours. The solution was concentrated in vacuo and the residue was purified by flash chromatography on SiO2 (eluent: 0 to 100% EtOAc in petroleum ether, then 0 to 10% MeOH in EtOAc) to give 0.264 g of a colorless oil. Yield = 82% LC-MS: [M+H] = 321 11H NMR (300 MHz, CDCl3) δ ppm: 7.35 (s, 10H), 5.07 (dd, J = 11.7, 9 Hz, 2H), 4.97 (dd, J = 11.7, 5.7 Hz, 2H), 3.65 (t, J = 5.7 Hz, 2H), 1.96 - 1.74 (m, 4H)

[0341] B) 3 - [Dibenzyloxy(dioxo)-lambda6 - phosphanyl]propyl 5 - bromo - 3 - [(Z)-1 - cyano - 2-(2 - cyano - 5 - methoxyphenyl)vinyl]indole - 1 - carboxylate Under N2, dibenzyl(3 - hydroxypropyl)phosphonate (0.042 g, 0.132 mmol) was dissolved in DCM (1 mL), and the solution was cooled to 0 °C. Triphosgene (14 mg, 0.049 mmol) and then DIPEA (0.023 mL, 0.132 mmol) were added. The solution was stirred for 1 h and then at room temperature for 1 h. In a separate flask, (Z)-3-(2-(5 - bromo - 1H - indol - 3 - yl)-2 - cyanovinyl)-4 - methoxybenzonitrile (0.05 g, 0.132 mmol) was dissolved in DMF (1 mL) under N2 and cooled to 0 °C. HNa (0.063 g, 0.158 mmol) was added, and the solution was stirred at room temperature for 1 h. The chloroformate solution was added dropwise to the indole solution at room temperature. The reaction mixture was stirred at room temperature for 2 h and then quenched by the addition of saturated NH4Cl aqueous solution (5 mL) and extracted with EtOAc (20 mL). The organic layer was washed with saturated NH4Cl aqueous solution (3 × 5 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by flash chromatography on SiO2 column (eluent: 0 to 100% EtOAc in petroleum ether) to give 0.041 g of a pale yellow solid. Yield = 43% 11H NMR (300 MHz, DMSO) δ ppm: 8.27 (d, J = 1.8 Hz, 1H), 8.17 (d, J = 1.8 Hz, 1H), 8.14 (s, 1H), 8.11 (s, 1H), 8.0 (dd, J = 1.8, 8.7 Hz, 1H), 7.92 (s, 1H), 7.61 (dd, J = 1.8, 9.0 Hz, 1H), 7.44 - 7.25 (m, 11H), 5.11 - 4.93 (m, 4H), 4.45 (t, J = 5.7 Hz, 2H), 3.97 (s, 3H) 2.15 - 1.90 (m, 4H)

[0342] C) 3 - [Hydroxy(dioxo)-λ6 - phosphanyl]propyl 5 - bromo - 3 - [(Z)-1 - cyano - 2 - (2 - cyano - 5 - methoxyphenyl)vinyl]indole - 1 - carboxylate 3 - [Dibenzyloxy(dioxo)-λ6 - phosphanyl]propyl 5 - bromo - 3 - [(Z)-1 - cyano - 2 - (2 - cyano - 5 - methoxyphenyl)vinyl]indole - 1 - carboxylate (0.348 g, 0.48 mmol) was dissolved in 4M HCl (9.5 mL) in dioxane and the solution was stirred at 90 °C for 2 h. The solution was concentrated under vacuum and the residue was triturated in DCM to precipitate the product. The solid was filtered and dried under vacuum to give 0.21 g of the desired compound as a white solid. Yield = 80% LC - MS: [M - H] = 542 1 1H NMR (400 MHz, DMSO) δ ppm: 8.26 (s, 1H), 8.14 (m, 3H), 7.99 (d, J = 8.4 Hz, 1H), 7.92 (s, 1H), 7.64 (d, J = 9.0 Hz, 1H), 7.36 (d, J = 8.7 Hz, 1H), 4.49 (t, J = 6.3 Hz, 2H), 3.98 (s, 3H), 2.01 (m, 2H), 1.70 (q, J = 8.2 Hz, 2H) 31P NMR (162 MHz, DMSO d6, δ ppm: 25.44

[0343] (Example 18) 4-[Hydroxy(dioxo)-lambda6-phosphanyl]butyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate

[0344] [Chemical formula]

[0345] A) 2-(4-Bromobutoxy)tetrahydro-2H-pyran To ice-cooled 4-bromobutanol (0.153 g, 1 mmol) in DCM (2 mL), DHP (0.091 mL, 1.1 mmol) and PTSA (0.002 g, 0.01 mmol) were added successively. The solution was stirred at room temperature for 2 h, diluted with DCM (15 mL), washed with saturated NaHCO3 (10 mL), brine (10 mL), dried and concentrated to dryness. The residue was purified by flash chromatography on SiO2 column (eluent 0 to 10% EtOAc in petroleum ether) to give 0.145 g of a clear liquid. Yield = 61% 1 H NMR (300 MHz, CDCl3) δ ppm: 4.68-4.54 (m, 1H), 3.96-3.66 (m, 2H), 3.64-3.32 (m, 4H), 2.12-1.44 (m, 10H)

[0346] B) Dibenzyl (4-(tetrahydropyran-2-yloxy)butyl)phosphonate Dibenzyl phosphite (1.56 mL, 7 mmol) was added to a stirred suspension of HNa (0.35 g, 8.75 mmol) in dry DMF (10.5 mL) at 0 °C under nitrogen. After 15 minutes at room temperature, the solution was cooled to 0 °C, and then 2-(4-bromobutoxy)tetrahydro-2H-pyran (1.67 g, 7 mmol) in dry DMF (4 mL) was added. The reaction mixture was stirred at room temperature for 3 hours. The solution was diluted with water (20 mL) and extracted with diethyl ether (2 × 70 mL). The combined extracts were washed with aqueous NH4Cl solution (4 × 20 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by flash chromatography on SiO2 column (eluent: 10 to 100% EtOAc in petroleum ether) to give 1.71 g of a colorless liquid. Yield = 58% 1 H NMR (300 MHz, CDCl3) δ ppm: 7.35 (s, 10H), 5.06 (dd, J = 11.7, 8.7 Hz, 2H), 4.97 (dd, J = 11.7, 8.1 Hz, 2H), 4.54 (t, J = 3.6 Hz, 1H), 3.88 - 3.77 (m, 1H), 3.75 - 3.65 (m, 1H), 3.54 - 3.43 (m, 1H), 3.39 - 3.28 (m, 1H), 1.94 - 1.40 (m, 12H)

[0347] C) Dibenzyl (4-hydroxybutyl)phosphonate PPTS (0.086 g, 0.34 mmol) was added to a stirred solution of the previous compound (1.44 g, 3.44 mmol) in EtOH (34 mL), and the solution was heated at 55 °C for 4 hours. The solution was concentrated in vacuo, and the residue was purified by flash chromatography on SiO2 column (eluent: 0 to 10% MeOH in DCM) to give 1.1 g of a colorless liquid. Yield = 95%

[0348] D) 4-[Dibenzyloxy(dioxo)-lambda6-phosphanyl]butyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate Under N2, dibenzyl(4-hydroxybutyl)phosphonate (0.265 g, 0.796 mmol) was dissolved in DCM (6 mL), and the solution was cooled to 0 °C. Triphosgene (87 mg, 0.295 mmol) and then DIPEA (0.139 mL, 0.796 mmol) were added. The solution was stirred for 1 hour and then at room temperature for 1 hour. In a separate flask, (Z)-3-(2-(5-bromo-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile (0.3 g, 0.796 mmol) was dissolved in dry DMF (6 mL) under N2 and cooled to 0 °C. HNa (0.038 g, 0.955 mmol) was added, and the solution was stirred at room temperature for 1 hour. The chloroformate solution was added dropwise to the indole solution at room temperature. The reaction mixture was stirred at room temperature overnight and then quenched by the addition of saturated aqueous NH4Cl (30 mL) and extracted with EtOAc (120 mL). The organic layer was washed with saturated aqueous NH4Cl (3 × 30 mL), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by flash chromatography on SiO2 column (eluent: 0 to 100% EtOAc in petroleum ether) to give 0.342 g of a pale yellow solid. Yield = 43%

[0349] E) 4-[Hydroxy(dioxo)-lambda6-phosphanyl]butyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate 4-[Dibenzyloxy(dioxo)-lambda6-phosphanyl]butyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate (0.334 g, 0.452 mmol) was dissolved in 4M HCl in dioxane (9.5 mL), and the solution was stirred at 90 °C for 2 hours. The solution was concentrated under vacuum, and the residue was triturated in DCM to precipitate the product. The solid was collected by filtration and dried under vacuum to give 0.222 g of the desired compound as a white solid. Yield = 88% LC-MS: [M-H] = 557.8 11H NMR (300 MHz, DMSO) δ ppm: 8.26 (s, 1H), 8.12 (m, 3H), 7.99 (d, J = 8.4 Hz, 1H), 7.92 (s, 1H), 7.63 (d, J = 8.7 Hz, 1H), 7.36 (d, J = 9.0 Hz, 1H), 4.46 (t, J = 5.4 Hz, 2H), 3.98 (s, 3H), 1.90 (m, 2H), 1.63 (m, 4H) 31 31P NMR (162 MHz, DMSO, δ ppm: 26.18

[0350] II. L1 Prodrug

[0351] (Example 19) (2S,3S,4S,5R,6S)-6-[4-[[5-Bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenoxy]-3,4,5-trihydroxytetrahydropyran-2-carboxylic acid

[0352]

Chem.

[0353] 1) Methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-[4-(hydroxymethyl)phenoxy]tetrahydropyran-2-carboxylate The compound was synthesized in two steps according to Yu-Ling Leu et al., J. Med. Chem. 2008, 51, 1470 - 1746.

[0354]

Chem.

[0355] a) Methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-(4-formylphenoxy)tetrahydropyran-2-carboxylate In a 100 mL round-bottom flask equipped with magnetic stirring, methyl acetobromo-α-D-glucuronate acetate (2.00 g, 5.04 mmol) was dissolved in CH3CN (40 mL), and then 4-hydroxybenzaldehyde (584 mg, 4.78 mmol) and Ag2O (1.11 g, 4.78 mmol) were slowly added. The mixture was stirred at room temperature for 18 hours. The insoluble material was filtered off. The filtrate was evaporated under reduced pressure to give a dark brown oil. The crude oil was triturated with EtOH (10 mL) to give 1.64 g of a brown solid. The solid was dissolved in DCM (50 mL) and purified by chromatography on a SiO2 cake, eluting from DCM 100% and then from DCM / MeOH (98 / 2) to give 1.10 g of methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-(4-formylphenoxy)tetrahydropyran-2-carboxylate as an off-white solid (yield = 50%). LCMS-ESI: [M+H+18] + =456

[0356] b) Methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-[4-(hydroxymethyl)phenoxy]tetrahydropyran-2-carboxylate In a 250 mL round-bottom flask, methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-(4-formylphenoxy)tetrahydro-2H-pyran-2-carboxylate (1.10 g, 2.52 mmol) was dissolved in CHCl3 (94 mL) and 2-propanol (16 mL). SiO2 (5.50 g) was added. Then, NaBH4 (95 mg, 2.52 mmol) was added slowly at 5 °C. The mixture was stirred at +5 °C for 2 h, hydrolyzed with 200 mL of H2O, filtered to remove SiO2, and extracted with DCM. The organic layer was dried over MgSO4 and evaporated under reduced pressure to give a colorless oil, which was triturated with ethanol (5 mL) to give 1.00 g of methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-[4-(hydroxymethyl)phenoxy]tetrahydro-2H-pyran-2-carboxylate as a white solid (yield = 90%). LCMS-ESI: [M+H+18] + =458

[0357] 2) Methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-[4-[(4-nitrophenoxy)carbonyloxymethyl]phenoxy]tetrahydro-2H-pyran-2-carboxylate

[0358] [Chemical formula]

[0359] In a 100 mL eggplant flask equipped with magnetic stirring, methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-[4-(hydroxymethyl)phenoxy]tetrahydro-2H-pyran-2-carboxylate (1.00 g, 2.27 mmol) was dissolved in dry THF (30 mL) and DCM (15 mL). 4-Nitrophenyl chloroformate (687 mg, 3.41 mmol) was slowly added at room temperature. After 5 minutes, pyridine (286 μL, 3.41 mmol) was added and the reaction mixture was stirred at 20 °C for 2 hours. Ethyl acetate (150 mL) and an aqueous solution of 10% citric acid (150 mL) were added to the reaction mixture. The organic layer was separated, washed with water and brine, dried over magnesium sulfate, and the solvent was removed under vacuum to obtain 1.6 g of a pale yellow oil. The crude oil was triturated with EtOH (5 mL) to obtain 1.64 g of methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-[4-[(4-nitrophenoxy)carbonyloxymethyl]phenoxy]tetrahydro-2H-pyran-2-carboxylate as a pale yellow oil (yield = 92%). LCMS-ESI: [M+H+18] + =623

[0360] 3) [4-[(2S,3R,4S,5S,6S)-3,4,5-Triacetoxy-6-methoxycarbonyltetrahydro-2H-pyran-2-yl]oxyphenyl]methyl 5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate

[0361]

Chemical Structure

[0362] In a 15 mL reactor equipped with magnetic stirring, (Z)-3-(2-(5-bromo-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile (250 mg, 0.66 mmol) was charged into THF (10 mL). At +5 °C, HNa 60% (26 mg, 0.66 mmol) was slowly added under a nitrogen atmosphere. The reaction mixture was stirred at +5 °C for 10 minutes (mn). In a separate 100 ml round-bottom flask equipped with magnetic stirring, under an inert atmosphere, methyl (2S,3S,4S,5R,6S)-3,4,5-triacetoxy-6-[4-[(4-nitrophenoxy)carbonyloxymethyl]phenoxy]tetrahydropyran-2-carboxylate (400 mg, 0.66 mmol) was dissolved in THF (25 mL). To this solution, the previous solution of sodium indole in THF was slowly added at 5 °C. Then, the reaction mixture was stirred at room temperature for 2 hours. An aqueous solution of citric acid (150 mL) was slowly added, and the mixture was stirred for 18 hours. The precipitated solid was filtered and washed with water to obtain 580 mg of a yellow solid. The crude solid was purified directly by flash chromatography on a SiO2 column (24 g), eluting with a gradient from 100% heptane to 100% AcOEt, to give 377 mg of [4-[(2S,3R,4S,5S,6S)-3,4,5-triacetoxy-6-methoxycarbonyltetrahydropyran-2-yl]oxyphenyl]methyl 5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate as an off-white solid (yield = 67%). LCMS-ESI: [M+H+18] + =863 11H NMR (400 MHz, DMSO-d6) δ ppm: 8.25 (s, 1H), 8.18 - 8.10 (m, 3H), 7.99 (d, 1H), 7.93 (s, 1H), 7.63 (d, 1H), 7.54 (d, 2H), 7.34 (d, 1H), 7.05 (d, 2H), 5.70 (d, 1H), 5.46 (brs, 3H), 5.10 - 5.03 (m, 2H), 4.71 (d, 1H), 3.96 (s, 3H), 3.62 (s, 3H), 2.01 (s, 9H)

[0363] 4) (2S,3S,4S,5R,6S)-6-[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenoxy]-3,4,5-trihydroxytetrahydropyran-2-carboxylic acid

[0364]

Chem.

[0365] In a 10 mL reactor equipped with magnetic stirring, [4-[(2S,3R,4S,5S,6S)-3,4,5-triacetoxy-6-methoxycarbonyltetrahydropyran-2-yl]oxyphenyl]methyl 5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate (150 mg, 0.169 mmol) was charged into acetonitrile (1.7 mL). Concentrated HCl (1.7 mL, 16.9 mmol) was added and the reaction mixture was heated at 40 °C for 2 days. The mixture was concentrated under reduced pressure and the residue was purified by flash chromatography on a SiO2 cake, eluting with 100% DCM then DCM / MeOH (95 / 5), then AcOEt / MeOH / AcCN / H2O (7 / 1 / 1 / 1) to give a yellow solid which was further purified by elution on a SiO2 cake with a mixture of AcOEt / MeOH / AcCN / H2O (7 / 1 / 1 / 1) to give 12 mg of (2S,3S,4S,5R,6S)-6-[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenoxy]-3,4,5-trihydroxytetrahydropyran-2-carboxylic acid as a yellow solid (yield = 10%). LCMS-ESI: [M+H+18] + =723

[0366] (Example 22) [4-[[(2S)-2-[[(2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino](peg4)propanoylamino]-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate

[0367]

Chemical Structure

[0368] 1) Example 20: [4-[[(2S)-2-[[(2S)-2-(tert-Butoxycarbonylamino)-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate

[0369]

Chem.

[0370] In a 50 mL eggplant-shaped flask under an argon atmosphere, (Z)-3-(2-(5-bromo-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile (140 mg, 0.37 mmol) was solubilized in dry THF (6 mL). HNa60% (15.6 mg, 0.39 mmol) was added at 5 °C to obtain an orange-red solution, and then the solution was stirred at room temperature for 10 minutes. The solution was cooled again to 5 °C, and then Boc-Val-Cit-PABA-PNP (Iris Biotech, 240 mg, 0.37 mmol) dissolved in 15 mL of THF was added. After 30 minutes, an orange gel appeared with a yellow precipitate below. After 3 hours, water was slowly added, and the aqueous solution was extracted with ethyl acetate. The organic layer was washed with water and brine, dried over magnesium sulfate, and concentrated to dryness to obtain 400 mg of a crude product. The crude product was suspended in water and heated at 50 °C for 15 minutes. 4-Nitrophenol was filtered off, and the filtrate was concentrated under vacuum. The resulting solid was suspended in MeOH (20 mL), heated at 50 °C for 15 minutes, and filtered to obtain 230 mg of a pale yellow solid. The solvent of the filtrate was removed, and the resulting solid residue was triturated with a small amount of MeOH to obtain a second batch of 50 mg of [4-[[(2S)-2-[[(2S)-2-(tert-Butoxycarbonylamino)-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate as a pale yellow solid (total yield = 85%). LCMS-ESI: [M+H-Boc] + =783 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 10.19 (s, 1H), 8.27 (s, 1H), 8.20 (s, 1H), 8.14 (d, J = 12 Hz, 2H), 8.04 (t, J = 12 Hz, 2H), 8.0 (s, 1H), 7.69 (t, 3H), 7.52 (d, 2H), 7.38 (d, J = 8 Hz, 1H), 6.78 (d, J = 12 Hz, 1H), 5.99 (brs, 1H), 5.45 (d, J = 12 Hz, 4H), 4.46 (brs, 1H), 3.99 (s, 3H), 3.85 (brs, 1H), 3.0 (brd, 2H), 1.97 (m, 1H), 1.65 (m, 2H), 1.39 (m, 11H), 0.85 (dd, J = 8, 16 Hz, 6H)

[0371] 2) Example 21: [4-[[(2S)-2-[[(2S)-2-Aminopropanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate hydrochloride

[0372]

Chemical Structure

[0373] In a 50 mL eggplant flask, under an argon atmosphere, a solution of the previous compound (380 mg, 0.43 mmol) in 20 mL of THF was added to a solution of 4N HCl in dioxane (3.23 mL, 12.92 mmol). The reaction mixture was stirred at 20 °C for 18 hours. The solvent was removed under vacuum to obtain a solid, which was triturated with THF (2 mL) and filtered to give 210 mg of [4-[[(2S)-2-[[(2S)-2-Aminopropanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate hydrochloride as a pale yellow powder (yield = 92%). LCMS-ESI: [M+H] + =783 1 H NMR (400 MHz, DMSO-d6) δ ppm: 10.39 (s, 1H), 8.76 (d, J = 8 Hz, 1H), 8.19 - 7.94 (m, 8H), 7.66 (t, 3H), 7.52 (d, 2H), 7.36 (d, J = 12 Hz, 1H), 5.46 (s, 2H), 4.52 (brs, 1H), 4.11 (brs, 3H), 3.97 (s, 3H), 3.68 (brs, 1H), 3.01 (brd, 2H), 2.09 (brs, 1H), 1.65 (brs, 2H), 1.47 (brs, 2H), 0.95 (d, J = 8 Hz, 6H)

[0374] 3) Example 22: [4-[[(2S)-2-[[(2S)-2-[3-[2-[3-(2,5-Dioxopyrrol-1-yl)propanoylamino]ethoxy](peg4)propanoylamino]-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl 5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate

[0375]

Chemical Structure

[0376] In a 15 mL reactor equipped with magnetic stirring, 19-maleimido-17-oxo-4,7,10,13-tetraoxa-16-azanonadecanoic acid (122 mg, 0.29 mmol) and HBTU (185 mg, 0.49 mmol) were charged into DMF (4 mL). At +5 °C, DIPEA (148 μL, 0.85 mmol) was added. The solution was stirred at +5 °C for 20 minutes, and [4-[[(2S)-2-[[(2S)-2-amino-propanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate hydrochloride (200 mg, 0.24 mmol) was added. The reaction was stirred at room temperature for 18 hours. The mixture was slowly poured into water (25 mL) with stirring, and the precipitate was filtered and washed with water to give a brown solid. The crude solid was purified directly by flash chromatography on a SiO2 column (12 g) with solid loading, eluting with a gradient from 100% DCM to DCM / MeOH (8 / 2) to give 85 mg of [4-[[(2S)-2-[[(2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy](peg4)propanoylamino]-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate as a yellow solid (yield = 29%). LCMS-ESI: [M+H] + =1184 1H NMR (400 MHz, DMSO-d6) δ 10.09 (s, 1H), 8.18 (s, 1H), 8.15 - 7.88 (m, 7H), 7.66 (t, 3H), 7.50 (d, 2H), 7.36 (d, 1H), 7.00 (s, 2H), 5.98 (brs, 1H), 5.43 (d, 4H), 4.37 (brs, 1H), 4.22 (t, 1H), 3.96 (s, 3H), 3.58 (t, 4H), 3.48 (s, 12H), 3.14 (m, 2H), 2.97 (dl, 2H), 2.32 (m, 2H), 1.95 (m, 1H), 1.60 (dl, 2H), 1.43 (dl, 2H), 0.84 (dd, 6H)

[0377] (Example 23) [4 - [[(2S)-2 - [[(2S)-2 - [3 - [2 - [3 - (2,5 - dioxopyrrol - 1 - yl)propanoylamido](peg24)propanoylamino]-3 - methylbutanoyl]amino]-5 - ureidopentanoyl]amino]phenyl]methyl 5 - bromo - 3 - [(Z)-1 - cyano - 2 - (5 - cyan - 2 - methoxyphenyl)vinyl]indole - 1 - carboxylate

[0378] [Chemical formula]

[0379] In a 15 mL reactor equipped with magnetic stirring, [4-[[(2S)-2-[[(2S)-2-aminopropanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate hydrochloride (140 mg, 0.17 mmol) and diisopropylethylamine (59 μL, 0.34 mmol) were added to DMF (2 mL). Mal-PEG(24)-NHS (Iris Biotech, 262 mg, 0.19 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was triturated 3 times with DIPE (8 mL) to remove DMF, and the resulting oil was suspended in 0.2N HCl (20 mL) and extracted with DCM (4 × 15 mL). The organic phase was washed with water (15 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain 350 mg of a yellow oil. The crude oil was purified directly 2 times by flash chromatography on a SiO2 column with a solid load, eluting with a gradient from 100% DCM to 8 / 2 DCM / MeOH, to obtain 100 mg of [4-[[(2S)-2-[[(2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamide](peg24)propanoylamino]-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate (yield = 28%). MS (MALDI-TOF): [M+Na] + =2083.91

[0380] (Example 26) (2R)-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino][peg4]propanoylamino]-3-oxopropane-1-sulfonic acid

[0381]

Chem.

[0382] 1) Example 24: 3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-(tert-butoxycarbonylamino)-3-oxopropane-1-sulfonic acid

[0383]

Chem.

[0384] In a 15 mL reactor equipped with magnetic stirring, BOC-cysteic acid, TEA (151 mg, 0.41 mmol) and HBTU (248 mg, 0.65 mmol) were charged into DMF (1 mL). Diisopropylethylamine (198 μL, 1.14 mmol) was added. The solution was stirred at room temperature for 30 minutes. [4-[[(2S)-2-[[(2S)-2-Aminopropanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate, hydrochloride (268 mg, 0.33 mmol) was added at room temperature and the reaction was stirred at room temperature for 18 hours. The reaction mixture was poured into water, extracted with DCM, the organic layer was washed with water, dried over Na2SO4 and concentrated under reduced pressure to give a brown oil. The crude oil was purified by flash chromatography on a SiO2 column (24 g), eluting with a gradient from 100% DCM to 3 / 1 DCM / MeOH to give 190 mg of 3-[[(1S)-1-[[(1S)-1-[[4-[[5-Bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-(tert-butoxycarbonylamino)-3-oxopropane-1-sulfonic acid as a pale yellow solid (yield = 56%). LCMS-ESI: [M-H] - =1034 11H NMR (400 MHz, DMSO) δ ppm: 9.49 (s, 1H), 8.25 (s, 1H), 8.16 (s, 1H), 8.12 (m, 2H), 7.98 (d, 1H), 7.93 (s, 1H), 7.79 (d, 2H), 7.63 (d, 1H), 7.47 (d, 2H), 7.34 (d, 1H), 6.94 (d, 1H), 5.93 (brs, 1H), 5.46 (s, 2H), 5.42( s, 2H), 4.37 (brs, 1H), 4.16 (brs, 1H), 3.97 (brs, 4H), 2.98 (brd, 4H), 2.13 (brs, 1H), 1.71 (brs, 2H), 1.33 - 1.35 (m, 11H), 1.22 (brs, 2H), 0.89 (d, J = 8 Hz, 6H)

[0385] 2) Example 25: 2 - Amino - 3 - [[(1S)-1 - [[(1S)-1 - [[4 - [[5 - Bromo - 3 - [(Z)-1 - cyano - 2 - (5 - cyano - 2 - methoxyphenyl)vinyl]indole - 1 - carbonyl]oxymethyl]phenyl]carbamoyl]-4 - ureidobutyl]carbamoyl]-2 - methylpropyl]amino]-3 - oxopropane - 1 - sulfonic acid

[0386]

Chemical formula

[0387] In a 15 mL reactor equipped with magnetic stirring, the previous compound (120 mg, 0.12 mmol) was charged into THF (2 mL). 4N HCl (870 μL, 3.48 mmol) in dioxane was added. The solution was heated to reflux for 5 minutes, cooled, the precipitate was filtered, and washed with THF to obtain 65 mg of 2-amino-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-3-oxopropane-1-sulfonic acid as a yellow solid (yield = 58%). LCMS-ESI: [M-H] - =934 1 H NMR (400 MHz, DMSO) δ ppm: 9.93 (s, 1H), 8.77 (d J = 8 Hz, 1H), 8.25 (s, 2H), 8.14 (m, 2H), 7.98 (d, 2H), 7.94 (s, 1H), 7.70 (d, 2H), 7.50 (d, 2H), 7.34 (d, 1H), 6.05 (brs, 1H), 5.45 (s, 2H), 4.32 - 4.15 (m, 3H), 3.96 (s, 3H), 3.56 (brs, 4H), 2.98 (m, 4H), 2.13 (brs, 1H), 1.75 (brs, 2H), 1.35-1.25 (m, 2H), 0.87 (d, J = 8 Hz, 6H)

[0388] 3) Example 26: (2R)-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy]propanoylamino]-3-oxopropane-1-sulfonic acid

[0389] [Chem.]

[0390] In a 15 mL reactor equipped with magnetic stirring, 19-maleimido-17-oxo-4,7,10,13-tetraoxa-16-azanonadecanoic acid (77 mg, 0.19 mmol) and HBTU (117 mg, 0.31 mmol) were charged into DMF (1.5 mL). Then, DIPEA (94 μL, 0.54 mmol) was added. The solution was stirred at room temperature for 20 minutes, and subsequently, 2-amino-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-3-oxopropane-1-sulfonic acid (150 mg, 0.15 mmol) was added. The reaction mixture was stirred at room temperature for 18 hours. The solution was poured into 0.2 N HCl, extracted with DCM, the organic layer was washed with water, dried over Na2SO4, and concentrated under reduced pressure. The crude oil was purified directly by flash chromatography on a SiO2 column, eluting with a gradient from 100% DCM to 8 / 2 DCM / MeOH to give 90 mg of (2R)-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy]propanoylamino]-3-oxopropane-1-sulfonic acid as a yellow solid (yield = 44%). LCMS-ESI: [M+H] + = 1334

[0391] III. Solubility A. Protocol: For each compound, three solutions were prepared at 500 μM in 0.01 N HCl (pH 1.0) and three solutions were prepared at 500 μM in phosphate-buffered saline (pH 7.4). The solutions were prepared by dissolving each compound in the appropriate solvent in opaque tubes and stirred at room temperature for 24 hours. After 24 hours, for each solution, an aliquot was centrifuged at 3500 rpm for 10 minutes to separate the insoluble fraction from the soluble fraction. The supernatant was collected, diluted (1 / 20) in H2O, and then injected into LC-MSMS. After HPLC separation, the analyte was detected and quantified by mass spectrometry. A standard curve was performed for each test substance. The solubility of each test substance was determined by calculating the concentration of each supernatant according to the standard curve.

[0392] B: Results The solubility results are presented in Table 1 (Table 2) below:

[0393]

Table 2A

[0394]

Table 2B

[0395] IV. In Vitro Evaluation: Cathepsin B-Mediated Drug Release Protocol

[0396] (Example 27) Synthesis of N-acetyl-S-(1-((R)-2-(((S)-1-(((S)-1-((4-(((5-bromo-3-((Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indole-1-carbonyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-4,20-dioxo-1-sulfo-7,10,13,16-tetraoxa-3,19-diazadocosan-22-yl)-2,5-dioxopyrrolidin-3-yl)-L-cysteine (quenched malprodrug)

[0397] [Chemical formula]

[0398] A solution of Example 26 (crude material from the previous step) (60 mg, 0.045 mmol) in 0.8 ml of DMSO was treated with NAcCys (8 mg, 0.049 mmol) in 0.2 mL of DMSO, and DIPEA (16 μL, 0.09 mmol) was added to the mixture at room temperature. The mixture was stirred for 1 hour: UPLC indicated complete conversion to the title compound. HCOOH (0.1 mmol, 4 μL) was added and the pH was checked (4, orange with test paper Ph1 - 12). The crude product was purified via preparative RP-HPLC. Eluent: acetonitrile (+0.1% HCOOH;) and water (+0.1% HCOOH;). Column: XBridge PROTEIN, 150×19 mm, 300 Å, 5 μm. Flow rate: 30 ml / min. The fractions containing the title compound were collected and freeze-dried. 16 mg of a white solid was recovered. UPLC-MS (M3): Rt = 3.86 peak observed m / z (ES+) = 1497.2 [MH]+ which is in agreement with the predicted value (MW: 1496.42); purity (peak area %) 93%

[0399] Drug release of (Z)-3-(2-(5-bromo-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile from Example 27 was carried out using cathepsin B enzyme.

[0400] Cathepsin B (50 units / mL) was pre-incubated in an activation buffer containing 25 mM sodium acetate (pH 5.5), 30 mM DTT and 15 mM EDTA at 37 °C for 15 minutes and then added as a 20% (v / v) addition to initiate each reaction.

[0401] The final in vitro cathepsin B reaction mixture will contain 25 mM sodium acetate (pH 5.5), 10 units / mL activated cathepsin B, 30 mM DTT and 15 mM EDTA, and 0.5 μg / mL of Example 27 in a 2.5% (v / v) incubation sample background.

[0402] Incubation will occur at 37 °C for 2 hours (0 - 5 - 10 - 15 - 30 - 60 - 120 minutes).

[0403] The reaction was terminated by the addition of an organic solvent and the supernatant was analyzed by LC-MS to quantify the released free drug.

[0404] The results in Figure 1 show efficient drug release of (Z)-3-(2-(5-bromo-1H-indol-3-yl)-2-cyanovinyl)-4-methoxybenzonitrile.

Claims

1. A compound of formula (I): 【Chemical 1】 [wherein, - X represents a nitrogen atom, a C-CN unit or an N⁺-O⁻ unit, - R 1 and R 1 ' each independently represents hydrogen, halogen, (C 1 -C 6 ) alkoxy, or -SO 2 -CH 3 group, provided that when R 1 or R 1 ' is (C 1 -C 6 ) alkoxy, R 2 is not halogen, - R 2 is ・(C 1 ~C 6 ) alkoxy, (C 3 ~C 6 ) cycloalkoxy, (C 3 ~C 6 ) heterocycloalkoxy, aryloxy, heteroaryloxy, (C 1 ~C 6 ) alkylaryloxy, (C 1 ~C 6 ) alkylheteroaryloxy group, which is optionally substituted by at least one halogen, said group, or thio(C 1 ~C 6 ) alkyl, thioaryl, thioheteroaryl, thio(C 1 ~C 6 ) alkylaryl or thio(C 1 ~C 6 )-alkylheteroaryl group, which is optionally substituted by at least one halogen or by a (C 1 ~C 6 ) alkoxy group, said group, ・-NR 4 R 5 unit, O-(C 1 ~C 6 )alkyl-NR 4 R 5 unit or S-(C 1 ~C 6 )alkyl-NR 4 R 5 unit (wherein R 4 and R 5 represent H, (C 1 ~C 6 )alkyl group, or R 4 and R 5 together form a 3- to 7-membered ring optionally interrupted by one or several heteroatoms, provided that at least one of R 4 and R 5 is not H), ・NHCOR 6 unit (wherein R 6 represents a (C 1 -C 6 ) alkyl group), ・At least one halogen, trifluoromethyl group or (C 1 -C 3 ) alkoxy group, optionally substituted, aryl or heteroaryl group, or - halogen represents, - R 3 represents hydrogen, a (C 1 to C 3 ) alkyl group, a (C 1 to C 3 ) alkoxy group or a halogen, - Ra is, - a group of formula (A): 【Chemical 2】 or a group of formula (A'): [Chemical 3] (wherein n is an integer included between 1 and 12), and - a group of formula (B): (wherein, [Chemical Formula 4] - m is an integer equal to 0 or 1) is a group selected from the group consisting of] ・L 1 is a cleavable group selected from a pH-sensitive group, a photoinducible cleavable group, a bioreductive cleavable group, and an enzymatically cleavable group, ・L 2 is a tert-butoxycarbonyl group or a conjugated protein linker, or a pharmaceutically acceptable salt thereof.

2. - A compound of formula (I) which represents a heteroaryl group, preferably a furanyl or triazolyl group . R 2 is ・(C 1 ~C 6 ) alkoxy group, preferably a methoxy group, ・(C 3 ~C 6 ) heterocyclic alkoxy group, preferably an oxetanoxy group, or

3. - Wherein X represents a C-CN unit, a compound of formula (I) according to claim 1 or 2.

4. - R 1 represents a halogen, preferably bromine, and - R 1 ' represents hydrogen, and - R 2 represents a (C 1 to C 6 ) alkoxy group, preferably a methoxy group, - R 3 represents hydrogen, - Wherein X represents a C-CN unit, a compound of formula (I) according to claim 1 or 2.

5. - R 1 represents hydrogen, and - R 1 ' represents an (C 1 ~C 6 ) alkoxy group, preferably a methoxy group, - R 2 represents a (C 1 to C 6 ) alkoxy group, preferably a methoxy group, - R 3 represents hydrogen, Wherein Ra is a group of formula (A): or a group of formula (A'): (wherein n is an integer included between 1 and 12, preferably between 2 and 10, more preferably between 2 and 8, between 2 and 6, and even more preferably, n is 2, 3, 4, 5, or 6) 【Chemical Formula 5】 is a compound of formula (I) according to any one of claims 1 to 4. 【Chemical Formula 6】

6. The pharmaceutically acceptable salt is selected from sodium salt, disodium salt, lysine salt, dilysine salt, arginine salt or diarginine salt, preferably sodium salt or disodium salt, and more preferably sodium salt, a compound of formula (I) according to any one of claims 1 to 5.

7. - Example 1: (Z)-5-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonic acid, - Example 2: (Z)-3-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid, - Example 3: (Z)-7-(5-Bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-7-oxoheptylphosphonic acid, - Example 4: (Z)-5-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-5-oxopentylphosphonic acid, ​ ​ ​ - Example 5: (Z)-7-(3-(1-Cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-7-oxoheptylphosphonic acid, - Example 6: Sodium (Z)-5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate, - Example 7: Sodium (Z)-3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonate, - Example 8: Sodium (Z)-4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonate, - Example 9: Sodium (Z)-6-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxohexylphosphonate, - Example 10: Sodium (Z)-7-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-7-oxoheptylphosphonate, - Example 11: Sodium (Z)-5-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-5-oxopentylphosphonate, - Example 12: Sodium (Z)-7-(3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-6-methoxy-1H-indol-1-yl)-7-oxoheptylphosphonate, - Example 13: Disodium (Z)-4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonate, - Example 14: Disodium (Z)-5-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-5-oxopentylphosphonate, - Example 15: (S)-2,6-diaminohexanoic acid compound and (Z)-3-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-3-oxopropylphosphonic acid (2:1), - Example 16: (S)-2,6-diaminohexanoic acid compound and (Z)-4-(5-bromo-3-(1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl)-1H-indol-1-yl)-4-oxobutylphosphonic acid (2:1), - Example 17: 3-[hydroxy(dioxo)-λ6-phosphanyl]propyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate, and - Example 18: 4-[hydroxy(dioxo)-λ6-phosphanyl]butyl 5-bromo-3-[(Z)-1-cyano-2-(2-cyano-5-methoxyphenyl)vinyl]indole-1-carboxylate The compound of formula (I) according to claim 1 or 5, selected from the group consisting of

8. Ra is a group of formula (B): 【Chemical Formula 7】 [wherein, ・L 1 is a cleavable group selected from among a pH-sensitive group, a photoinducible cleavable group, a bioreductive cleavable group, and an enzymatically cleavable group, ・L 2 is a tert-butoxycarbonyl group or a conjugated protein linker, ・m is an integer equal to 0 or 1] The compound of formula (I) according to any one of claims 1 to 4.

9. L 1 The compound of formula (I) according to claim 8, wherein L is an enzymatically cleavable group cleaved by protease, peptidase, esterase, beta-glucuronidase, glycosidase, phosphodiesterase, phosphatase, pyrophosphatase, tubulin tyrosine ligase or lipase.

10. L 1 is p-aminobenzyloxycarbonyl-AA1 w -AA2 x -AA3 y -AA4 z group, where AA1, AA2, AA3, and AA4 independently represent an amino acid selected from the group consisting of alanine, valine, citrulline, phenylalanine, lysine, glycine, aspartic acid, asparagine, glutamic acid, and derivatives thereof, preferably citrulline, valine, cysteic acid, glycine, and glutamic acid, more preferably citrulline, valine, and cysteic acid, and w, x, y, and z are independently integers equal to 0 or 1, a compound of formula (I) according to claim 8 or 9.

11. m is 1, and L 2 is given by the following formula: 【Chemical Formula 8】 [wherein, r is an integer included between 1 and 36, preferably between 1 and 24, more preferably an integer equal to 2, 4, 8, 12, 16, 20, or 24] The compound of formula (I) according to any one of claims 8 to 10, which is a conjugate protein having

12. - Example 19: (2S,3S,4S,5R,6S)-6-[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenoxy]-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid, - Example 20: [4-[[(2S)-2-[[(2S)-2-(tert-butoxycarbonylamino)-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl 5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate, - Example 21: [4-[[(2S)-2-[[(2S)-2-Aminopropanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate hydrochloride, - Example 22: [4-[[(2S)-2-[[(2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy](peg4)propanoylamino]-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate, - Example 23: [4-[[(2S)-2-[[(2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamido](peg24)propanoylamino]-3-methylbutanoyl]amino]-5-ureidopentanoyl]amino]phenyl]methyl-5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carboxylate, - Example 24: 3-[[(1S)-1-[[(1S)-1-[[4-[[5-Bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-(tert-butoxycarbonylamino)-3-oxopropane-1-sulfonic acid, - Example 25: 2-Amino-3-[[(1S)-1-[[(1S)-1-[[4-[[5-Bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-3-oxopropane-1-sulfonic acid, and - Example 26: (2R)-3-[[(1S)-1-[[(1S)-1-[[4-[[5-bromo-3-[(Z)-1-cyano-2-(5-cyano-2-methoxyphenyl)vinyl]indole-1-carbonyl]oxymethyl]phenyl]carbamoyl]-4-ureidobutyl]carbamoyl]-2-methylpropyl]amino]-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy]propanoylamino]-3-oxopropane-1-sulfonic acid A compound of formula (I) according to claim 1 or 8, selected from the group consisting of **Claim 13** Conjugate of formula (II): 【Chemical Formula 9】 [wherein, - X, R 1 , R 1' , R 2 , R 3 , and L 1 is as defined in any one of claims 1 to 4 and 8 to 10, - L 2 is a conjugate protein conjugate, - P is a peptide or protein capable of binding to a target of interest, preferably an antibody, an antibody fragment, or an antigen-binding fragment, - v is an integer from 1 to 10]. **Claim 14** A pharmaceutical composition comprising a compound according to any one of claims 1 to 13 and a pharmaceutically acceptable excipient. **Claim 15** Preferably, the pharmaceutical composition according to claim 14 for use in the treatment of cancer, selected from among leukemia, acute myeloid leukemia, lymphoma, breast cancer, pancreatic cancer, lung cancer or colon cancer.

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