Antibody conjugates comprising toll-like receptor agonist
Provided herein are antibody conjugates comprising toll-like receptor agonists and the use of such conjugates for the treatment of cancer. In some embodiments, the conjugates comprise anti-HER2 antibodies.
1. A conjugate of Formula (II), or pharmaceutically acceptable salt thereof:
wherein:
R 50 is
where the * indicates the point of attachment to Ab;
Ab is an antibody or antigen binding fragment thereof that specifically binds to human HER2, wherein Ab is selected from any of the following:
(a) an antibody that comprises:
a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 1;
a heavy chain complementary determining region 2 (HCDR2) comprising the amino acid sequence of SEQ ID NO: 2;
a heavy chain complementary determining region 3 (HCDR3) comprising the amino acid sequence of SEQ ID NO: 3;
a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 11;
a light chain complementary determining region 2 (LCDR2) comprising the amino acid sequence of SEQ ID NO: 12; and
a light chain complementary determining region 3 (LCDR3) comprising the amino acid sequence of SEQ ID NO: 13;
(b) an antibody that comprises:
a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4;
a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5;
a HCDR3 comprising the amino acid sequence of SEQ ID NO: 3;
a LCDR1 comprising the amino acid sequence of SEQ ID NO: 14;
a LCDR2 comprising the amino acid sequence of SEQ ID NO: 15; and
a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16;
(c) an antibody that comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 17;
(d) an antibody that comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9, and a light chain comprising the amino acid sequence of SEQ ID NO: 19;
(e) an antibody that comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 21, and a light chain comprising the amino acid sequence of SEQ ID NO: 19;
(f) an antibody that comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 23, and a light chain comprising the amino acid sequence of SEQ ID NO: 19;
(g) an antibody that comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 32, and a light chain comprising the amino acid sequence of SEQ ID NO: 19; or
(h) pertuzumab;
and wherein
R 1 is —NHR 2 or —NHCHR 2 R 3 ;
R 2 is —C 3 -C 6 alkyl or —C 4 -C 6 alkyl;
R 3 is L 1 OH;
L 1 is —(CH 2 ) m —;
L 2 is —(CH 2 ) n —, —((CH 2 ) n O) t (CH 2 ) n —, —(CH 2 ) n X 1 (CH 2 ) n —, —(CH 2 ) n NHC(═O)(CH 2 ) n —, —(CH 2 ) n NHC(═O)(CH 2 ) n C(═O)NH(CH 2 ) n —, —((CH 2 ) n O) t (CH 2 ) n NHC(═O)(CH 2 ) n , —C(═O)(CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n C(═O)NH(CH 2 ) n —, —C(═O)NH((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)X 2 X 3 C(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)X 2 X 3 C(═O)(CH 2 ) n —, —C(═O)X 2 C(═O)(CH 2 ) n NHC(═O)(CH 2 ) n —, —C(═O)X 2 C(═O)(CH 2 ) n NHC(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)(CH 2 ) n C(R 7 ) 2 —, —C(═O)(CH 2 ) n C(R 7 ) 2 SS(CH 2 ) n NHC(═O)(CH 2 ) n —, —(CH 2 ) n X 2 C(═O)(CH 2 ) n NHC(═O)((CH 2 ) n O) t (CH 2 ) n — or —C(═O)(CH 2 ) n C(═O)NH(CH 2 ) n ;
R 40 is
—S—, —NHC(═O)CH 2 —, —S(═O) 2 CH 2 CH 2 —, —(CH 2 ) 2 S(═O) 2 CH 2 CH 2 —, —NHS(═O) 2 CH 2 CH 2 , —NHC(═O)CH 2 CH 2 —, —CH 2 NHCH 2 CH 2 —, —NHCH 2 CH 2 —,
X 1 is
X 2 is
X 3 is
each R 7 is independently selected from H and C 1 -C 6 alkyl;
each R 8 is independently selected from H, C 1 -C 6 alkyl, F, Cl, and —OH;
each R 9 is independently selected from H, C 1 -C 6 alkyl, F, Cl, —NH 2 , —OCH 3 , —OCH 2 CH 3 , —N(CH 3 ) 2 , —CN, —NO 2 and —OH;
each R 10 is independently selected from H, C 1-6 alkyl, fluoro, benzyloxy substituted with —C(═O)OH, benzyl substituted with —C(═O)OH, C 1-4 alkoxy substituted with —C(═O)OH and C 1-4 alkyl substituted with —C(═O)OH;
R 12 is H, methyl or phenyl;
each m is independently selected from 1, 2, 3, and 4;
each n is independently selected from 1, 2, 3, and 4;
each t is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 and 18, and
y is an integer from 1 to 16.
2. The conjugate of claim 1 , wherein Ab comprises a modified Fc region compared to a wild-type constant region.
3. The conjugate of claim 1 , wherein Ab comprises cysteine at one or more of the following positions (all positions by EU numbering):
(a) positions 152, 360 and 375 of the antibody heavy chain, and
(b) positions 107, 159, and 165 of the antibody light chain.
4. The conjugate of claim 1 , wherein Ab comprises cysteines at positions 152 and 375 of the antibody heavy chains (all positions by EU numbering).
5. The conjugate of claim 1 , wherein the antibody conjugate of Formula (II) comprises the structure of Formula (IIa):
wherein:
R 1 is —NHR 2 ;
R 2 is —C 4 -C 6 alkyl;
L 2 is —(CH 2 ) n —, —((CH 2 ) n O) t (CH 2 ) n —, —(CH 2 ) n X 1 (CH 2 ) n —, —C(═O)(CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)NH((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)X 2 X 3 C(═O)((CH 2 ) n O) t (CH 2 ) n — or —C(═O)X 2 C(═O)(CH 2 ) n NHC(═O)(CH 2 ) n —;
R 40 is
X 1 is
X 3 is
each n is independently selected from 1, 2, 3, and 4;
each t is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 and 18, and
y is an integer from 1 to 16.
6. The conjugate of claim 1 , wherein
R 1 is —NHR 2 ;
R 2 is —C 4 -C 6 alkyl;
L 2 is —(CH 2 ) n — or —C(═O)(CH 2 ) n ;
R 40 is
and
each n is independently selected from 1, 2, 3, and 4, and
y is an integer from 1 to 16.
7. The conjugate of claim 1 , wherein the conjugate has a hydrophobicity index of 0.8 or greater, as determined by hydrophobic interaction chromatography.
8. The conjugate of claim 1 , wherein the Ab is trastuzumab, or margetuximab.
9. A pharmaceutical composition comprising one or more conjugates of claim 1 and a pharmaceutically acceptable carrier.
10. A conjugate comprising any of the following formulas:
wherein Ab is an antibody or antigen binding fragment thereof that specifically binds to human HER2, and y is an integer from 1 to 4, wherein Ab is selected from any of the following:
(a) an antibody that comprises:
a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 1;
a heavy chain complementary determining region 2 (HCDR2) comprising the amino acid sequence of SEQ ID NO: 2;
a heavy chain complementary determining region 3 (HCDR3) comprising the amino acid sequence of SEQ ID NO: 3;
a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 11;
a light chain complementary determining region 2 (LCDR2) comprising the amino acid sequence of SEQ ID NO: 12; and
a light chain complementary determining region 3 (LCDR3) comprising the amino acid sequence of SEQ ID NO: 13;
(b) an antibody that comprises:
a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4;
a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5;
a HCDR3 comprising the amino acid sequence of SEQ ID NO: 3;
a LCDR1 comprising the amino acid sequence of SEQ ID NO: 14;
a LCDR2 comprising the amino acid sequence of SEQ ID NO: 15; and
a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16;
(c) an antibody that comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 17;
(d) an antibody that comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9, and a light chain comprising the amino acid sequence of SEQ ID NO: 19;
(e) an antibody that comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 21, and a light chain comprising the amino acid sequence of SEQ ID NO: 19;
(f) an antibody that comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 23, and a light chain comprising the amino acid sequence of SEQ ID NO: 19;
(g) an antibody that comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 32, and a light chain comprising the amino acid sequence of SEQ ID NO: 19; or
(h) pertuzumab.
11. The conjugate of claim 10 , wherein Ab comprises cysteine at one or more of the following positions (all positions by EU numbering):
(a) positions 152, 360 and 375 of the antibody heavy chain, and
(b) positions 107, 159, and 165 of the antibody light chain.
12. The conjugate of claim 10 , wherein Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9, and a light chain comprising the amino acid sequence of SEQ ID NO: 19.
13. The conjugate of claim 11 , wherein the compound is attached to cysteines at positions 152 and 375 of the antibody heavy chain (all positions by EU numbering).
14. The conjugate of claim 10 , wherein y is about 3 to 4.
15. The conjugate of claim 10 , wherein the conjugate has a hydrophobicity index of 0.8 or greater, as determined by hydrophobic interaction chromatography.
16. The conjugate of claim 10 , wherein the Ab is trastuzumab, or margetuximab.
17. A pharmaceutical composition comprising one or more conjugates of claim 10 and a pharmaceutically acceptable carrier.
18. A method of treating a HER2-positive cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the conjugate of claim 1 , wherein the HER2-positive cancer is selected from gastric cancer, breast cancer, and ovarian cancer, and wherein the HER2-positive cancer has an IHC score of 2+ or 3+.
19. The method of claim 18 , wherein the conjugate is capable of suppressing the HER2-positive cancer for a sustained period and/or reducing recurrence of the HER2-positive cancer, when compared to an anti-HER2 antibody alone.
20. The method of claim 18 , wherein the conjugate is administered to the subject intravenously, intratumorally, or subcutaneously.
21. The method of claim 18 , wherein the conjugate is administered at a dose of about 0.01-20 mg per kg of body weight.
22. The method of claim 18 further comprising administering a second agent to the subject.
23. A method of treating a HER2-positive cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the conjugate of claim 10 , wherein the HER2-positive cancer is selected from gastric cancer, breast cancer, and ovarian cancer, and wherein the HER2-positive cancer has an IHC score of 2+ or 3+.
24. The method of claim 23 , wherein the conjugate is capable of suppressing the HER2-positive cancer for a sustained period and/or reducing recurrence of the HER2-positive cancer, when compared to an anti-HER2 antibody alone.
25. The method of claim 23 , wherein the conjugate is administered to the subject intravenously, intratumorally, or subcutaneously.
26. The method of claim 23 , wherein the conjugate is administered at a dose of about 0.01-20 mg per kg of body weight.
27. The method of claim 23 further comprising administering a second agent to the subject.
28. A compound having the structure of Formula (I), or the pharmaceutically acceptable salts thereof:
wherein:
R D is
and R E is H;
R 1 is —NHR 2 or —NHCHR 2 R 3 ;
R 2 is —C 3 -C 6 alkyl or —C 4 -C 6 alkyl;
R 3 is L 1 OH;
L 1 is —(CH 2 ) m —;
L 2 is —(CH 2 ) n —, —((CH 2 ) n O) t (CH 2 ) n —, —(CH 2 ) n X 1 (CH 2 ) n —, —(CH 2 ) n NHC(═O)(CH 2 ) n —, —(CH 2 ) n NHC(═O)(CH 2 ) n C(═O)NH(CH 2 ) n —, —((CH 2 ) n O) t (CH 2 ) n NHC(═O)(CH 2 ) n , —C(═O)(CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n C(═O)NH(CH 2 ) n —, —C(═O)NH((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)X 2 X 3 C(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)X 2 X 3 C(═O)(CH 2 ) n —, —C(═O)X 2 C(═O)(CH 2 ) n NHC(═O)(CH 2 ) n —, —C(═O)X 2 C(═O)(CH 2 ) n NHC(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)(CH 2 ) n C(R 7 ) 2 —, —C(═O)(CH 2 ) n C(R 7 ) 2 SS(CH 2 ) n NHC(═O)(CH 2 ) n —, —(CH 2 ) n X 2 C(═O)(CH 2 ) n NHC(═O)((CH 2 ) n O) t (CH 2 ) n — or —C(═O)(CH 2 ) n C(═O)NH(CH 2 ) n ;
R 4 is
—ONH 2 , —NH 2 ,
—N 3 , - -C≡CH, —NHC(═O)CH═CH 2 , —SH, —SR 7 , —OH —SSR 6 , —S(═O) 2 (CH═CH 2 ), —(CH 2 ) 2 S(═O) 2 (CH═CH 2 ), —NHS(═O) 2 (CH═CH 2 ), —NHC(═O)CH 2 Br, —NHC(═O)CH 2 I, —C(O)NHNH 2 ,
—CO 2 H, —C(O)NHNH 2 ,
R 5 is
X 1 is
X 2 is
X 3 is
R 6 is 2-pyridyl or 4-pyridyl;
each R 7 is independently selected from H and C 1 -C 6 alkyl;
each R 8 is independently selected from H, C 1 -C 6 alkyl, F, Cl, and —OH;
each R 9 is independently selected from H, C 1 -C 6 alkyl, F, Cl, —NH 2 , —OCH 3 , —OCH 2 CH 3 , —N(CH 3 ) 2 , —CN, —NO 2 and —OH;
each R 10 is independently selected from H, C 1-6 alkyl, fluoro, benzyloxy substituted with —C(═O)OH, benzyl substituted with —C(═O)OH, C 1-4 alkoxy substituted with —C(═O)OH and C 1-4 alkyl substituted with —C(═O)OH;
each m is independently selected from 1, 2, 3, and 4;
each n is independently selected from 1, 2, 3, and 4;
and
each t is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 and 18.