# US 9,434,748 B2 · Cytotoxic benzodiazepine derivatives

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- Number: US 9,434,748 B2
- Document ID: US9434748B2
- Status: Granted
- Kind: B2
- Application number: 14/849,270
- Filed: 2015-09-09
- Published: 2016-04-21
- Granted: 2016-09-06
- Earliest claimed: 2011-02-15
- Assignees: ImmunoGen, Inc.
- Applicants: ImmunoGen, Inc.
- Inventors: Wei Li; Michael Louis Miller; Nathan Elliott Fishkin; Ravi V. J. Chari
- Web page: https://patentgrep.com/patent/US9434748B2

## CPC

- A61K 31/5517: condensed with five-membered rings having nitrogen as a ring hetero atom, e.g. imidazobenzodiazepines, triazolam
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61K 47/545: {Heterocyclic compounds (A61K47/558 takes precedence)}
- A61K 47/68035: {the drug being a pyrrolobenzodiazepine}
- A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant}
- A61K 47/6851: {the antibody targeting a determinant of a tumour cell}
- A61K 47/6867: {the tumour determinant being from a cell of a blood cancer}
- A61P 35/00: Antineoplastic agents
- C07D 487/04: Ortho-condensed systems
- C07D 519/00: Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
- C07K 16/30: from tumour cells
- A61K 2039/505: {comprising antibodies}
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- C07K 2317/94: Stability, e.g. half-life, pH, temperature or enzyme-resistance
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The invention relates to novel benzodiazepine derivatives with antiproliferative activity and more specifically to novel benzodiazepine compounds of formula (I)-(VII). The invention also provides conjugates of the benzodiazepine compounds linked to a cell-binding agent. The invention further provides compositions and methods useful for inhibiting abnormal cell growth or treating a proliferative disorder in a mammal using the compounds or conjugates of the invention.

## Claims

### Claim 1

A method of treating a cancer in a mammal comprising administering to said mammal a therapeutically effective amount of a conjugate comprising a cytotoxic compound and a cell binding agent (CBA), wherein the cytotoxic compound comprises a linking group which covalently links the cytotoxic compound to the CBA, and wherein said cytotoxic compound represented by the following structural formula:
or a pharmaceutically acceptable salt thereof, wherein:
the double line between N and C represents a single bond or a double bond, provided that when it is a double bond X is absent and Y is —H, or a linear or branched alkyl having 1 to 4 carbon atoms, and when it is a single bond, X is —H, the linking group, or an amine protecting moiety;
Y is a leaving group selected from —OR, —OCOR′, —OCOOR′, —OCONR′R″, —NR′R″, —NR′COR″, —NR′NR′R″, an optionally substituted 5- or 6-membered nitrogen-containing heterocycle, a guanidinum represented by —NR′(C═NH)NR′R″, an amino acid, or a peptide represented by —NRCOP′, wherein P′ is an amino acid or a polypeptide containing between 2 to 20 amino acid units, —SR, —SOR′, —SO2M, —SO3M, —OSO3M, halogen, cyano and an azido;
M is —H or a cation;
R, for each occurrence, is independently selected from the group consisting of —H, an optionally substituted linear, branched or cyclic alkyl, alkenyl or alkynyl having from 1 to 10 carbon atoms, a polyethylene glycol unit —(CH2CH2O)n—Rc, an optionally substituted aryl having 6 to 18 carbon atoms, an optionally substituted 5- to 18-membered heteroaryl ring containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an optionally substituted 3- to 18-membered heterocyclic ring containing 1 to 6 heteroatoms independently selected from O, S, N and P;
R′ and R″ are each independently selected from —H, —OH, —OR, —NHR, —NR2, —COR, an optionally substituted linear, branched or cyclic alkyl, alkenyl or alkynyl having from 1 to 10 carbon atoms, a polyethylene glycol unit —(CH2CH2O)n—Rc, and an optionally substituted 3-18-membered heterocyclic ring having 1 to 6 heteroatoms independently selected from O, S, N and P;
Rc is —H or an optionally substituted linear or branched alkyl having 1 to 4 carbon atoms, or the linking group;
n is an integer from 1 to 24;
X′ is selected from —H, an amine-protecting group, the linking group, an optionally substituted linear, branched or cyclic alkyl, alkenyl or alkynyl having from 1 to 10 carbon atoms, a polyethylene glycol unit —(CH2CH2O)n—Rc, an optionally substituted aryl having 6 to 18 carbon atoms, an optionally substituted 5- to 18-membered heteroaryl ring containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an optionally substituted 3- to 18-membered heterocyclic ring containing 1 to 6 heteroatoms independently selected from O, S, N and P;
Y′ is selected from —H, an oxo group, the linking group, an optionally substituted linear, branched or cyclic alkyl, alkenyl or alkynyl having from 1 to 10 carbon atoms, an optionally substituted 6- to 18-membered aryl, an optionally substituted 5- to 18-membered heteroaryl ring containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur, an optionally substituted 3 to 18-membered heterocyclic ring having 1 to 6 heteroatoms;
R6 is —H, —R, —OR, —SR, —NR′R″, —NO2, halogen or the linking group;
A and A′ are the same or different, and are independently selected from —O—, oxo (—C(═O)—), —CRR′O—, —CRR′—, —S—, —CRR′S—, —N(R5)— and —CRR′N(R5)—,
R5 for each occurrence is independently —H or an optionally substituted linear or branched alkyl having 1 to 10 carbon atoms;
L′, L″, and L′″ are the same or different, and are independently selected from —H, an optionally substituted linear, branched or cyclic alkyl, alkenyl or alkynyl having from 1 to 10 carbon atoms, a polyethylene glycol unit —(OCH2CH2)n—Rc, halogen, guanidinium [—NH(C═NH)NH2], —OR, —NR′R″, —NO2, —NR′COR″, —SR, a sulfoxide represented by —SOR′, a sulfone represented by —SO2R′, a sulfonate —SO3M, a sulfate —OSO3M, a sulfonamide represented by —SO2NR′R″, cyano, an azido, —COR′, —OCOR′, —OCONR′R″ and the linking group, provided only one of L′, L″, and L′″ is the linking group; and
G is selected from —CH— or —N—,
provided that the compound is not any one of the following compounds:
wherein the cancer is selected from the group consisting of leukemia, lymphoma, multiple myeloma, cervical cancer, non-small cell lung cancer, small-cell lung cancer, head and neck cancer and colon cancer; and
the optional substituent described above is a linear, branched or cyclic alkyl, alkenyl or alkynyl having from 1 to 10 carbon atoms, aryl, heteroaryl, heterocycyclyl, halogen, —NH(C═NH)NH2, —OR100, NR101R102, —NO2, —NR101COR102, —SR100, —SOR101, —SO2R101, —SO3M, —OSO3M, —SO2NR101R102, cyano, an azido, —COR101, —OCOR101, —OCONR101R102 and a polyethylene glycol unit (—OCH2CH2)nR101 wherein M is H or a cation; R100, R101, R102 and R103 are each independently selected from the group consisting of H, linear, branched or cyclic alkyl, alkenyl or alkynyl having from 1 to 10 carbon atoms, a polyethylene glycol unit (—OCH2CH2)n—R104, an aryl having from 6 to 10 carbon atoms, a heterocyclic ring having from 3 to 10 carbon atoms and a heteroaryl having 5 to 10 carbon atoms; n is an integer from 1 to 24, and R104 is H or a linear or branched alkyl having 1 to 4 carbon atoms, wherein the alkyl, alkenyl, alkynyl, aryl, heteroaryl and heterocyclcyl in the groups represented by R100, R101, R102, R103 and R104 are optionally substituted with one or more substituents independently selected from the group consisting of halogen, —OH, —CN, —NO2 and unsubstituted linear or branched alkyl having 1 to 4 carbon atoms.

### Claim 2 (depends on 1)

The method of claim 1, wherein L′ is the linking group, and L″ and L′″ are —H.

### Claim 3 (depends on 2)

The method of claim 2, wherein A and A′ are both —O—, R6 is —OMe, and G is —CH—.

### Claim 4 (depends on 3)

The method of claim 3, wherein L′ is represented by the following formula:
—W′—Rx—V—Ry-J,
wherein:
W′ and V are the same or different, and are each independently absent, or selected from —CReRe′—, —O—, —O—C(═O)—, —C(═O)—O—, —S—, —SO—, —SO2—, —CH2—S—, —CH2O—, —CH2NRe—, —O—(C═O)O—, —O—(C═O)N(Re)—, —N(Re)—, —N(Re)—C(═O)—, —C(═O)—N(Re)—, —N(Re)—C(═O)O—, —N(C(═O)Re)C(═O)—, —N(C(═O)Re)—, —(O—CH2—CH2)n—, —SS—, or —C(═O)—, or an amino acid, or a peptide having 2 to 8 amino acids;
Rx and Ry are the same or different, and are each independently absent or an optionally substituted linear, branched or cyclic alkyl, alkenyl, or alkynyl having 1 to 10 carbon atoms, an aryl bearing 6 to 10 carbon atoms or a 3- to 8-membered heterocyclic ring bearing 1 to 3 heteroatoms selected from O, N or S;
Re and Re′ are the same or different, and are selected from —H, a linear, branched or cyclic alkyl, alkenyl, or alkynyl having 1 to 10 carbon atoms or —(CH2—CH2—O)n—Rk, wherein Rk is a —H, a linear, branched cyclic alkyl having 1 to 6 carbon atoms, optionally bearing a secondary amino (—NHR101) or tertiary amino (—NR101R102) group or a 5- or 6-membered nitrogen containing heterocycle, such as piperidine or morpholine, wherein R101 and R102 are each independently a linear, branched, or cyclic alkyl, alkenyl or alkynyl having 1 to 10 carbon atoms; preferably, R101 and R102 are each independently a linear or branched alkyl having 1 to 6 carbon atoms;
n is an integer from 1 to 24; and
J is covalently linked to the CBA, and is selected from a succinimide, a acetamido, —S—, —SS—, —CH2S—, —CH(Me)S—, —C(Me)2S—, —NRc1—, —CH2NRc1—, —NRc1N—, and —C(═O)—, wherein Rc1 is —H or an optionally substituted linear or branched alkyl having 1 to 4 carbon atoms.

### Claim 5 (depends on 4)

The method of claim 4, wherein:
W′ is —O—, —N(Re)— or —N(Re)—C(═O)—;
Re is —H, -Me, or —(CH2—CH2—O)n-Me;
n is an integer from 2 to 6;
Rx is linear or branched alkyl bearing 1 to 6 carbon atoms;
V and Ry are absent; and
J is —C(═O)—.

### Claim 6 (depends on 1)

The method of claim 1, wherein the compound is represented by the following formula:
or a pharmaceutically acceptable salt thereof, wherein:
W′ is absent, or selected from —O—, —N(Re)—, —N(Re)—C(═O)—, —N(C(═O)Re)—, —S—, —CH2—S—, or —CH2NRe—;
Rx is absent or selected from a linear, branched or cyclic alkyl having 1 to 10 carbon atoms;
Re is —H, a linear, branched or cyclic alkyl, alkenyl or alkynyl having 1 to 10 carbon atoms or —(CH2—CH2—O)n—Rk, wherein Rk is a —H, a linear, branched cyclic alkyl having 1 to 6 carbon atoms, optionally bearing a secondary amino (—NHR101) or tertiary amino (—NR101R102) group or a 5 or 6-membered nitrogen containing heterocycle, such as piperidine or morpholine, wherein R101 and R102 are each independently a linear, branched, or cyclic alkyl, alkenyl or alkynyl having 1 to 10 carbon atoms;
n is an integer from 2 to 6;
Zs is linked to the CBA, and is selected from:
a bond;
wherein:
q is an integer from 1 to 5; and,
M is —H or a cation, such as Na+ or K+.

### Claim 7 (depends on 6)

The method of claim 6, wherein W′ is —N(Re)—.

### Claim 8 (depends on 7)

The method of claim 7, wherein Re is —(CH2—CH2—O)n—Rk, wherein Rk is a —H, a linear, branched cyclic alkyl having 1 to 6 carbon atoms.

### Claim 9 (depends on 8)

The method of claim 8, wherein Rx is a linear or branched alkyl having 1 to 6 carbon atoms.

### Claim 10 (depends on 9)

The method of claim 9, wherein Rx is —(CH2)p—(CRfRg)—, wherein Rf and Rg are each independently selected from H or a linear or branched alkyl having 1 to 4 carbon atoms; and p is 0, 1, 2 or 3.

### Claim 11 (depends on 10)

The method of claim 10, wherein Rf and Rg are the same or different, and are selected from H and Me; and p is 1.

### Claim 12 (depends on 6)

The method of claim 6, wherein:
the double line between N and C represents a single bond or a double bond, provided that when it is a double bond X is absent and Y is —H, and when it is a single bond, X is —H; Y is —OH or —SO3M;
M is —H or a pharmaceutically acceptable cation;
X′ and Y′ are both —H;
A and A′ are both —O—;
R6 is —OMe; and
Rx is a linear or branched alkyl having 1 to 6 carbon atoms.

### Claim 13 (depends on 1)

The method of claim 1, wherein the conjugate is represented by the following formula:
or a pharmaceutically acceptable salt thereof,
wherein r is an integer from 1 to 10, Y is —SO3M, and M is —H or a pharmaceutically acceptable cation.

### Claim 14 (depends on 13)

The method of claim 13, wherein the conjugate is represented by the following formula:
or a pharmaceutically acceptable salt thereof.

### Claim 15 (depends on 14)

The method of claim 14, wherein M is —H, Na+ or K+.

### Claim 16 (depends on 14)

The method of claim 14, wherein the antibody is huMy9-6.

### Claim 17 (depends on 1)

The method of claim 1, wherein the conjugate comprises 1-10 cytotoxic compounds, each cytotoxic compound comprising the linking group linking the cytotoxic compound to the CBA, and each cytotoxic compound on the conjugate is the same.

### Claim 18 (depends on 1)

The method of claim 1, wherein the cell-binding agent binds to target cells selected from tumor cells, virus infected cells, microorganism infected cells, parasite infected cells, autoimmune cells, activated cells, myeloid cells, activated T-cells, B cells, or melanocytes; cells expressing the CD4, CD6, CD19, CD20, CD22, CD30, CD33, CD37, CD38, CD40, CD44, CD56, EpCAM, CanAg, CALLA, or Her-2 antigens; Her-3 antigens; or cells expressing insulin growth factor receptor, epidermal growth factor receptor, and folate receptor.

### Claim 19 (depends on 1)

The method of claim 1, wherein the cell-binding agent is an antibody, a single chain antibody, an antibody fragment that specifically binds to the target cell, a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment that specifically binds to a target cell, a chimeric antibody, a chimeric antibody fragment that specifically binds to the target cell, a domain antibody, a domain antibody fragment that specifically binds to the target cell, a lymphokine, a hormone, a vitamin, a growth factor, a colony stimulating factor, or a nutrient-transport molecule.

### Claim 20 (depends on 1)

The method of claim 1, wherein the antibody is a resurfaced antibody, a resurfaced single chain antibody, or a resurfaced antibody fragment.

### Claim 21 (depends on 1)

The method of claim 1, wherein the antibody is a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment thereof.

### Claim 22 (depends on 1)

The method of claim 1, wherein the antibody is a humanized antibody, a humanized single chain antibody, or a humanized antibody fragment.

### Claim 23 (depends on 1)

The method of claim 1, wherein the cancer is acute myelogenous leukemia (AML).

### Claim 24 (depends on 23)

The method of claim 23, wherein the conjugate is represented by the following formula:
or a pharmaceutically acceptable salt thereof.

### Claim 25 (depends on 24)

The method of claim 24, wherein the antibody is huMy9-6.