Quinone-containing conjugates
Conjugates comprise a drug or probe, a cell binding agent, and a quinone-containing linker. The quinone-containing linker may be reduced intracellularly to trigger release of the drug or probe. These conjugates may selectively deliver a drug or probe to a site of action of interest for local release of the active drug or probe.
1. A conjugate of formula (I), or a salt thereof,
Cb-(G) p (I)
wherein
Cb is a cell binding moiety;
p is a number from 1 to 20;
G is formula (II), (III), or (IV)
R 9 and R 10 are each independently selected from C 1-4 alkyl;
R 11 , R 12 , and R 13 are each independently selected from the group consisting of H, C 1-4 alkyl, C 1-4 alkoxy, bromo, chloro, and amino, or R 11 and R 12 in formula (II) or (IV), together with the atoms to which they are attached form a fused phenyl ring;
L 1 is a first linker moiety;
L 2 is a second linker moiety;
G 1 is -A 2 ,
G 2 is
X 1 and X 2 are each an antibody-linking moiety;
Y is O or NR 15 ;
R 14 and R 5 are each independently H, C 1-30 alkyl optionally substituted with 1-8 halogens, —C 1-30 alkylene-OH, —C 1-30 alkylene-C 1-4 alkoxy, —C 1-30 alkylene-COOH, or —C 1-30 alkylene-amido;
R 16 , at each occurrence, is independently H, halogen, CH 3 , OCH 3 , or NO 2 ;
R 17 , at each occurrence, is independently H or C 1-4 alkyl or both R 17 together with the carbon to which they are attached form a cycloalkyl ring having from 3-7 carbons;
m is an integer from 0-2;
n is an integer from 0-2;
A 1 is a payload moiety bonded through a substitutable oxygen or sulfur atom; and
A 2 is a payload moiety bonded through a substitutable oxygen, sulfur, or nitrogen atom.
2. The conjugate of claim 1 , or a salt thereof, wherein:
X 1 or X 2 is
—C(O)CH 2 —, —NHC(S)—, —NHC(O)—, —C(O)—, —CH(SO 3 H)—C(O)—,
—NH—, or —N(C 1-6 alkyl)-.
3. The conjugate of claim 1 , or a salt thereof, wherein G is formula (II) or (IV);
L 1 is L 1a or L 1a -L 1b , wherein L 1b is bonded to X 1 ;
L 1a is —C 1-12 alkylene-, -(C 2-6 alkylene-O) x —C 1-6 alkylene-, or C 3-8 cycloalkylene;
L 1b comprises one or more covalently bonded divalent members, the one or more divalent members being selected from the group consisting of —C 1-6 alkylene-, —C 2-6 alkylene-O-, C 3-8 cycloalkylene, —C(O)—, —O—, —S—, —S(O)—, —S(O) 2 —, —NR 20 —, —C(R 21 )═N—NH—, —CH(CO 2 H)—,
an amino acid moiety, and phenylene;
wherein the C 3-8 cycloalkylene and phenylene of L 1a and/or L 1b is optionally independently substituted with 1-4 substituents independently selected from the group consisting of C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, halo, cyano, and hydroxy;
R 20 and R 21 at each occurrence are independently hydrogen or C 1-4 alkyl; and
x is an integer from 1 to 20.
4. The conjugate of claim 3 , or a salt thereof, wherein
L 1b is —C(O)NR 20 -L 1c -, -cit-val-C(O)C 1-6 alkylene-, —NR 20 C(O)—, —NR 20 C(O)-L 1c -, —NR 20 C(O)O-L 1c -, -ala-val-C(O)C 1-6 alkylene-, -cit-val-C(O)O-C 2-6 alkylene-O-C 1-6 alkylene-, -ala-val-C(O)O-C 2-6 alkylene-(—C 1-6 alkylene-, —S—S-C 1-6 alkylene-, —NH—N═C(R 21 )-phenylene-O-C 1-6 alkylene-,
C 1-6 alkylene-, —C 2-6 alkylene-O—, or C 3-8 cycloalkylene, and
L 1c is —C 1-6 alkylene-, —C 1-6 alkylene-NR 20 C(O)—C 1-6 alkylene-S—S-C 1-6 alkylene-, —C 1-6 alkylene-NR 20 C(O)NH-N═C(R 21 )-phenylene-O-C 1-6 alkylene-, C 3-8 cycloalkylene, -(C 2-6 alkylene-O)—C 1-6 alkylene-, —C 1-6 alkylene-C 3-8 cycloalkylene-, —C 3-8 cycloalkylene-C 1-6 alkylene-, —C 1-6 alkylene-C 3-8 cycloalkylene-C 1-6 alkylene-, —C 1-6 alkylene-S—S-C 1-6 alkylene-, or
5. The conjugate of claim 3 , or a salt thereof, wherein
L 1a is -(CH 2 CH 2 O) 1-2 —CH 2 CH 2 —;
L 1b is —C(O)NR 20 -L 1c -, -ala-val-C(O)(CH 2 ) 5 —, -cit-val-C(O)(CH 2 ) 5 —, or
and
L 1c is —CH 2 CH 2 —, —CH 2 CH 2 —NR 20 C(O)—CH 2 CH 2 —S—S—CH(CH 3 )CH 2 CH 2 —, —CH 2 CH 2 —NR 20 C(O)NH—N═C(CH 3 )-1,4-phenylene-O—CH 2 CH 2 CH 2 —, or
6. The conjugate of claim 4 , or a salt thereof, wherein:
L 1b -X 1 is
—C(O)NR 20 —C 1-6 alkylene-NR 20 C(O)—C 1-6 alkylene-S—S—C 1-6 alkylene-C(O)—, —C(O)NR 20 —C 1-6 alkylene-NR 20 C(O)—C 1-6 alkylene-S—S—C 1-6 alkylene-CH(SO 3 H)—C(O)—, —C(O)NR 20 —C 1-6 alkylene-NR 20 C(O)NH—N═C(R 21 )-phenylene-O—C 1-6 alkylene-C(O)—, or
7. The conjugate of claim 3 , or a salt thereof, wherein L 1 -X 1 is:
8. The conjugate of claim 1 , or a salt thereof, wherein G is formula (III);
L 2 is L 2a or L 2a -L 2b , wherein L 2b is bonded to X 2 ;
L 2a is —C 2-6 alkylene-;
L 2b comprises one or more covalently bonded divalent members, the one or more divalent members being selected from the group consisting of —C 1-6 alkylene-, —C 2-6 alkylene-O—, C 3-8 cycloalkylene, —C(O)—, —O—, —S—, —S(O)—, —S(O) 2 —, —NR 30 —, —C(R 31 )═N—N—, —CH(CO 2 H)—,
an amino acid moiety, and phenylene;
wherein the C 3-8 cycloalkylene and phenylene of L 1b are optionally independently substituted with 1-4 substituents independently selected from the group consisting of C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, halo, cyano, and hydroxy; and
R 30 and R 31 at each occurrence are independently hydrogen or C 1-4 alkyl.
9. The conjugate of claim 8 , or a salt thereof, wherein:
L 2b is —C(O)NR 30 -L 2c -, -cit-val-C(O)C 1-6 alkylene-, —NR 30 C(O)—, —NR 30 C(O)-L 2c -, —NR 30 C(O)O-L 2c -, -ala-val-C(O)C 1-6 alkylene-, -cit-val-C(O)O-C 2-6 alkylene-O-C 1-6 alkylene-, -ala-val-C(O)O-C 2-6 alkylene-O-C 1-6 alkylene-, —S—S-C 1-6 alkylene-, —NH—N═C(R 31 )-phenylene-O-C 1-6 alkylene-,
C 1-6 alkylene-, —C 2-6 alkylene-O—, or C 3-8 cycloalkylene, and
L 2c is —C 1-6 alkylene-, —C 1-6 alkylene-NR 30 C(O)—C 1-6 alkylene-S—S-C 1-6 alkylene-, —C 1-6 alkylene-NR 30 C(O)NH—N═C(R 31 )-phenylene-O-C 1-6 alkylene-, —C 3-8 cycloalkylene, —(C 2-6 alkylene-O)—C 1-6 alkylene-, —C 1-6 alkylene-C 3-8 cycloalkylene-, —C 3-8 cycloalkylene-C 1-6 alkylene-, —C 1-6 alkylene-C 3-8 cycloalkylene-C 1-6 alkylene-, —C 1-6 alkylene-S—S-C 1-6 alkylene, or
10. The conjugate of claim 9 , or a salt thereof, wherein:
L 2b -X 2 is
—C(O)NR 30 —C 1-6 alkylene-NR 30 C(O)—C 1-6 alkylene-S—S—C 1-6 alkylene-C(O)—, —C(O)NR 30 —C 1-6 alkylene-NR 30 C(O)—C 1-6 alkylene-S—S—C 1-6 alkylene-CH(SO 3 H)—C(O)—, —C(O)NR 30 —C 1-6 alkylene-NR 30 C(O)NH—N═C(R 31 )-phenylene-O—C 1-6 alkylene-C(O)—, or
11. The conjugate of claim 1 , or a pharmaceutically acceptable salt thereof, wherein A 1 and A 2 are a cytotoxic moiety.
12. The conjugate of claim 1 , or a pharmaceutically acceptable salt thereof, wherein A 1 and A 2 are an anticancer drug moiety derived from a duocarmycin, a pyrrolobenzodiazepine, an auristatin, a doxorubicin, SN-38, or a maytansinoid.
13. The conjugate of claim 1 , or a salt thereof, wherein the cell binding moiety is an antibody moiety, Ab.
14. The conjugate of claim 1 , or a salt thereof, wherein the cell binding moiety is a small molecule drug moiety, and p is 1.
15. A pharmaceutical composition comprising the conjugate of claim 1 , or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
16. A method of treating a proliferative disorder comprising administering a therapeutically effective amount of the conjugate of claim 1 , or a pharmaceutically acceptable salt thereof, to a patient in need thereof.
17. A kit comprising the conjugate of claim 1 , or a pharmaceutically acceptable salt thereof, and instructions for use.
18. The conjugate of claim 1 , or a salt thereof, wherein A 1 and A 2 are a reporter moiety.
19. The conjugate of claim 18 , or a salt thereof, wherein the reporter moiety comprises a bioluminescent reporter moiety, fluorescent reporter moiety, or colorimetric reporter moiety.
20. The conjugate of claim 19 , or a salt thereof, wherein the bioluminescent reporter moiety comprises a substrate for a luciferase.
21. The conjugate of claim 19 , or a salt thereof, wherein the fluorescent reporter moiety comprises a fluorophore.
22. The conjugate of claim 21 , or a salt thereof, wherein the fluorophore is coumarin, R110, fluorescein, DDAO, resorufin, cresyl violet, silyl xanthene, or carbopyronine.
23. A method for evaluating cellular uptake of an agent, the method comprising:
a) contacting a sample with a conjugate of claim 18 , or a salt thereof, wherein the sample comprises a cell; and
b) detecting light emission,
whereby the detection of light emission indicates cellular uptake of the agent.
24. The method of claim 23 , wherein the cellular uptake of the agent results in the reduction of the conjugate and the generation of a released reporter moiety.
25. The method of claim 24 , wherein the released reporter moiety comprises a fluorescent reporter moiety and wherein detecting light emission comprises exposing the sample to a wavelength of light and detecting fluorescence level in the sample, wherein an increase in fluorescence or a change in fluorescence wavelength as compared to the fluorescence or fluorescence wavelength of a control sample indicates cellular uptake of the agent.
26. The method of claim 25 , wherein the control sample is a sample that is not contacted with a labeled agent or a sample that is incompetent for uptake of the agent.
27. The method of claim 24 , wherein the reporter moiety comprises a bioluminescent reporter moiety.
28. The method of claim 27 , wherein the bioluminescent reporter moiety comprises a substrate for a luciferase and detecting light emission comprises detecting the luminescence produced by the luciferase utilizing the released reporter moiety.
29. The method of claim 27 , wherein the cell comprises a luciferase.
30. The method of claim 27 , further comprising contacting the sample with a luciferase, and wherein detecting light emission comprises detecting the luminescence produced by the luciferase utilizing the released reporter moiety.
31. The method of claim 23 , wherein the light emission is detected inside or outside the cell.