Claims for Patent: 9,814,784

Summary for Patent: 9,814,784

Title:	Antibody-linker-drug conjugate, preparation method therefor, and anticancer drug composition containing same
Abstract:	The present invention relates to an antibody-linker-drug conjugate in which an antibody and a cytotoxic drug are conjugated through an enzyme cleavable peptide linker capable of directly binding to a lysine residue of an antibody, a preparation method therefor, and an anticancer drug composition containing the same as an active ingredient.
Inventor(s):	Park; Young Jun (Incheon, KR), Jeong; Jin-kyo (Incheon, KR), Choi; Young Mi (Incheon, KR), Lee; Minseob (Incheon, KR), Kim; Yeon Jung (Incheon, KR), Kim; Kyoung Suk (Incheon, KR), Choi; Joon hun (Incheon, KR), Lee; Jin Seo (Incheon, KR), Cho; Eun Joo (Incheon, KR), Song; Hyunnam (Incheon, KR), Park; Sung Jun (Incheon, KR), Lee; Jong-hyoup (Incheon, KR), Lee; Matthew Sangyup (Incheon, KR), Lee; In-Suk (Incheon, KR), Kim; Joon woo (Incheon, KR), Hong; Seung Suh (Incheon, KR)
Assignee:	CELLTRION, INC. (Incheon, KR)
Application Number:	14/655,357
Patent Claims:	1. An antibody-linker-drug conjugate, wherein the linker-drug moiety is directly bonded to a lysine residue of the antibody, represented by Chemical Formula I-1, or a pharmaceutically acceptable salt thereof: ##STR00214## wherein, Ab is an antibody, X is C.sub.1-C.sub.8 alkyl or C.sub.3-C.sub.6 cycloalkyl, R.sub.1 is a hydroxy, C.sub.1-C.sub.4 alkoxy, or oxo (.dbd.O), R.sub.2 is a hydrogen atom or C.sub.1-C.sub.4 alkyl, R.sub.3 is a hydrogen atom, hydroxy, C.sub.1-C.sub.4 alkoxy, amino, oxo (.dbd.O), or hydroxyimino (.dbd.N--OH), R.sub.4 is aryl, and n is an integer ranging from 1 to 5. 2. The antibody-linker-drug conjugate of claim 1, or a pharmaceutically acceptable salt thereof, wherein: Ab is an antibody immunospecifically binding to a cancer cell antigen, X is C.sub.1-C.sub.8 alkyl or C.sub.3-C.sub.6 cycloalkyl, R.sub.1 is a hydroxy, or C.sub.1-C.sub.4 alkoxy, R.sub.2 is a hydrogen atom or C.sub.1-C.sub.4 alkyl, R.sub.3 is a hydrogen atom, oxo (.dbd.O) or hydroxyimino (.dbd.N--OH), R.sub.4 is phenyl unsubstituted or substituted with at least one substituent selected from the group consisting of C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy, and a halogen atom. 3. The antibody-linker-drug conjugate of claim 1, or a pharmaceutically acceptable salt thereof, wherein: Ab is trastuzumab, X is methyl, n-propyl, n-pentyl, n-octyl, or cyclopropyl, R.sub.1 is a, hydroxy, or methoxy, R.sub.2 is a hydrogen atom or methyl, R.sub.3 is a hydrogen atom, oxo (.dbd.O) or hydroxyimino (.dbd.N--OH), and R.sub.4 is phenyl unsubstituted or substituted with at least one substituent selected from the group consisting of methyl, methoxy, and a halogen atom. 4. The antibody-linker-drug conjugate of claim 1, being selected from the group consisting of the following compounds, or a pharmaceutically acceptable salt thereof: ##STR00215## wherein, n is an integer ranging from 1 to 5. 5. The antibody-linker-drug conjugate of claim 1, or a pharmaceutically acceptable salt thereof, wherein the antibody-linker-drug conjugate has a normal antibody structure at a rate of 98% or higher. 6. An antibody-linker-drug conjugate, wherein the linker-drug moiety is directly bonded to a lysine residue of the antibody, represented by Chemical Formula I-3, or a pharmaceutically acceptable salt thereof: ##STR00216## wherein, Ab is an antibody, X is C.sub.1-C.sub.8 alkyl, or C.sub.3-C.sub.6 cycloalkyl, R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are each independently a hydrogen atom, or C.sub.1-C.sub.4 alkyl, R.sub.5 is a hydrogen atom, hydroxy, C.sub.1-C.sub.4 alkoxy, amino, oxo (.dbd.O), or hydroxyimino (.dbd.N--OH), Ar is aryl, Y is a nitrogen atom or an oxygen atom, R.sub.6 is a hydrogen atom or C.sub.1-C.sub.4 alkyl when Y is a nitrogen atom; or is absent when Y is an oxygen atom, and n is an integer ranging from 1 to 5. 7. The antibody-linker-drug conjugate of claim 6, or a pharmaceutically acceptable salt thereof, wherein Ab is an antibody immunospecifically binding to a cancer cell antigen, X is C.sub.1-C.sub.8 alkyl or C.sub.3-C.sub.6 cycloalkyl, R.sub.1, R.sub.2, and R.sub.4 are each independently a hydrogen atom or C.sub.1-C.sub.4 alkyl, R.sub.3 is C.sub.1-C.sub.4 alkyl, R.sub.5 is a hydrogen atom, amino, oxo (.dbd.O), or hydroxyimino (.dbd.N--OH), Ar is phenyl or naphthyl unsubstituted or substituted with at least one substituent selected from the group consisting of C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy, and a halogen atom, Y is a nitrogen atom or an oxygen atom, and R.sub.6 is a hydrogen atom or C.sub.1-C.sub.4 alkyl when Y is a nitrogen atom, or is absent when Y is an oxygen atom. 8. The antibody-linker-drug conjugate of claim 6, or a pharmaceutically acceptable salt thereof, wherein: Ab is trastuzumab, X is methyl, n-propyl, n-pentyl, n-octyl, or cyclopropyl, R.sub.1, R.sub.2, and R.sub.3 are each methyl, R.sub.4 is a hydrogen atom, or methyl, R.sub.5 is a hydrogen atom, oxo (.dbd.O), or hydroxyimino (.dbd.N--OH), Ar is phenyl or naphthyl unsubstituted or substituted with at least one substituent selected from the group consisting of methyl, methoxy, and a halogen atom, Y is a nitrogen atom or an oxygen atom, and R.sub.6 is a hydrogen atom, or methyl when Y is a nitrogen atom, or is absent when Y is an oxygen atom. 9. The antibody-linker-drug conjugate of claim 6, being selected from the group consisting of the following compounds, or a pharmaceutically acceptable salt thereof: ##STR00217## ##STR00218## ##STR00219## wherein, n is an integer ranging from 1 to 5. 10. The antibody-linker-drug conjugate of claim 6, or a pharmaceutically acceptable salt thereof, wherein the antibody-linker-drug conjugate has a normal antibody structure at a rate of 90% or higher. 11. A method for preparing an antibody-linker-drug conjugate represented by Chemical Formula I-1, comprising the steps of: (i) condensating a compound of the following Chemical Formula II-1 with a dolastatin 10derivative of the following Chemical Formula III-1 to give a compound of the following Chemical Formula IV-1; (ii) subjecting the compound of the following Chemical Formula IV-1 to an addition reaction with a compound of the following Chemical Formula V to give a compound of the following Chemical Formula VI-1; (iii) condensating the compound of the following Chemical Formula VI-1 with N-hydroxysuccinimide to give a compound of the following Chemical Formula VII-1; and (iv) reacting the compound of the following Chemical Formula VII-1 with an antibody: ##STR00220## ##STR00221## wherein, Ab is an antibody, X is C.sub.1-C.sub.8 alkyl or C.sub.3-C.sub.6 cycloalkyl, R.sub.1 is, hydroxy, C.sub.1-C.sub.4 alkoxy or oxo (.dbd.O), R.sub.2 is a hydrogen atom or C.sub.1-C.sub.4 alkyl, R.sub.3 is a hydrogen atom, hydroxy, C.sub.1-C.sub.4 alkoxy, amino, oxo (.dbd.O), or hydroxyimino (.dbd.N--OH), R.sub.4 is aryl, and n is an integer ranging from 1 to 5. 12. The method of claim 11, wherein the condensation reaction of step (i) is carried out in presence of a condensing agent selected from the group consisting of hydroxybenzotriazole (HOBt), hydroxyazabenzotriazole (HOAt), and hydroxysuccinimide (HOSu), and an organic base selected from the group consisting of pyridine and diisopropylethylamine. 13. The method of claim 11, wherein the condensation reaction of step (iii) is carried out in presence of a condensing agent selected from the group consisting of dicyclohexylcarbodiimide (DCC), and (3-dimethylaminopropyl)carbodiimide (EDC). 14. The method of claim 11, wherein the reaction of step (iv) is carried out in a phosphate buffer having a pH of 6.0 to 8.0 as a reaction solvent. 15. The method of claim 11, wherein the reaction of step (iv) is carried out in a mixed reaction solvent of a phosphate buffer having a pH of 6.0 to 8.0 and an organic solvent. 16. The method of claim 15, wherein the organic solvent amounts to 50% or less of the mixed solvent. 17. The method of claim 11, wherein the compound of Chemical Formula VII-1is used in an amount of 3 to 25 equivalents, based on the antibody, in step (iv). 18. A method for preparing an antibody-linker-drug conjugate represented by Chemical Formula I -3, comprising the steps of: (i) condensating a compound of the following Chemical Formula II-1 with a dolastatin 10 derivative of the following Chemical Formula III-3 to give a compound of the following Chemical Formula IV-3; (ii) subjecting the compound of the following Chemical Formula IV-3 to an addition reaction with a compound of the following Chemical Formula V to give a compound of the following Chemical Formula VI-3; (iii) condensating the compound of the following Chemical Formula VI-3 with N-hydroxysuccinimide to give a compound of the following Chemical Formula VII-3; and (iv) reacting the compound of the following Chemical Formula VII-3 with an antibody: ##STR00222## ##STR00223## wherein, Ab is an antibody, X is C.sub.1-C.sub.8 alkyl, or C.sub.3-C.sub.6 cycloalkyl, R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are each independently a hydrogen atom, or C.sub.1-C.sub.4 alkyl, R.sub.5 is a hydrogen atom, hydroxy, C.sub.1-C.sub.4 alkoxy, amino, oxo (.dbd.O), or hydroxyimino (.dbd.N--OH), Ar is aryl, Y is a nitrogen atom or an oxygen atom, R.sub.6 is a hydrogen atom or C.sub.1-C.sub.4 alkyl when Y is a nitrogen atom; or is absent when Y is an oxygen atom, and n is an integer ranging from 1 to 5. 19. The method of claim 18, wherein the condensation reaction of step (i) is carried out in presence of a condensing agent selected from the group consisting of hydroxybenzotriazole (HOBt), hydroxyazabenzotriazole (HOAt), and hydroxysuccinimide (HOSu), and an organic base selected from the group consisting of pyridine and diisopropylethylamine. 20. The method of claim 18, wherein the condensation reaction of step (iii) is carried out in presence of a condensing agent selected from the group consisting of dicyclohexylcarbodiimide (DCC), and (3-dimethylaminopropyl)carbodiimide (EDC). 21. The method of claim 18, wherein the reaction of step (iv) is carried out in a phosphate buffer having a pH of 6.0 to 8.0 as a reaction solvent. 22. The method of claim 18, wherein the reaction of step (iv) is carried out in a mixed reaction solvent of a phosphate buffer having a pH of 6.0 to 8.0 and an organic solvent. 23. The method of claim 22, wherein the organic solvent amounts to 50% or less of the mixed solvent. 24. The method of claim 18, wherein the compound of Chemical Formula VII-3 is used in an amount of 3 to 25 equivalents, based on the antibody, in step (iv).

Details for Patent 9,814,784

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	09/25/1998		2033-01-03
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	02/10/2017		2033-01-03
Genentech, Inc.	HERCEPTIN HYLECTA	trastuzumab and hyaluronidase-oysk	Injection	761106	02/28/2019		2033-01-03