Conjugation linkers, cell binding molecule-drug conjugates containing the linkers, methods of making and uses such conjugates with the linkers

Patent metadata

Field	Value
Publication number	US-12364767-B2
Application number	US-201616348749-A
Country	US
Kind code	B2
Filing date	Nov 14, 2016
Priority date	Nov 14, 2016
Publication date	Jul 22, 2025
Grant date	Jul 22, 2025

Abstract

The present invention relates to linkers having a group of propiolyl, substituted acryl (acryloyl), or disubstituted propanoyl, and using such linkers for the conjugation of compounds, in particular, cytotoxic agents to a cell-binding molecule.

First claim

What is claimed is:

1. A cell-binding agent-drug conjugate compound of Formula (VIII):

wherein Formula (VIII) is Formula (VIII′):

wherein:
n is 1-20; and
T is N—C(O)—CH 2 —N—N or N—C(O)—CH 2 CH 2 —N—N,
Cb represents a cell-binding agent of an antibody or a functional fragment thereof;
Drug, Drug′, and Drug″ represent same or different of, a cytotoxic agent, or a therapeutic drug, an antibody or an antibody fragment, or siRNA, miRNA, mRNA, piRNA, or DNA, which is linked to the cell-binding agent via the linker through R 1 , R 1 ′, and R 1 ″ respectively containing a C 1 -C 8 alkane; C 2 -C 8 alkylene, alkenylene, alkynylene, aromatic, ether, polyoxyalkylene, ester, amine, imine, polyamine, hydrazine, hydrazone, amide, urea, semicarbazide, carbazide, alkoxyamine, urethanes, amino acid, peptide, acyloxylamine, hydroxamic acid, disulfide, thioether, thioester, carbamate, carbonate, heterocyclic ring, heteroalkyl, heteroaromatic, or alkoxime; or a combination thereof, wherein at least one of Drug, Drug′ and Drug″ is a tubulysin molecule;
“ ” represents either single bond or double bond;
m 1 , m 1′ , and m 1″ are independently an integer from 1 to 10;
m 2 , m 2′ , m 2″ and m 4 are 1;
R 1 , R 1′ , and R 1″ , are same or different, and are C 1 -C 8 alkyl; C 2 -C 8 heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; or C 2 -C 8 ester, ether, or amide; or polyethyleneoxy unit of formula (OCH 2 CH 2 ) p or (OCH 2 CH(CH 3 )) p , wherein p is an integer from 0 to about 1000, or a combination thereof,
X 1 , X 1′ , L 2 , L 2′ and L 2′″ are absent;
L 1 , L 1′ , and L 1″ are same or different and independently O, NH, S, NHNH, N(R 3 ), N(R 3 )N(R 3′ ), polyethyleneoxy unit of formula (OCH 2 CH 2 ) p OR 3 , or (OCH 2 CH(CH 3 )) p OR 3 , or NH(CH 2 CH 2 O) p R 3 , or NH(CH 2 CH(CH 3 )O) p R 3 , or N[(CH 2 CH 2 O) p R 3 ][(CH 2 CH 2 O) p′ R 3′ ], or (OCH 2 CH 2 ) p C(═O)X 1 R 3 , or CH 2 CH 2 (OCH 2 CH 2 ) p C(═O)X 1 R 3 , wherein p and p′ are independently an integer selected from 1 to about 1000, or a combination thereof; C 1 -C 8 alkyl; C 2 -C 8 heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; wherein X 1 , R 3 and R 3 ′ are defined above;
or L 1 , L 1′ , or L 1″ contains a group of self-immolative or a non-self-immolative component, peptidic units (1-8 of natural or unnatural amino acids), a hydrazone bond, a disulfide, an ester, an oxime, an amide, or a thioether bond;
the self-immolative unit comprises an aromatic compound selected from the group consisting of para-aminobenzylcarbamoyl (PAB) groups, 2-aminoimidazol-5-methanol compounds, heterocyclic PAB compounds, beta-glucuronide, and ortho or para-aminobenzylacetals; the self-immolative unit has one of following structures:

wherein the (*) atom is the point of attachment of additional spacer or releasable linker units, or a cytotoxic agent, and/or a binding molecule (CBA); X 1 , Y 1 , Z 2 and Z 3 are independently NH, O, or S; Z 1 is independently H, NHR 1 , OR 1 , SR 1 , COX 1 R 1 , wherein X 1 and R 1 are defined above; v is 0 or 1; U 1 is independently H, OH, C 1 -C 6 alkyl, (OCH 2 CH 2 ) n , F, Cl, Br, I, OR 5 , SR 5 , NR 5 R 5 ′, N═NR 5 , N═R 5 , NR 5 R 5 ′, NO 2 , SOR 5 R 5 ′, SO 2 R 5 , SO 3 R 5 , OSO 3 R 5 , PR 5 R 5 ′, POR 5 R 5 ′, PO 2 R 5 R 5 ′, OPO(OR 5 )(OR 5 ′), or OCH 2 PO(OR 5 (OR 5 ′), wherein R 5 and R 5 ′ are independently H, C 1 ˜C 8 alkyl; C 2 ˜C 8 alkenyl, alkynyl, heteroalkyl, or aminocaine; C 3 ˜C 8 aryl, heterocyclic, carbocyclic, cycloalkyl, heterocycloalkyl, heteroaralkyl, alkylcarbonyl, or glycoside; or pharmaceutical cation salts;
the non-self-immolative linker component is one of following structures:

wherein the (*) atom is a point of attachment of additional spacer or releasable linker, a cytotoxic agent, and/or a binding molecule; X 1 , Y 1 , U 1 , R 5 , R 5 ′ are defined as above; r is 0-100; m and n are 0˜6 independently;
or L 1 , L 1′ , L 1″ and L 1′″ are independently absent;
or L 1 , L 1′ , L 1″ , or L 1′″ are composed of one or more linker components, selected from 6-maleimidocaproyl (“MC”), maleimidopropanoyl (“MP”), valine-citrulline (“val-cit” or “vc”), alanine-phenylalanine (“ala-phe” or “af”), p-aminobenzyloxycarbonyl (“PAB”), 4-thiopentanoate (“SPP”), 4-(N-maleimidomethyl)-cyclohexane-1 carboxylate (“MCC”), (4-acetyl)aminobenzoate (“SIAB”), 4-thio-butyrate (SPDB), 4-thio-2-hydroxysulfonyl-butyrate (2-Sulfo-SPDB), ethyleneoxy —CH 2 CH 2 O— as one or more repeating units (“EO” or “PEO”).

2. The conjugate compound according to claim 1 , wherein a pair of thiols of the cell-binding agent which are linked to the linker are the inter chain disulfide atoms of the cell-binding agent that are reduced by a reduction agent of dithiothreitol (DTT), dithioerythritol (DTE), dithiolbutylamine (DTBA), L-glutathione (GSH), tris (2-carboxyethyl) phosphine (TCEP), 2-mercaptoethylamine (P-MEA), or/and beta mercaptoethanol (β-ME, 2-ME).

3. The conjugate compound according to claim 1 , wherein Drug, Drug′, or Drug″ is a chromophore molecule.

4. The conjugate compound according to claim 1 , wherein one, two or all of the Drug, Drug′, or Drug″ are independently a poly(ethylene glycol) (PEGs), poly(propylene glycol), or a copolymer of ethylene oxide or propylene oxide.

5. The conjugate compound according to claim 1 , wherein one, two or all of the Drug, Drug′, or Drug″ are independently a cell-binding ligand or a cell receptor agonist, or a cell receptor binding molecule.

6. The conjugate compound according to claim 1 , wherein Drug, Drug′, or Drug″ is selected from the group consisting of tubulysins, calicheamicins, auristatins, maytansinoids, CC-1065 compounds, daunorubicin and doxorubicin compounds, taxanoids (taxanes), cryptophycins, epothilones, benzodiazepine dimers (selected from the group consisting of dimmers of pyrrolobenzodiazepine (PBD), tomaymycins, anthramycins, indolinobenzodiazepines, imidazobenzothiadiazepines, or oxazolidinobenzodiazepines), calicheamicins and the enediyne antibiotics, actinomycins, amanitins, azaserines, bleomycins, epirubicins, tamoxifen, idarubicin, dolastatins/auristatins (selected from the group consisting of monomethyl auristatin E, MMAE, MMAF, auristatin PYE, auristatin TP, Auristatins 2-AQ, 6-AQ, EB (AEB), and EFP (AEFP)), duocarmycins, geldanamycins, methotrexates, thiotepa, vindesines, vincristines, hemiasterlins, nazumamides, microginins, radiosumins, alterobactins, microsclerodermins, theonellamides, esperamicins, siRNA, miRNA, piRNA, nucleolytic enzymes, and/or pharmaceutically acceptable salts, or acids of any of the above molecules.

7. The conjugate compound according to claim 1 , wherein Cb is selected from the group consisting of an antibody, a full-length antibody (polyclonal antibody, monoclonal antibody, antibody dimer, antibody multimer, or multispecific antibody (select

Assignees

• Hangzhou Dac Biotech Co Ltd

Inventors

• Zhao Yongxin Robert
• Yang Qingliang
• Huang Yuanyuan
• Gai Shun
• Ye Hangbo
• Zhao Linyao
• Yang Chengyu
• Guo Huihui
• Zhou Xiaomai
• Xie Hongsheng
• Zhu Haifeng
• Xu Yifang
• Tong Qianqian
• Jia Junxiang
• Cao Minjun
• Li Wenjun
• Gao Shuihong
• Guo Zhixiang
• Bai Lu
• Li Chen
• Yang Yanlei
• Wang Chunyan
• Ye Zhichang

Classifications

• A61K47/68035

  the drug being a pyrrolobenzodiazepine · CPC title

• A61K47/68031

  the drug being an auristatin · CPC title

• A61K47/6863

  the tumour determinant being from stomach or intestines cancer cell · CPC title

• A61K47/6855

  the tumour determinant being from breast cancer cell · CPC title

• A61K47/6829

  Bacterial toxins, e.g. diphteria toxins or Pseudomonas exotoxin A · CPC title