WO2020056194A1 - Benzazepine compounds, conjugates, and uses thereof - Google Patents

Benzazepine compounds, conjugates, and uses thereof Download PDF

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Publication number
WO2020056194A1
WO2020056194A1 PCT/US2019/050896 US2019050896W WO2020056194A1 WO 2020056194 A1 WO2020056194 A1 WO 2020056194A1 US 2019050896 W US2019050896 W US 2019050896W WO 2020056194 A1 WO2020056194 A1 WO 2020056194A1
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compound
salt
optionally substituted
alkyl
independently selected
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PCT/US2019/050896
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French (fr)
Inventor
Sean Wesley Smith
Craig Alan Coburn
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Silverback Therapeutics, Inc.
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Publication of WO2020056194A1 publication Critical patent/WO2020056194A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • cancer One of the leading causes of death in the United States is cancer.
  • the conventional methods of cancer treatment like chemotherapy, surgery, or radiation therapy, tend to be either highly toxic or nonspecific to a cancer, or both, resulting in limited efficacy and harmful side effects.
  • the immune system has the potential to be a powerful, specific tool in fighting cancers. In many cases tumors can specifically express genes whose products are required for inducing or maintaining the malignant state. These proteins may serve as antigen markers for the development and establishment of more specific anti-cancer immune response. The boosting of this specific immune response has the potential to be a powerful anti-cancer treatment that can be more effective than conventional methods of cancer treatment and can have fewer side effects.
  • the present disclosure provides a compound represented by the structure of Formula (II A):
  • L 10 is -X 10 -;
  • L 2 is selected from -X 2 -, -X 2 -C I-6 alkylene-X 2 -, -X 2 -C 2-6 alkenyl ene-X 2 -, and - X 2 -C 2-6 alkynylene-X 2 -, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R 12 ;
  • X 10 is selected from -C(O)-, and -C(0)N(R 10 )-*, wherein * represents where X 10 is bound to R 5 ;
  • X 2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R 10 )-, -C(O)-,
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
  • the compound of Formula (IIA) is represented by Formula (IIB):
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen, -OH, -OR 10 , -N0 2 , -CN, and C1-10 alkyl.
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen, -OH, -OR 10 , -N0 2 , -CN, and C1-10 alkyl.
  • R 21 , R 22 , and R 23 may be each hydrogen. In certain embodiments, R 21 is halogen. In certain embodiments, R 21 is hydrogen. In certain embodiments, R 21 is -OR 10 . For example, R 21 may be - OCH3.
  • R 24 and R 25 are independently selected from hydrogen, halogen, -OH, -N0 2 , -CN, and C1-10 alkyl, or R 24 and R 25 taken together form an optionally substituted saturated C3-7 carbocycle. In certain embodiments, R 24 and R 25 are each hydrogen.
  • R 2 is an amine masking group that is enzymatically-labile.
  • R 2 is represented by the formula:
  • R 101 is selected from an amino acid, a peptide, -0-(Ci-C 6 alkyl) and -Ci-C 6 alkyl, wherein alkyl of -0-(Ci-C 6 alkyl) and -Ci-C 6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -N(R 10 ) 2 , -C(0)R 10 , - C(O)N(R 10 ) 2 , -N0 2 , -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle; and
  • R 101 of R 2 is a peptide selected from a dipeptide, tripeptide and tetrapeptide.
  • L 10 is selected from -C(0)N(R 10 )-*.
  • R 10 of -C(0)N(R 10 )-* is selected from hydrogen and Ci- 6 alkyl.
  • L 10 may be -C(0)NH-*.
  • R 5 is an optionally substituted bicyclic carbocycle. In certain embodiments, R 5 is an optionally
  • R 5 may be selected from:
  • R 5 is selected from any one of which is optionally substituted.
  • the R 5 is selected from any one of which is optionally substituted.
  • the R 5 is selected from any one of which is optionally substituted.
  • R 5 is an optionally substituted unsaturated C 4-8 carbocycle. In certain embodiments, R 5 is an optionally substituted unsaturated C 4-6 carbocycle. In certain embodiments, R 5 is an optionally substituted unsaturated C 4-6 carbocycle with one or more substituents independently selected from optionally substituted C 3 -i 2 carbocycle, and optionally substituted 3- to l2-membered heterocycle.
  • R 5 may be an optionally substituted unsaturated C 4-6 carbocycle with one or more substituents independently selected from optionally substituted phenyl, optionally substituted 3- to 12- heterocycle, optionally substituted Ci-io alkyl, optionally substituted C 2-i o alkenyl, and halogen.
  • R 5 is selected from an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle.
  • R 5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle with one or more substituents independently selected from -C(0)OR 10 , -N(R 10 ) 2 , - OR 10 , and optionally substituted Ci-io alkyl.
  • R 5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle substituted with -C(0)OR 10 .
  • R 5 is an optionally substituted fused 6-6 bicyclic heterocycle.
  • the fused 6-6 bicyclic heterocycle may be an optionally substituted pyridine-piperidine.
  • L 10 is bound to a carbon atom of the pyridine of the fused pyridine- piperidine.
  • R 5 is selected from tetrahydroquinoline,
  • R 5 may be an optionally substituted
  • R 5 is selected from
  • R 5 is not substituted.
  • L 2 is selected from -C(0)-, and -C(0)NR 10 -.
  • L 2 is -C(0)-.
  • L 2 is selected from -C(0)NR 10 -.
  • R 10 of -C(0)NR 10 - may be selected from hydrogen and Ci -6 alkyl.
  • L 2 may be -C(0)NH-.
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ; Ci-io alkyl, C 2 -io alkenyl, C 2 -io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , - N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -OC(0)R 10 , -N0
  • R 4 is - N(R 10 ) 2 .
  • R 10 of -N(R 10 ) 2 may be independently selected at each occurrence from optionally substituted Ci -6 alkyl.
  • R 10 of -N(R 10 ) 2 is independently selected at each occurrence from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted.
  • R 4 may certain embodiments
  • R 12 is
  • the present disclosure provides a compound represented by the structure of F ormul a (HI A) :
  • L 11 is -X 11 -;
  • L 2 is selected from -X 2 -, -X 2 -C I-6 alkylene-X 2 -, -X 2 -C 2-6 alkenyl ene-X 2 -, and - X 2 -C 2-6 alkynylene-X 2 -, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R 12 ;
  • X 11 is selected from -C(O)- and -C(0)N(R 10 )-*, wherein * represents where X 11 is bound to R 6 ;
  • X 2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R 10 )-, -C(O)-,
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , - S(0)R 10 , and -S(0) 2 R 10 ; Ci - l o alkyl, C 2-i o alkenyl, C 2-i o alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR 10 ,
  • R 7 is selected from -C(0)NHNH 2 , -C(0)NH-Ci -3 alkylene-NH(R 10 ), -C(0)CH 3 , - Ci- 3 alkyl ene-NHC(0)0R 11 , -Ci -3 alkylene-NHC(0)R 10 , -Ci -3 alkylene-NHC(0)NHR 10 , -Ci- 3 alkylene-NHC(0)-Ci -3 alkylene-R 10 , and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R 12 ;
  • R 11 is selected from C 3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from R 12 ;
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
  • the compound of Formula (IIIA) is represented by Formula (IIIB):
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen,
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen, -OH, -N0 2 , -CN, and C 1-10 alkyl.
  • R 20 , R 21 , R 22 , and R 23 are each hydrogen.
  • R 24 and R 25 are independently selected from hydrogen, halogen, -OH, -N0 2 , -CN, and C 1-10 alkyl, or R 24 and R 25 taken together form an optionally substituted saturated C 3-7 carbocycle.
  • R 24 and R 25 are each hydrogen.
  • R 24 and R 25 taken together form an optionally substituted saturated C 3-5 carbocycle.
  • R 2 is represented by the formula:
  • R 101 is selected from an amino acid, a peptide, -0-(Ci-C 6 alkyl) and -Ci-C 6 alkyl, wherein alkyl of -0-(Ci-C 6 alkyl) and -Ci-C 6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -N(R 10 ) 2 , -C(0)R 10 , - C(O)N(R 10 ) 2 , -N0 2 , -CN, C 3-13 carbocycle, and 3- to l2-membered heterocycle; and
  • R 101 of R 2 is a peptide selected from a dipeptide, tripeptide and tetrapeptide.
  • L 11 is selected from
  • R 10 of -C(0)N(R 10 )-* is selected from hydrogen and Ci -6 alkyl.
  • L 11 may be -C(0)NH-*.
  • R 6 is phenyl substituted with R 7 and R 6 is further optionally substituted with one or more additional substituents independently selected from R 12 .
  • R 6 is selected from phenyl substituted with one or more substituents independently selected from -C(0)NHNH 2 , -C(0)NH- Ci.3alkylene-NH(R 10 ), -Ci.
  • R 6 may be selected from:
  • R 6 is selected from a 5- and 6-membered heteroaryl substituted with one or more substituents independently selected from R 7 , and R 6 is further optionally substituted with one or more additional substituents selected from R 12 .
  • R 6 is selected from 5- and 6-membered heteroaryl substituted with one or more substituents independently selected from -C(0)CH 3 , -Ci-3alkylene- NHC(0)OR 10 , -Ci- 3 alkylene-NHC(0)R 10 , -Ci- 3 alkylene-NHC(0)NHR 10 , and -Ci -3 alkylene- NHC(O) -Ci-3alkylene-(R 10 ); and 3- to l2-membered heterocycle, which is optionally substituted with one or more substituents selected from -OH, -N(R 10 ) 2 , -NHC(0)(R 10 ), -NHC(0)0(R 10 ), - NHC(O)N(R 10 ) 2 , -C(0)R 10 , -C(O)N(R 10 ) 2 , -C(0) 2 R 10 , and -Ci-3alkylene-(R 10 ), and R 6 is optionally further substituted
  • R 6 may be selected from substituted pyridine, pyrazine, pyrimidine, pyridazine, furan, pyran, oxazole, thiazole, imidazole, pyrazole, oxadiazole, oxathiazole, and triazole, and R 6 is optionally further substituted with one or more additional substituents independently selected from R 12 .
  • R 6 is substituted pyridine and R 6 is optionally further substituted with one or more additional substituents independently selected from R 12 .
  • R 6 may be selected from substituted pyridine, pyrazine, pyrimidine, pyridazine, furan, pyran, oxazole, thiazole, imidazole, pyrazole, oxadiazole, oxathiazole, and triazole, and R 6 is optionally further substituted with one or more additional substituents independently selected from R 12 .
  • R 6 may be selected from substituted pyr
  • R 6 is substituted pyridine, and wherein R 7 is -Ci -3 alkylene-NHC(0)-Ci -3 alkylene-R 1 °. In certain embodiments, R 7 is -Cialkylene-NHC(0)-Cialkylene-R 10 . In certain embodiments, R 7 is -Cialkylene-NHC(O)-
  • R 6 is selected from:
  • L 2 is selected from -C(O)-, and -C(0)NR 10 -.
  • L 2 is selected from -C(0)NR 10 -.
  • R 10 of - C(0)NR 10 - may be selected from hydrogen and Ci -6 alkyl.
  • L 2 may be -C(0)NH-.
  • L 2 is -C(O)-.
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ; Ci-io alkyl, C 2 -io alkenyl, C 2 -io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , - N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -OC(0)R 10 , -N0 2
  • R 4 is -N(R 10 ) 2.
  • R 10 of -N(R 10 ) 2 may be independently selected at each occurrence from optionally substituted Ci -6 alkyl.
  • R 10 of -N(R 10 ) 2 is independently selected at each occurrence from methyl, ethyl, propyl, and butyl, any of which are optionally substituted.
  • R 4 may be v .
  • -L 2 -R 4 is CHs .
  • the present disclosure provides a compound represented by the structure of Formula (IA):
  • L 1 is selected from -X 1 -, -X 2 -C I-6 alkyl ene-X 2 -Ci -6 alkylene-, -X 2 -C 2-6 alkenylene- X 2 -, and -X 2 -C 2-6 alkynyl ene-X 2 -, each of which is optionally substituted on alkylene, alkenylene or alkynyl ene with one or more R 12 ;
  • L 2 is selected from -X 2 -, -X 2 -C I-6 alkylene-X 2 -, -X 2 -C 2-6 alkenyl ene-X 2 -, and -X 2 -C 2-6 alkynyl ene-X 2 -, each of which is optionally substituted on alkylene, alkenylene or alkynylene with one or more R 12 ;
  • X 1 is selected from -S-*, -N(R 10 )-*, -C(0)0-*, -OC(O)-*, -0C(0)0-*, - C(O)N(R 10 )C(O)- *, -C(O)N(R 10 )C(O)N(R 10 )*, -N(R 10 )C(O)-*, -CR 10 2 N(R 10 )C (O)-*, -N(R 10 )C(O)N(R 10 )-*, -N(R 10 )C(O)O-*, -OC(0)N(R 10 )-*, -C(NR 10 )-*, -N(R 10 )C(NR 10 )-*, -C(NR 10 )N(R 10 )-*, -N(R 10 )C(NR 10 )N(R 10 )-*, -S(0) 2 -*, -0S
  • X 2 is independently selected at each occurrence from -O- , -S-, -N(R 10 )-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R 10 ), -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(0)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )N(R 10 )-, -S(0) 2 -, -OS(O)-,
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , - S(0)R 10 , and -S(0) 2 R 10 ; Ci - l o alkyl, C 2 -io alkenyl, C 2 -io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 ,
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
  • the compound of Formula (IA) is represented by Formula (IB):
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen, -
  • R 24 and R 25 are independently selected from hydrogen, halogen, -OR 10 , -SR 10 , -
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen, -OH, -N0 2 , -CN, and C1-10 alkyl. In certain embodiments, R 20 , R 21 , R 22 , and R 23 are each hydrogen.
  • R 24 and R 25 are independently selected from hydrogen, halogen, -OH, -N0 2 , -CN, and C 1-10 alkyl, or R 24 and R 25 taken together form an optionally substituted saturated C3-7 carbocycle.
  • R 24 and R 25 are each hydrogen.
  • R 24 and R 25 taken together form an optionally substituted saturated C3-5 carbocycle.
  • R 2 is represented by the formula:
  • R 101 is selected from an amino acid, a peptide, -0-(Ci-C 6 alkyl) and -Ci-C 6 alkyl, wherein alkyl of -0-(Ci-C 6 alkyl) and -Ci-C 6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -N(R 10 ) 2 , -C(0)R 10 , - C(O)N(R 10 ) 2 , -N0 2 , -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle; and
  • R 101 is a peptide selected from a dipeptide, tripeptide and tetrapeptide.
  • L 1 is selected from -N(R 10 )C(O)-*, -S(0) 2 N(R 10 )-*, -CR 10 2 N(R 10 )C (0)-*and -X 2 -CI -6 alkylene-X 2 -Ci -6 alkyl ene-. In some embodiments, L 1 is selected from -N(R 10 )C(O)-*. In certain embodiments,
  • R 10 of -N(R 10 )C(O)-* is selected from hydrogen and Ci -6 alkyl.
  • L 1 may be - NHC(O)-*.
  • L 1 is selected from -S(0) 2 N(R 10 )-*.
  • R 10 of -S(0) 2 N(R 10 )-* is selected from hydrogen and Ci -6 alkyl.
  • L 1 is -S(0) 2 NH-*.
  • L 1 is -CR 10 2N(R 10 )C(O)-*.
  • L 1 is selected from -CH 2 N(H)C(0)-* and -CH(CH 3 )N(H)C(0)-*.
  • R 3 is selected from optionally substituted C 3-12 carbocycle, and optionally substituted 3- to l2-membered
  • R 3 is selected from an optionally substituted aryl and an optionally substituted heteroaryl.
  • R 3 is an optionally substituted heteroaryl.
  • R 3 is selected from an optionally substituted 6-membered heteroaryl.
  • R 3 may be an optionally substituted pyridine.
  • R 3 is an optionally substituted aryl.
  • R 3 may be an optionally substituted phenyl.
  • R 3 is selected from pyridine, phenyl, tetrahydronaphthalene, tetrahydroquinoline, tetrahydroisoquinoline, indane, cyclopropylbenzene, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is
  • R 3 may be selected from:
  • R 3 may be selected from:
  • L 2 is selected from -C(O)-, and -C(0)NR 10 -.
  • L 2 is -C(O)-.
  • L 2 is selected from -C(0)NR 10 -.
  • R 10 of -C(0)NR 10 - may be selected from hydrogen and Ci -6 alkyl.
  • L 2 may be -C(0)NH-.
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ; Ci-io alkyl, C 2 -io alkenyl, C 2 -io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , - N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -OC(0)R 10 , -N0 2
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ; Ci-io alkyl, C 2 -io alkenyl, C 2 -io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , - N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -OC(0)R 10 , -N0 2
  • R 4 is -N(R 10 ) 2 .
  • R 10 of -N(R 10 ) 2 may be independently selected at each occurrence from optionally substituted Ci -6 alkyl.
  • R 10 of -N(R 10 ) 2 is independently selected at each occurrence from methyl, ethyl, propyl, and butyl, any
  • R 4 may certain
  • R 12 is
  • the present disclosure provides a compound represented by the structure of Formula (IVA):
  • L 12 is selected from -X 3 -, -X 3 -C I-6 alkylene-X 3 -, -X 3 -C 2-6 alkenylene-X 3 -, and - X 3 -C 2-6 alkynylene-X 3 -, each of which is optionally substituted on alkyl ene, alkenylene, or alkynylene with one or more substituents independently selected from R 12 ;
  • L 22 is independently selected from -X 4 -, -X 4 -C I-6 alkylene-X 4 -, -X 4 -C 2-6 alkenylene-X 4 -, and -X 4 -C 2-6 alkynylene-X 4 -, each of which is optionally substituted on alkyl ene, alkenylene, or alkynylene with one or more substituents independently selected from R 10 ;
  • X 3 and X 4 are independently selected at each occurrence from a bond, -0-, -S-, -N(R 10 )-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R 10 )-, -C(O)N(R 10 )C(O)-,
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • L 3 is a linker moiety, wherein at least one of R 10 is L 3 or at least one substituent on a group selected from R 4 , R 8 , X 3 and X 4 is L 3 ;
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
  • the compound of Formula (IV A) is represented by Formula (IVB):
  • I 24 , and R 25 are independently selected from hydrogen, halogen, -OR 10 , -SR 10 , -N(R 10 ) 2 , -
  • L 12 is - C(0)N(R 10 )-.
  • R 10 of -C(0)N(R 10 )- is selected from hydrogen, Ci -6 alkyl, and L 3 .
  • L 12 may be -C(0)NH-.
  • R 8 is an optionally substituted 5- or 6-membered heteroaryl.
  • R 8 may be an optionally substituted 5- or 6- membered heteroaryl, substituted with L 3 .
  • R 8 is an optionally substituted pyridine, substituted with L 3 .
  • L 22 is selected from -C(O)-, and -C(0)NR 10 -.
  • L 22 is -C(O)-.
  • L 22 is -C(0)NR 10 -.
  • R 10 of -C(0)NR 10 - may be selected from hydrogen, Ci -6 alkyl, and -L 3 .
  • L 22 may be -C(0)NH-.
  • R 4 is selected from: -OR 10 , and -N(R 10 ) 2 ; and C1-10 alkyl, C 2 -io alkenyl, C 2 -io alkynyl, C 3 -i 2 carbocycle, 3- to 12- membered heterocycle, aryl, and heteroaryl, each of which is optionally substituted with one or more substituents independently selected from L 3 , halogen, -OR 10 , -SR 10 , -N(R 10 ) 2 , -S(0)R 10 , -
  • R 4 is -N(R 10 ) 2 and R 10 of -N(R 10 ) 2 is selected from L 3 and hydrogen, and wherein at least one R 10 of -N(R 10 ) 2 is L 3 .
  • the compound of any one of Formula (IA), (IB), (IIA), (IIB), (IIIA), and (PIB), is further covalently bound to a linker, L 3 .
  • L 3 is a noncleavable linker.
  • L 3 is a cleavable linker.
  • L 3 may be cleavable by a lysosomal enzyme.
  • the compound is covalently attached to an antibody construct.
  • the compound is covalently attached to a targeting moiety, optionally through the linker.
  • the targeting moiety or antibody construct specifically binds to a tumor antigen.
  • the antibody construct or targeting moiety further comprises a target binding domain.
  • L 3 is represented by the formula:
  • L 4 represents the C-terminus of the peptide and L 5 is selected from a bond, alkylene and heteroalkylene, wherein L 5 is optionally substituted with one or more groups independently selected from R 32 , and RX is a reactive moiety;
  • RX comprises a leaving group.
  • RX comprises a maleimide.
  • L 3 is further covalently bound to an antibody construct.
  • the antibody construct is directed against a tumor antigen.
  • the antibody construct further comprises target binding domain.
  • L 3 is represented by the formula:
  • L 4 represents the C-terminal of the peptide and L 5 is selected from a bond, alkylene and heteroalkylene, wherein L 5 is optionally substituted with one or more groups independently selected from R 32 ;
  • RX * comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein * on RX* represents the point of attachment to the residue of the antibody construct;
  • the peptide of L 3 comprises Val— Cit or Val— Ala.
  • L 3 is represented by the formula:
  • RX comprises a reactive moiety
  • n 0 9
  • RX comprises a leaving group.
  • RX comprises a
  • L 3 is represented as follows:
  • RX * comprises a succinamide moiety and is bound to a cysteine residue of an antibody construct. In some embodiments, RX * comprises a hydrolyzed succinamide moiety and is bound to a cysteine residue of an antibody construct. In some embodiments, the antibody construct comprises a HER2 antigen binding domain. In some embodiments, the antibody construct comprises a TROP2 antigen binding domain. In some embodiments, the antibody construct is pertuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is trastuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is sacituzumab or an antigen binding fragment thereof.
  • the present disclosure provides a conjugate represented by the formula:
  • Antibody is an antibody construct
  • D is a compound or salt of any one of Formula (IA), (IB), (IIA), (IIB), (IIIA), and (IPB)
  • L 3 is a linker moiety.
  • the present disclosure provides a conjugate represented by the formula:
  • Antibody is an antibody construct and D-L 3 is a compound or salt of any one of Formula (IV A), and (IVB).
  • the antibody construct comprises a HER2 antigen binding domain. In some embodiments, the antibody construct comprises a TROP2 antigen binding domain. In some embodiments, the antibody construct is pertuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is trastuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is sacituzumab or an antigen binding fragment thereof.
  • the present disclosure provides a pharmaceutical composition
  • the present disclosure provides a pharmaceutical composition, comprising the conjugate disclosed herein and at least one pharmaceutically acceptable excipient.
  • the average DAR of the conjugate is from about 2 to about 8, or about 1 to about 3, or about 3 to about 5.
  • the present disclosure provides a method of killing tumor cells in vivo, comprising contacting a tumor cell population with a conjugate disclosed herein. In some aspects, the present disclosure provides a method for treatment, comprising administering to a subject a conjugate disclosed herein. In some aspects, the present disclosure provides a method for treatment, comprising administering to a subject in need thereof a compound or salt disclosed herein.
  • the present disclosure provides a method for treating cancer, comprising administering to a subject in need thereof a conjugate disclosed herein. In some aspects, the present disclosure provides a method for treating cancer, comprising administering to a subject in need thereof a compound or salt disclosed herein. In some aspects, the present disclosure provides a compound or salt disclosed herein for use in a method of treatment of a subject’s body by therapy.
  • the present disclosure provides a compound or salt disclosed herein for use in a method of treating cancer.
  • the present disclosure provides a conjugate disclosed herein for use in a method of treatment of a subject’s body by therapy.
  • the present disclosure provides a conjugate disclosed herein for use in a method of treating cancer.
  • the present disclosure provides a method of delivering a benzazepine compound to a cancer cell in a subject comprising administering a conjugate disclosed herein to a subject in need thereof.
  • the present disclosure provides a method for the treatment of cancer, comprising administering an effective amount of an antibody conjugate, wherein the antibody conjugate comprises an antibody construct covalently bound through a linker to a TLR8 agonist, wherein the TLR8 agonist has a K d for TLR7 that is two times or greater than two times the K d for TLR8, and wherein the antibody conjugate comprises from 1 to 20 TLR8 agonists per antibody construct, preferably from 1-8, 3-5 or 1-3.
  • the present disclosure provides a method for the treatment of cancer, comprising administering an effective amount of an antibody conjugate, wherein the antibody conjugate comprises an antibody construct covalently bound through a linker to a TLR8 agonist, wherein the TLR8 agonist agonizes TLR8 with at least an ECso of an order of magnitude less than the amount of the same compound required to show an agonizing effect on TLR7, and wherein the antibody conjugate comprises from 1 to 20 TLR8 agonists per antibody construct, preferably from 1-8, 3-5 or 1-3.
  • the present disclosure provides a method for the treatment of cancer, comprising administering an effective amount of an antibody conjugate, wherein the antibody conjugate comprises an antibody construct covalently bound through a linker to a TLR8 agonist, wherein the TLR8 agonist comprises a benzazepine substituted with a bicyclic heterocycle, and wherein the antibody conjugate comprises from 1 to 20 TLR8 agonists per antibody construct, preferably from 1-8, 3-5 or 1-3.
  • the present disclosure provides a method of preparing an antibody
  • L 3 -D is selected from a compound or salt disclosed herein comprising contacting L 3 -D with an antibody construct.
  • the antibody construct comprises a HER2 antigen binding domain. In some embodiments, the antibody construct comprises a TROP2 antigen binding domain. In some embodiments, the antibody construct is pertuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is trastuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is sacituzumab or an antigen binding fragment thereof. In some embodiments, the methods further comprising purifying the antibody conjugate. BRIEF DESCRIPTION OF THE DRAWINGS
  • FIG. 1 depicts an increase production of a pro-inflammatory cytokine, murine TNFa, from bone marrow-derived murine macrophages administered TLR7 small molecules with and without amine masking groups.
  • the present disclosure provides compounds, conjugates and pharmaceutical compositions for use in the treatment of disease.
  • the compounds of the disclosure are TLR8 modulators.
  • the compounds are TLR8 agonists.
  • Toll-like receptors are a family of membrane-spanning receptors that are expressed on cells of the immune system like dendritic cells, macrophages, monocytes, T cells, B cells, NK cells and mast cells but also on a variety of non-immune cells such as endothelial cells, epithelial cells and even tumor cells.
  • TLRs can have many isoforms, including TLR4, TLR7 and TLR8.
  • compounds or conjugates of the disclosure are administered in a form suitable to attenuate or eliminate immune-modulatory activity until the compound or conjugate reaches a desired target and the active site amine is unmasked. While not wishing to be bound by a mechanistic theory, the modification of compounds to attenuate or eliminate immune- modulatory activity may prevent undesired off-target immune-stimulatory activity, e.g., immune- stimulation in healthy tissue.
  • a compound such as a TLR8 agonist is modified with a masking group, such that the TLR8 agonist has limited activity or is inactive until it reaches an
  • the TLR8 agonist is covalently modified at an amine involved in binding to the active site of a TLR8 receptor such that the compound is unable to bind the active site of the receptor in its modified form.
  • the masking group may be removed under physiological conditions, e.g., enzymatic or acidic conditions, specific to the site of delivery, e.g., intracellular or extracellular adjacent to target cells.
  • Masking groups may be removed from the amine of the compound or salt described herein due to the action of lysosomal proteases, e.g., cathepsin and plasmin. These proteases can be present at elevated levels in certain tumor tissues.
  • the masking group may be removed by a lysosomal enzyme.
  • the lysosomal enzyme can be, for example, cathepsin B, cathepsin S, b-glucuronidase, or b-galactosidase.
  • the amine masking group inhibits binding of the amine group of the compound with residues of a TLR8 receptor.
  • the amine masking group may be removable under physiological conditions within a cell but remains covalently bound to the amine outside of a cell.
  • Masking groups that may be used to inhibit or attenuate binding of an amine group of a compound with residues of a TLR8 receptor include, for example, peptides and carbamates.
  • TLR8 receptors are localized to the endolysosomal/phagosomal compartment and predominantly found to be expressed by cells of the myeloid lineage. TLR ligation leads to activation of NF-kB and IRF-dependent pathways with the specific activation sequence and response with respect to the specific TLR and cell type. While TLR7 is mainly expressed in all dendritic cells subtypes (DC and here highly in pDC, plasmacytoid DC) and can be induced in B cells upon IFNa stimulation, TLR8 expression is rather restricted to monocytes, macrophages and myeloid DC. TLR8 signaling via MyD88 can be activated by bacterial single stranded RNA, small molecule agonists and microRNAs.
  • TLR8 results in the production of various pro-inflammatory cytokines such as IL-6, IL-12 and TNF-a as well as enhanced expression of co-stimulatory molecules, such as CD80, CD86, and chemokine receptors.
  • TLR8 activation can induce type I interferon (IRNb) in primary human monocytes.
  • TLR agonists can range from simple molecules to complex macromolecules. Likewise, the sizes of TLR agonists can range from small to large. TLR agonists can be synthetic or biosynthetic agonists. TLR agonists can also be Pathogen-Associated Molecular Pattern molecules (PAMPs).
  • PAMPs Pathogen-Associated Molecular Pattern molecules
  • the compounds of the present disclosure may be useful for the treatment and prevention, e.g., vaccination, of cancer, autoimmune diseases, inflammation, sepsis, allergy, asthma, graft rejection, graft-versus-host disease, immunodeficiencies, and infectious diseases.
  • the compounds have utility in the treatment of cancer either as single agents or in combination therapy.
  • the compounds have utility as single agent immunomodulators, vaccine adjuvants and in combination with conventional cancer therapies.
  • the compounds are incorporated into a conjugate that can be utilized, for example, to enhance an immune response.
  • the disclosure provides antibody construct-benzazepine compound conjugates and their use for treating cancer.
  • an“amine masking group” refers to any moiety covalently bound to the nitrogen of an amine, e.g., primary amine, which attenuates the interaction or activity, or blocks the amine from interacting with a TLR8 receptor and that is removable from the amine.
  • Exemplary amine masking groups include enzymatically-cleavable moieties such as amino acids, and peptides.
  • the term“antibody” refers to an immunoglobulin molecule that specifically binds to, or is immunologically reactive toward, a specific antigen.
  • the term antibody can include, for example, polyclonal, monoclonal, genetically engineered, and antigen binding fragments thereof.
  • An antibody can be, for example, murine, chimeric, humanized, heteroconjugate, bispecific, a diabody, a triabody, or a tetrabody.
  • the antigen binding fragment can include, for example, Fab', F(ab') 2 , Fab, Fv, rlgG, and scFv.
  • an“antigen binding domain” refers to a region of a molecule that specifically binds to an antigen.
  • An antigen binding domain may be a domain that can
  • An antigen binding domain can be an antigen-binding portion of an antibody or an antibody fragment.
  • An antigen binding domain can be one or more fragments of an antibody that retain the ability to specifically bind to an antigen.
  • An antigen binding domain can be an antigen binding fragment.
  • an antigen binding domain can recognize a single antigen.
  • an antigen binding domain can recognize, for example, two, three, four, five, six, seven, eight, nine, ten, or more than one antigen.
  • an“antibody construct” refers to a molecule, e.g., a protein, peptide, antibody or portion thereof, that contains an antigen binding domain and an Fc domain.
  • An antibody construct can recognize, for example, multiple antigens.
  • amino acids are conventional and can be as follows: alanine (A, Ala); arginine (R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine (C, Cys); glutamic acid (E, Glu); glutamine (Q, Gln); glycine (G, Gly); histidine (H, His); isoleucine (I, Ile); leucine (L, Leu); lysine (K, Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro); serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y, Tyr); valine (V, Val).
  • Other amino acids include citrulline (Cit); homocysteine (Hey); hydroxyproline (Hyp); ornithine (Orn); and th
  • “Conjugate”, as used herein, refers to an antibody construct that is linked, e.g., covalently linked, either directly or through a linker to a compound or compound-linker described herein, e.g., a compound or salt of any one of the Formulas described herein.
  • an“Fc domain” can be an Fc domain from an antibody or from a non antibody that can bind to an Fc receptor.
  • a“target binding domain” refers to a construct that contains an antigen binding domain from an antibody or from a non-antibody that can bind to the antigen.
  • a“tumor antigen” is an antigenic substance associated with a tumor or cancer cell, and can trigger an immune response in a host.
  • salts or“pharmaceutically acceptable salt” refers to salts derived from a variety of organic and inorganic counter ions well known in the art.
  • Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids.
  • Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.
  • Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, / oluenesulfonic acid, salicylic acid, and the like.
  • Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.
  • Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like.
  • Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine.
  • the pharmaceutically acceptable base addition salt is chosen from ammonium, potassium, sodium, calcium, and magnesium salts.
  • C x-y when used in conjunction with a chemical moiety, such as alkyl, alkenyl, or alkynyl is meant to include groups that contain from x to y carbons in the chain.
  • the term“Ci ⁇ alkyl” refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups that contain from 1 to 6 carbons.
  • the term -C x-y alkyl ene- refers to a substituted or unsubstituted alkylene chain with from x to y carbons in the alkylene chain.
  • -Ci -6 alkylene- may be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any one of which is optionally substituted.
  • C x-y alkenyl and“C x-y alkynyl” refer to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively.
  • the term -C x-y alkenylene- refers to a substituted or unsubstituted alkenylene chain with from x to y carbons in the alkenyl ene chain.
  • -C 2-6 alkenylene- may be selected from ethenylene, propenylene, butenylene, pentenylene, and hexenylene, any one of which is optionally substituted.
  • An alkenylene chain may have one double bond or more than one double bond in the alkenylene chain.
  • the term -C x-y alkynylene- refers to a substituted or unsubstituted alkynylene chain with from x to y carbons in the alkenylene chain.
  • -C 2-6 alkenylene- may be selected from ethynylene, propynylene, butynylene, pentynylene, and hexynylene, any one of which is optionally substituted.
  • An alkynylene chain may have one triple bond or more than one triple bond in the alkynylene chain.
  • Alkylene refers to a divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation, and preferably having from one to twelve carbon atoms, for example, methylene, ethylene, propylene, butylene, and the like.
  • the alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond.
  • the points of attachment of the alkylene chain to the rest of the molecule and to the radical group are through the terminal carbons respectively.
  • an alkylene comprises one to five carbon atoms (i.e., C1-C5 alkylene).
  • an alkylene comprises one to four carbon atoms
  • an alkylene comprises one to three carbon atoms
  • an alkylene comprises one to two carbon atoms
  • an alkylene comprises one carbon atom (i.e., Ci alkylene). In other embodiments, an alkylene comprises five to eight carbon atoms (i.e., Cs-Cx alkylene). In other embodiments, an alkylene comprises two to five carbon atoms (i.e., C 2 -C 5 alkylene). In other embodiments, an alkylene comprises three to five carbon atoms (i.e., C3-C5 alkylene). Unless stated otherwise specifically in the specification, an alkylene chain is optionally substituted by one or more substituents such as those substituents described herein.
  • Alkenylene refers to a divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon double bond, and preferably having from two to twelve carbon atoms.
  • the alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond.
  • the points of attachment of the alkenylene chain to the rest of the molecule and to the radical group are through the terminal carbons respectively.
  • an alkenylene comprises two to five carbon atoms (i.e., C 2 -C 5 alkenylene).
  • an alkenylene comprises two to four carbon atoms (i.e., C 2 -C 4 alkenylene).
  • C 2 -C 5 alkenylene i.e., C 2 -C 5 alkenylene
  • an alkenylene comprises two to four carbon atoms (i.e., C 2 -C 4 alkenylene).
  • an alkenylene comprises two to three carbon atoms (i.e., C 2 -C 3 alkenylene). In other embodiments, an alkenylene comprises two carbon atom (i.e., C 2 alkenylene). In other embodiments, an alkenylene comprises five to eight carbon atoms (i.e., Cs-Cx alkenylene). In other embodiments, an alkenylene comprises three to five carbon atoms (i.e., C 3 -C 5 alkenylene). Unless stated otherwise specifically in the specification, an alkenylene chain is optionally substituted by one or more substituents such as those substituents described herein.
  • Alkynylene refers to a divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond, and preferably having from two to twelve carbon atoms.
  • the alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond.
  • the points of attachment of the alkynylene chain to the rest of the molecule and to the radical group are through the terminal carbons respectively.
  • an alkynylene comprises two to five carbon atoms (i.e., C 2 -C 5 alkynylene).
  • an alkynylene comprises two to four carbon atoms (i.e., C 2 -C 4 alkynylene).
  • an alkynylene comprises two to three carbon atoms (i.e., C 2 -C 3 alkynylene). In other embodiments, an alkynylene comprises two carbon atom (i.e., C 2 alkynylene). In other embodiments, an alkynylene comprises five to eight carbon atoms (i.e., Cs-Cx alkynylene). In other embodiments, an alkynylene comprises three to five carbon atoms (i.e., C 3 -C 5 alkynylene). Unless stated otherwise specifically in the specification, an alkynylene chain is optionally substituted by one or more substituents such as those substituents described herein.
  • Heteroalkylene refers to a divalent hydrocarbon chain including at least one heteroatom in the chain, containing no unsaturation, and preferably having from one to twelve carbon atoms and from one to 6 heteroatoms, e.g., -0-, -NH-, -S-.
  • the heteroalkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond.
  • the points of attachment of the heteroalkyl ene chain to the rest of the molecule and to the radical group are through the terminal atoms of the chain.
  • a heteroalkylene comprises one to five carbon atoms and from one to three heteroatoms.
  • a heteroalkylene comprises one to four carbon atoms and from one to three heteroatoms. In other embodiments, a heteroalkylene comprises one to three carbon atoms and from one to two heteroatoms. In other embodiments, a heteroalkylene comprises one to two carbon atoms and from one to two heteroatoms. In other embodiments, a heteroalkylene comprises one carbon atom and from one to two heteroatoms. In other embodiments, a heteroalkylene comprises five to eight carbon atoms and from one to four heteroatoms. In other embodiments, a heteroalkylene comprises two to five carbon atoms and from one to three heteroatoms.
  • a heteroalkylene comprises three to five carbon atoms and from one to three heteroatoms. Unless stated otherwise specifically in the specification, a heteroalkylene chain is optionally substituted by one or more substituents such as those substituents described herein.
  • Carbocycle refers to a saturated, unsaturated or aromatic ring in which each atom of the ring is carbon.
  • Carbocycle includes 3- to lO-membered monocyclic rings, 6- to l2-membered bicyclic rings, and 6- to l2-membered bridged rings.
  • Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings.
  • an aromatic ring e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene.
  • a bicyclic carbocycle includes any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits.
  • a bicyclic carbocycle includes any combination of ring sizes such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems.
  • Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl.
  • unsaturated carbocycle refers to carbocycles with at least one degree of unsaturation and excluding aromatic carbocycles.
  • unsaturated carbocycles include cyclohexadiene, cyclohexene, and cyclopentene.
  • heterocycle refers to a saturated, unsaturated or aromatic ring comprising one or more heteroatoms.
  • exemplary heteroatoms include N, O, Si, P, B, and S atoms.
  • Heterocycles include 3- to lO-membered monocyclic rings, 6- to l2-membered bicyclic rings, and 6- to l2-membered bridged rings.
  • a bicyclic heterocycle includes any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits.
  • an aromatic ring e.g., pyridyl
  • a saturated or unsaturated ring e.g., cyclohexane, cyclopentane, morpholine, piperidine or cyclohexene.
  • a bicyclic heterocycle includes any combination of ring sizes such as 4-5 fused ring systems, 5-5 fused ring systems, 5- 6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems.
  • the term“unsaturated heterocycle” refers to heterocycles with at least one degree of unsaturation and excluding aromatic heterocycles.
  • unsaturated heterocycles include dihydropyrrole, dihydrofuran, oxazoline, pyrazoline, and dihydropyridine.
  • heteroaryl includes aromatic single ring structures, preferably 5- to 7- membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms.
  • heteroaryl also includes polycyclic ring systems having two or more rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other rings can be aromatic or non-aromatic carbocyclic, or
  • heterocyclic heterocyclic.
  • Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.
  • substitution refers to moieties having substituents replacing a hydrogen on one or more carbons or substitutable heteroatoms, e.g., an NH or NH 2 of a compound. It will be understood that“substitution” or“substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, i.e., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.
  • substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as substituting the two hydrogen atoms on a single carbon with an oxo, imino or thioxo group.
  • substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as substituting the two hydrogen atoms on a single carbon with an oxo, imino or thioxo group.
  • the term“substituted” is contemplated to include all permissible substituents of organic compounds.
  • the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds.
  • the permissible substituents can be one or more and the same or different for appropriate organic compounds.
  • phrases“parenteral administration” and“administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
  • phrases“pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • phrases“pharmaceutically acceptable excipient” or“pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material.
  • Each carrier must be“acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
  • materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide;
  • targeting moiety refers to a structure that has a selective affinity for a target molecule relative to other non-target molecules.
  • the targeting moiety binds to a target molecule.
  • a targeting moiety may include, for example, an antibody, a peptide, a ligand, a receptor, or a binding portion thereof.
  • the target biological molecule may be a biological receptor or other structure of a cell such as a tumor antigen.
  • antibodies that may be used together with compounds of the disclosure.
  • compounds of the disclosure are linked, e.g., covalently linked, either directly or through a linker to a compound of the disclosure forming conjugates.
  • conjugates of the disclosure are represented by the following formula:
  • A is an antibody construct
  • L 3 is a linker
  • D is a compound or salt of any one of
  • Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB) or (IIIC), or L 3 -D is a compound or salt of any one of Formulas (IV A), (IVB) or (IVC), and n is from 1 to 20.
  • n is from 1 to 10, such as from 1 to 9, such as from 1 to 8, such as from 2 to 8, such as from 1 to 6, such as from 3 to 5, or such as from 1 to 3.
  • n is 4.
  • each D or L 3 -D are independently selected from Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB) or (IIIC), or Formulas (IVA), (IVB) or (IVC), respectively.
  • a compound or salt of the disclosure e.g., a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC)
  • a drug D
  • a benzazepine compound an immune-stimulatory compound
  • ISC ISC
  • a payload particularly when referenced as part of a conjugate.
  • “LP”,“linker- payload”,“L 3 -D”, or“compound-linker” may be used herein to refer to a compound or salt of the disclosure bound to a linker.
  • An antibody construct may contain, for example, two, three, four, five, six, seven, eight, nine, ten, or more antigen binding domains.
  • An antibody construct may contain two antigen binding domains in which each antigen binding domain can recognize the same antigen.
  • An antibody construct may contain two antigen binding domains in which each antigen binding domain can recognize different antigens.
  • An antigen binding domain may be in a scaffold, in which a scaffold is a supporting framework for the antigen binding domain.
  • An antigen binding domain may be in a non-antibody scaffold.
  • An antigen binding domain may be in an antibody scaffold.
  • An antibody construct may comprise an antigen binding domain in a scaffold.
  • the antibody construct may comprise a Fc fusion protein. In some embodiments, the antibody construct is a Fc fusion protein.
  • An antigen binding domain may specifically bind to a tumor antigen.
  • An antigen binding domain may specifically bind to an antigen that is at least 80%, at least 90%, at least 95%, at least 99%, or 100% homologous to a tumor antigen.
  • An antigen binding domain may specifically bind to an antigen on an antigen presenting cell (APC).
  • An antigen binding domain may specifically bind to an antigen that is at least 80%, at least 90%, at least 95%, at least 99%, or 100% homologous to an antigen on an antigen presenting cell (APC).
  • An antigen binding domain of an antibody may comprise one or more light chain (LC) CDRs and one or more heavy chain (HC) CDRs.
  • an antibody binding domain of an antibody may comprise one or more of the following: a light chain complementary determining region 1 (LC CDR1), a light chain complementary determining region 2 (LC CDR2), or a light chain complementary determining region 3 (LC CDR3).
  • an antibody binding domain may comprise one or more of the following: a heavy chain complementary determining region 1 (HC CDR1), a heavy chain complementary determining region 2 (HC CDR2), or a heavy chain complementary determining region 3 (HC CDR3).
  • an antibody binding domain of an antibody may comprise one or more of the following: LC CDR1, LC CDR2, LC CDR3, HC CDR1, HC CDR2, and HC CDR3.
  • the antigen binding domain of an antibody construct may be selected from any domain that binds the antigen including, but not limited to, from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or an antigen binding fragment thereof, for example, a heavy chain variable domain (V H ) and a light chain variable domain (V L ), or from a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a T cell receptor, or a recombinant T cell receptor.
  • the antigen binding domain of an antibody construct may be selected from any domain that binds the antigen including, but not limited to, from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or an antigen binding fragment thereof, for example, a heavy chain variable domain (V H ) and a light chain variable domain (V L ).
  • V H heavy chain variable domain
  • V L light chain variable domain
  • the antigen binding domain of an antibody construct may be at least 80% identical to an antigen binding domain selected from, but not limited to, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (V H ) and a light chain variable domain (V L ), or to a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a cytokine, a ligand, an immunocytokine, a T cell receptor, or a recombinant T cell receptor.
  • V H heavy chain variable domain
  • V L light chain variable domain
  • an antigen binding domain of an antibody construct may be at least 80% identical to an antigen binding domain selected from, but not limited to, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (V H ) and a light chain variable domain (V L ).
  • an antibody construct comprises an Fc domain that may further comprise an Fc domain, in which the Fc domain may be the part of an Fc region that interacts with Fc receptors.
  • the Fc domain of an antibody construct may interact with Fc-receptors (FcRs) found on immune cells.
  • FcRs Fc-receptors
  • the Fc domain may also mediate the interaction between effector molecules and cells, which can lead to activation of the immune system.
  • the Fc domain may be derived from IgG, IgA, or IgD antibody isotypes, and may comprise two identical protein fragments, which are derived from the second and third constant domains of the antibody’s heavy chains.
  • the Fc region may comprise a highly-conserved N-glycosylation site, which may be essential for FcR-mediated downstream effects.
  • the Fc domain may be derived from IgM or IgE antibody isotypes, in which the Fc domain may comprise three heavy chain constant domains.
  • An Fc domain may interact with different types of FcRs.
  • the different types of FcRs may include, for example, FcyRI, FcyRIIA, FcyRIIB, FcyRI 11 A, FcyRIIIB, FcaRI, FcpR, FceRI, FceRII, and FcRn.
  • FcRs may be located on the membrane of certain immune cells including, for example, B lymphocytes, natural killer cells, macrophages, neutrophils, follicular dendritic cells, eosinophils, basophils, platelets, and mast cells.
  • the FcR may initiate functions including, for example, clearance of an antigen-antibody complex via receptor-mediated endocytosis, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody dependent cell-mediated phagocytosis (ADCP), and ligand-triggered transmission of signals across the plasma membrane that can result in alterations in secretion, exocytosis, and cellular metabolism.
  • FcRs may deliver signals when FcRs are aggregated by antibodies and multivalent antigens at the cell surface.
  • ITAMs immunoreceptor tyrosine- based activation motifs
  • SRC family tyrosine kinases may sequentially activate SRC family tyrosine kinases and SYK family tyrosine kinases.
  • ITAM comprises a twice-repeated YxxL sequence flanking seven variable residues.
  • the SRC and SYK kinases may connect the transduced signals with common activation pathways.
  • An antibody may consist of two identical light protein chains and two identical heavy protein chains, all held together covalently by disulfide linkages. The N-terminal regions of the light and heavy chains together may form the antigen recognition site of an antibody.
  • various functions of an antibody may be confined to discrete protein domains.
  • the sites that can recognize and can bind antigen may consist of three complementarities determining regions (CDRs) that may lie within the variable heavy chain region and variable light chain region at the N-terminal end of the heavy chain and the light chain.
  • CDRs complementarities determining regions
  • the constant domains may provide the general framework of the antibody and may not be involved directly in binding the antibody to an antigen, but may be involved in various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity, and may bind Fc receptors.
  • the constant domains may include an Fc region.
  • the constant domains may include an Fc domain.
  • the domains of natural light and heavy chains may have the same general structures, and each domain may comprise four framework regions, whose sequences can be somewhat conserved, connected by three hyper-variable regions or CDRs.
  • the four framework regions (FR) may largely adopt a b-sheet conformation and the CDRs can form loops connecting, and in some aspects forming part of, the b -sheet structure.
  • the CDRs in each chain may be held in close proximity by the framework regions and, with the CDRs from the other chain, may contribute to the formation of the antigen binding site.
  • An antibody construct may comprise a light chain of an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten modifications and in certain embodiments, not more than 40, 35, 30, 25, 20, 15 or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence.
  • An antibody construct may comprise a heavy chain of an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten modifications and in certain embodiments, not more than 40, 35, 30, 25, 20, 15 or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence.
  • An antibody of an antibody construct may include an antibody of any type, which may be assigned to different classes of immunoglobins, e.g ., IgA, IgD, IgE, IgG, and IgM. Several different classes may be further divided into isotypes, e.g. , IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. An antibody may further comprise a light chain and a heavy chain, often more than one chain.
  • the heavy-chain constant regions (Fc) that corresponds to the different classes of immunoglobulins may be a, d, e, g, and m, respectively.
  • the light chains may be one of either kappa (K) or lambda (l), based on the amino acid sequences of the constant domains.
  • the Fc region may contain an Fc domain.
  • An Fc receptor may bind an Fc domain.
  • Antibody constructs may also include any fragment or recombinant forms thereof, including but not limited to, single chain variable fragments (scFvs),‘T-bodies’, anti-calins, centyrins, affibodies, domain antibodies, or peptibodies.
  • An antibody construct may comprise an antibody fragment.
  • An antibody fragment may include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH , CL and CHI domains; (ii) a F(ab') 2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; and (iii) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody.
  • the two domains of the Fv fragment, VL and VH may be coded for by separate genes, they may be linked by a synthetic linker to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules.
  • An antibody may include an Fc region comprising multiple Fc domains.
  • the Fc domain of an antibody may interact with FcRs found on immune cells.
  • the Fc domain may also mediate the interaction between effector molecules and cells, which may lead to activation of the immune system.
  • the Fc region may comprise two identical protein fragments, which can be derived from the second and third constant domains of the antibody’s heavy chains.
  • the Fc regions may comprise three heavy chain constant domains.
  • the Fc regions may comprise a highly-conserved N-glycosylation site, which may be important for FcR-mediated downstream effects.
  • An antibody used herein may be chimeric or“humanized.” Chimeric and humanized forms of non-human (e.g, murine) antibodies can be chimeric immunoglobulins,
  • the humanized antibody may comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the framework regions are those of a human immunoglobulin sequence.
  • the humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin consensus sequence.
  • an antibody described herein may be a human antibody.
  • “human antibodies” can include antibodies having, for example, the amino acid sequence of a human immunoglobulin and may include antibodies isolated from human immunoglobulin libraries or from animals transgenic for one or more human immunoglobulins that do not express
  • Human antibodies may be produced using transgenic mice which are incapable of expressing functional endogenous immunoglobulins, but which may express human immunoglobulin genes.
  • Completely human antibodies that recognize a selected epitope may be generated using guided selection.
  • a selected non-human monoclonal antibody e.g ., a mouse antibody, may be used to guide the selection of a completely human antibody recognizing the same epitope.
  • An antibody described herein may be a bispecific antibody or a dual variable domain antibody (DVD).
  • Bispecific and DVD antibodies may be monoclonal, often human or humanized, antibodies that can have binding specificities for at least two different antigens.
  • An antibody described herein may be a derivatized antibody.
  • derivatized antibodies may be modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein.
  • an antibody described herein may have a sequence that has been modified to alter at least one constant region-mediated biological effector function relative to the corresponding wild type sequence.
  • the antibody can be modified to reduce at least one constant region-mediated biological effector function relative to an unmodified antibody, e.g. , reduced binding to the Fc receptor (FcR).
  • FcR binding may be reduced by, for example, mutating the immunoglobulin constant region segment of the antibody at particular regions necessary for FcR interactions.
  • An antibody or Fc domain as described herein may be modified to acquire or improve at least one constant region-mediated biological effector function relative to an unmodified antibody or Fc domain, e.g. , to enhance FcyR interactions.
  • an antibody with a constant region that binds FcyRIIA, FcyRIIB and/or FcyRIIIA with greater affinity than the corresponding wild type constant region may be produced according to the methods described herein.
  • An Fc domain that binds FcyRIIA, FcyRIIB and/or FcyRIIIA with greater affinity than the corresponding wild type Fc domain may be produced according to the methods described herein.
  • the antibody construct comprises an antigen binding domain and an IgG Fc domain, wherein a K d for binding of the antigen binding domain to a first antigen in a presence of the immune-stimulatory compound is less than about 100 nM and no greater than about 100 times a K d for binding of the antigen binding domain to the first antigen in the absence of the immune-stimulatory compound.
  • the antibody construct comprises a K d for binding of the IgG Fc domain to an Fc receptor in the presence of the immune- stimulatory compound is no greater than about 100 times a K d for binding the IgG Fc domain to the Fc receptor in the absence of the immune-stimulatory compound.
  • the first antigen is selected from CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD40, CD45, CAMPATH-l, BCMA, CS-l, PD-L1, B7-H3, B7-DC, HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, folate-binding protein, A33, G250, prostate-specific membrane antigen (PSMA), GD2, GD3, GM2, Le y , CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein, tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, avB3, WT1, LMP2, HPV E6, HPV E7, Her-2/neu, MAGE A3, p53 nonmutant, NY-ESO-l, MelanA
  • an antigen binding domain specifically binds to an antigen, such as those selected from CD5, CD25, CD37, CD33, CD45, BCMA, CS-l, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM,
  • an antigen such as those selected from CD5, CD25, CD37, CD33, CD45, BCMA, CS-l, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM,
  • the antibody construct comprises a human antibody or a humanized antibody or an antigen binding portion thereof, e.g., a humanized CD40, humanized HER2 or humanized TROP2 antibody.
  • the antibody construct comprises a TROP2 antibody, e.g., sacituzumab, or an antigen binding portion thereof.
  • the antibody construct comprises the heavy and light chain variable region sequences of sacituxumab (SEQ ID NOs:3 and 4).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sacituzumab (SEQ ID NO:4), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sacituzumab (SEQ ID NO:3), as determined by the Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sacituzumab (SEQ ID NO:4), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sacituzumab (SEQ ID NO:3), as determined by IMGT (ImMunoGeneTics).
  • the antibody construct comprises a HER2 antibody, e.g., pertuzumab, trastuzumab, or an antigen binding portion thereof.
  • the antibody construct comprises the heavy and light chain variable region sequences of pertuzumab (SEQ ID NOs: l and 2).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of pertuzumab (SEQ ID NO:2), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of pertuzumab (SEQ ID NO: 1), as determined by the Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of pertuzumab (SEQ ID NO:2), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of pertuzumab (SEQ ID NO: 1), as determined by IMGT.
  • the antibody construct comprises the heavy and light chain variable region sequences of trastuzumab (SEQ ID NOs:7 and 8). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of trastuzumab (SEQ ID NO: 8), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of trastuzumab (SEQ ID NO: 7), as determined by the Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of trastuzumab (SEQ ID NO: 8), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of trastuzumab (SEQ ID NO:7), as determined by IMGT.
  • the antibody construct comprises a CD40 antibody or an antigen binding portion thereof.
  • the antibody construct comprises the heavy and light chain variable region sequence of sacituxumab (SEQ ID NO:3 and 4).
  • the antibody construct comprises the LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sacituzumab (SEQ ID NO:4), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sacituzumab (SEQ ID NO:3), as determined by Rabat index.
  • the antibody construct comprises a Liv-l antibody, e.g., ladiratuzumab, huLivl-l4 (WO 2012078688), Livl-l.7A4 (US 2011/0117013), huLivl-22 (WO 2012078688) or an antigen binding portion thereof.
  • the antibody construct comprises the heavy and light chain variable region sequences of ladiratuzumab (SEQ ID NOs:5 and 6).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of ladiratuzumab (SEQ ID NO:6), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of ladiratuzumab (SEQ ID NO: 5), as determined by Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of
  • the antibody construct comprises the heavy and light chain variable region sequences of huLivl-l4 (SEQ ID NOs: l7 and 18).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of huLivl-l4 (SEQ ID NO: 18), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of huLivl-l4 (SEQ ID NO: 17), as determined by Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of huLivl-l4 (SEQ ID NO: 18), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of huLivl-l4 (SEQ ID NO: 17), as determined by IMGT.
  • the antibody construct comprises the heavy and light chain variable region sequences of Livl-l.7A4 (SEQ ID NOs: 19 and 20).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of Livl-l.7A4 (SEQ ID NO:20), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of Livl-l .7A4 (SEQ ID NO: 19), as determined by Rabat index.
  • the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of Livl-l.7A4 (SEQ ID NO:20), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of Livl-l.7A4 (SEQ ID NO: 19), as determined by Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of Livl-l.7A4 (SEQ ID NO:20), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of Livl-l.7A4 (SEQ ID NO: 19), as determined by IMGT.
  • the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of Livl-l.7A4 (SEQ ID NO:20), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of Livl-l.7A4 (SEQ ID NO: 19), as determined by IMGT.
  • the antibody construct comprises the heavy and light chain variable region sequences of huLivl-22 (SEQ ID NOs:2l and 22).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of huLivl-22 (SEQ ID NO:22), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of huLivl-22 (SEQ ID NO:2l), as determined by Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of huLivl-22 (SEQ ID NO:22), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of huLivl-22 (SEQ ID NO:2l), as determined by IMGT.
  • the antibody construct comprises a MUC16 antibody, e.g., sofituzumab, 4H11 (US2013/0171152), 4H5 (US2013/0171152) or an antigen binding portion thereof.
  • the antibody construct comprises the heavy and light chain variable region sequences of sofituzumab (SEQ ID NOs:23 and 24).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sofituzumab (SEQ ID NO:24), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sofituzumab (SEQ ID NO:23), as determined by Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sofituzumab (SEQ ID NO:24), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sofituzumab (SEQ ID NO:23), as determined by IMGT.
  • the antibody construct comprises the heavy and light chain variable region sequences of antibody 4H11 (SEQ ID NOs: 13 and 14).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4H11 (SEQ ID NO: 14), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of antibody 4H11 (SEQ ID NO: 13), as determined by Rabat index.
  • the antibody construct comprises a humanized antibody comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4H11 (SEQ ID NO: 14), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of antibody 4H11 (SEQ ID NO: 13), as determined by Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4H11 (SEQ ID NO: 14), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of 4H11 (SEQ ID NO: 13), as determined by IMGT.
  • the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4H11 (SEQ ID NO: 14), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of 4H11 (SEQ ID NO: 13), as determined by IMGT.
  • the antibody construct comprises the heavy and light chain variable region sequences of antibody 4A5 (SEQ ID NOs: 15 and 16). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4A5 (SEQ ID NO: 16), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of 4A5 (SEQ ID NO: 15), as determined by Rabat index. In certain
  • the antibody construct comprises a humanized antibody or an antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4A5 (SEQ ID NO: 16), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of antibody 4A5 (SEQ ID NO: 15), as determined by Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4A5 (SEQ ID NO: 16), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of antibody 4A5 (SEQ ID NO: 15), as determined by IMGT.
  • the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of 4A5 (SEQ ID NO: 16), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of 4A5 (SEQ ID NO: 15), as determined by IMGT.
  • the antibody construct comprises a PD-L1 antibody, e.g., atezolizumab, MDX-l 105 (WO 2007/005874) or an antigen binding portion thereof.
  • the antibody construct comprises the heavy and light chain variable region sequences of atezolizumab (SEQ ID NOs: 11 and 12).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of atezolizumab (SEQ ID NO: 12), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of atezolizumab (SEQ ID NO: 11), as determined by Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of atezolizumab (SEQ ID NO: 12), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of atezolizumab (SEQ ID NO: 11), as determined by IMGT.
  • the antibody construct comprises the heavy and light chain variable region sequences of MDX-l 105 (SEQ ID NOs:9 and 10).
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of MDX-l 105 (SEQ ID NO: 10), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of MDX-l 105 (SEQ ID NO: 9), as determined by Rabat index.
  • the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of MDX-l 105 (SEQ ID NO: 10), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of MDX-l 105 (SEQ ID NO: 9), as determined by Rabat index.
  • the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of MDX-l 105 (SEQ ID NO: 10), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of MDX-l 105 (SEQ ID NO:9), as determined by IMGT.
  • the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of MDX-l 105 (SEQ ID NO: 10), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of MDX-l 105 (SEQ ID NO: 9), as determined by IMGT.
  • An antibody construct may comprise an antibody with modifications of at least one amino acid residue. Modifications may be substitutions, additions, mutations, deletions, or the like. An antibody modification can be an insertion of an unnatural amino acid.
  • VH sequences and VL sequences are illustrated in Table A below.
  • An antibody construct may further comprise a target binding domain.
  • a target binding domain may comprise a domain that binds to a target.
  • a target may be an antigen.
  • a target binding domain may comprise an antigen binding domain.
  • a target binding domain may be a domain that can specifically bind to an antigen.
  • a target binding domain may be an antigen binding portion of an antibody or an antibody fragment.
  • a target binding domain may be one or more fragments of an antibody that can retain the ability to specifically bind to an antigen.
  • a target binding domain may be any antigen binding fragment.
  • a target binding domain may be in a scaffold, in which a scaffold is a supporting framework for the antigen binding domain.
  • a target binding domain may comprise an antigen binding domain in a scaffold.
  • a target binding domain may comprise an antigen binding domain which can refer to a portion of an antibody comprising the antigen recognition portion, z.e., an antigenic determining variable region of an antibody sufficient to confer recognition and binding of the antigen recognition portion to a target, such as an antigen, z.e., the epitope.
  • a target binding domain may comprise an antigen binding domain of an antibody.
  • a target binding domain is a CD40 agonist.
  • An Fv can be the minimum antibody fragment which contains a complete antigen- recognition and antigen-binding site. This region may consist of a dimer of one heavy chain and one light chain variable domain in tight, non-covalent association. In this configuration, the three hypervariable regions of each variable domain may interact to define an antigen-binding site on the surface of the VH-VL dimer. A single variable domain (or half of an Fv comprising only three hypervariable regions specific for an antigen) can recognize and bind antigen, although at a lower affinity than the entire binding site.
  • a target binding domain may be at least 80% homologous to an antigen binding domain selected from, but not limited to, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (VH) and a light chain variable domain (VL), a single chain variable fragment (scFv), a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a cytokine, a ligand, an immunocytokine, a T cell receptor, or a recombinant T cell receptor.
  • VH heavy chain variable domain
  • VL light chain variable domain
  • scFv single chain variable fragment
  • DARPin an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor
  • a target binding domain may be attached to an antibody construct.
  • an antibody construct may be fused with a target binding domain to create an antibody construct target binding domain fusion.
  • the antibody construct-target binding domain fusion may be the result of the nucleic acid sequence of the target binding domain being expressed in frame with the nucleic acid sequence of the antibody construct.
  • the antibody construct- target binding domain fusion may be the result of an in-frame genetic nucleotide sequence or a contiguous peptide sequence encoding the antibody construct with the target binding domain.
  • a target binding domain may be linked to an antibody construct.
  • a target binding domain may be linked to an antibody construct by a chemical conjugation.
  • a target binding domain may be attached to a terminus of an Fc region.
  • a target binding domain may be attached to a terminus of an Fc region.
  • a target binding domain may be attached to a terminus of an antibody construct.
  • a target binding domain may be attached to a terminus of an antibody.
  • a target binding domain may be attached to a light chain of an antibody.
  • a target binding domain may be attached to a terminus of a light chain of an antibody.
  • a target binding domain may be attached to a heavy chain of an antibody.
  • a target binding domain may be attached to terminus of a heavy chain of an antibody.
  • the terminus may be a C-terminus.
  • An antibody construct may be attached to 1, 2, 3, and/or 4 target binding domains. The target binding domain may direct the antibody construct to, for example, a particular cell or cell type.
  • a target binding domain of an antibody construct may be selected in order to recognize an antigen, e.g., an antigen expressed on an immune cell.
  • An antigen can be a peptide or fragment thereof.
  • An antigen may be expressed on an antigen-presenting cell.
  • An antigen may be expressed on a dendritic cell, a macrophage, or a B cell.
  • an antigen may be a tumor antigen.
  • the tumor antigen may be any tumor antigen described herein.
  • an antibody construct described herein specifically binds a second antigen.
  • the target binding domain is linked, e.g., covalently bound, to the antibody construct at a C-terminal end of the Fc domain.
  • the present disclosure provides a compound represented by the structure of Formula (IA):
  • L 1 is selected from -X 1 -, -X 2 -CI -6 alkyl ene-X 2 -Ci -6 alkylene-, -X 2 -C 2 -6 alkenylene- X 2 -, and -X 2 -C 2-6 alkynylene-X 2 -, each of which is optionally substituted on alkylene, alkenylene or alkynylene with one or more R 12 ;
  • L 2 is selected from -X 2 -, -X 2 -C I-6 alkylene-X 2 -, -X 2 -C 2-6 alkenyl ene-X 2 -, and -X 2 -C 2-6 alkynylene-X 2 -, each of which is optionally substituted on alkylene, alkenylene or alkynylene with one or more R 12 ;
  • X 1 is selected from -S-*, -N(R 10 )-*, -C(0)0-*, -OC(O)-*, -0C(0)0-*, - C(O)N(R 10 )C(O)- *, -C(O)N(R 10 )C(O)N(R 10 )*, -N(R 10 )C(O)-*, -CR 10 2 N(R 10 )C
  • X 2 is independently selected at each occurrence from a bond, -O- , -S-, -N(R 10 )-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R 10 ), -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(0)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )N(R 10 )-, -S(0) 2 -, -OS(
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , - S(0)R 10 , and -S(0) 2 R 10 ; Ci - l o alkyl, C 2-i o alkenyl, C 2-i o alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 ,
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
  • X 2 is independently selected at each occurrence from -O-
  • L 1 is independently selected from -X 1 -, -X 2 -C I-6 alkyl-X 2 -, -X 2 -C 2-6 alkenyl-X 2 -, and
  • L 2 is independently selected from -X 2 -, -X 2 -C I-6 alkyl-X 2 -, -X 2 -C 2-6 alkenyl-X 2 -, and
  • X 1 at each occurrence is independently selected
  • X 2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R 10 )-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(0)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )N(R 10 )-, -S(0) 2 -, -
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 3 is selected from optionally substituted C 3-i2 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents are independently selected at each occurrence from: halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , - N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 ,
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , - S(0)R 10 , and -S(0) 2 R 10 ; Ci - l o alkyl, C 2 -io alkenyl, C 2 -io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, - OR 10 , -SR 10 ,
  • each C 3 -i 2 carbocycle, and 3- to l2-membered heterocycle in R 4 is independently optionally substituted with one or more substituents selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , - N(R 10 )C(O)R 10 ,
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )2, -S(0)R 10 , - S(0) 2 R 10 ,
  • Benzazepine core refers to both a benzazepine and a 4,5-dihydrobenzazepine core ring system.
  • L 2 can be attached at C2, C3, C4 or C5 of the benzazepine core, wherein the numbering of the benzazepine is as follows:
  • L 2 is attached to the benzazepine core at C4.
  • L 2 is attached to the benzazepine core at C4.
  • L 1 can be attached at C6, C7, C8 or C9. In certain embodiments, for a compound or salt of Formula (IA), L 1 is attached to the benzazepine core at C8. In certain embodiments for a compound or salt of Formula (IA), represents a double bond, L 2 is attached to the benzazepine core at C4 and L 1 is attached to the benzazepine core at C8.
  • the substitutable carbon on the benzazepine core is selected from C2, C3, C4, C5, C6, C7, C8, and C9.
  • the compound of Formula (IA) is represented by Formula (IB):
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen, -
  • R 24 and R 25 are independently selected from hydrogen, halogen, -OR 10 , -SR 10 , -
  • the compound of Formula (IA) is represented by Formula (IC):
  • L 1 is selected from
  • L 1 is selected from -N(R 10 )C(O)-*.
  • R 10 of -N(R 10 )C(O)-* is selected from hydrogen and Ci -6 alkyl.
  • L 1 is -NHC(O)-*.
  • L 1 is selected from
  • R 10 of -S(0) 2 N(R 10 )-* is selected from hydrogen and Ci -6 alkyl.
  • L 1 is selected from -S(0) 2 NH-*.
  • L 1 is selected from -CR 10 2N(R 10 )C(O)-*. In certain embodiments, L 1 is selected
  • R 3 is selected from optionally substituted C 3-i2 carbocycle, and optionally substituted 3-
  • R 3 is selected from an optionally substituted aryl and an optionally substituted heteroaryl, such as R 3 is an optionally substituted heteroaryl.
  • R 3 is an optionally substituted heteroaryl substituted with one or more substituents independently selected from halogen, -
  • R 3 may be selected from an optionally substituted 5- or 6-membered
  • R 3 may be selected from
  • R 3 is selected from an optionally substituted 6- membered heteroaryl, e.g., R 3 is optionally substituted pyridine.
  • R 3 may be selected from an optionally substituted phenyl and an optionally substituted naphthyl. In certain embodiments, R 3 is an optionally substituted phenyl.
  • R 3 is selected from pyridine, phenyl, tetrahydronaphthalene, tetrahydroquinoline,
  • R 3 may be selected from:
  • R 3 may be selected from:
  • L 1 is selected from -N(R 10 )C(O)-*, -S(0) 2 N(R 10 )-*, -CR 10 2 N(R 10 )C(O)-*and -X 2 -C I-6 alkyl ene-X 2 -C i-6 alkylene-;
  • L 2 is -C(O)-
  • X 2 at each occurrence is independently selected from -O- , -S-, -N(R 10 )-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)N(R 10 )-, and -N(R 10 )C(O)O-;
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • the compound may further comprise a linker (L 3 ).
  • the linker may be covalently bound to any position, valence permitting, on a compound or salt of Formula (IA), (IB) or (IC).
  • the linker may be bound to R 4 , R 3 , X 1 or X 2 .
  • the linker is bound to a nitrogen or oxygen atom of a compound or salt of Formula (IA), (IB) or (IC).
  • the linker may comprise a reactive moiety, e.g., an electrophile that can react to form a covalent bond with a moiety of an antibody, e.g., a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue of an antibody.
  • a compound or salt of Formula (IA), (IB) or (IC) may be covalently bound through the linker to an antibody.
  • the present disclosure provides a compound represented by the structure of Formula (II A):
  • L 10 is -X 10 -;
  • L 2 is selected from -X 2 -, -X 2 -C I-6 alkylene-X 2 -, -X 2 -C2-6 alkenyl ene-X 2 -, and - X 2 -C2-6 alkynylene-X 2 -, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R 12 ;
  • X 10 is selected from -C(O)-, and -C(0)N(R 10 )-*, wherein * represents where X 10 is bound to R 5 ;
  • X 2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R 10 )-, -C(O)-,
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
  • L 10 is independently selected from -X 10 -;
  • L 2 is independently selected from -X 2 -, -X 2 -C I-6 alkyl-X 2 -, -X 2 -C 2-6 alkenyl-X 2 -, and -X 2 -C2-6 alkynyl-X 2 -, each of which is optionally substituted at each occurrence with one or more R 10 ;
  • X 10 at each occurrence is independently selected from -C(O)-, and -C(0)N(R 10 )-*, wherein * represents where X 10 is bound to R 5 ;
  • X 2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R 10 )-, -C(O)-,
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , - S(0)R 10 , and -S(0) 2 R 10 ; Ci - l o alkyl, C 2 -io alkenyl, C 2 -io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, - OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 ) 2 , -
  • each C3-0 carbocycle, and 3- to l2-membered heterocycle in R 5 is independently optionally substituted with one or more substituents selected from halogen, - OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 ,
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )2, -S(0)R 10 , - S(0) 2 R 10 ,
  • L 2 can be attached at C2, C3, C4, or C5 of the benzazepine core, wherein the numbering of the benzazepine is as follows:
  • L 2 is attached to the benzazepine core at C4. In certain embodiments represents a double bond and L 2 is attached to the benzazepine core at C4.
  • L 10 can be attached at C6, C7, C8 or C9.
  • L 10 is attached to the benzazepine core at C8.
  • L 2 is attached to the benzazepine core at
  • C4 and L 10 is attached to the benzazepine core at C8.
  • the substitutable carbon on the benzazepine core is selected from C2, C3, C4, C5, C6, C7, C8, and C9.
  • the compound of Formula (IIA) is represented by Formula (IIB):
  • R 24 , and R 25 are independently selected from hydrogen, halogen, -OR 10 , -SR 10 , -
  • the compound of Formula (IIA) is represented by Formula (IIC):
  • L 10 is -C(O)-. In certain embodiments, L 10 is selected from -C(0)N(R 10 )-*. In certain embodiments, R 10 of -C(0)N(R 10 )-* is selected from hydrogen and Ci -6 alkyl. In certain embodiments, L 10 is -C(0)NH-*.
  • R 5 is an optionally substituted 8- to l2-membered bicyclic carbocycle.
  • R 5 is an optionally substituted bicyclic carbocycle, e.g., an optionally substituted 8- to 12- membered bicyclic carbocycle, such as indane, naphthalene, dihydronaphthalene, tetrahydronaphthalene.
  • R 5 is an optionally substituted 8- to 12- membered bicyclic carbocycle with one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -N(R 10 ) 2 , -
  • R 5 is an optionally substituted indane or an optionally substituted
  • R 5 is selected from: and , either of which is optionally substituted.
  • R 5 is selected from:
  • R 5 is an optionally substituted unsaturated C 4-8 carbocycle.
  • R 5 is an optionally substituted unsaturated C 4-6 carbocycle, e.g., cyclopentene, cyclopentadiene, cyclohexene, cyclobutene.
  • R 5 is an optionally substituted unsaturated C 4-6 carbocycle with one or more substituents independently selected from optionally substituted C3-12 carbocycle, and optionally substituted 3- to l2-membered heterocycle.
  • R 5 is an optionally substituted unsaturated C 4-6 carbocycle with one or more substituents
  • R 5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle.
  • R 5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle with one or more substituents independently selected from -C(0)OR 10 , -N(R 10 )2, - OR 10 , and optionally substituted Ci-io alkyl.
  • R 5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle substituted with -C(0)OR 10 .
  • R 5 is an optionally substituted fused 6-6 bicyclic heterocycle.
  • the fused 6-6 bicyclic heterocycle may be an optionally substituted pyridine-piperidine.
  • L 10 is bound to a carbon atom of the pyridine of the fused pyridine-piperidine.
  • R 5 is selected from tetrahydroquinoline, tetrahydroisoquinoline,
  • R 5 is an optionally substituted
  • R 5 is selected from:
  • R 5 is selected from tetrahydronaphthalene, tetrahydroquinoline, tetrahydroisoquinoline, indane, cyclopropylbenzene, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is optionally substituted.
  • R 5 is selected from tetrahydronaphthalene, tetrahydroquinoline, tetrahydroisoquinoline, indane, cyclopropylbenzene, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is optionally substituted.
  • R 5 is selected from:
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen, -OH, -OR 10 , -N0 2 , -CN, and Ci-10 alkyl. In some embodiments, R 20 , R 21 , R 22 , and R 23 are each hydrogen. In some
  • R 21 is halogen, e.g. fluorine.
  • R 21 is hydrogen.
  • R 21 is -OR 10 .
  • R 21 may be -OCH3.
  • L 10 is selected from -C(0)N(R 10 )-* and -C(O)-;
  • L 2 is -C(O)-
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 5 is selected from optionally substituted unsaturated C 4-8 carbocycle, optionally substituted bicyclic carbocycle, and optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle wherein substituents on R 5 are independently selected at each occurrence from: halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -N(R 10 ) 2 , -N(R 10 ) 2 , -
  • each C 3 -i 2 carbocycle, and 3- to l2-membered heterocycle in R 5 is independently optionally substituted with one or more substituents selected from halogen, - OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 ,
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 .
  • L 10 is selected from -C(0)N(R 10 )-* and -C(O)-;
  • L 2 is -C(O)-
  • R 1 is hydrogen;
  • R 2 is an amine masking group;
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ;
  • R 5 is selected from optionally substituted optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle wherein substituents are independently selected at each occurrence from: halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 ) 2 ,
  • each C 3 -i 2 carbocycle, and 3- to l2-membered heterocycle in R 5 is independently optionally substituted with one or more substituents selected from halogen, - OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 ,
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 .
  • L 10 is selected from -C(0)N(R 10 )-*, such as -C(0)NH-*;
  • L 2 is -C(O)-
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 , such as R 4 is -N(R 10 ) 2 ;
  • R 5 is selected from optionally substituted optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle, such as R 5 is selected from a tetrahydroquinoline,
  • R 5 substituents on R 5 are independently selected at each occurrence from: halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , -
  • the compound may further comprise a linker (L 3 ).
  • the linker may be covalently bound to any position, valence permitting, on a compound or salt of Formula (IIA), (IIB) or (IIC).
  • the linker may be bound to R 4 , R 5 , X 10 or X 2 .
  • the linker may be bound to a nitrogen atom, or an oxygen atom of a compound or salt of Formula (IIA), (IIB) or (IIC).
  • the linker may comprise a reactive moiety, e.g., an electrophile that can react to form a covalent bond with a moiety of an antibody, e.g., lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue of the antibody.
  • a compound or salt of Formula (IIA), (IIB) or (IIC) may be covalently bound through the linker to an antibody construct.
  • the present disclosure provides a compound represented by the structure of F ormul a (HI A) :
  • L 11 is -X 11 -;
  • L 2 is selected from -X 21 -, -X 21 -C I-6 alkylene-X 21 -, -X 21 -C 2-6 alkenylene-X 21 -, and - X 2 -C 2-6 alkynylene-X 2 -, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R 12 ;
  • X 11 is selected from -C(O)- and -C(0)N(R 10 )-*, wherein * represents where X 11 is bound to R 6 ;
  • X 21 at each occurrence is independently selected from a bond, -O- , -S-, -N(R 10 )-, -C(O)-,
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , - S(0)R 10 , and -S(0) 2 R 10 ; Ci - l o alkyl, C 2-i o alkenyl, C 2-i o alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR 10 ,
  • R 6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R 7 and R 6 is further optionally substituted by one or more additional substituents independently selected from R 12 ;
  • R 7 is selected from -C(0)NHNH 2 , -C(0)NH-Ci -3 alkylene-NH(R 10 ), -C(0)CH 3 , - Ci- 3 alkyl ene-NHC(0)0R 11 , -Ci -3 alkylene-NHC(0)R 10 , -Ci -3 alkylene-NHC(0)NHR 10 , -Ci- 3 alkylene-NHC(0)-Ci -3 alkylene-R 10 , and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R 12 ;
  • R 10 is independently selected at each occurrence from hydrogen, -NH 2 , - C(0)0CH2C 6 H5; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH
  • R 11 is selected from C 3-12 carbocycle and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from R 12 ;
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
  • L 11 is independently selected from -X 11 -;
  • L 2 is independently selected from -X 2 -, -X 2 -C I-6 alkylene-X 2 -, -X 2 -Ci- ( , alkenyl ene-X 2 -, and -X 2 -Ci- ( , alkynylene-X 2 -, each of which is optionally substituted at each occurrence with one or more R 10 ;
  • X 11 at each occurrence is independently selected from -C(O)-, and -C(0)N(R 10 )-*, wherein * represents where X 11 is bound to R 6 ;
  • X 2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R 10 )-, -C(O)-, -C(0)0-, -0C(0)-, -0C(0)0-, -C(0)N(R 10 )-, -C(O)N(R 10 )C(O)-, - (O)N(R 10 )C(O)N(R 10 ), -N(R 10 )C(O)-,
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , - S(0)R 10 , and -S(0) 2 R 10 ; Ci - l o alkyl, C 2 -io alkenyl, C 2 -io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, - OR 10 ,
  • each C 3-i2 carbocycle, and 3- to l2-membered heterocycle in R 4 is independently optionally substituted with one or more substituents selected from halogen, - OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 ,
  • R 6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R 7 ;
  • R 7 is selected from -C(0)NHNH 2 , -C(0)NH-Ci -3 alkylene-NH(R 10 ), -C(0)CH 3 , - Ci -3 alkylene-NHC(0)OR 10 , -Ci -3 alkylene-NHC(0)R 10 , -Ci -3 alkylene-NHC(0)NHR 10 , and -Ci- 3 alkylene-NHC(0) -Ci -3 alkylene-(R 10 ) 2 ; and an optionally substituted 3- to l2-membered heterocycle;
  • L 2 can be attached at C2, C3, C4, or C5 of the benzazepine core, wherein the numbering of the benzazepine is as follows:
  • L 2 is attached to the benzazepine core at C4.
  • L 2 is attached to the benzazepine core at C4.
  • L 11 can be attached at C6, C7, C8 or C9. In certain embodiments, for a compound or salt of Formula (IIIA), L 11 is attached to the benzazepine core at C8. In certain embodiments for a compound or salt of Formula (IIIA), represents a double bond, L 2 is attached to the benzazepine core at C4 and L 11 is attached to the benzazepine core at C8.
  • the substitutable carbon on the benzazepine core is selected from C2, C3, C4, C5, C6, C7, C8, and C9.
  • the benzazepine core for a compound or salt of Formula (IIIA) can be optionally substituted by a substituent selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -S(0)R 10 , -S(0) 2 R 10 - C(0)R 10 , -C(0)OR 10 , -OC(0)R 10 ,
  • a moiety at any one of C2, C3, C4, C5, C6, C7, C8, and C9 of the benzazepine core is independently selected from hydrogen, halogen, -OR 10 , -SR 10 ,
  • the compound of Formula (IIIA) is represented by Formula (IIIB):
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen,
  • the compound of Formula (IIIA) is represented by Formula (IIIC):
  • L 11 is selected from -C(O)-, and -C(0)N(R 10 )-*, wherein * represents where L 11 is bound to R 6 .
  • L 11 is -C(O)-.
  • L 11 is selected from -C(0)N(R 10 )-*.
  • R 10 or L 11 is selected from hydrogen and Ci -6 alkyl.
  • R 6 is further optionally substituted by one or more additional substituents independently selected from R 12 .
  • R 7 is selected from -C(O)N(R 10 )N(R 10 ) 2 , -C(O)N(R 10 )-C I-3 alkylene-N(R 10 ) 2 , -C(0)CH 3 , -Ci -3 alkylene- N(R 10 )C(O)OR u , -Ci -3 alkylene-N(R 10 )C(O)R 10 , -Ci -3 alkylene-N(R 10 )C(O)N(R 10 ) 2 , -Ci- 3 alkylene-N(R 10 )C(O)-Ci -3 alkylene-R 10 , and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R 12 .
  • R 11 may be selected from C3-12 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from R 12 .
  • R 7 is selected from -C(0)NHNH 2 , - C(0)NH-CI -3 alkylene-NH(R 10 ), -C(0)CH 3 , -C1.3 alkylene-NHC
  • R 7 is selected from -C(0)NHNH 2 , -C(0)NH-C I-3 alkyl ene-NH(R 10 ), -C(0)CH 3 , -C1.3 alkylene-NHC(0)OR u , -Ci- 3 alkylene-NHC(0)NHR 10 , -Ci- 3 alkylene-NHC(0)-Ci- 3 alkylene-R 10 , and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R 12 .
  • R 7 is a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R 12 .
  • R 6 is selected from an optionally substituted phenyl and an optionally substituted 5-6 membered heteroaryl, wherein R 6 is substituted with one or more substituents independently selected from R 7 .
  • R 6 is selected from an optionally substituted phenyl, wherein the phenyl is substituted with one or more substituents independently selected from R 7 .
  • R 6 is selected from an optionally substituted phenyl, wherein the phenyl is substituted with one or more substituents independently selected from -C(0)NHNH 2 , -C(0)NH- Ci.3alkylene-NH(R 10 ), and -C(0)CH 3 ; and 3- to l2-membered heterocycle, which is optionally substituted with one or more substituents selected from -OH, -N(R 10 ) 2 , -NHC(0)(R 10 ), - NHC(0)0(R 10 ), -NHC(O)N(R 10 ) 2 , -C(O)R 10 ,-C(O) 2 R 10 , and -Ci-3alkylene-(R 10 ), and optionally further substituted with one or more additional substituents.
  • R 6 is selected from:
  • R 6 is selected from an optionally substituted 5-6 membered heteroaryl, wherein the heteroaryl is substituted with one or more substituents independently selected from R 7 .
  • R 6 may be selected from a 5- and 6-membered heteroaryl substituted with one or more substituents independently selected from R 7 , and R 6 may be further optionally substituted with one or more additional substituents selected from R 12 .
  • R 6 is selected from an optionally substituted 5-6 membered heteroaryl, wherein the heteroaryl is substituted with one or more substituents independently selected from
  • R 6 is selected from substituted pyridine, pyrazine, pyrimidine, pyridazine, furan, pyran, oxazole, thiazole, imidazole, pyrazole, oxadiazole, oxathiazole, and triazole, and R 6 is optionally further substituted with one or more additional substituents independently selected from R 12 .
  • R 6 may be an optionally substituted pyridine and R 6 is optionally further substituted with one or more additional substituents independently selected from R 12 .
  • R 6 may be represented as follows: some embodiments, R is substituted pyridine, and wherein R 7 is -Ci -3 alkylene-NHC(0)-Ci -3 alkylene-R 1 °. R 7 may be -Cialkylene-NHC(0)-Cialkylene-R 10 . R 7 may be -Cialkylene-NHC(0)-Cialkylene-NH 2 . In some embodiments for a compound or salt
  • R 6 is selected from:
  • the compound may further comprise a linker (L 3 ).
  • the linker may be covalently bound to any position, valence permitting, on a compound or salt of Formula (IIIA), (IIIB) or (IIIC).
  • the linker may be bound to R 4 , R 6 , X 11 or X 2 .
  • the linker is bound to a nitrogen or oxygen atom of a compound or salt of Formula (IIIA), (IIIB) or (IIIC).
  • the linker may comprise a reactive moiety, e.g., an electrophile, that can react to form a covalent bond with a moiety of an antibody construct, a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue of an antibody.
  • a compound or salt of Formula (IIIA), (IIIB) or (IIIC) may be covalently bound through the linker moiety to an antibody construct.
  • L 11 is -C(0)N(R 10 )- *, such as L 11 is -C(0)NH-;
  • L 2 is selected from -C(O)- and -C(0)NR 10 -;
  • R 1 is hydrogen;
  • R 2 is an amine masking group;
  • R 6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R 7 ;
  • R 7 is selected from -C(0)NHNH 2 , -C(0)NH-Ci -3 alkylene-NH(R 10 ), -C(0)CH 3 , -Ci- 3 alkylene-NHC(0)OR u , -Ci -3 alkylene-NHC(0)R 10 , -Ci -3 alkylene-NHC(0)NHR 10 , and -Ci- 3 alkylene-NHC(0) -Ci -3 alkylene-(R 10 ) 2 ; and a 3 - to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R 12 ;
  • R 11 is selected from C 3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from R 12 ; and wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 .
  • L 11 is -C(0)N(R 10 )- *
  • L 2 is -C(O)-
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ;
  • R 6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R 7 ;
  • R 7 is selected from -C(0)NHNH 2 , -C(0)NH-Ci -3 alkylene-NH(R 10 ), -C(0)CH 3 , -Ci- 3 alkylene-NHC(0)OR u , -Ci -3 alkylene-NHC(0)R 10 , -Ci -3 alkylene-NHC(0)NHR 10 , and -Ci- 3 alkylene-NHC(0) -Ci -3 alkylene-(R 10 ) 2 ;
  • R 10 is independently selected at each occurrence from hydrogen, -NH 2 , -C(0)0CH 2 C 6 H 5 ; and Ci-io alkyl, C 2 -io alkenyl, C 2 -io alkynyl, C 3 -i 2 carbocycle, and 3- to l2-membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -CN, -N0 2 , -NH
  • R 11 is selected from C 3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from R 12 ; and wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 .
  • L 11 is -C(0)N(R 10 )- *, such as L 11 is -C(0)NH-;
  • L 2 is -C(O)-
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ;
  • R 6 is selected from phenyl and 5- or 6- membered heteroaryl, e.g., pyridine, any one of which is substituted with one or more substituents selected from R 7 ;
  • R 7 is selected from -C(0)NHNH 2 , -C(0)NH-Ci -3 alkylene-NH(R 10 ), -C(0)CH 3 , -Ci- 3 alkylene-NHC(0)OR u , -Ci -3 alkylene-NHC(0)NHR 10 , and -Ci -3 alkylene-NHC(0) -Ci- 3 alkylene-(R 10 ) 2 ; and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R 12 ;
  • R 11 is selected from C 3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from R 12 ; and wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 .
  • R 20 , R 21 , R 22 , and R 23 are independently selected from hydrogen, halogen, -OH, -N0 2 , -CN, and Ci-io alkyl.
  • R 20 , R 21 , R 22 , and R 23 are each hydrogen.
  • R 24 and R 25 are independently selected from hydrogen, halogen, -OH,
  • R 24 and R 25 are each hydrogen.
  • R 24 and R 25 taken together form an optionally substituted saturated C3-5 carbocycle, wherein substituents may be selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , - N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -OC(0)R 10 , -N0 2 , 0,
  • R 2 is an amine masking group selected from an acid-labile moiety or an enzymatically-labile moiety.
  • R 2 is selected from a moiety covalently bound to an amine that is selectively cleaved under intracellular conditions.
  • R 2 together with the nitrogen to which it is attached forms a carbamate or an amide.
  • R 2 is represented by the formula:
  • R 101 is selected from an amino acid, a peptide, -0-(Ci-C 6 alkyl) and -Ci-C 6 alkyl, wherein alkyl of -0-(Ci-C 6 alkyl) and -Ci-C 6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -N(R 10 ) 2 , -C(0)R 10 , - C(O)N(R 10 ) 2 , -N0 2 , -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle and R 10 is as previously defined; and
  • R 101 is selected from -0-(Ci-C 4 alkyl) and -Ci-C 4 alkyl, wherein alkyl of -0-(Ci-C 4 alkyl) and -Ci-C 4 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR 10 , -SR 10 , -N(R 10 ) 2 , -C(0)R 10 , -C(O)N(R 10 ) 2 , -N0 2 , - CN, C3-13 carbocycle, and 3- to l2-membered heterocycle.
  • R 2 is selected from 9-fluorenylmethylcarbonyl-, /c/V-butoxy carbonyl-, benzyloxy carbonyl-, acetyl-, and trifluoroacetyl-.
  • the amino acid of R 101 is selected from any natural or non natural amino acid.
  • the amino acid may be selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan.
  • the amino acid may be selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan.
  • the amino acid is an L-amino acid.
  • the peptide of R 101 includes amino acids each independently selected from any natural or non-natural amino acid.
  • the first amino acid may each be independently selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan.
  • the amino acids are each independently L-amino acids or D-amino acids.
  • the peptide is a dipeptide, tripeptide or tetrapeptide.
  • each amino acid of a dipeptide, tripeptide or tetrapeptide is independently selected from a D- and L-amino acid.
  • the amino acid immediately attached to the amine is an L-amino acid, e.g., R 101 is represented by the formula: -aal-aa2, or - aal-aa2-aa3, where aal is an L-amino acid and aa2 and aa3 are independently selected from D- and L-amino acids.
  • the first amino acid (including R 100 ) is an L-amino acid selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan and the remaining amino acids are D or L amino acids selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan.
  • an amine masking group selected from arginine, histidine,
  • L 2 is independently selected from -X 2 -, -X 2 -C I-6
  • L 2 is selected from -C(O)- , and -C(0)NR 10 -.
  • a compound of any one of Formulas (IA), (IB) is selected from -C(O)- , and -C(0)NR 10 -.
  • L 2 is selected from -C(O)-.
  • L 2 is selected from -C(0)NR 10 -.
  • R 10 of -C(0)NR 10 - is selected from hydrogen and Ci -6 alkyl.
  • L 2 is -C(0)NH-.
  • X 2 at each occurrence is independently selected from a bond, -0-,
  • X 2 is selected from -C(O)-.
  • X 2 is selected from -C(0)NR 10 -.
  • R 10 of -C(0)NR 10 - is selected from hydrogen and Ci- 6 alkyl.
  • X 2 is -C(0)NH-.
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , - C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ; Ci-io alkyl, C 2-i o alkenyl, C 2-i o alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 ) 2 , - N(R 10 )
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , - C(0)R 10 ,
  • R 4 is - N(R 10 ) 2 .
  • R 4 is -N(R 10 )2, and each R 10 is independently selected from optionally substituted Ci -6 alkyl.
  • R 4 is -N(R 10 ) 2, and each R 10 is independently selected from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted.
  • R 10 is independently selected from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted.
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R 12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
  • the disclosure provides compounds that include a linker, L 3 .
  • a compound bound to a linker is represented by the structure of Formula (IV A):
  • L 12 is independently selected from -X 3 -, -X 3 -C I-6 alkyl-X 3 -, -X 3 -C 2-6 alkenyl-X 3 -, and
  • L 22 is independently selected from -X 4 -, -X 4 -C I-6 alkyl-X 4 -, -X 4 -C 2-6 alkenyl-X 4 -, and
  • X 3 and X 4 are independently selected at each occurrence from a bond, -O- , -S-, -N(R 10 )-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)-, -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(0)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )
  • R 1 is hydrogen
  • R 2 is an amine masking group
  • L 3 is a linker moiety, wherein at least one of R 10 is L 3 or at least one substituent on R 4
  • any substitutable carbon on the benzazepine core is optionally substituted by a substituent selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 -, -N(R 10 ) 2 , -S(0)R 10 , - S(0) 2 R 10 ,
  • Ci-io alkyl, C2-10 alkenyl, and C2-10 alkynyl, or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
  • L 22 can be attached at C2, C3, C4, or C5 of the benzazepine core, wherein the numbering of the benzazepine is as
  • L 22 is attached to the benzazepine core at C4.
  • L 22 is attached to the benzazepine core at
  • L 12 can be attached at C6, C7, C8 or C9. In certain embodiments, for a compound or salt of Formula (IV A), L 12 is attached to the benzazepine core at C8. In certain embodiments for a compound or salt of Formula (IV A), represents a double bond, L 22 is attached to the benzazepine core at C4 and L 12 is attached to the benzazepine core at C8.
  • the substitutable carbon on the benzazepine core is selected from C2, C3, C4, C5, C6, C7, C8, and C9.
  • the benzazepine core for a compound or salt of Formula (IV A) can be optionally substituted by a substituent selected from halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 -, -N(R 10 ) 2 , -S(0)R 10 , -S(0) 2 R 10 , - C(0)R 10 , -C(0)OR 10 , -OC(0)R 10 ,
  • a moiety at any one of C2, C3, C4, C5, C6, C7, C8, and C9 of the benzazepine core is independently selected from hydrogen, halogen, -OR 10 , -SR 10 ,
  • the compound of Formula (IV A) is represented by Formula (IVB):
  • the compound of Formula (IV A) is represented by Formula (IVC):
  • L 12 is selected from -X 3 -, -X 3 -C I-6 alkylene-X 3 -, -X 3 -C 2-6 alkenylene-X 3 -, and -X 3 -C 2-6 alkynyl ene-X 3 -, each of which is optionally substituted on the alkylene, alkenylene and alkyneylene with one or more R 12 .
  • L 22 is selected from -X 4 -, -X 4 -C I-6 alkylene-X 4 -, -X 4 -C 2-6 alkenylene-X 4 -, and -X 4 -C 2-6 alkynylene-X 4 -, optionally substituted on the alkylene, alkenylene and alkynylene with one or more R 12 .
  • L 12 is independently selected from -X 3 -, -X 3 -C I-6 alkyl-X 3 -, -X 3 -C 2-6 alkenyl-X 3 -, and -X 3 -C 2-6 alkynyl-X 3 -, each of which is optionally substituted at each occurrence with one or more R 10 .
  • L 12 is selected from -C(O)-, and -C(0)NR 10 -.
  • L 12 is -C(0)N(R 10 )-.
  • R 10 of -C(0)N(R 10 )- is selected from hydrogen, Ci -6 alkyl, and -L 3 .
  • L 12 is -C(0)NH-.
  • L 12 is - C(0)N(L 3 )-.
  • R 4 and R 8 are independently selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , - S(0)R 10 , and -S(0) 2 R 10 ; Ci - l o alkyl, C 2-i o alkenyl, C 2-i o alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from L 3 , halogen, -OR 10 , -SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 ) 2 , - N(R 10 ) 2 , -C(0)R 10 , -C(0)
  • R 8 is an optionally substituted 5-6 membered heteroaryl. In some embodiments, R 8 is an optionally substituted 5-6 membered heteroaryl, substituted with -L 3 . R 8 may be an optionally substituted pyridine, substituted with -L 3 .
  • L 22 is independently selected from -X 4 -, -X 4 -C I-6 alkyl-X 4 -, -X 4 -C 2-6 alkenyl-X 4 -, and -X 4 -C 2-6 alkynyl-X 4 -, each of which is optionally substituted at each occurrence with one or more R 10 .
  • L 22 is selected from -C(O)-, and -C(0)NR 10 -. L 22 may be -C(O)-.
  • L 22 is -C(0)NR 10 -, wherein, R 10 of -C(0)NR 10 - may be selected from hydrogen, Ci -6 alkyl, and -L 3 .
  • L 22 is -C(0)NH-.
  • L 12 is -C(0)N(L 3 )-.
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ; Ci-io alkyl, C 2 -io alkenyl, C 2 -io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from -L 3 , halogen, -OR 10 , - SR 10 , -C(O)N(R 10 ) 2 , -N(RIO)C(0)RIO,
  • R 10 of -N(R 10 ) 2 is selected from -L 3 and hydrogen, and wherein at least one R 10 of -N(R 10 ) 2 is -L 3 .
  • R 4 is selected from: -OR 10 , -N(R 10 ) 2 , -C(O)N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 , -S(0)R 10 , and -S(0) 2 R 10 ; and Ci-io alkyl, C 2-i o alkenyl, and C 2-i o alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from L 3 , halogen, -OR 10 , - SR 10 , -C(O)N(R 10 ) 2 , -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 ) 2 , -N(R 10 ) 2 , -C(0)R 10 , -C(0)OR 10 ,
  • L 3 is a noncleavable linker.
  • L 3 is cleavable linker such as a linker cleavable by a lysosomal enzyme.
  • a moiety described herein includes the symbol which indicates the point of attachment, e.g., the point of attachment of a chemical moiety to the remainder of the compound, the point of attachment of a linker to a compound of the disclosure, or the point of attachment of a linker to an antibody construct, as described herein.
  • linkers (L 3 ) are described in the following paragraphs and additional linkers are described in the subsequent section entitled“Linkers”.
  • peptide is a group comprising from one to ten
  • L 4 represents the C-terminal of the peptide and L 5 is selected from a bond, alkylene and heteroalkylene, wherein L 5 is optionally substituted with one or more groups independently selected from R 32 ;
  • L 4 represents the C-terminal of the peptide and L 5 is selected from a bond, alkyl ene and heteroalkylene, wherein L 5 is optionally substituted with one or more groups independently selected from R 32 ;
  • RX * is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein Antibody is an antibody construct;
  • L 4 and L 5 are independently selected from a bond, an alkylene and a heteroalkylene, each of which is optionally substituted with one or more groups independently selected from R 12 ;
  • * 3 ⁇ 4 ⁇ on the left represents the point of attachment to the remainder of the compound,
  • RX * is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety attached at the on the right to a residue of an antibody construct;
  • peptide is a group comprising from one to 10 amino acids.
  • the compound is:
  • n 0-9 and ⁇ represents the point of attachment to the remainder of the compound.
  • n 0-9 and ⁇ on the left represents the point of attachment to the remainder of the compound.
  • Chemical entities having carbon-carbon double bonds or carbon-nitrogen double bonds may exist in Z- or E- form (or cis- or trans- form). Furthermore, some chemical entities may exist in various tautomeric forms. Unless otherwise specified, compounds described herein are intended to include all Z-, E- and tautomeric forms as well.
  • a "tautomer” refers to a molecule wherein a proton shift from one atom of a molecule to another atom of the same molecule is possible.
  • the compounds disclosed herein are used in different enriched isotopic forms, e.g., enriched in the content of 2 H, 3 ⁇ 4, U C, 13 C and/or 14 C.
  • the compound is deuterated in at least one position.
  • deuterated forms can be made by the procedure described in U.S. Patent Nos. 5,846,514 and 6,334,997.
  • deuteration can improve the metabolic stability and or efficacy, thus increasing the duration of action of drugs.
  • compounds described herein are intended to include compounds which differ only in the presence of one or more isotopically enriched atoms.
  • compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by 13 C- or 14 C-enriched carbon are within the scope of the present disclosure.
  • the compounds of the present disclosure optionally contain unnatural proportions of atomic isotopes at one or more atoms that constitute such compounds.
  • the compounds may be labeled with isotopes, such as for example, deuterium ( 2 H), tritium (3 ⁇ 4), iodine-l25 ( 125 I) or carbon-l4 ( 14 C). Isotopic substitution with 2 H, U C, 13 C, 14 C, 15 C, 12 N, 13 N,
  • the compounds disclosed herein have some or all of the 3 ⁇ 4 atoms replaced with 2 H atoms.
  • the methods of synthesis for deuterium-containing compounds are known in the art and include, by way of non-limiting example only, the following synthetic methods.
  • Deuterium substituted compounds are synthesized using various methods such as described in: Dean, Dennis C.; Editor. Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [In: Curr., Pharm. Des., 2000; 6(10)] 2000, 110 pp; George W.; Varma, Rajender S.
  • Deuterated starting materials are readily available and are subjected to the synthetic methods described herein to provide for the synthesis of deuterium-containing compounds. Large numbers of deuterium-containing reagents and building blocks are available commercially from chemical vendors, such as Aldrich Chemical Co.
  • Compounds of the present invention also include crystalline and amorphous forms of those compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof.
  • the present disclosure provides a method for treating cancer.
  • the present disclosure provides a method comprising administering a conjugate, compound or salt of Formula (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IV A),
  • the compounds of the disclosure exhibit selective binding or agonizing properties for one receptor over another receptor.
  • a compound described herein selectively binds or modulates the activity of one toll-like receptor over another, e.g., TLR8 and TLR7.
  • IVC has greater activity for one toll-like receptor over another, e.g., TLR8 and TLR7.
  • a compound of the disclosure agonizes TLR8 with an EC so of 500 nM or less while the same compound agonizes TLR7 with an EDso of greater than 1 mM.
  • a compound of the disclosure agonizes TLR8 with at least an ECso of an order of magnitude or even two orders of magnituge less than the amount of the same compound required to show an agonizing effect on TLR7.
  • salts particularly pharmaceutically acceptable salts, of the compounds described herein.
  • the compounds of the present disclosure that possess a sufficiently acidic, a sufficiently basic, or both functional groups can react with any of a number of inorganic bases, and inorganic and organic acids, to form a salt.
  • compounds that are inherently charged such as those with a quaternary nitrogen, can form a salt with an appropriate counterion, e.g., a halide such as bromide, chloride, or fluoride, particularly bromide.
  • the compounds described herein may in some cases exist as diastereomers, enantiomers, or other stereoisomeric forms.
  • the compounds presented herein include all diastereomeric, enantiomeric, and epimeric forms as well as the appropriate mixtures thereof. Separation of stereoisomers may be performed by chromatography or by forming diastereomers and separating by recrystallization, or chromatography, or any combination thereof.
  • Stereoisomers may also be obtained by stereoselective synthesis.
  • compositions described herein include the use of amorphous forms as well as crystalline forms (also known as polymorphs).
  • the compounds described herein may be in the form of pharmaceutically acceptable salts.
  • active metabolites of these compounds having the same type of activity are included in the scope of the present disclosure.
  • the compounds described herein can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like.
  • the solvated forms of the compounds presented herein are also considered to be disclosed herein.
  • compounds or salts of the compounds of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) may be prodrugs, e.g., wherein a hydroxyl in the parent compound is presented as an ester or a carbonate, or carboxylic acid present in the parent compound is presented as an ester.
  • prodrug is intended to encompass compounds which, under physiologic conditions, are converted into pharmaceutical agents of the present disclosure.
  • One method for making a prodrug is to include one or more selected moieties which are hydrolyzed under physiologic conditions to reveal the desired molecule.
  • the prodrug is converted by an enzymatic activity of the host animal such as specific target cells in the host animal.
  • esters or carbonates e.g., esters or carbonates of alcohols or carboxylic acids and esters of phosphonic acids
  • esters or carbonates are preferred prodrugs of the present disclosure.
  • Prodrug forms of the herein described compounds wherein the prodrug is metabolized in vivo to produce a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB), or (IVC) as set forth herein are included within the scope of the claims.
  • some of the herein-described compounds may be a prodrug for another derivative or active compound.
  • Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent is not. Prodrugs may help enhance the cell permeability of a compound relative to the parent drug. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. Prodrugs may be designed as reversible drug derivatives, for use as modifiers to enhance drug transport to site-specific tissues or to increase drug residence inside of a cell.
  • the design of a prodrug increases the lipophilicity of the pharmaceutical agent. In some embodiments, the design of a prodrug increases the effective water solubility. See, e.g., Fedorak e/ a/., Am. J Physiol ., 269:G2l0-2l8 (1995); McLoed e/ a/., Gastroenterol , 106:405-413 (1994); Hochhaus et al., Biomed. Chrom., 6:283-286 (1992); J. Larsen and H. Bundgaard, Int. J. Pharmaceutics, 37, 87 (1987); J. Larsen et al., Int. J.
  • the present disclosure provides methods of producing the above-defined compounds.
  • the compounds may be synthesized using conventional techniques.
  • these compounds are conveniently synthesized from readily available starting materials.
  • the compounds and salts described herein may be bound to a linker also referred to herein as L 3 , e.g., a peptide linker.
  • the linker is also bound to an antibody construct and referred to as an antibody construct conjugate or conjugate.
  • Linkers of the conjugates described herein may not affect the binding of active portions of a conjugate, e.g, the antigen binding domains, Fc domains, target binding domains, antibodies, agonists or the like, to a target, which can be a cognate binding partner such as an antigen.
  • a conjugate can comprise multiple linkers, each having one or more compounds attached. These linkers can be the same linkers or different linkers.
  • a linker can be short, flexible, rigid, cleavable, non-cleavable, hydrophilic, or hydrophobic.
  • a linker can contain segments that have different characteristics, such as segments of flexibility or segments of rigidity.
  • the linker can be chemically stable to extracellular environments, for example, chemically stable in the blood stream, or may include linkages that are not stable or selectively stable.
  • the linker can include linkages that are designed to cleave and/or immolate or otherwise breakdown specifically or non- specifically inside cells.
  • a cleavable linker can be sensitive to enzymes.
  • a cleavable linker can be cleaved by enzymes such as proteases.
  • a cleavable linker may comprise a valine-citrulline linker or a valine-alanine peptide.
  • a valine-citrulline- or valine-alanine-containing linker can contain a pentafluorophenyl group.
  • a valine-citrulline- or valine-alanine-containing linker can contain a maleimide or succinimide group.
  • a valine-citrulline- or valine-alanine-containing linker can contain a para aminobenzyl alcohol (PABA) group or para-aminobenzyl carbamate (PABC).
  • PABA para aminobenzyl alcohol
  • PABC para-aminobenzyl carbamate
  • a valine-citrulline- or valine-alanine-containing linker can contain a PABA group and a pentafluorophenyl group.
  • a valine-citrulline- or valine-alanine-containing linker can contain a PABA group and a maleimide or succinimide group.
  • a non-cleavable linker can be protease insensitive.
  • a non-cleavable linker can be maleimidocaproyl linker.
  • a maleimidocaproyl linker can comprise N- maleimidomethylcyclohexane-l-carboxylate.
  • a maleimidocaproyl linker can contain a succinimide group.
  • a maleimidocaproyl linker can contain pentafluorophenyl group.
  • a linker can be a combination of a maleimidocaproyl group and one or more polyethylene glycol molecules.
  • a linker can be a maleimide-PEG4 linker.
  • a linker can be a combination of a maleimidocaproyl linker containing a succinimide group and one or more polyethylene glycol molecules.
  • a linker can be a combination of a maleimidocaproyl linker containing a
  • a linker can contain maleimides linked to polyethylene glycol molecules in which the polyethylene glycol can allow for more linker flexibility or can be used lengthen the linker.
  • a linker can be a
  • a linker can be a linker suitable for attachment to an engineered cysteine (THIOMAB).
  • a THIOMAB linker can be a (maleimidocaproyl)-(valine-citrulline)-(para-aminobenzyloxycarbonly)-linker.
  • a linker can also comprise alkylene, alkenylene, alkynylene, polyether, polyester, polyamide group(s) and also, polyamino acids, polypeptides, cleavable peptides, or
  • a linker can contain a maleimide at one end and an N- hydroxysuccinimidyl ester at the other end.
  • a linker can contain a lysine with an N-terminal amine acetylated, and a valine-citrulline cleavage site.
  • a linker can be a link created by a microbial transglutaminase, wherein the link can be created between an amine-containing moiety and a moiety engineered to contain glutamine as a result of the enzyme catalyzing a bond formation between the acyl group of a glutamine side chain and the primary amine of a lysine chain.
  • a linker can contain a reactive primary amine.
  • a linker can be a Sortase A linker.
  • a Sortase A linker can be created by a Sortase A enzyme fusing an LXPTG recognition motif to an N-terminal GGG motif to regenerate a native amide bond. The linker created can therefore link a moiety attached to the LXPTG recognition motif with a moiety attached to the N-terminal GGG motif.
  • a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB), or (IVC) is linked to the antibody construct by way of a linker(s), also referred to herein as L 3 .
  • L 3 as used herein, may be selected from any of the linker moieties discussed herein.
  • the linker linking the compound or salt to the antibody construct of a conjugate may be short, long, hydrophobic, hydrophilic, flexible or rigid, or may be composed of segments that each independently have one or more of the above- mentioned properties such that the linker may include segments having different properties.
  • the linkers may be polyvalent such that they covalently link more than one compound or salt to a single site on the antibody construct, or monovalent such that covalently they link a single compound or salt to a single site on the antibody construct.
  • Linkers of the disclosure may have from about 10 to about 500 atoms in a linker, such as from about 10 to about 400 atoms, such as about 10 to about 300 atoms in a linker. In certain embodiments, linkers of the disclosure have from about 30 to about 400 atoms, such as from about 30 to about 300 atoms in the linker.
  • the linkers may link a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) to the antibody construct by a covalent linkages between the linker and the antibody construct and compound.
  • linker is intended to include (i) unconjugated forms of the linker that include a functional group capable of covalently linking the linker to a benzazepine compound(s) and a functional group capable of covalently linking the linker to an antibody construct; (ii) partially conjugated forms of the linker that include a functional group capable of covalently linking the linker to an antibody construct and that is covalently linked to a compound(s) or salt(s) of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (MB), (IIIC), (IV A), (IVB), or (IVC), or vice versa; and (iii) fully conjugated forms of the linker that is covalently linked to both a compound(s) or salt(s) of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIA), (III
  • One embodiment pertains to a conjugate formed by contacting an antibody construct that binds a cell surface receptor or tumor associated antigen expressed on a tumor cell with a linker-compound described herein under conditions in which the linker-compound covalently links to the antibody construct.
  • One embodiment pertains to a method of making a conjugate formed by contacting a linker-compound described herein under conditions in which the linker-compound covalently links to the antibody construct.
  • One embodiment pertains to a method of stimulating immune activity in a cell that expresses CD40, comprising contacting the cell with a conjugate described herein that is capable of binding the cell, under conditions in which the conjugate binds the cell.
  • Exemplary polyvalent linkers that may be used to link many benzazepine compounds to an antibody construct are described.
  • Fleximer® linker technology has the potential to enable high-DAR conjugates with good physicochemical properties.
  • the Fleximer® linker technology is based on incorporating drug molecules into a solubilizing poly acetal backbone via a sequence of ester bonds. The methodology renders highly-loaded conjugates (DAR up to 20) whilst maintaining good physicochemical properties. This methodology could be utilized with a benzazepine compound as shown in the Scheme below.
  • an aliphatic alcohol can be present or introduced into the benzazepine compound.
  • the alcohol moiety is then conjugated to an alanine moiety, which is then synthetically incorporated into the Fleximer® linker. Liposomal processing of the conjugate in vitro releases the parent alcohol-containing drug.
  • Cleavable linkers can be cleavable in vitro and in vivo.
  • Cleavable linkers can include chemically or enzymatically unstable or degradable linkages.
  • Cleavable linkers can rely on processes inside the cell to liberate a benzazepine compound, such as reduction in the cytoplasm, exposure to acidic conditions in the lysosome, or cleavage by specific proteases or other enzymes within the cell.
  • Cleavable linkers can incorporate one or more chemical bonds that are either chemically or enzymatically cleavable while the remainder of the linker can be non cleavable.
  • a linker can contain a chemically labile group such as hydrazone and/or disulfide groups.
  • Linkers comprising chemically labile groups can exploit differential properties between the plasma and some cytoplasmic compartments.
  • the intracellular conditions that can facilitate benzazepine compound release for hydrazone containing linkers can be the acidic environment of endosomes and lysosomes, while the disulfide containing linkers can be reduced in the cytosol, which can contain high thiol concentrations, e.g., glutathione.
  • the plasma stability of a linker containing a chemically labile group can be increased by introducing steric hindrance using substituents near the chemically labile group.
  • Acid-labile groups such as hydrazone
  • This pH dependent release mechanism can be associated with nonspecific release of the drug.
  • the linker can be varied by chemical modification, e.g., substitution, allowing tuning to achieve more efficient release in the lysosome with a minimized loss in circulation.
  • Hydrazone-containing linkers can contain additional cleavage sites, such as additional acid-labile cleavage sites and/or enzymatically labile cleavage sites.
  • Antibody construct benzazepine compound conjugates including exemplary hydrazone-containing linkers can include, for example, the following structures:
  • D is a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC) and Ab is an antibody construct, respectively, and n represents the number of compound-bound linkers (LP) bound to the antibody construct.
  • linkers such as linker (la)
  • the linker can comprise two cleavable groups, a disulfide and a hydrazone moiety.
  • Linkers such as (lb) and (Ic) can be effective with a single hydrazone cleavage site.
  • linkers include c/.s-aconityl -containing linkers.
  • c .s-Aconityl chemistry can use a carboxylic acid juxtaposed to an amide bond to accelerate amide hydrolysis under acidic conditions.
  • Cleavable linkers can also include a disulfide group.
  • Disulfides can be
  • thermodynamically stable at physiological pH can be designed to release the benzazepine compound upon internalization inside cells, wherein the cytosol can provide a significantly more reducing environment compared to the extracellular environment.
  • Scission of disulfide bonds can require the presence of a cytoplasmic thiol cofactor, such as (reduced) glutathione (GSH), such that disulfide-containing linkers can be reasonably stable in circulation, selectively releasing the benzazepine compound in the cytosol.
  • GSH reduced glutathione
  • the intracellular enzyme protein disulfide isomerase, or similar enzymes capable of cleaving disulfide bonds can also contribute to the preferential cleavage of disulfide bonds inside cells.
  • GSH can be present in cells in the concentration range of 0.5-10 mM compared with a significantly lower concentration of GSH or cysteine, the most abundant low-molecular weight thiol, in circulation at approximately 5 mM.
  • Tumor cells where irregular blood flow can lead to a hypoxic state, can result in enhanced activity of reductive enzymes and therefore even higher glutathione concentrations.
  • the in vivo stability of a disulfide-containing linker can be enhanced by chemical modification of the linker, e.g., use of steric hindrance adjacent to the disulfide bond.
  • Antibody construct benzazepine compound conjugates including exemplary disulfide- containing linkers can include the following structures:
  • n represents the number of compounds bound to linkers (L 3 ) bound to the antibody construct and R is independently selected at each occurrence from hydrogen or alkyl, for example.
  • R is independently selected at each occurrence from hydrogen or alkyl, for example.
  • Increasing steric hindrance adjacent to the disulfide bond can increase the stability of the linker. Structures such as (Ila) and (lie) can show increased in vivo stability when one or more R groups is selected from a lower alkyl such as methyl.
  • linker that is specifically cleaved by an enzyme.
  • the linker can be cleaved by a lysosomal enzyme.
  • Such linkers can be peptide-based or can include peptidic regions that can act as substrates for enzymes. Peptide based linkers can be more stable in plasma and extracellular milieu than chemically labile linkers.
  • Peptide bonds can have good serum stability, as lysosomal proteolytic enzymes can have very low activity in blood due to endogenous inhibitors and the unfavorably high pH value of blood compared to lysosomes. Release of a benzazepine compound from an antibody construct can occur due to the action of lysosomal proteases, e.g., cathepsin and plasmin. These proteases can be present at elevated levels in certain tumor tissues.
  • the linker can be cleavable by a lysosomal enzyme.
  • the lysosomal enzyme can be, for example, cathepsin B, cathepsin S, b- glucuronidase, or b-galactosidase.
  • the cleavable peptide can be selected from tetrapeptides such as Gly-Phe-Leu-Gly, Ala- Leu-Ala-Leu or dipeptides such as Val-Cit, Val-Ala, and Phe-Lys. Dipeptides can have lower hydrophobicity compared to longer peptides.
  • a variety of dipeptide-based cleavable linkers can be used in the antibody constructs benzazepine compound conjugates described herein.
  • Enzymatically cleavable linkers can include a self-immolative spacer to spatially separate the benzazepine compound from the site of enzymatic cleavage.
  • the direct attachment of a benzazepine compound to a peptide linker can result in proteolytic release of the benzazepine compound or of an amino acid adduct of the benzazepine compound, thereby impairing its activity.
  • the use of a self-immolative spacer can allow for the elimination of the fully active, chemically unmodified benzazepine compound upon amide bond hydrolysis.
  • One self-immolative spacer can be a bifunctional para- aminobenzyl alcohol group, which can link to the peptide through the amino group, forming an amide bond, while amine containing benzazepine compounds can be attached through carbamate functionalities to the benzylic hydroxyl group of the linker (to give a / amidobenzyl carbarn ate, PABC).
  • the resulting pro-benzazepine compound can be activated upon protease- mediated cleavage, leading to a 1,6- elimination reaction releasing the unmodified benzazepine compound, carbon dioxide, and remnants of the linker.
  • the following scheme depicts the fragmentation of p- amidobenzyl carbamate and release of the benzazepine compound:
  • X-D represents the unmodified benzazepine compound.
  • the enzymatically cleavable linker can be a B-glucuronic acid-based linker. Facile release of the benzazepine compound can be realized through cleavage of the B-glucuronide glycosidic bond by the lysosomal enzyme B-glucuronidase. This enzyme can be abundantly present within lysosomes and can be overexpressed in some tumor types, while the enzyme activity outside cells can be low.
  • B- Glucuronic acid-based linkers can be used to circumvent the tendency of an antibody construct benzazepine compound conjugate to undergo aggregation due to the hydrophilic nature of B-glucuronides.
  • B-glucuronic acid-based linkers can link an antibody construct to a hydrophobic benzazepine compound.
  • the following scheme depicts the release of a benzazepine compound (D) from an antibody construct benzazepine compound conjugate containing a B-glucuronic acid-based linker:
  • cleavable b-glucuronic acid-based linkers useful for linking drugs such as auristatins, camptothecin and doxorubicin analogues, CBI minor-groove binders, and psymberin to antibodies have been described. These b-glucuronic acid-based linkers may be used in the conjugates described herein.
  • the enzymatically cleavable linker is a b- galactoside-based linker.
  • b-Galactoside is present abundantly within lysosomes, while the enzyme activity outside cells is low.
  • benzazepine compounds containing a phenol group can be covalently bonded to a linker through the phenolic oxygen.
  • One such linker relies on a methodology in which a diamino-ethane "Space Link” is used in conjunction with traditional "PABO”-based self- immolative groups to deliver phenols.
  • Cleavable linkers can include non-cleavable portions or segments, and/or cleavable segments or portions can be included in an otherwise non-cleavable linker to render it cleavable.
  • polyethylene glycol (PEG) and related polymers can include cleavable groups in the polymer backbone.
  • a polyethylene glycol or polymer linker can include one or more cleavable groups such as a disulfide, a hydrazone or a dipeptide.
  • linkers can include ester linkages formed by the reaction of PEG carboxylic acids or activated PEG carboxylic acids with alcohol groups on a benzazepine compound, wherein such ester groups can hydrolyze under
  • Hydrolytically degradable linkages can include, but are not limited to, carbonate linkages; imine linkages resulting from reaction of an amine and an aldehyde; phosphate ester linkages formed by reacting an alcohol with a phosphate group; acetal linkages that are the reaction product of an aldehyde and an alcohol; orthoester linkages that are the reaction product of a formate and an alcohol; and oligonucleotide linkages formed by a phosphoramidite group, including but not limited to, at the end of a polymer, and a 5' hydroxyl group of an oligonucleotide.
  • a linker can contain an enzymatically cleavable peptide moiety, for example, a linker comprising structural formula (Ilia), (Mb), (Me), or (Hid):
  • “peptide” represents a peptide (illustrated in N C orientation, wherein peptide includes the amino and carboxy“termini”) that is cleavable by a lysosomal enzyme
  • T represents a polymer comprising one or more ethylene glycol units or an alkylene chain, or combinations thereof
  • R a is selected from hydrogen, alkyl, sulfonate and methyl sulfonate
  • R y is hydrogen or Ci -4 alkyl-(0) r -(Ci-4 alkylene) s -G 1 or Ci -4 alkyl-(N)-[(Ci -4 alkylene)- G3 ⁇ 4
  • R z is Ci-4 alkyl-(0) r -(Ci -4 alkylene) s -G 2
  • G 1 is SO3H, C0 2 H, PEG 4-32, or a sugar moiety
  • G 2 is SO3H, CO2H, or a PEG 4-32 moiety
  • r is
  • the peptide can be selected from natural amino acids, unnatural amino acids or combinations thereof.
  • the peptide can be selected from a tripeptide or a dipeptide.
  • the dipeptide can comprise L-amino acids and be selected from: Val-Cit; Cit-Val; Ala-Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala; Phe- Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu- Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Tr
  • linkers according to structural formula (Ilia) are illustrated below (as illustrated, the linkers include a reactive group suitable for covalently linking the linker to an antibody construct):
  • linkers according to structural formula (Illb), (IIIc), or (Hid) that can be included in the conjugates described herein can include the linkers illustrated below (as illustrated, the linkers can include a reactive group suitable for covalently linking the linker to an antibody construct):
  • ⁇ indicates an attachment site to a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
  • the linker can contain an enzymatically cleavable sugar moiety, for example, a linker comprising structural formula (IVa), (IVb), (IVc), (IVd), or (IVe):
  • linkers according to structural formula (IVa) that may be included in the antibody construct benzazepine compound conjugates described herein can include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
  • » represents the point of attachment of the linker (L 3 ) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
  • linkers according to structural formula (IVb) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
  • linkers according to structural formula (IVc) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
  • linkers according to structural formula (IVd) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct): wherein represents the point of attachment of the linker (L 3 ) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
  • linkers according to structural formula (IVe) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
  • » represents the point of attachment of the linker (L 3 ) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
  • cleavable linkers can provide certain advantages, the linkers comprising the conjugate described herein need not be cleavable.
  • the benzazepine compound release may not depend on the differential properties between the plasma and some cytoplasmic compartments. The release of the benzazepine compound can occur after
  • benzazepine compound conjugate via antigen-mediated endocytosis and delivery to lysosomal compartment, where the antibody construct can be degraded to the level of amino acids through intracellular proteolytic degradation.
  • This process can release a benzazepine compound derivative, which is formed by the benzazepine compound, the linker, and the amino acid residue or residues to which the linker was covalently attached.
  • the benzazepine compound derivative from antibody construct benzazepine compound conjugates with non-cleavable linkers can be more hydrophilic and less membrane permeable, which can lead to less bystander effects and less nonspecific toxicities compared to antibody construct benzazepine compound conjugates with a cleavable linker.
  • Antibody construct benzazepine compound conjugates with non-cleavable linkers can have greater stability in circulation than antibody construct benzazepine compound conjugates with cleavable linkers.
  • Non-cleavable linkers can include alkylene chains, or can be polymeric, such as, for example, based upon polyalkylene glycol polymers, amide polymers, or can include segments of alkylene chains, polyalkylene glycols and/or amide polymers.
  • the linker can contain a polyethylene glycol segment having from 1 to 6 ethylene glycol units.
  • the linker can be non-cleavable in vivo , for example, a linker according to the formulations below:
  • R a is selected from hydrogen, alkyl, sulfonate and methyl sulfonate
  • R x is a reactive moiety including a functional group capable of covalently linking the linker to an antibody construct; and represents the point of attachment of the linker (L 3 ) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB), or (IVC).
  • linkers according to structural formula (Va)-(Vf) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct, and represents the point of attachment of the linker (L 3 ) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC):
  • Attachment groups that are used to attach the linkers to an antibody construct can be electrophilic in nature and include, for example, maleimide groups, alkynes, alkynoates, allenes and allenoates, activated disulfides, active esters such as NHS esters and HOBt esters, haloformates, acid halides, alkyl, and benzyl halides such as haloacetamides.
  • maleimide groups alkynes, alkynoates, allenes and allenoates
  • activated disulfides such as NHS esters and HOBt esters
  • haloformates acid halides
  • alkyl alkyl
  • benzyl halides such as haloacetamides
  • Maleimide groups are frequently used in the preparation of conjugates because of their specificity for reacting with thiol groups of, for example, cysteine groups of the antibody of a conjugate.
  • the reaction between a thiol group of an antibody and a drug with a linker including a maleimide group proceeds according to the following scheme:
  • succinimide may also take place.
  • This reverse reaction is undesirable as the maleimide group may subsequently react with another available thiol group such as other proteins in the body having available cysteines. Accordingly, the reverse reaction can undermine the specificity of a conjugate.
  • One method of preventing the reverse reaction is to incorporate a basic group into the linking group shown in the scheme above. Without wishing to be bound by theory, the presence of the basic group may increase the nucleophilicity of nearby water molecules to promote ring opening hydrolysis of the succinimide group. The hydrolyzed form of the attachment group is resistant to deconjugation in the presence of plasma proteins. So-called“self-stabilizing” linkers provide conjugates with improved stability. A representative schematic is shown below:
  • the identity of the base as well as the distance between the base and the maleimide group can be modified to tune the rate of hydrolysis of the thio- substituted succinimide group and optimize the delivery of a conjugate to a target by, for example, improving the specificity and stability of the conjugate.
  • Bases suitable for inclusion in a linker described herein may facilitate hydrolysis of a nearby succinimide group formed after conjugation of the antibody construct to the linker.
  • Bases may include, for example, amines (e.g., -N(R 26 )(R 27 ), where R 26 and R 27 are independently selected from H and Ci -6 alkyl), nitrogen-containing heterocycles (e.g., a 3- to l2-membered heterocycle including one or more nitrogen atoms and optionally one or more double bonds), amidines, guanidines, and carbocycles or heterocycles substituted with one or more amine groups (e.g., a 3- to l2-membered aromatic or non-aromatic cycle optionally including a heteroatom such as a nitrogen atom and substituted with one or more amines of the type - N(R 26 )(R 27 ), where R 26 and R 27 are independently selected from H or Ci -6 alkyl).
  • amines e.g., -N(R 26 )(R 27 ), where R 26 and R 27 are independently selected from H and Ci -6 alkyl
  • nitrogen-containing heterocycles e.g
  • a basic unit may be separated from a maleimide group by, for example, an alkylene chain of the form - (CH 2 ) m -, where m is an integer from 0 to 10.
  • An alkylene chain may be optionally substituted with other functional groups as described herein.
  • Self-stabilizing linkers may also include aryl, e.g., phenyl, or heteroaryl, e.g., pyridine, groups optionally substituted with electron withdrawing groups such as those described herein.
  • a linker of the disclosure (L 3 ) comprises a stabilizing linker
  • the bottom structure may be referred to as (maleimido)- DPR-Val-Cit-PAB, where DPR refers to diaminopropinoic acid, Val refers to valine, Cit refers to citrulline, and PAB refers to para-aminobenzylcarbonyl.
  • ⁇ represents the point of attachment to compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (MC), (IV A), (IVB), or (IVC).
  • a method for bridging a pair of sulfhydryl groups derived from reduction of a native hinge disulfide bond has been disclosed and is depicted in the schematic below.
  • An advantage of this methodology is the ability to synthesize homogenous DAR4 conjugates by full reduction of IgGs (to give 4 pairs of sulfhydryls from interchain disulfides) followed by reaction with 4 equivalents of the alkylating agent.
  • Conjugates containing "bridged disulfides” are also claimed to have increased stability.
  • a linker of the disclosure, L 3 can contain the following structural formulas (Via), (VIb), or (Vic):
  • R q is H or-0-(CH2CH20)n-CH3; x is 0 or 1; y is 0 or 1; G 2 is- CH2CH2CH2SO3H or-CH 2 CH20-(CH2CH 2 0)ii-CH3; R w is-0-CH 2 CH 2 S03H or-NH(CO)- CH2CH20-(CH2CH20)i2-CH3; and * represents the point of attachment to the remainder of the linker.
  • linkers according to structural formula (Via) and (VIb) that can be included in the conjugates described herein can include the linkers illustrated below (as illustrated, the linkers can include a group suitable for covalently linking the linker to an antibody construct):
  • > represents the point of attachment of the linker (L 3 ) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
  • linkers according to structural formula (Vic) that can be included in the antibody construct benzazepine compound conjugates described herein can include the linkers illustrated below (as illustrated, the linkers can include a group suitable for covalently linking the linker to an antibody construct):
  • » represents the point of attachment of the linker (L 3 ) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
  • the linker selected for a particular conjugate may be influenced by a variety of factors, including but not limited to, the site of attachment to the antibody construct (e.g., lys, cys or other amino acid residues), structural constraints of the drug pharmacophore and the lipophilicity of the drug.
  • the specific linker selected for a conjugate should seek to balance these different factors for the specific antibody construct/drug
  • conjugates have been observed to effect killing of bystander antigen negative cells present in the vicinity of the antigen-positive tumor cells.
  • the mechanism of bystander cell killing by conjugates has indicated that metabolic products formed during intracellular processing of the conjugates may play a role.
  • Neutral cytotoxic metabolites generated by metabolism of the conjugates in antigen-positive cells appear to play a role in bystander cell killing while charged metabolites may be prevented from diffusing across the membrane into the medium, or from the medium across the membrane, and therefore cannot affect bystander killing.
  • the linker is selected to attenuate the bystander killing effect caused by cellular metabolites of the conjugate.
  • the linker is selected to increase the bystander killing effect.
  • linker may also impact aggregation of the conjugate under conditions of use and/or storage.
  • conjugates reported in the literature contain no more than 3-4 drug molecules per antibody molecule. Attempts to obtain higher drug- to-antibody ratios (“DAR”) often failed, particularly if both the drug and the linker were hydrophobic, due to aggregation of the conjugate. In many instances, DARs higher than 3-4 could be beneficial as a means of increasing potency. In instances where the benzazepine compound is more hydrophobic in nature, it may be desirable to select linkers that are relatively hydrophilic as a means of reducing conjugate aggregation, especially in instances where DARs greater than 3-4 are desired.
  • the linker incorporates chemical moieties that reduce aggregation of the conjugates during storage and/or use.
  • a linker may incorporate polar or hydrophilic groups such as charged groups or groups that become charged under physiological pH to reduce the aggregation of the conjugates.
  • a linker may incorporate charged groups such as salts or groups that deprotonate, e.g., carboxylates, or protonate, e.g., amines, at physiological pH.
  • the aggregation of the conjugates during storage or use is less than about 40% as determined by size-exclusion chromatography (SEC). In particular embodiments, the aggregation of the conjugates during storage or use is less than 35%, such as less than about 30%, such as less than about 25%, such as less than about 20%, such as less than about 15%, such as less than about 10%, such as less than about 5%, such as less than about 4%, or even less, as determined by size-exclusion chromatography (SEC).
  • SEC size-exclusion chromatography
  • the conjugates described herein may comprise a linker, e.g., a cleavable linker such as a peptide linker or a noncleavable linker.
  • Linkers of the conjugates and methods described herein may not affect the binding of active portions of a conjugate (e.g., active portions include antigen binding domains, Fc domains, target binding domains, antibodies, compounds or salts of the disclosed, e.g., Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (MB) or (IIIC), or linker- compounds of Formulas (IV A), (IVB), or (IVC) to a target, which can be a cognate binding partner such as an antigen.
  • a linker e.g., a cleavable linker such as a peptide linker or a noncleavable linker.
  • Linkers of the conjugates and methods described herein may not affect the binding
  • a linker sequence can form a linkage between different parts of a conjugate, e.g., between an antibody construct and a compound or salt of the disclosure.
  • a conjugate comprises multiple linkers.
  • the linkers may be the same linkers or different linkers.
  • a linker may be bound to an antibody construct by a bond between the antibody construct and the linker.
  • a linker may be bound to an anti-tumor antigen antibody construct by a bond between the anti-tumor antigen antibody construct and the linker.
  • a linker may be bound to a terminus of an amino acid sequence of an antibody construct, or could be bound to a side chain modification to the antibody construct, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue.
  • a linker may be bound to a terminus of an amino acid sequence of an Fc domain or Fc region of an antibody construct, or may be bound to a side chain modification of an Fc domain or Fc region of an antibody construct, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue.
  • a linker may be bound to a terminus of an amino acid sequence of an Fc domain of an antibody construct, or may be bound to a side chain modification of an Fc domain of an antibody construct, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue.
  • a linker may be bound to an antibody construct at a hinge cysteine.
  • a linker may be bound to an antibody construct at a light chain constant domain lysine.
  • a linker may be bound to an antibody construct at a heavy chain constant domain lysine.
  • a linker may be bound to an antibody construct at an engineered cysteine in the light chain.
  • a linker may be bound to an antibody construct at an Fc region lysine.
  • a linker may be bound to an antibody construct at an Fc domain lysine.
  • a linker may be bound to an antibody construct at an Fc region cysteine.
  • a linker may be bound to an antibody construct at an Fc domain cysteine.
  • a linker may be bound to an antibody construct at a light chain glutamine, such as an engineered glutamine.
  • a linker may be bound to an antibody construct at a heavy chain glutamine, such as an engineered glutamine.
  • a linker may be bound to an antibody construct at an unnatural amino acid engineered into the light chain.
  • a linker may be bound to an antibody construct at an unnatural amino acid engineered into the heavy chain.
  • Amino acids can be engineered into an amino acid sequence of an antibody construct, for example, a linker of a conjugate.
  • Engineered amino acids may be added to a sequence of existing amino acids.
  • Engineered amino acids may be substituted for one or more existing amino acids of a sequence of amino acids.
  • a linker may be conjugated to an antibody construct via a sulfhydryl group on the antibody construct.
  • a linker may be conjugated to an antibody construct via a primary amine on the antibody construct.
  • a linker may be conjugated to an antibody construct via a residue of an unnatural amino acid on an antibody construct, e.g., a ketone moiety.
  • an Fc domain of the antibody construct can bind to Fc receptors.
  • an antibody construct bound through a linker or an antibody construct bound to a linker bound to a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB) or (IIIC), or bound through a compound-linker of Formulas (IV A), (IVB) or (IVC) retains the ability of the Fc domain of the antibody to bind to Fc receptors.
  • the antigen binding domain of an antibody construct bound to a linker or an antibody construct bound through a linker bound to a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB) or (IIIC), or bound through a compound-linker of Formulas (IV A), (IVB), or (IVC) can bind its antigen.
  • a linker when a linker is connected to an antibody construct at the sites described herein, a target binding domain of an antibody construct bound to a linker or an antibody construct bound to a linker bound to a compound or salt of any one of Formulas
  • (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) can bind its antigen.
  • the engineered cysteine residue is at one or more of positions HC S239C, LC V205C, LC A114C, HC A140C, LC K149C, LC S168C, LC S153C, LC A127C, HC T116C, and HC Sl 15C, where HC refers to heavy chain, LC refers t light chain and the numbering of amino acid residues in the Fc region is according to the EU index as in Kabat.
  • a compound or a compound-linker of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) disclosed herein may not be attached to an amino acid residue of an IgG Fc domain disclosed herein selected from: 221, 222, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 246, 247, 249,
  • An antibody construct can be conjugated to a linker via lysine-based bioconjugation.
  • An antibody construct can be exchanged into an appropriate buffer, for example, phosphate, borate, PBS, Tris-Acetate, Tris-Glycine, HEPES, MOPS, MES, EPS, HEPPS, Histidine, or HEPBS at a concentration of about 2 mg/mL to about 10 mg/mL.
  • An appropriate number of equivalents of a compound-linker e.g., a compound or salt of Formula (IV A), (IVB), or (IVC), described herein can be added as a solution with stirring.
  • a co-solvent can be introduced prior to the addition of the benzazepine-linker construct to facilitate solubility.
  • the reaction can be stirred at room temperature for 2 hours to about 12 hours depending on the observed reactivity. The progression of the reaction can be monitored by LC-MS. Once the reaction is deemed complete, the remaining benzazepine-linker constructs can be removed by applicable methods and the antibody construct- benzazepine conjugate can be exchanged into the desired formulation buffer.

Abstract

Benzazepine compounds, conjugates, and pharmaceutical compositions for use in the treatment of disease, such as cancer, are disclosed herein. The disclosed benzazepine compounds are useful, among other things, in the treating of cancer and modulating TLR8. Additionally, benzazepine compounds incorporated into a conjugate with an antibody construct are described herein.

Description

BENZAZEPINE COMPOUNDS, CONJUGATES, AND USES THEREOF
RELATED APPLICATION INFORMATION
[0001] This application claims the benefit of U.S. Provisional Application No. 62/730,468 filed September 12, 2018, and U.S. Provisional Application No. 62/730,469 filed September 12, 2018, the contents of each of which are incorporated herein by reference in their entirety.
BACKGROUND OF THE INVENTION
[0002] One of the leading causes of death in the United States is cancer. The conventional methods of cancer treatment, like chemotherapy, surgery, or radiation therapy, tend to be either highly toxic or nonspecific to a cancer, or both, resulting in limited efficacy and harmful side effects. However, the immune system has the potential to be a powerful, specific tool in fighting cancers. In many cases tumors can specifically express genes whose products are required for inducing or maintaining the malignant state. These proteins may serve as antigen markers for the development and establishment of more specific anti-cancer immune response. The boosting of this specific immune response has the potential to be a powerful anti-cancer treatment that can be more effective than conventional methods of cancer treatment and can have fewer side effects.
SUMMARY OF THE INVENTION
[0003] In some aspects, the present disclosure provides a compound represented by the structure of Formula (II A):
Figure imgf000002_0001
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L10 is -X10-;
L2 is selected from -X2-, -X2-CI-6 alkylene-X2-, -X2-C2-6 alkenyl ene-X2-, and - X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R12; X10 is selected from -C(O)-, and -C(0)N(R10)-*, wherein * represents where X10 is bound to R5;
X2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R10)-, -C(O)-,
-C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)
-C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(O), -0S(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-,
and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-0 carbocycle, and 3- to l2-membered heterocycle in R4 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R5 is selected from unsaturated C4-8 carbocycle; bicyclic carbocycle; and fused 5- 5, fused 5-6, and fused 6-6 bicyclic heterocycle, wherein R5 is optionally substituted and wherein substituents are independently selected at each occurrence from: halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-0 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-0 carbocycle, and 3- to l2-membered heterocycle in R5 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, C MO alkyl, -Ci-io haloalkyl, -O-Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
[0004] In some embodiments, the compound of Formula (IIA) is represented by Formula (IIB):
Figure imgf000004_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen, - OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-10 alkyl, C2-io alkenyl, and C2-io alkynyl; and R24, and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, - N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
[0005] In some embodiments for a compound or salt of Formula (IIB), R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -OH, -OR10, -N02, -CN, and C1-10 alkyl. R20,
R21, R22, and R23 may be each hydrogen. In certain embodiments, R21 is halogen. In certain embodiments, R21 is hydrogen. In certain embodiments, R21 is -OR10. For example, R21 may be - OCH3.
[0006] In some embodiments for a compound or salt of Formula (IIB), R24 and R25 are independently selected from hydrogen, halogen, -OH, -N02, -CN, and C1-10 alkyl, or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle. In certain embodiments, R24 and R25 are each hydrogen. In other embodiments, R24 and R25 taken together form an optionally substituted saturated C3-5 carbocycle, wherein substituents are selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-0 carbocycle, and 3- to l2-membered heterocycle.
[0007] In some embodiments for a compound or salt of Formula (IIA) or (IIB), R2 is an amine masking group that is enzymatically-labile. In certain embodiments, R2 is represented by the formula:
Figure imgf000005_0001
wherein:
R101 is selected from an amino acid, a peptide, -0-(Ci-C6 alkyl) and -Ci-C6 alkyl, wherein alkyl of -0-(Ci-C6 alkyl) and -Ci-C6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N02, -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle; and
R100 is C(=0), wherein when R101 is selected from an amino acid or peptide R100 is the C- terminus of the amino acid or peptide.
[0008] In certain embodiments, R101 of R2 is a peptide selected from a dipeptide, tripeptide and tetrapeptide. [0009] In some embodiments for a compound or salt of Formula (IIA) or (IIB), L10 is selected from -C(0)N(R10)-*. In certain embodiments, R10 of -C(0)N(R10)-* is selected from hydrogen and Ci-6 alkyl. For example, L10 may be -C(0)NH-*.
[0010] In some embodiments for a compound or salt of Formula (IIA) or (IIB), R5 is an optionally substituted bicyclic carbocycle. In certain embodiments, R5 is an optionally
substituted 8- to 12- membered bicyclic carbocycle. R5 may be an optionally substituted 8- to 12- membered bicyclic carbocycle substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, -CN, Ci- 6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. In certain embodiments, R5 is an optionally substituted 8- to 12 membered bicyclic carbocycle substituted with one or more substituents independently selected from -OR10, -N(R10)2, and =0. In some embodiments, R5 is an optionally substituted
indane, and optionally substituted tetrahydronaphthalene. R5 may be selected from: (
Figure imgf000006_0001
any one of which is optionally substituted. For example, the R5 is selected
Figure imgf000006_0002
[0011] In some embodiments for a compound or salt of Formula (IIA) or (IIB), R5 is an optionally substituted unsaturated C4-8 carbocycle. In certain embodiments, R5 is an optionally substituted unsaturated C4-6 carbocycle. In certain embodiments, R5 is an optionally substituted unsaturated C4-6 carbocycle with one or more substituents independently selected from optionally substituted C3-i2 carbocycle, and optionally substituted 3- to l2-membered heterocycle. R5 may be an optionally substituted unsaturated C4-6 carbocycle with one or more substituents independently selected from optionally substituted phenyl, optionally substituted 3- to 12- heterocycle, optionally substituted Ci-io alkyl, optionally substituted C2-io alkenyl, and halogen.
[0012] In some embodiments for a compound or salt of Formula (IIA) or (IIB), R5 is selected from an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle. In certain embodiments, R5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle with one or more substituents independently selected from -C(0)OR10, -N(R10)2, - OR10, and optionally substituted Ci-io alkyl. In certain embodiments, R5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle substituted with -C(0)OR10. In certain embodiments, R5 is an optionally substituted fused 6-6 bicyclic heterocycle. For example, the fused 6-6 bicyclic heterocycle may be an optionally substituted pyridine-piperidine. In some embodiments, L10 is bound to a carbon atom of the pyridine of the fused pyridine- piperidine. In certain embodiments, R5 is selected from tetrahydroquinoline,
tetrahydroisoquinoline, tetrahydronaphthyridine, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is optionally substituted. R5 may be an optionally substituted
tetrahydronaphthyridine. In some embodiments, R5 is selected from
Figure imgf000007_0001
Figure imgf000007_0002
[0013] In some embodiments for a compound or salt of Formula (IIA) or (IIB), when R5 is substituted, substituents on R5 are independently selected at each occurrence from: halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2- io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. In certain embodiments, the substituents on R5 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle. In certain embodiments, the substituents on R5 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, and -CN; and Ci-io alkyl optionally substituted with one or more substituents independently selected from halogen, - OR10, -SR10, -N(R10)2, -C(0)R10, -C(0)OR10, -N02, =0, and -CN. In some embodiments, R5 is not substituted.
[0014] In some embodiments for a compound or salt of Formula (IIA) or (IIB), L2 is selected from -C(0)-, and -C(0)NR10-. In some embodiments, L2 is -C(0)-. In some embodiments, L2 is selected from -C(0)NR10-. R10 of -C(0)NR10- may be selected from hydrogen and Ci-6 alkyl. For example, L2 may be -C(0)NH-.
[0015] In some embodiments for a compound or salt of Formula (IIA) or (IIB), R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle and 3- to 12- membered, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. In some embodiments, R4 is selected from: -OR10, and -N(R10)2; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, - OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl. In certain embodiments, R4 is - N(R10)2. R10 of -N(R10)2 may be independently selected at each occurrence from optionally substituted Ci-6 alkyl. In certain embodiments, R10 of -N(R10)2 is independently selected at each occurrence from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted. For
example, R4 may
Figure imgf000009_0001
certain embodiments,
Figure imgf000009_0002
[0016] In some embodiments for a compound or salt of Formula (IIA) or (IIB), R12 is
independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -
S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. In certain embodiments, R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -
S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle.
[0017] In some aspects, the present disclosure provides a compound represented by the structure of F ormul a (HI A) :
Figure imgf000010_0001
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L11 is -X11-;
L2 is selected from -X2-, -X2-CI-6 alkylene-X2-, -X2-C2-6 alkenyl ene-X2-, and - X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R12;
X11 is selected from -C(O)- and -C(0)N(R10)-*, wherein * represents where X11 is bound to R6;
X2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R10)-, -C(O)-,
-C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-,
-C(O)N(R10)C(O)N(R10)-, -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(O)-, -0S(0)2-, -S(0)20- , -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-,
-N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10,
-SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; R6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R7 and R6 is further optionally substituted by one or more additional substituents independently selected from R12;
R7 is selected from -C(0)NHNH2, -C(0)NH-Ci-3 alkylene-NH(R10), -C(0)CH3, - Ci-3 alkyl ene-NHC(0)0R11 , -Ci-3alkylene-NHC(0)R10, -Ci-3alkylene-NHC(0)NHR10, -Ci- 3alkylene-NHC(0)-Ci-3alkylene-R10, and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R12;
R10 is independently selected at each occurrence from hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, -CI-IO alkyl, -Ci-io haloalkyl, -O-Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to l2-membered heterocycle;
R11 is selected from C3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from R12;
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C3-io carbocycle and 3- to lO-membered heterocycle; and C3-io carbocycle and 3- to lO-membered heterocycle, wherein each C3-io carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle. In some embodiments, the compound of Formula (IIIA) is represented by Formula (IIIB):
Figure imgf000012_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen,
-OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; and
R24, and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, - N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
[0018] In some embodiments for a compound or salt of Formula (MB), R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -OH, -N02, -CN, and C1-10 alkyl. In certain embodiments, R20, R21, R22, and R23 are each hydrogen. In some embodiments, R24 and R25 are independently selected from hydrogen, halogen, -OH, -N02, -CN, and C1-10 alkyl, or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle. In certain embodiments, R24 and R25 are each hydrogen. In certain embodiments, R24 and R25 taken together form an optionally substituted saturated C3-5 carbocycle.
[0019] In certain embodiments for a compound or salt of Formula (IIIA) or (IIIB), R2 is represented by the formula:
Figure imgf000012_0002
wherein:
R101 is selected from an amino acid, a peptide, -0-(Ci-C6 alkyl) and -Ci-C6 alkyl, wherein alkyl of -0-(Ci-C6 alkyl) and -Ci-C6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N02, -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle; and
R100 is C(=0), wherein when R101 is selected from an amino acid or peptide R100 is the C- terminus of the amino acid or peptide.
[0020] In certain embodiments, R101 of R2 is a peptide selected from a dipeptide, tripeptide and tetrapeptide. [0021] In some embodiments for a compound or salt of Formula (IIIA) or (IIIB), L11 is selected from
-C(0)N(R10)- *. In some embodiments, R10 of -C(0)N(R10)-* is selected from hydrogen and Ci-6 alkyl. For example, L11 may be -C(0)NH-*.
[0022] In some embodiments for a compound or salt of Formula (IIIA) or (IIIB), R6 is phenyl substituted with R7 and R6 is further optionally substituted with one or more additional substituents independently selected from R12. In some embodiments, R6 is selected from phenyl substituted with one or more substituents independently selected from -C(0)NHNH2, -C(0)NH- Ci.3alkylene-NH(R10), -Ci.3alkylene-NHC(0)R10, and -C(0)CH3; and 3- to l2-membered heterocycle, which is optionally substituted with one or more substituents selected from -OH, - N(R10)2, -NHC(0)(R10), -NHC(0)0(R10), -NHC(O)N(R10)2, -C(0)R10, -C(O)N(R10)2, - C(0)2R10, and -Ci-3alkylene-(R10) and R6 is further optionally substituted with one or more additional substituents independently selected from R12. For example, R6 may be selected from:
Figure imgf000013_0001
[0023] In some embodiments for a compound or salt of Formula (IIIA) or (IIIB), R6 is selected from a 5- and 6-membered heteroaryl substituted with one or more substituents independently selected from R7, and R6 is further optionally substituted with one or more additional substituents selected from R12. In certain embodiments, R6 is selected from 5- and 6-membered heteroaryl substituted with one or more substituents independently selected from -C(0)CH3, -Ci-3alkylene- NHC(0)OR10, -Ci-3alkylene-NHC(0)R10, -Ci-3alkylene-NHC(0)NHR10, and -Ci-3alkylene- NHC(O) -Ci-3alkylene-(R10); and 3- to l2-membered heterocycle, which is optionally substituted with one or more substituents selected from -OH, -N(R10)2, -NHC(0)(R10), -NHC(0)0(R10), - NHC(O)N(R10)2, -C(0)R10, -C(O)N(R10)2, -C(0)2R10, and -Ci-3alkylene-(R10), and R6 is optionally further substituted with one or more additional substituents independently selected from R12. R6 may be selected from substituted pyridine, pyrazine, pyrimidine, pyridazine, furan, pyran, oxazole, thiazole, imidazole, pyrazole, oxadiazole, oxathiazole, and triazole, and R6 is optionally further substituted with one or more additional substituents independently selected from R12. In some embodiments, R6 is substituted pyridine and R6 is optionally further substituted with one or more additional substituents independently selected from R12. R6 may be
represented as follows:
Figure imgf000014_0001
some embodiments, R6 is substituted pyridine, and wherein R7 is -Ci-3alkylene-NHC(0)-Ci-3alkylene-R1°. In certain embodiments, R7 is -Cialkylene-NHC(0)-Cialkylene-R10. In certain embodiments, R7 is -Cialkylene-NHC(O)-
Cialkylene-Nkh. In some embodiments, R6 is selected from:
Figure imgf000014_0002
Figure imgf000014_0003
Figure imgf000015_0001
[0024] In some embodiments for a compound or salt of Formula (IIIA) or (IIIB), L2 is selected from -C(O)-, and -C(0)NR10-. In some embodiments, L2 is selected from -C(0)NR10-. R10 of - C(0)NR10- may be selected from hydrogen and Ci-6 alkyl. For example, L2 may be -C(0)NH-. In some embodiments, L2 is -C(O)-.
[0025] In some embodiments for a compound or salt of Formula (IIIA) or (IIIB), R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle and 3- to 12- membered, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. In some embodiments, R4 is selected from: -OR10 and -N(R10)2; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10 -C(0)R10, -C(0)OR10, -OC(0)R10, - N02, =0, =S, =N(R10), -CN, Ci -lo alkyl, C2-io alkenyl, and C2-io alkynyl. In certain embodiments, R4 is -N(R10)2. R10 of -N(R10)2 may be independently selected at each occurrence from optionally substituted Ci-6 alkyl. In some embodiments, R10 of -N(R10)2 is independently selected at each occurrence from methyl, ethyl, propyl, and butyl, any of which are optionally substituted. For
example, R4 may be v . In some embodiments, -L2-R4 is CHs .
[0026] In some embodiments for a compound or salt of Formula (IIIA) or (IIIB), R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-10 alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -
S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. In certain embodiments, R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -
S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle.
[0027] In some aspects, the present disclosure provides a compound represented by the structure of Formula (IA):
Figure imgf000016_0001
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L1 is selected from -X1-, -X2-CI-6 alkyl ene-X2-Ci-6 alkylene-, -X2-C2-6 alkenylene- X2-, and -X2-C2-6 alkynyl ene-X2-, each of which is optionally substituted on alkylene, alkenylene or alkynyl ene with one or more R12;
L2 is selected from -X2-, -X2-CI-6 alkylene-X2-, -X2-C2-6 alkenyl ene-X2-, and -X2-C2-6 alkynyl ene-X2-, each of which is optionally substituted on alkylene, alkenylene or alkynylene with one or more R12;
X1 is selected from -S-*, -N(R10)-*, -C(0)0-*, -OC(O)-*, -0C(0)0-*, - C(O)N(R10)C(O)- *, -C(O)N(R10)C(O)N(R10)*, -N(R10)C(O)-*, -CR10 2N(R10)C (O)-*, -N(R10)C(O)N(R10)-*, -N(R10)C(O)O-*, -OC(0)N(R10)-*, -C(NR10)-*, -N(R10)C(NR10)-*, -C(NR10)N(R10)-*, -N(R10)C(NR10)N(R10)-*, -S(0)2-*, -0S(0)-*, -S(0)0-*, -S(0), -0S(0)2- *, -S(0)20*, -N(R10)S(O)2-*, -S(0)2N(R10)-*, -N(R10)S(O)-*, -S(0)N(R10)-*, -N(R10)S(0)2N(R1 °)-*, and -N(R10)S(O)N(R10)-*, wherein * represents where X1 is bound to R3;
X2 is independently selected at each occurrence from -O- , -S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(0), -0S(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R3 is selected from optionally substituted C3-0 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents on R3 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), and -CN; C1-10 alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10,
-N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to 12- membered heterocycle, wherein each C3-0 carbocycle, and 3- to l2-membered heterocycle in R3 is optionally substituted with one or more substituents independently selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10,
-OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-0 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-0 carbocycle, and 3- to l2-membered heterocycle in R4 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from: hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, CI-IO alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C 1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle. In some embodiments, the compound of Formula (IA) is represented by Formula (IB):
Figure imgf000018_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -
OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; and
R24 and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, -
N(R10)2,
-S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
[0028] In some embodiments for a compound or salt of Formula (IB), R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -OH, -N02, -CN, and C1-10 alkyl. In certain embodiments, R20, R21, R22, and R23 are each hydrogen.
[0029] In some embodiments for a compound or salt of Formula (IB), R24 and R25 are independently selected from hydrogen, halogen, -OH, -N02, -CN, and C 1-10 alkyl, or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle. In some embodiments, R24 and R25 are each hydrogen. In some embodiments, R24 and R25 taken together form an optionally substituted saturated C3-5 carbocycle.
[0030] In certain embodiments for a compound or salt of Formula (IA) or (IB), R2 is represented by the formula:
Figure imgf000019_0001
wherein:
R101 is selected from an amino acid, a peptide, -0-(Ci-C6 alkyl) and -Ci-C6 alkyl, wherein alkyl of -0-(Ci-C6 alkyl) and -Ci-C6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N02, -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle; and
R100 is C(=0), wherein when R101 is selected from an amino acid or peptide R100 is the C- terminus of the amino acid or peptide.
[0031] In certain embodiments, R101 is a peptide selected from a dipeptide, tripeptide and tetrapeptide.
[0032] In some embodiments for a compound or salt of Formula (IA) or (IB), L1 is selected from -N(R10)C(O)-*, -S(0)2N(R10)-*, -CR10 2N(R10)C (0)-*and -X2-CI-6 alkylene-X2-Ci-6 alkyl ene-. In some embodiments, L1 is selected from -N(R10)C(O)-*. In certain embodiments,
R10 of -N(R10)C(O)-* is selected from hydrogen and Ci-6 alkyl. For example, L1 may be - NHC(O)-*. In some embodiments, L1 is selected from -S(0)2N(R10)-*. In certain embodiments, R10 of -S(0)2N(R10)-* is selected from hydrogen and Ci-6 alkyl. For example, L1 is -S(0)2NH-*. In some embodiments, L1 is -CR102N(R10)C(O)-*. In certain embodiments, L1 is selected from -CH2N(H)C(0)-* and -CH(CH3)N(H)C(0)-*.
[0033] In some embodiments for a compound or salt of Formula (IA) or (IB), R3 is selected from optionally substituted C3-12 carbocycle, and optionally substituted 3- to l2-membered
heterocycle, wherein substituents on R3 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2- 10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, C3-12 carbocycle, and 3- to l2-membered heterocycle; and C3-12 carbocycle, and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. In certain embodiments, R3 is selected from optionally substituted C3-12 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents on R3 are
independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0,
=S, =N(R10), and -CN; Ci -lo alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -NO2, =0, =S, =N(R10), -CN, C3-12 carbocycle, and 3- to l2-membered heterocycle.
[0034] In some embodiments for a compound or salt of Formula (IA) or (IB), R3 is selected from an optionally substituted aryl and an optionally substituted heteroaryl. In some embodiments, R3 is an optionally substituted heteroaryl. R3 may be an optionally substituted heteroaryl substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
In certain embodiments, R3 is selected from an optionally substituted 6-membered heteroaryl.
For example, R3 may be an optionally substituted pyridine. In some embodiments, R3 is an optionally substituted aryl. In certain embodiments, R3 is an optionally substituted aryl substituted with one or more substituents independently selected from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. R3 may be an optionally substituted phenyl. In certain embodiments, R3 is selected from pyridine, phenyl, tetrahydronaphthalene, tetrahydroquinoline, tetrahydroisoquinoline, indane, cyclopropylbenzene, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is
optionally substituted. R3 may be selected from:
Figure imgf000021_0001
Figure imgf000021_0002
any one of which is optionally substituted. For example, R3 may be selected from:
Figure imgf000021_0003
Figure imgf000022_0001
[0035] In some embodiments for a compound or salt of Formula (IA) or (IB), L2 is selected from -C(O)-, and -C(0)NR10-. In certain embodiments, L2 is -C(O)-. In certain embodiments, L2 is selected from -C(0)NR10-. R10 of -C(0)NR10- may be selected from hydrogen and Ci-6 alkyl. For example, L2 may be -C(0)NH-.
[0036] In some embodiments for a compound or salt of Formula (IA) or (IB), R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. [0037] In some embodiments for a compound or salt of Formula (IA) or (IB), R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle. In some embodiments, R4 is selected from: -OR10, and -N(R10)2; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl. In certain embodiments, R4 is -N(R10)2. R10 of -N(R10)2 may be independently selected at each occurrence from optionally substituted Ci-6 alkyl. In certain embodiments, R10 of -N(R10)2 is independently selected at each occurrence from methyl, ethyl, propyl, and butyl, any
one of which is optionally substituted. For example, R4 may
Figure imgf000023_0001
certain
embodiments,
Figure imgf000023_0002
[0038] In some embodiments for a compound or salt of Formula (IA) or (IB), R12 is
independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -
S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl. In some embodiments, R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -
S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle.
[0039] In some aspects, the present disclosure provides a compound represented by the structure of Formula (IVA):
Figure imgf000024_0001
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L12 is selected from -X3-, -X3-CI-6 alkylene-X3-, -X3-C2-6 alkenylene-X3-, and - X3-C2-6 alkynylene-X3-, each of which is optionally substituted on alkyl ene, alkenylene, or alkynylene with one or more substituents independently selected from R12;
L22 is independently selected from -X4-, -X4-CI-6 alkylene-X4-, -X4-C2-6 alkenylene-X4-, and -X4-C2-6 alkynylene-X4-, each of which is optionally substituted on alkyl ene, alkenylene, or alkynylene with one or more substituents independently selected from R10;
X3 and X4 are independently selected at each occurrence from a bond, -0-, -S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-,
-C(O)N(R10)C(O)N(R10)-, -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -0S(0)-, -S(O) 0-, -S(O)-, -0S(0)2-, -S(0)20-, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 and R8 are independently selected from: -OR10, -N(R10)2, -C(O)N(R10)2, - C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from L3, halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-12 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-12 carbocycle, and 3- to l2-membered heterocycle in R4 and R8 is optionally substituted with one or more substituents independently selected from L3, halogen, -
OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -
C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from L3, hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -NO2, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, CI-IO alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
L3 is a linker moiety, wherein at least one of R10 is L3 or at least one substituent on a group selected from R4, R8, X3 and X4 is L3; and
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(0)N(R10)2, -N(R10)C(0)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle. In some embodiments, the compound of Formula (IV A) is represented by Formula (IVB):
Figure imgf000026_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen, - OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S,
=N(R10; 1, -CN, Ci-10 alkyl, C2-io alkenyl, and C2-io alkynyl; and
I24, and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, -N(R10)2, -
S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl,
C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
[0040] In some embodiments for a compound or salt of Formula (IV A) or (IVB), L12 is - C(0)N(R10)-. In some embodiments, R10 of -C(0)N(R10)- is selected from hydrogen, Ci-6 alkyl, and L3. For example, L12 may be -C(0)NH-.
[0041] In some embodiments for a compound or salt of Formula (IV A) or (IVB), R8 is an optionally substituted 5- or 6-membered heteroaryl. R8 may be an optionally substituted 5- or 6- membered heteroaryl, substituted with L3. In some embodiments, R8 is an optionally substituted pyridine, substituted with L3.
[0042] In some embodiments for a compound or salt of Formula (IV A) or (IVB), L22 is selected from -C(O)-, and -C(0)NR10-. In certain embodiments, L22 is -C(O)-. In certain embodiments, L22 is -C(0)NR10-. R10 of -C(0)NR10- may be selected from hydrogen, Ci-6 alkyl, and -L3. For example, L22 may be -C(0)NH-.
[0043] In some embodiments for a compound or salt of Formula (IV A) or (IVB), R4 is selected from: -OR10, and -N(R10)2; and C1-10 alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to 12- membered heterocycle, aryl, and heteroaryl, each of which is optionally substituted with one or more substituents independently selected from L3, halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, -
S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO¾ =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl. In some embodiments, R4 is -N(R10)2 and R10 of -N(R10)2 is selected from L3 and hydrogen, and wherein at least one R10 of -N(R10)2 is L3.
[0044] In some embodiments, the compound of any one of Formula (IA), (IB), (IIA), (IIB), (IIIA), and (PIB), is further covalently bound to a linker, L3. In some embodiments for a compound or salt of any one of Formula (IA), (IB), (IIA), (IIB), (IIIA), (IIIB), (IV A), and (IVB), L3 is a noncleavable linker. In some embodiments for a compound or salt of any one of Formula (IA), (IB), (IIA), (IIB), (IIIA), (IIIB), (IV A), and (IVB), L3 is a cleavable linker. L3 may be cleavable by a lysosomal enzyme. In some embodiments, the compound is covalently attached to an antibody construct. In some embodiments, the compound is covalently attached to a targeting moiety, optionally through the linker. In some embodiments, the targeting moiety or antibody construct specifically binds to a tumor antigen. In some embodiments, the antibody construct or targeting moiety further comprises a target binding domain.
[0045] In some embodiments for a compound or salt of any one of Formula (IA), (IB), (IIA), (IIB), (IIIA), (IIIB), (IV A), and (IVB), L3 is represented by the formula:
Figure imgf000027_0001
, wherein:
L4 represents the C-terminus of the peptide and L5 is selected from a bond, alkylene and heteroalkylene, wherein L5 is optionally substituted with one or more groups independently selected from R32, and RX is a reactive moiety; and
R32 is independently selected at each occurrence from halogen, -OH, -CN, -O-alkyl, -SH, =0, =S, -NH2, -NO2; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -O- alkyl, -SH, =0, =S, -NH2, -NO2. In some embodiments, RX comprises a leaving group. In some embodiments, RX comprises a maleimide. In some embodiments, L3 is further covalently bound to an antibody construct. In some embodiments, the antibody construct is directed against a tumor antigen. In some embodiments, the antibody construct further comprises target binding domain.
[0046] In some embodiments for a compound or salt of any one of Formula (IA), (IB), (IIA), (IIB), (PIA), (IIIB), (IV A), and (IVB), L3 is represented by the formula:
Figure imgf000027_0002
, wherein L4 represents the C-terminal of the peptide and L5 is selected from a bond, alkylene and heteroalkylene, wherein L5 is optionally substituted with one or more groups independently selected from R32 ; RX* comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein * on RX* represents the point of attachment to the residue of the antibody construct; and,
R32 is independently selected at each occurrence from halogen, -OH, -CN, -O-alkyl, -SH, =0, =S, -NH2, -N02; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -O- alkyl, -SH, =0, =S, -NH2, -N02. In some embodiments, the peptide of L3 comprises Val— Cit or Val— Ala.
[0047] In some embodiments for a compound or salt of any one of Formula (IA), (IB), (IIA), (IIB), (IIIA), (IIIB), (IV A), and (IVB), L3 is represented by the formula:
Figure imgf000028_0001
, wherein RX comprises a reactive moiety, and n = 0 9 In some embodiments, RX comprises a leaving group. In some embodiments, RX comprises a
maleimide. In some embodiments, L3 is represented as follows:
Figure imgf000028_0002
wherein RX* comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein
Figure imgf000028_0003
on RX* represents the point of attachment to the residue of the antibody construct, and n = 0 9
[0048] In some embodiments, RX* comprises a succinamide moiety and is bound to a cysteine residue of an antibody construct. In some embodiments, RX* comprises a hydrolyzed succinamide moiety and is bound to a cysteine residue of an antibody construct. In some embodiments, the antibody construct comprises a HER2 antigen binding domain. In some embodiments, the antibody construct comprises a TROP2 antigen binding domain. In some embodiments, the antibody construct is pertuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is trastuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is sacituzumab or an antigen binding fragment thereof.
[0049] In some aspects, the present disclosure provides a conjugate represented by the formula:
Antibody
Figure imgf000028_0004
, wherein Antibody is an antibody construct, D is a compound or salt of any one of Formula (IA), (IB), (IIA), (IIB), (IIIA), and (IPB), and L3 is a linker moiety. In some aspects, the present disclosure provides a conjugate represented by the formula:
Antibody
Figure imgf000029_0001
1-8 , wherein Antibody is an antibody construct and D-L3 is a compound or salt of any one of Formula (IV A), and (IVB).
[0050] In some embodiments, the antibody construct comprises a HER2 antigen binding domain. In some embodiments, the antibody construct comprises a TROP2 antigen binding domain. In some embodiments, the antibody construct is pertuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is trastuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is sacituzumab or an antigen binding fragment thereof.
[0051] In some aspects, the present disclosure provides a pharmaceutical composition
comprising the compound or salt disclosed herein and at least one pharmaceutically acceptable excipient. In some aspects, the present disclosure provides a pharmaceutical composition, comprising the conjugate disclosed herein and at least one pharmaceutically acceptable excipient. In some embodiments, the average DAR of the conjugate is from about 2 to about 8, or about 1 to about 3, or about 3 to about 5.
[0052] In some aspects, the present disclosure provides a method of killing tumor cells in vivo, comprising contacting a tumor cell population with a conjugate disclosed herein. In some aspects, the present disclosure provides a method for treatment, comprising administering to a subject a conjugate disclosed herein. In some aspects, the present disclosure provides a method for treatment, comprising administering to a subject in need thereof a compound or salt disclosed herein.
[0053] In some aspects, the present disclosure provides a method for treating cancer, comprising administering to a subject in need thereof a conjugate disclosed herein. In some aspects, the present disclosure provides a method for treating cancer, comprising administering to a subject in need thereof a compound or salt disclosed herein. In some aspects, the present disclosure provides a compound or salt disclosed herein for use in a method of treatment of a subject’s body by therapy.
[0054] In some aspects, the present disclosure provides a compound or salt disclosed herein for use in a method of treating cancer. In some aspects, the present disclosure provides a conjugate disclosed herein for use in a method of treatment of a subject’s body by therapy. In some aspects, the present disclosure provides a conjugate disclosed herein for use in a method of treating cancer. [0055] In some aspects, the present disclosure provides a method of delivering a benzazepine compound to a cancer cell in a subject comprising administering a conjugate disclosed herein to a subject in need thereof.
[0056] In some aspects, the present disclosure provides a method for the treatment of cancer, comprising administering an effective amount of an antibody conjugate, wherein the antibody conjugate comprises an antibody construct covalently bound through a linker to a TLR8 agonist, wherein the TLR8 agonist has a Kd for TLR7 that is two times or greater than two times the Kd for TLR8, and wherein the antibody conjugate comprises from 1 to 20 TLR8 agonists per antibody construct, preferably from 1-8, 3-5 or 1-3.
[0057] In some aspects, the present disclosure provides a method for the treatment of cancer, comprising administering an effective amount of an antibody conjugate, wherein the antibody conjugate comprises an antibody construct covalently bound through a linker to a TLR8 agonist, wherein the TLR8 agonist agonizes TLR8 with at least an ECso of an order of magnitude less than the amount of the same compound required to show an agonizing effect on TLR7, and wherein the antibody conjugate comprises from 1 to 20 TLR8 agonists per antibody construct, preferably from 1-8, 3-5 or 1-3.
[0058] In some aspects, the present disclosure provides a method for the treatment of cancer, comprising administering an effective amount of an antibody conjugate, wherein the antibody conjugate comprises an antibody construct covalently bound through a linker to a TLR8 agonist, wherein the TLR8 agonist comprises a benzazepine substituted with a bicyclic heterocycle, and wherein the antibody conjugate comprises from 1 to 20 TLR8 agonists per antibody construct, preferably from 1-8, 3-5 or 1-3.
[0059] In some aspects, the present disclosure provides a method of preparing an antibody
Antibody
Figure imgf000030_0001
conjugate of the formula 1 8 , wherein Antibody is an antibody construct, and
L3-D is selected from a compound or salt disclosed herein comprising contacting L3-D with an antibody construct.
[0060] In some embodiments, the antibody construct comprises a HER2 antigen binding domain. In some embodiments, the antibody construct comprises a TROP2 antigen binding domain. In some embodiments, the antibody construct is pertuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is trastuzumab or an antigen binding fragment thereof. In some embodiments, the antibody construct is sacituzumab or an antigen binding fragment thereof. In some embodiments, the methods further comprising purifying the antibody conjugate. BRIEF DESCRIPTION OF THE DRAWINGS
[0061] FIG. 1 depicts an increase production of a pro-inflammatory cytokine, murine TNFa, from bone marrow-derived murine macrophages administered TLR7 small molecules with and without amine masking groups.
INCORPORATION BY REFERENCE
[0062] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.
DETAILED DESCRIPTION OF THE INVENTION
[0063] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.
[0064] The present disclosure provides compounds, conjugates and pharmaceutical compositions for use in the treatment of disease. In certain embodiments, the compounds of the disclosure are TLR8 modulators. In certain embodiments, the compounds are TLR8 agonists. Toll-like receptors (TLRs) are a family of membrane-spanning receptors that are expressed on cells of the immune system like dendritic cells, macrophages, monocytes, T cells, B cells, NK cells and mast cells but also on a variety of non-immune cells such as endothelial cells, epithelial cells and even tumor cells. TLRs can have many isoforms, including TLR4, TLR7 and TLR8.
[0065] In certain aspects, compounds or conjugates of the disclosure are administered in a form suitable to attenuate or eliminate immune-modulatory activity until the compound or conjugate reaches a desired target and the active site amine is unmasked. While not wishing to be bound by a mechanistic theory, the modification of compounds to attenuate or eliminate immune- modulatory activity may prevent undesired off-target immune-stimulatory activity, e.g., immune- stimulation in healthy tissue.
[066] In certain embodiments, a compound such as a TLR8 agonist is modified with a masking group, such that the TLR8 agonist has limited activity or is inactive until it reaches an
environment where the masking group is removed to reveal the active compound. For example, the TLR8 agonist is covalently modified at an amine involved in binding to the active site of a TLR8 receptor such that the compound is unable to bind the active site of the receptor in its modified form. In such an example, the masking group may be removed under physiological conditions, e.g., enzymatic or acidic conditions, specific to the site of delivery, e.g., intracellular or extracellular adjacent to target cells. Masking groups may be removed from the amine of the compound or salt described herein due to the action of lysosomal proteases, e.g., cathepsin and plasmin. These proteases can be present at elevated levels in certain tumor tissues. The masking group may be removed by a lysosomal enzyme. The lysosomal enzyme can be, for example, cathepsin B, cathepsin S, b-glucuronidase, or b-galactosidase.
[067] In certain embodiments, the amine masking group inhibits binding of the amine group of the compound with residues of a TLR8 receptor. The amine masking group may be removable under physiological conditions within a cell but remains covalently bound to the amine outside of a cell. Masking groups that may be used to inhibit or attenuate binding of an amine group of a compound with residues of a TLR8 receptor include, for example, peptides and carbamates.
[0068] TLR8 receptors are localized to the endolysosomal/phagosomal compartment and predominantly found to be expressed by cells of the myeloid lineage. TLR ligation leads to activation of NF-kB and IRF-dependent pathways with the specific activation sequence and response with respect to the specific TLR and cell type. While TLR7 is mainly expressed in all dendritic cells subtypes (DC and here highly in pDC, plasmacytoid DC) and can be induced in B cells upon IFNa stimulation, TLR8 expression is rather restricted to monocytes, macrophages and myeloid DC. TLR8 signaling via MyD88 can be activated by bacterial single stranded RNA, small molecule agonists and microRNAs. The activation of TLR8 results in the production of various pro-inflammatory cytokines such as IL-6, IL-12 and TNF-a as well as enhanced expression of co-stimulatory molecules, such as CD80, CD86, and chemokine receptors. In addition, TLR8 activation can induce type I interferon (IRNb) in primary human monocytes.
[0069] Several agonists targeting activation of different TLRs can be used in various
immunotherapies, including vaccine adjuvants and in cancer immunotherapies. TLR agonists can range from simple molecules to complex macromolecules. Likewise, the sizes of TLR agonists can range from small to large. TLR agonists can be synthetic or biosynthetic agonists. TLR agonists can also be Pathogen-Associated Molecular Pattern molecules (PAMPs).
[0070] The compounds of the present disclosure may be useful for the treatment and prevention, e.g., vaccination, of cancer, autoimmune diseases, inflammation, sepsis, allergy, asthma, graft rejection, graft-versus-host disease, immunodeficiencies, and infectious diseases.
[0071] In certain embodiments, the compounds have utility in the treatment of cancer either as single agents or in combination therapy. In certain embodiments, the compounds have utility as single agent immunomodulators, vaccine adjuvants and in combination with conventional cancer therapies. In certain embodiments, the compounds are incorporated into a conjugate that can be utilized, for example, to enhance an immune response. In certain embodiments, the disclosure provides antibody construct-benzazepine compound conjugates and their use for treating cancer.
Definitions
[0072] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs.
All patents and publications referred to herein are incorporated by reference.
[0073] As used in the specification and claims, the singular form“a”,“an” and“the” includes plural references unless the context clearly dictates otherwise.
[0074] As used herein, an“amine masking group” refers to any moiety covalently bound to the nitrogen of an amine, e.g., primary amine, which attenuates the interaction or activity, or blocks the amine from interacting with a TLR8 receptor and that is removable from the amine.
Exemplary amine masking groups include enzymatically-cleavable moieties such as amino acids, and peptides.
[0075] As used herein, the term“antibody” refers to an immunoglobulin molecule that specifically binds to, or is immunologically reactive toward, a specific antigen. The term antibody can include, for example, polyclonal, monoclonal, genetically engineered, and antigen binding fragments thereof. An antibody can be, for example, murine, chimeric, humanized, heteroconjugate, bispecific, a diabody, a triabody, or a tetrabody. The antigen binding fragment can include, for example, Fab', F(ab')2, Fab, Fv, rlgG, and scFv.
[0076] As used herein, an“antigen binding domain” refers to a region of a molecule that specifically binds to an antigen. An antigen binding domain may be a domain that can
specifically bind to an antigen. An antigen binding domain can be an antigen-binding portion of an antibody or an antibody fragment. An antigen binding domain can be one or more fragments of an antibody that retain the ability to specifically bind to an antigen. An antigen binding domain can be an antigen binding fragment. In some embodiments, an antigen binding domain can recognize a single antigen. In some embodiments, an antigen binding domain can recognize, for example, two, three, four, five, six, seven, eight, nine, ten, or more than one antigen.
[0077] As used herein, an“antibody construct” refers to a molecule, e.g., a protein, peptide, antibody or portion thereof, that contains an antigen binding domain and an Fc domain. An antibody construct can recognize, for example, multiple antigens.
[0078] As used herein, the abbreviations for amino acids are conventional and can be as follows: alanine (A, Ala); arginine (R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine (C, Cys); glutamic acid (E, Glu); glutamine (Q, Gln); glycine (G, Gly); histidine (H, His); isoleucine (I, Ile); leucine (L, Leu); lysine (K, Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro); serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y, Tyr); valine (V, Val). Other amino acids include citrulline (Cit); homocysteine (Hey); hydroxyproline (Hyp); ornithine (Orn); and thyroxine (Thx).
[0079]“Conjugate”, as used herein, refers to an antibody construct that is linked, e.g., covalently linked, either directly or through a linker to a compound or compound-linker described herein, e.g., a compound or salt of any one of the Formulas described herein.
[0080] As used herein, an“Fc domain” can be an Fc domain from an antibody or from a non antibody that can bind to an Fc receptor.
[0081] As used herein,“recognize” with regard to antibody interactions refers to the specific association or specific binding between an antigen binding domain or portion thereof and an antigen.
[0082] As used herein, a“target binding domain” refers to a construct that contains an antigen binding domain from an antibody or from a non-antibody that can bind to the antigen.
[0083] As used herein, a“tumor antigen” is an antigenic substance associated with a tumor or cancer cell, and can trigger an immune response in a host.
[0084] The term“salt” or“pharmaceutically acceptable salt” refers to salts derived from a variety of organic and inorganic counter ions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, / oluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is chosen from ammonium, potassium, sodium, calcium, and magnesium salts. [0085] The term“Cx-y” when used in conjunction with a chemical moiety, such as alkyl, alkenyl, or alkynyl is meant to include groups that contain from x to y carbons in the chain. For example, the term“Ci^alkyl” refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups that contain from 1 to 6 carbons. The term -C x-y alkyl ene- refers to a substituted or unsubstituted alkylene chain with from x to y carbons in the alkylene chain. For example -Ci-6alkylene- may be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any one of which is optionally substituted.
[0086] The terms“Cx-yalkenyl” and“Cx-yalkynyl” refer to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively. The term -Cx-yalkenylene- refers to a substituted or unsubstituted alkenylene chain with from x to y carbons in the alkenyl ene chain. For example, -C2-6alkenylene- may be selected from ethenylene, propenylene, butenylene, pentenylene, and hexenylene, any one of which is optionally substituted. An alkenylene chain may have one double bond or more than one double bond in the alkenylene chain. The term -Cx-yalkynylene- refers to a substituted or unsubstituted alkynylene chain with from x to y carbons in the alkenylene chain. For example, -C2-6alkenylene- may be selected from ethynylene, propynylene, butynylene, pentynylene, and hexynylene, any one of which is optionally substituted. An alkynylene chain may have one triple bond or more than one triple bond in the alkynylene chain.
[0087] "Alkylene" refers to a divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation, and preferably having from one to twelve carbon atoms, for example, methylene, ethylene, propylene, butylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group are through the terminal carbons respectively. In other embodiments, an alkylene comprises one to five carbon atoms (i.e., C1-C5 alkylene). In other embodiments, an alkylene comprises one to four carbon atoms
(i.e., C1-C4 alkylene). In other embodiments, an alkylene comprises one to three carbon atoms
(i.e., C1-C3 alkylene). In other embodiments, an alkylene comprises one to two carbon atoms
(i.e., Ci-C2 alkylene). In other embodiments, an alkylene comprises one carbon atom (i.e., Ci alkylene). In other embodiments, an alkylene comprises five to eight carbon atoms (i.e., Cs-Cx alkylene). In other embodiments, an alkylene comprises two to five carbon atoms (i.e., C2-C5 alkylene). In other embodiments, an alkylene comprises three to five carbon atoms (i.e., C3-C5 alkylene). Unless stated otherwise specifically in the specification, an alkylene chain is optionally substituted by one or more substituents such as those substituents described herein.
[0088] "Alkenylene" refers to a divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon double bond, and preferably having from two to twelve carbon atoms. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group are through the terminal carbons respectively. In other embodiments, an alkenylene comprises two to five carbon atoms (i.e., C2-C5 alkenylene). In other embodiments, an alkenylene comprises two to four carbon atoms (i.e., C2-C4 alkenylene). In other
embodiments, an alkenylene comprises two to three carbon atoms (i.e., C2-C3 alkenylene). In other embodiments, an alkenylene comprises two carbon atom (i.e., C2 alkenylene). In other embodiments, an alkenylene comprises five to eight carbon atoms (i.e., Cs-Cx alkenylene). In other embodiments, an alkenylene comprises three to five carbon atoms (i.e., C3-C5 alkenylene). Unless stated otherwise specifically in the specification, an alkenylene chain is optionally substituted by one or more substituents such as those substituents described herein.
[0089] " Alkynylene" refers to a divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond, and preferably having from two to twelve carbon atoms. The alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to the radical group are through the terminal carbons respectively. In other embodiments, an alkynylene comprises two to five carbon atoms (i.e., C2-C5 alkynylene). In other embodiments, an alkynylene comprises two to four carbon atoms (i.e., C2-C4 alkynylene). In other
embodiments, an alkynylene comprises two to three carbon atoms (i.e., C2-C3 alkynylene). In other embodiments, an alkynylene comprises two carbon atom (i.e., C2 alkynylene). In other embodiments, an alkynylene comprises five to eight carbon atoms (i.e., Cs-Cx alkynylene). In other embodiments, an alkynylene comprises three to five carbon atoms (i.e., C3-C5 alkynylene). Unless stated otherwise specifically in the specification, an alkynylene chain is optionally substituted by one or more substituents such as those substituents described herein.
[0090] "Heteroalkylene" refers to a divalent hydrocarbon chain including at least one heteroatom in the chain, containing no unsaturation, and preferably having from one to twelve carbon atoms and from one to 6 heteroatoms, e.g., -0-, -NH-, -S-. The heteroalkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the heteroalkyl ene chain to the rest of the molecule and to the radical group are through the terminal atoms of the chain. In other embodiments, a heteroalkylene comprises one to five carbon atoms and from one to three heteroatoms. In other embodiments, a heteroalkylene comprises one to four carbon atoms and from one to three heteroatoms. In other embodiments, a heteroalkylene comprises one to three carbon atoms and from one to two heteroatoms. In other embodiments, a heteroalkylene comprises one to two carbon atoms and from one to two heteroatoms. In other embodiments, a heteroalkylene comprises one carbon atom and from one to two heteroatoms. In other embodiments, a heteroalkylene comprises five to eight carbon atoms and from one to four heteroatoms. In other embodiments, a heteroalkylene comprises two to five carbon atoms and from one to three heteroatoms. In other embodiments, a heteroalkylene comprises three to five carbon atoms and from one to three heteroatoms. Unless stated otherwise specifically in the specification, a heteroalkylene chain is optionally substituted by one or more substituents such as those substituents described herein.
[0091] The term“carbocycle” as used herein refers to a saturated, unsaturated or aromatic ring in which each atom of the ring is carbon. Carbocycle includes 3- to lO-membered monocyclic rings, 6- to l2-membered bicyclic rings, and 6- to l2-membered bridged rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. A bicyclic carbocycle includes any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits. A bicyclic carbocycle includes any combination of ring sizes such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl. The term“unsaturated carbocycle” refers to carbocycles with at least one degree of unsaturation and excluding aromatic carbocycles. Examples of unsaturated carbocycles include cyclohexadiene, cyclohexene, and cyclopentene.
[0092] The term“heterocycle” as used herein refers to a saturated, unsaturated or aromatic ring comprising one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to lO-membered monocyclic rings, 6- to l2-membered bicyclic rings, and 6- to l2-membered bridged rings. A bicyclic heterocycle includes any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits. In an exemplary embodiment, an aromatic ring, e.g., pyridyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, morpholine, piperidine or cyclohexene. A bicyclic heterocycle includes any combination of ring sizes such as 4-5 fused ring systems, 5-5 fused ring systems, 5- 6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. The term“unsaturated heterocycle” refers to heterocycles with at least one degree of unsaturation and excluding aromatic heterocycles.
Examples of unsaturated heterocycles include dihydropyrrole, dihydrofuran, oxazoline, pyrazoline, and dihydropyridine.
[0093] The term“heteroaryl” includes aromatic single ring structures, preferably 5- to 7- membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms.
The term“heteroaryl” also includes polycyclic ring systems having two or more rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other rings can be aromatic or non-aromatic carbocyclic, or
heterocyclic. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.
[0094] The term“substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons or substitutable heteroatoms, e.g., an NH or NH2 of a compound. It will be understood that“substitution” or“substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, i.e., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. In certain embodiments, substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as substituting the two hydrogen atoms on a single carbon with an oxo, imino or thioxo group. As used herein, the term“substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds.
[0095] In some embodiments, substituents may include any substituents described herein, for example: halogen, hydroxy, oxo (=0), thioxo (=S), cyano (-CN), nitro (-N02), imino (=N-H), oximo (=N-OH), hydrazino (=N-
NH2), -Rb-ORa, -Rb-OC(0)-Ra, -Rb-OC(0)-ORa, -Rb-OC(0)-N(Ra)2, -Rb-N(Ra)2, -Rb-C(0)Ra, -R b-C(0)ORa, -Rb-C(0)N(Ra)2, -Rb-0-Rc-C(0)N(Ra)2, -Rb-N(Ra)C(0)ORa, -Rb-N(Ra)C(0)Ra, -Rb- N(Ra)S(0)tRa (where t is 1 or 2), -Rb-S(0)tRa (where t is 1 or 2), -Rb-S(0)tORa (where t is 1 or 2), and -Rb-S(0)tN(Ra)2 (where t is 1 or 2); and alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl any of which may be optionally substituted by alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=0), thioxo (=S), cyano (-CN), nitro (-N02), imino (=N-H), oximo (=N-OH), hydrazine (=N-
NH2), -Rb-ORa, -Rb-0C(0)-Ra, -Rb-0C(0)-0Ra, -Rb-0C(0)-N(Ra)2, -Rb-N(Ra)2, -Rb-C(0)Ra, -R b-C(0)0Ra, -Rb-C(0)N(Ra)2, -Rb-0-Rc-C(0)N(Ra)2, -Rb-N(Ra)C(0)0Ra, -Rb-N(Ra)C(0)Ra, -Rb- N(Ra)S(0)tRa (where t is 1 or 2), -Rb-S(0)tRa (where t is 1 or 2), -Rb-S(0)tORa (where t is 1 or 2) and -Rb-S(0)tN(Ra)2 (where t is 1 or 2); wherein each Ra is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl,
heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, wherein each Ra, valence permitting, may be optionally substituted with alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=0), thioxo (=S), cyano (-CN), nitro (-N02), imino (=N-H), oximo (=N-OH), hydrazine (=N-
NH2), -Rb-ORa, -Rb-0C(0)-Ra, -Rb-0C(0)-0Ra, -Rb-0C(0)-N(Ra)2, -Rb-N(Ra)2, -Rb-C(0)Ra, -R b-C(0)0Ra, -Rb-C(0)N(Ra)2, -Rb-0-Rc-C(0)N(Ra)2, -Rb-N(Ra)C(0)0Ra, -Rb-N(Ra)C(0)Ra, -Rb- N(Ra)S(0)tRa (where t is 1 or 2), -Rb-S(0)tRa (where t is 1 or 2), -Rb-S(0)t0Ra (where t is 1 or 2) and -Rb-S(0)tN(Ra)2 (where t is 1 or 2); and wherein each Rb is independently selected from a direct bond or a straight or branched alkylene, alkenylene, or alkynylene chain, and each Rc is a straight or branched alkylene, alkenylene or alkynylene chain.
[0096] It will be understood by those skilled in the art that substituents can themselves be substituted, if appropriate. Unless specifically stated as“unsubstituted,” references to chemical moieties herein are understood to include substituted variants. For example, reference to a “heteroaryl” group or moiety implicitly includes both substituted and unsubstituted variants, unless specified otherwise.
[0097] The phrases“parenteral administration” and“administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
[0098] The phrase“pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
[0099] The phrase“pharmaceutically acceptable excipient” or“pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be“acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen- free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.
[0100] The phrase“targeting moiety” refers to a structure that has a selective affinity for a target molecule relative to other non-target molecules. The targeting moiety binds to a target molecule. A targeting moiety may include, for example, an antibody, a peptide, a ligand, a receptor, or a binding portion thereof. The target biological molecule may be a biological receptor or other structure of a cell such as a tumor antigen.
Antibody Construct
[0101] Disclosed herein are antibody constructs that may be used together with compounds of the disclosure. In certain embodiments, compounds of the disclosure are linked, e.g., covalently linked, either directly or through a linker to a compound of the disclosure forming conjugates.
In certain embodiments, conjugates of the disclosure are represented by the following formula:
Figure imgf000040_0001
wherein A is an antibody construct, L3 is a linker, D is a compound or salt of any one of
Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB) or (IIIC), or L3-D is a compound or salt of any one of Formulas (IV A), (IVB) or (IVC), and n is from 1 to 20. In certain
embodiments, n is from 1 to 10, such as from 1 to 9, such as from 1 to 8, such as from 2 to 8, such as from 1 to 6, such as from 3 to 5, or such as from 1 to 3. In certain embodiments, n is 4. In certain embodiments, each D or L3-D are independently selected from Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB) or (IIIC), or Formulas (IVA), (IVB) or (IVC), respectively.
[0102] In certain embodiments, a compound or salt of the disclosure, e.g., a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC), may be referred to herein as a drug, D, a benzazepine compound, an immune-stimulatory compound, an ISC, or a payload, particularly when referenced as part of a conjugate.“LP”,“linker- payload”,“L3-D”, or“compound-linker” may be used herein to refer to a compound or salt of the disclosure bound to a linker.
[0103] An antibody construct may contain, for example, two, three, four, five, six, seven, eight, nine, ten, or more antigen binding domains. An antibody construct may contain two antigen binding domains in which each antigen binding domain can recognize the same antigen. An antibody construct may contain two antigen binding domains in which each antigen binding domain can recognize different antigens. An antigen binding domain may be in a scaffold, in which a scaffold is a supporting framework for the antigen binding domain. An antigen binding domain may be in a non-antibody scaffold. An antigen binding domain may be in an antibody scaffold. An antibody construct may comprise an antigen binding domain in a scaffold. The antibody construct may comprise a Fc fusion protein. In some embodiments, the antibody construct is a Fc fusion protein. An antigen binding domain may specifically bind to a tumor antigen. An antigen binding domain may specifically bind to an antigen that is at least 80%, at least 90%, at least 95%, at least 99%, or 100% homologous to a tumor antigen. An antigen binding domain may specifically bind to an antigen on an antigen presenting cell (APC). An antigen binding domain may specifically bind to an antigen that is at least 80%, at least 90%, at least 95%, at least 99%, or 100% homologous to an antigen on an antigen presenting cell (APC).
[0104] An antigen binding domain of an antibody may comprise one or more light chain (LC) CDRs and one or more heavy chain (HC) CDRs. For example, an antibody binding domain of an antibody may comprise one or more of the following: a light chain complementary determining region 1 (LC CDR1), a light chain complementary determining region 2 (LC CDR2), or a light chain complementary determining region 3 (LC CDR3). For another example, an antibody binding domain may comprise one or more of the following: a heavy chain complementary determining region 1 (HC CDR1), a heavy chain complementary determining region 2 (HC CDR2), or a heavy chain complementary determining region 3 (HC CDR3). As an additional example, an antibody binding domain of an antibody may comprise one or more of the following: LC CDR1, LC CDR2, LC CDR3, HC CDR1, HC CDR2, and HC CDR3. [0105] The antigen binding domain of an antibody construct may be selected from any domain that binds the antigen including, but not limited to, from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or an antigen binding fragment thereof, for example, a heavy chain variable domain (VH) and a light chain variable domain (VL), or from a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a T cell receptor, or a recombinant T cell receptor.
[0106] In certain embodiments, the antigen binding domain of an antibody construct may be selected from any domain that binds the antigen including, but not limited to, from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or an antigen binding fragment thereof, for example, a heavy chain variable domain (VH) and a light chain variable domain (VL).
[0107] The antigen binding domain of an antibody construct may be at least 80% identical to an antigen binding domain selected from, but not limited to, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (VH) and a light chain variable domain (VL), or to a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a cytokine, a ligand, an immunocytokine, a T cell receptor, or a recombinant T cell receptor. In certain embodiments, an antigen binding domain of an antibody construct may be at least 80% identical to an antigen binding domain selected from, but not limited to, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (VH) and a light chain variable domain (VL).
[0108] In certain embodiments, an antibody construct comprises an Fc domain that may further comprise an Fc domain, in which the Fc domain may be the part of an Fc region that interacts with Fc receptors. The Fc domain of an antibody construct may interact with Fc-receptors (FcRs) found on immune cells. The Fc domain may also mediate the interaction between effector molecules and cells, which can lead to activation of the immune system. The Fc domain may be derived from IgG, IgA, or IgD antibody isotypes, and may comprise two identical protein fragments, which are derived from the second and third constant domains of the antibody’s heavy chains. In an Fc domain derived from an IgG antibody isotype, the Fc region may comprise a highly-conserved N-glycosylation site, which may be essential for FcR-mediated downstream effects. The Fc domain may be derived from IgM or IgE antibody isotypes, in which the Fc domain may comprise three heavy chain constant domains.
[0109] An Fc domain may interact with different types of FcRs. The different types of FcRs may include, for example, FcyRI, FcyRIIA, FcyRIIB, FcyRI 11 A, FcyRIIIB, FcaRI, FcpR, FceRI, FceRII, and FcRn. FcRs may be located on the membrane of certain immune cells including, for example, B lymphocytes, natural killer cells, macrophages, neutrophils, follicular dendritic cells, eosinophils, basophils, platelets, and mast cells. Once the FcR is engaged by the Fc domain, the FcR may initiate functions including, for example, clearance of an antigen-antibody complex via receptor-mediated endocytosis, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody dependent cell-mediated phagocytosis (ADCP), and ligand-triggered transmission of signals across the plasma membrane that can result in alterations in secretion, exocytosis, and cellular metabolism. FcRs may deliver signals when FcRs are aggregated by antibodies and multivalent antigens at the cell surface. The aggregation of FcRs with immunoreceptor tyrosine- based activation motifs (ITAMs) may sequentially activate SRC family tyrosine kinases and SYK family tyrosine kinases. ITAM comprises a twice-repeated YxxL sequence flanking seven variable residues. The SRC and SYK kinases may connect the transduced signals with common activation pathways.
[0110] An antibody may consist of two identical light protein chains and two identical heavy protein chains, all held together covalently by disulfide linkages. The N-terminal regions of the light and heavy chains together may form the antigen recognition site of an antibody.
Structurally, various functions of an antibody may be confined to discrete protein domains. The sites that can recognize and can bind antigen may consist of three complementarities determining regions (CDRs) that may lie within the variable heavy chain region and variable light chain region at the N-terminal end of the heavy chain and the light chain. The constant domains may provide the general framework of the antibody and may not be involved directly in binding the antibody to an antigen, but may be involved in various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity, and may bind Fc receptors. The constant domains may include an Fc region. The constant domains may include an Fc domain. The domains of natural light and heavy chains may have the same general structures, and each domain may comprise four framework regions, whose sequences can be somewhat conserved, connected by three hyper-variable regions or CDRs. The four framework regions (FR) may largely adopt a b-sheet conformation and the CDRs can form loops connecting, and in some aspects forming part of, the b -sheet structure. The CDRs in each chain may be held in close proximity by the framework regions and, with the CDRs from the other chain, may contribute to the formation of the antigen binding site.
[0111] An antibody construct may comprise a light chain of an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten modifications and in certain embodiments, not more than 40, 35, 30, 25, 20, 15 or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence. An antibody construct may comprise a heavy chain of an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten modifications and in certain embodiments, not more than 40, 35, 30, 25, 20, 15 or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence.
[0112] An antibody of an antibody construct may include an antibody of any type, which may be assigned to different classes of immunoglobins, e.g ., IgA, IgD, IgE, IgG, and IgM. Several different classes may be further divided into isotypes, e.g. , IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. An antibody may further comprise a light chain and a heavy chain, often more than one chain. The heavy-chain constant regions (Fc) that corresponds to the different classes of immunoglobulins may be a, d, e, g, and m, respectively. The light chains may be one of either kappa (K) or lambda (l), based on the amino acid sequences of the constant domains. The Fc region may contain an Fc domain. An Fc receptor may bind an Fc domain. Antibody constructs may also include any fragment or recombinant forms thereof, including but not limited to, single chain variable fragments (scFvs),‘T-bodies’, anti-calins, centyrins, affibodies, domain antibodies, or peptibodies.
[0113] An antibody construct may comprise an antibody fragment. An antibody fragment may include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; and (iii) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody. Although the two domains of the Fv fragment, VL and VH, may be coded for by separate genes, they may be linked by a synthetic linker to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules.
[0114] An antibody may include an Fc region comprising multiple Fc domains. The Fc domain of an antibody may interact with FcRs found on immune cells. The Fc domain may also mediate the interaction between effector molecules and cells, which may lead to activation of the immune system. In the IgG, IgA, and IgD antibody isotypes, the Fc region may comprise two identical protein fragments, which can be derived from the second and third constant domains of the antibody’s heavy chains. In the IgM and IgE antibody isotypes, the Fc regions may comprise three heavy chain constant domains. In the IgG antibody isotype, the Fc regions may comprise a highly-conserved N-glycosylation site, which may be important for FcR-mediated downstream effects.
[0115] An antibody used herein may be chimeric or“humanized.” Chimeric and humanized forms of non-human (e.g, murine) antibodies can be chimeric immunoglobulins,
immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')2 or other target- binding subdomains of antibodies), which may contain minimal sequences derived from non human immunoglobulin. In general, the humanized antibody may comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the framework regions are those of a human immunoglobulin sequence. The humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin consensus sequence.
[0116] An antibody described herein may be a human antibody. As used herein,“human antibodies” can include antibodies having, for example, the amino acid sequence of a human immunoglobulin and may include antibodies isolated from human immunoglobulin libraries or from animals transgenic for one or more human immunoglobulins that do not express
endogenous immunoglobulins. Human antibodies may be produced using transgenic mice which are incapable of expressing functional endogenous immunoglobulins, but which may express human immunoglobulin genes. Completely human antibodies that recognize a selected epitope may be generated using guided selection. In this approach, a selected non-human monoclonal antibody, e.g ., a mouse antibody, may be used to guide the selection of a completely human antibody recognizing the same epitope.
[0117] An antibody described herein may be a bispecific antibody or a dual variable domain antibody (DVD). Bispecific and DVD antibodies may be monoclonal, often human or humanized, antibodies that can have binding specificities for at least two different antigens.
[0118] An antibody described herein may be a derivatized antibody. For example, derivatized antibodies may be modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein.
[0119] An antibody described herein may have a sequence that has been modified to alter at least one constant region-mediated biological effector function relative to the corresponding wild type sequence. For example, in some embodiments, the antibody can be modified to reduce at least one constant region-mediated biological effector function relative to an unmodified antibody, e.g. , reduced binding to the Fc receptor (FcR). FcR binding may be reduced by, for example, mutating the immunoglobulin constant region segment of the antibody at particular regions necessary for FcR interactions.
[0120] An antibody or Fc domain as described herein may be modified to acquire or improve at least one constant region-mediated biological effector function relative to an unmodified antibody or Fc domain, e.g. , to enhance FcyR interactions. For example, an antibody with a constant region that binds FcyRIIA, FcyRIIB and/or FcyRIIIA with greater affinity than the corresponding wild type constant region may be produced according to the methods described herein. An Fc domain that binds FcyRIIA, FcyRIIB and/or FcyRIIIA with greater affinity than the corresponding wild type Fc domain may be produced according to the methods described herein.
[0121] In certain embodiments, the antibody construct comprises an antigen binding domain and an IgG Fc domain, wherein a Kd for binding of the antigen binding domain to a first antigen in a presence of the immune-stimulatory compound is less than about 100 nM and no greater than about 100 times a Kd for binding of the antigen binding domain to the first antigen in the absence of the immune-stimulatory compound. In certain embodiments, the antibody construct comprises a Kd for binding of the IgG Fc domain to an Fc receptor in the presence of the immune- stimulatory compound is no greater than about 100 times a Kd for binding the IgG Fc domain to the Fc receptor in the absence of the immune-stimulatory compound. In certain embodiments, the first antigen is selected from CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD40, CD45, CAMPATH-l, BCMA, CS-l, PD-L1, B7-H3, B7-DC, HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, folate-binding protein, A33, G250, prostate-specific membrane antigen (PSMA), GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein, tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, avB3, WT1, LMP2, HPV E6, HPV E7, Her-2/neu, MAGE A3, p53 nonmutant, NY-ESO-l, MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyronsinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B 1, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, sLe(animal), CYP1B1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-l, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TESL Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, VEGFR2, MAD-CT-l, PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MUC16, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, MUC15, CA6, NAPI2B, TROP2, CLDN6, CLDN16, CLDN18.2, CLorfl86, RON, LY6E, FRA, DLL3, PTK7, STRA6, TMPRSS3, TMPRSS4, TMEM238, UPK1B, VTCN1, LIV1, ROR1, or Fos-related antigen 1. In certain embodiments, an antigen binding domain specifically binds to an antigen, such as those selected from CD5, CD25, CD37, CD33, CD45, BCMA, CS-l, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM,
MUC1, folate-binding protein (FOLR1), A33, G250 (carbonic anhydrase IX), prostate-specific membrane antigen (PSMA), GD2, GD3, GM2, Ley, CA-125, CA19-9 (MUC1 sLe(a)), epidermal growth factor, HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), a tenascin, a metalloproteinase, endosialin, avB3, LMP2, EphA2, PAP, AFP, ALK, polysialic acid, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, sLe(a), GM3, BORIS, Tn, TF, GloboH, STn, CSPG4, ARAP-4, SSX2, Legumain, Tie 2, Tim 3, VEGFR2, PDGFR-B, ROR2, TRAIL 1, MUC16, EGFR, CMET, HER3, MUC1, MUC15, CA6, NAPI2B, TROP2, CLDN18.2, RON, LY6E, FRAlpha, DLL3, PTK7, LIV1, ROR1, CLDN6, GPC3, ADAM 12, LRRC15, CDH6, TMEFF2, TMEM238, GPNMB, ALPPL2, UPK1B, UPK2, LAMP-l, LY6K, EphB2, STEAP, ENPP3, CDH3, Nectin4, LYPD3, EFNA4, GPA33, SLITRK6 or HA VCR 1. In certain embodiments, the first antigen is expressed on an immune cell. In certain embodiments, the first antigen is CD40, HER2 or TROP2. In certain embodiments, the first antigen is HER2 or TROP2.
[0122] In certain embodiments, the antibody construct comprises a human antibody or a humanized antibody or an antigen binding portion thereof, e.g., a humanized CD40, humanized HER2 or humanized TROP2 antibody. In certain embodiments, the antibody construct comprises a TROP2 antibody, e.g., sacituzumab, or an antigen binding portion thereof. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of sacituxumab (SEQ ID NOs:3 and 4). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sacituzumab (SEQ ID NO:4), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sacituzumab (SEQ ID NO:3), as determined by the Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sacituzumab (SEQ ID NO:4), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sacituzumab (SEQ ID NO:3), as determined by IMGT (ImMunoGeneTics). In certain embodiments, the antibody construct comprises a HER2 antibody, e.g., pertuzumab, trastuzumab, or an antigen binding portion thereof. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of pertuzumab (SEQ ID NOs: l and 2). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of pertuzumab (SEQ ID NO:2), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of pertuzumab (SEQ ID NO: 1), as determined by the Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of pertuzumab (SEQ ID NO:2), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of pertuzumab (SEQ ID NO: 1), as determined by IMGT. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of trastuzumab (SEQ ID NOs:7 and 8). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of trastuzumab (SEQ ID NO: 8), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of trastuzumab (SEQ ID NO: 7), as determined by the Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of trastuzumab (SEQ ID NO: 8), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of trastuzumab (SEQ ID NO:7), as determined by IMGT. In certain embodiments, the antibody construct comprises a CD40 antibody or an antigen binding portion thereof. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequence of sacituxumab (SEQ ID NO:3 and 4). In certain embodiments, the antibody construct comprises the LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sacituzumab (SEQ ID NO:4), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sacituzumab (SEQ ID NO:3), as determined by Rabat index.
[0123] In certain embodiments, the antibody construct comprises a Liv-l antibody, e.g., ladiratuzumab, huLivl-l4 (WO 2012078688), Livl-l.7A4 (US 2011/0117013), huLivl-22 (WO 2012078688) or an antigen binding portion thereof. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of ladiratuzumab (SEQ ID NOs:5 and 6). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of ladiratuzumab (SEQ ID NO:6), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of ladiratuzumab (SEQ ID NO: 5), as determined by Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of
ladiratuzumab (SEQ ID NO: 6), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of ladiratuzumab (SEQ ID NO:5), as determined by IMGT. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of huLivl-l4 (SEQ ID NOs: l7 and 18). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of huLivl-l4 (SEQ ID NO: 18), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of huLivl-l4 (SEQ ID NO: 17), as determined by Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of huLivl-l4 (SEQ ID NO: 18), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of huLivl-l4 (SEQ ID NO: 17), as determined by IMGT. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of Livl-l.7A4 (SEQ ID NOs: 19 and 20). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of Livl-l.7A4 (SEQ ID NO:20), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of Livl-l .7A4 (SEQ ID NO: 19), as determined by Rabat index. In certain embodiments, the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of Livl-l.7A4 (SEQ ID NO:20), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of Livl-l.7A4 (SEQ ID NO: 19), as determined by Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of Livl-l.7A4 (SEQ ID NO:20), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of Livl-l.7A4 (SEQ ID NO: 19), as determined by IMGT. In certain embodiments, the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of Livl-l.7A4 (SEQ ID NO:20), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of Livl-l.7A4 (SEQ ID NO: 19), as determined by IMGT. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of huLivl-22 (SEQ ID NOs:2l and 22). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of huLivl-22 (SEQ ID NO:22), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of huLivl-22 (SEQ ID NO:2l), as determined by Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of huLivl-22 (SEQ ID NO:22), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of huLivl-22 (SEQ ID NO:2l), as determined by IMGT.
[0124] In certain embodiments, the antibody construct comprises a MUC16 antibody, e.g., sofituzumab, 4H11 (US2013/0171152), 4H5 (US2013/0171152) or an antigen binding portion thereof. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of sofituzumab (SEQ ID NOs:23 and 24). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sofituzumab (SEQ ID NO:24), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sofituzumab (SEQ ID NO:23), as determined by Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of sofituzumab (SEQ ID NO:24), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of sofituzumab (SEQ ID NO:23), as determined by IMGT. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of antibody 4H11 (SEQ ID NOs: 13 and 14). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4H11 (SEQ ID NO: 14), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of antibody 4H11 (SEQ ID NO: 13), as determined by Rabat index. In certain embodiments, the antibody construct comprises a humanized antibody comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4H11 (SEQ ID NO: 14), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of antibody 4H11 (SEQ ID NO: 13), as determined by Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4H11 (SEQ ID NO: 14), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of 4H11 (SEQ ID NO: 13), as determined by IMGT. In certain embodiments, the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4H11 (SEQ ID NO: 14), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of 4H11 (SEQ ID NO: 13), as determined by IMGT. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of antibody 4A5 (SEQ ID NOs: 15 and 16). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4A5 (SEQ ID NO: 16), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of 4A5 (SEQ ID NO: 15), as determined by Rabat index. In certain
embodiments, the antibody construct comprises a humanized antibody or an antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4A5 (SEQ ID NO: 16), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of antibody 4A5 (SEQ ID NO: 15), as determined by Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of antibody 4A5 (SEQ ID NO: 16), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of antibody 4A5 (SEQ ID NO: 15), as determined by IMGT. In certain embodiments, the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of 4A5 (SEQ ID NO: 16), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of 4A5 (SEQ ID NO: 15), as determined by IMGT.
[0125] In certain embodiments, the antibody construct comprises a PD-L1 antibody, e.g., atezolizumab, MDX-l 105 (WO 2007/005874) or an antigen binding portion thereof. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of atezolizumab (SEQ ID NOs: 11 and 12). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of atezolizumab (SEQ ID NO: 12), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of atezolizumab (SEQ ID NO: 11), as determined by Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of atezolizumab (SEQ ID NO: 12), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of atezolizumab (SEQ ID NO: 11), as determined by IMGT. In certain embodiments, the antibody construct comprises the heavy and light chain variable region sequences of MDX-l 105 (SEQ ID NOs:9 and 10). In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of MDX-l 105 (SEQ ID NO: 10), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of MDX-l 105 (SEQ ID NO: 9), as determined by Rabat index. In certain embodiments, the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of MDX-l 105 (SEQ ID NO: 10), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of MDX-l 105 (SEQ ID NO: 9), as determined by Rabat index. In certain embodiments, the antibody construct comprises LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of MDX-l 105 (SEQ ID NO: 10), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of MDX-l 105 (SEQ ID NO:9), as determined by IMGT. In certain embodiments, the antibody construct comprises a humanized antibody or antigen binding fragment thereof comprising LC CDR1, LC CDR2 and LC CDR3 of the light chain variable region of MDX-l 105 (SEQ ID NO: 10), and HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable region of MDX-l 105 (SEQ ID NO: 9), as determined by IMGT.
[0126] An antibody construct may comprise an antibody with modifications of at least one amino acid residue. Modifications may be substitutions, additions, mutations, deletions, or the like. An antibody modification can be an insertion of an unnatural amino acid.
[0127] The exemplary antibody construct VH sequences and VL sequences are illustrated in Table A below.
Table A: Exemplary Antibody Construct VH sequences and VL sequences
Figure imgf000051_0001
Figure imgf000052_0001
Figure imgf000053_0001
Figure imgf000054_0001
Target binding domain of an Antibody Construct
[0128] An antibody construct may further comprise a target binding domain. A target binding domain may comprise a domain that binds to a target. A target may be an antigen. A target binding domain may comprise an antigen binding domain. A target binding domain may be a domain that can specifically bind to an antigen. A target binding domain may be an antigen binding portion of an antibody or an antibody fragment. A target binding domain may be one or more fragments of an antibody that can retain the ability to specifically bind to an antigen. A target binding domain may be any antigen binding fragment. A target binding domain may be in a scaffold, in which a scaffold is a supporting framework for the antigen binding domain. A target binding domain may comprise an antigen binding domain in a scaffold.
[0129] A target binding domain may comprise an antigen binding domain which can refer to a portion of an antibody comprising the antigen recognition portion, z.e., an antigenic determining variable region of an antibody sufficient to confer recognition and binding of the antigen recognition portion to a target, such as an antigen, z.e., the epitope. A target binding domain may comprise an antigen binding domain of an antibody. In certain embodiments, a target binding domain is a CD40 agonist.
[0130] An Fv can be the minimum antibody fragment which contains a complete antigen- recognition and antigen-binding site. This region may consist of a dimer of one heavy chain and one light chain variable domain in tight, non-covalent association. In this configuration, the three hypervariable regions of each variable domain may interact to define an antigen-binding site on the surface of the VH-VL dimer. A single variable domain (or half of an Fv comprising only three hypervariable regions specific for an antigen) can recognize and bind antigen, although at a lower affinity than the entire binding site.
[0131] A target binding domain may be at least 80% homologous to an antigen binding domain selected from, but not limited to, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (VH) and a light chain variable domain (VL), a single chain variable fragment (scFv), a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a cytokine, a ligand, an immunocytokine, a T cell receptor, or a recombinant T cell receptor.
[0132] A target binding domain may be attached to an antibody construct. For example, an antibody construct may be fused with a target binding domain to create an antibody construct target binding domain fusion. The antibody construct-target binding domain fusion may be the result of the nucleic acid sequence of the target binding domain being expressed in frame with the nucleic acid sequence of the antibody construct. The antibody construct- target binding domain fusion may be the result of an in-frame genetic nucleotide sequence or a contiguous peptide sequence encoding the antibody construct with the target binding domain. As another example, a target binding domain may be linked to an antibody construct. A target binding domain may be linked to an antibody construct by a chemical conjugation. A target binding domain may be attached to a terminus of an Fc region. A target binding domain may be attached to a terminus of an Fc region. A target binding domain may be attached to a terminus of an antibody construct. A target binding domain may be attached to a terminus of an antibody. A target binding domain may be attached to a light chain of an antibody. A target binding domain may be attached to a terminus of a light chain of an antibody. A target binding domain may be attached to a heavy chain of an antibody. A target binding domain may be attached to terminus of a heavy chain of an antibody. The terminus may be a C-terminus. An antibody construct may be attached to 1, 2, 3, and/or 4 target binding domains. The target binding domain may direct the antibody construct to, for example, a particular cell or cell type. A target binding domain of an antibody construct may be selected in order to recognize an antigen, e.g., an antigen expressed on an immune cell. An antigen can be a peptide or fragment thereof. An antigen may be expressed on an antigen-presenting cell. An antigen may be expressed on a dendritic cell, a macrophage, or a B cell. As another example, an antigen may be a tumor antigen. The tumor antigen may be any tumor antigen described herein. When multiple target binding domains are attached to an antibody construct, the target binding domains may bind to the same antigen. When multiple target binding domains are attached to an antibody construct, the target binding domains may bind different antigens.
[0133] In certain embodiments, an antibody construct described herein specifically binds a second antigen. In certain embodiments, the target binding domain is linked, e.g., covalently bound, to the antibody construct at a C-terminal end of the Fc domain.
Compounds
[0134] The following is a discussion of compounds and salts thereof that may be used in the methods of the disclosure. The compounds and salts described in Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC) may be covalently bound, to linkers, L3, which may further be covalently bound to antibody constructs. The compound and salts described in Formulas (IV A), (IVB), (IVC) are covalently bound to linkers, L3, which may further be covalently bound to antibody constructs.
[0135] In some aspects, the present disclosure provides a compound represented by the structure of Formula (IA):
Figure imgf000057_0001
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L1 is selected from -X1-, -X2-CI-6 alkyl ene-X2-Ci-6 alkylene-, -X2-C2-6 alkenylene- X2-, and -X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkylene, alkenylene or alkynylene with one or more R12;
L2 is selected from -X2-, -X2-CI-6 alkylene-X2-, -X2-C2-6 alkenyl ene-X2-, and -X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkylene, alkenylene or alkynylene with one or more R12;
X1 is selected from -S-*, -N(R10)-*, -C(0)0-*, -OC(O)-*, -0C(0)0-*, - C(O)N(R10)C(O)- *, -C(O)N(R10)C(O)N(R10)*, -N(R10)C(O)-*, -CR10 2N(R10)C
(O)-*, -N(R10)C(O)N(R10)-*, -N(R10)C(O)O-*, -OC(0)N(R10)-*, -C(NR10)-*, -N(R10)C(NR10)-*, -C(NR10)N(R10)-*, -N(R10)C(NR10)N(R10)-*, -S(0)2-*, -0S(0)-*, -S(0)0-*, -S(O), -0S(0)2- *, -S(0)20*, -N(R10)S(O)2-*, -S(0)2N(R10)-*, -N(R10)S(O)-*, -S(0)N(R10)-*, -N(R10)S(0)2N(R1 °)-*, and -N(R10)S(O)N(R10)-*, wherein * represents where X1 is bound to R3;
X2 is independently selected at each occurrence from a bond, -O- , -S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(O), -0S(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R3 is selected from optionally substituted C3-0 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents on R3 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), and -CN; C1-10 alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to 12- membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R3 is optionally substituted with one or more substituents independently selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10,
-OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from: hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, CI-IO alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C3-io carbocycle and 3- to lO-membered heterocycle; and C3-io carbocycle and 3- to lO-membered heterocycle, wherein each C3-io carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
[0136] In certain embodiments, for a compound or salt of Formula (IA), X2 is independently selected at each occurrence from -O-
, -S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, - C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(O), -OS(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-.
[0137] In certain embodiments, for a compound of Formula (IA):
represents an optional double bond;
L1 is independently selected from -X1-, -X2-CI-6 alkyl-X2-, -X2-C2-6 alkenyl-X2-, and
-X2-C2-6 alkynyl-X2, each of which is optionally substituted at each occurrence with one or more
R10;
L2 is independently selected from -X2-, -X2-CI-6 alkyl-X2-, -X2-C2-6 alkenyl-X2-, and
-X2-C2-6 alkynyl-X2-, each of which is optionally substituted at each occurrence with one or more
R10;
X1 at each occurrence is independently selected
from -S-*, -N(R10)-*, -C(0)0-*, -OC(O)-*, -0C(0)0-*, -C(O)N(R10)C(O)- *,
-C(O)N(R10)C(O)N(R10)-*, -N(R10)C(O)-*, -CR10 2N(R10)C
(O)-*, -N(R10)C(O)N(R10)-*, -N(R10)C(O)O-*, -OC(0)N(R10)-*, -C(NR10)-*, -N(R10)C(NR10)-*,
Figure imgf000059_0001
°)-*, and -N(R10)S(O)N(R10)-*, wherein * represents where X1 is bound to R3;
X2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10)-, -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -0S(0)-, -S(O) 0-, -S(0)-, -0S(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R3 is selected from optionally substituted C3-i2 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2,
-C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to 12- membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R3 is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10,
-C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, - OR10, -SR10,
-C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10,
-C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is independently optionally substituted with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10,
-N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from: hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to 12- membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -CN, -NO2, -NH2, =0, =S, - C(0)0CH2C6H5, -NHC(0)0CH2C6H5, Ci-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, 3- to l2-membered heterocycle, and haloalkyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)2, -S(0)R10, - S(0)2R10,
-C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -P(O)(OR10)2, -OP(O)(OR10)2, -CN, Ci-10 alkyl, C2-10 alkenyl, and C2-10 alkynyl, or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.“Benzazepine core” as used herein refers to both a benzazepine and a 4,5-dihydrobenzazepine core ring system.
[0138] In some embodiments for a compound or salt of Formula (IA), L2 can be attached at C2, C3, C4 or C5 of the benzazepine core, wherein the numbering of the benzazepine is as follows:
Figure imgf000061_0001
certain embodiments, for a compound or salt of Formula (IA), L2 is attached to the benzazepine core at C4. In certain embodiments for a compound or salt of Formula (IA), represents a double bond and L2 is attached to the benzazepine core at C4.
[0139] In some embodiments for a compound or salt of Formula (IA), L1 can be attached at C6, C7, C8 or C9. In certain embodiments, for a compound or salt of Formula (IA), L1 is attached to the benzazepine core at C8. In certain embodiments for a compound or salt of Formula (IA), represents a double bond, L2 is attached to the benzazepine core at C4 and L1 is attached to the benzazepine core at C8.
[0140] In some embodiments for a compound or salt of Formula (IA), the substitutable carbon on the benzazepine core is selected from C2, C3, C4, C5, C6, C7, C8, and C9. The benzazepine core for a compound or salt of Formula (IA), can be optionally substituted by a substituent selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)2, -S(0)R10, -S(0)2R10 - C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -P(O)(OR10)2, -OP(O)(OR10)2, -CN, Ci-10 alkyl, C2-10 alkenyl, and C2-10 alkynyl, or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle. In some embodiments for a compound or salt of Formula (IA), a moiety at any one of C2, C3, C4, C5, C6, C7, C8, and C9 of the benzazepine core is independently selected from hydrogen, halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10 -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl.
[0141] In one embodiment, the compound of Formula (IA) is represented by Formula (IB):
Figure imgf000062_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -
OR10, -SR10,
-N(R10)2, -S(0)R10, -S(0)2R10 -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; and
R24 and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, -
N(R10)2,
-S(0)R10, -S(0)2R10 -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
[0142] In certain embodiments, the compound of Formula (IA) is represented by Formula (IC):
Figure imgf000062_0002
or a pharmaceutically acceptable salt thereof.
[0143] In some embodiments, for a compound or salt of Formula (IA), (IB) or (IC), L1 is selected from
-N(R10)C(O)-*, -S(0)2N(R10)-*, CR10 2N(R10)C (O)-* and -X2-Ci.6 alkylene-X2-Ci-6 alkylene wherein each Ci-6 alkylene is optionally substituted with one or more R12. In some
embodiments, L1 is selected from -N(R10)C(O)-*. In some embodiments, R10 of -N(R10)C(O)-* is selected from hydrogen and Ci-6 alkyl. In some embodiments for a compound or salt of Formula (IA), (IB) or (IC), L1 is -NHC(O)-*. [0144] In some embodiments, for a compound or salt of Formula (IA), (IB) or (IC), L1 is selected from
-S(0)2N(R10)-*. In some embodiments, R10 of -S(0)2N(R10)-* is selected from hydrogen and Ci-6 alkyl. In some embodiments, L1 is selected from -S(0)2NH-*.
[0145] In some embodiments, for a compound or salt of Formula (IA), (IB) or (IC), L1 is selected from -CR102N(R10)C(O)-*. In certain embodiments, L1 is selected
from -CH2N(H)C(0)-* and -CH(CFF)N(H)C(0)-*.
[0146] In some embodiments, for a compound or salt of Formula (IA), (IB) or (IC), R3 is selected from optionally substituted C3-12 carbocycle, and optionally substituted 3- to 12- membered heterocycle, wherein substituents on R3 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(Ri0)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2- 10 alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, C3-12 carbocycle, and 3- to l2-membered heterocycle; and C3-12 carbocycle, and 3- to 12- membered heterocycle, wherein each C3-12 carbocycle, and 3- to l2-membered heterocycle in R3 is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
[0147] R3 may be selected from optionally substituted C3-12 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents on R3 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0,
=S, =N(R10), -CN, C3 -i2 carbocycle, and 3- to l2-membered heterocycle, and C3-12 carbocycle, and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. [0148] In certain embodiments, R3 is selected from optionally substituted C3-i2 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents on R3 are
independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0,
=S, =N(R10), and -CN; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle.
[0149] In some embodiments, for a compound or salt of Formula (IA), (IB) or (IC), R3 is selected from an optionally substituted aryl and an optionally substituted heteroaryl, such as R3 is an optionally substituted heteroaryl. In some embodiments, R3 is an optionally substituted heteroaryl substituted with one or more substituents independently selected from halogen, -
OR10, -SR10, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. R3 may be selected from an optionally substituted 5- or 6-membered
heteroaryl. For example, R3 may be selected from
Figure imgf000064_0001
Figure imgf000064_0002
any one of which is optionally substituted. In some embodiments, R3 is selected from an optionally substituted 6- membered heteroaryl, e.g., R3 is optionally substituted pyridine.
[0150] In some embodiments for a compound or salt of Formula (IA), (IB) or (IC), R3 is an optionally substituted aryl, such as R3 is an aryl optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. R3 may be selected from an optionally substituted phenyl and an optionally substituted naphthyl. In certain embodiments, R3 is an optionally substituted phenyl.
[0151] In some embodiments, for a compound or salt of Formula (IA), (IB) or (IC), R3 is selected from pyridine, phenyl, tetrahydronaphthalene, tetrahydroquinoline,
tetrahydroisoquinoline, indane, cyclopropylbenzene, cyclopentapyridine, and
dihydrobenzoxaborole, any one of which is optionally substituted. R3 may be selected from:
Figure imgf000064_0003
Figure imgf000065_0001
Figure imgf000065_0002
any one of which is optionally substituted. R3 may be selected from:
Figure imgf000065_0003
Figure imgf000066_0001
[0152] In some embodiments, for a compound of Formula (IA), (IB), or (IC) or a
pharmaceutically acceptable salt thereof:
L1 is selected from -N(R10)C(O)-*, -S(0)2N(R10)-*, -CR10 2N(R10)C(O)-*and -X2-CI-6 alkyl ene-X2-C i-6 alkylene-;
L2 is -C(O)-;
X2 at each occurrence is independently selected from -O- , -S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10)-, -N(R10)C(O)-, -N(R10)C(O)N(R10)-, and -N(R10)C(O)O-;
R1 is hydrogen;
R2 is an amine masking group;
R3 is selected from an optionally substituted phenyl, heteroaryl and 9- or lO-membered bicyclic carbocycle wherein substituents are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, - SR10, -N(R10)2, -C(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci -lo alkyl optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle;
R10 is independently selected at each occurrence from: hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -CN, -N02, -NH2, =0, =S, - C(0)0CH2C6H5, -NHC(0)0CH2C6H5, Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, 3- to l2-membered heterocycle, and haloalkyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted with one or more R12, wherein R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -NO2, =0, =S, =N(R10), and -CN; and C1-10 alkyl optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -NO2, =0, =s, =N(R10), -CN, C3 -lo carbocycle and 3- to lO-membered heterocycle.
[0153] In certain embodiments for a compound or salt of Formula (IA), (IB) or (IC), the compound may further comprise a linker (L3). The linker may be covalently bound to any position, valence permitting, on a compound or salt of Formula (IA), (IB) or (IC). For example, the linker may be bound to R4, R3, X1 or X2. In certain embodiments, the linker is bound to a nitrogen or oxygen atom of a compound or salt of Formula (IA), (IB) or (IC). The linker may comprise a reactive moiety, e.g., an electrophile that can react to form a covalent bond with a moiety of an antibody, e.g., a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue of an antibody. In some embodiments, a compound or salt of Formula (IA), (IB) or (IC), may be covalently bound through the linker to an antibody.
[0154] In some aspects, the present disclosure provides a compound represented by the structure of Formula (II A):
Figure imgf000067_0001
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L10 is -X10-;
L2 is selected from -X2-, -X2-CI-6 alkylene-X2-, -X2-C2-6 alkenyl ene-X2-, and - X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R12;
X10 is selected from -C(O)-, and -C(0)N(R10)-*, wherein * represents where X10 is bound to R5; X2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R10)-, -C(O)-,
-C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)
-C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(O), -0S(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-,
and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R5 is selected from unsaturated C4-8 carbocycle; bicyclic carbocycle; and fused 5- 5, fused 5-6, and fused 6-6 bicyclic heterocycle, wherein R5 is optionally substituted and wherein substituents are independently selected at each occurrence from: halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R5 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; R10 is independently selected at each occurrence from hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, C 1-10 alkyl, -C1-10 haloalkyl, -O-Ci-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
[0155] In certain embodiments, for a compound of Formula (IIA):
represents an optional double bond;
L10 is independently selected from -X10-;
L2 is independently selected from -X2-, -X2-CI-6 alkyl-X2-, -X2-C2-6 alkenyl-X2-, and -X2-C2-6 alkynyl-X2-, each of which is optionally substituted at each occurrence with one or more R10;
X10 at each occurrence is independently selected from -C(O)-, and -C(0)N(R10)-*, wherein * represents where X10 is bound to R5;
X2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R10)-, -C(O)-,
-C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10) -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -0S(0)-, -S(0)0-, -S(O), -0S(0)2- , -S(0)20,
-N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-,
and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -
N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10,
-N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R5 is selected from optionally substituted saturated and unsaturated C3-8 carbocycle, optionally substituted saturated and unsaturated 3- to 8-membered heterocycle, optionally substituted bicyclic carbocycle, and optionally substituted bicyclic heterocycle wherein substituents are independently selected at each occurrence from: halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), and -CN; C1-10 alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10,
-SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2,
-N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-0 carbocycle, and 3- to l2-membered heterocycle in R5 is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10,
-N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; R10 is independently selected at each occurrence from hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to 12- membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -CN, -NO2, -NH2, =0, =S, - C(0)0CH2C6H5, -NHC(0)0CH2C6H5, Ci-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, 3- to l2-membered heterocycle, and haloalkyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)2, -S(0)R10, - S(0)2R10,
-C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -P(O)(OR10)2, -OP(O)(OR10)2, -CN, Ci-10 alkyl, C2-10 alkenyl, and C2-10 alkynyl, or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
[0156] In some embodiments for a compound or salt of Formula (IIA), L2 can be attached at C2, C3, C4, or C5 of the benzazepine core, wherein the numbering of the benzazepine is as follows:
Figure imgf000071_0001
certain embodiments, L2 is attached to the benzazepine core at C4. In certain embodiments represents a double bond and L2 is attached to the benzazepine core at C4.
[0157] In some embodiments, for a compound or salt of Formula (IIA), L10 can be attached at C6, C7, C8 or C9. In certain embodiments, L10 is attached to the benzazepine core at C8. In certain embodiments represents a double bond, L2 is attached to the benzazepine core at
C4 and L10 is attached to the benzazepine core at C8.
[0158] In some embodiments, for a compound or salt of Formula (IIA), the substitutable carbon on the benzazepine core is selected from C2, C3, C4, C5, C6, C7, C8, and C9. The benzazepine core for a compound or salt of Formula (IIA), can be optionally substituted by a substituent selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)2, -S(0)R10, -S(0)2R10 - C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -P(O)(OR10)2, -OP(O)(OR10)2, -CN, Ci-10 alkyl, C2-10 alkenyl, and C2-10 alkynyl, or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle. In some embodiments for a compound or salt of Formula (IIA), a moiety at any one of C2, C3, C4, C5, C6, C7, C8, and C9 of the benzazepine core is independently selected from hydrogen, halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, - S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-10 alkenyl, and C2-10 alkynyl. In one embodiment, the compound of Formula (IIA) is represented by Formula (IIB):
Figure imgf000072_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -
Figure imgf000072_0002
N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =s,
=N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; and
R24, and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, -
N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN,
Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
[0159] In one embodiment, the compound of Formula (IIA) is represented by Formula (IIC):
Figure imgf000072_0003
or a pharmaceutically acceptable salt thereof.
[0160] In certain embodiments, for a compound or salt of Formula (IIA), (IIB), or (IIC), L10 is -C(O)-. In certain embodiments, L10 is selected from -C(0)N(R10)-*. In certain embodiments, R10 of -C(0)N(R10)-* is selected from hydrogen and Ci-6 alkyl. In certain embodiments, L10 is -C(0)NH-*.
[0161] In certain embodiments, for a compound or salt of Formula (IIA), (IIB), or (IIC), R5 is an optionally substituted 8- to l2-membered bicyclic carbocycle. In some embodiments, R5 is an optionally substituted bicyclic carbocycle, e.g., an optionally substituted 8- to 12- membered bicyclic carbocycle, such as indane, naphthalene, dihydronaphthalene, tetrahydronaphthalene. In some embodiments, R5 is an optionally substituted 8- to 12- membered bicyclic carbocycle with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -
C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. In some embodiments, R5 is an optionally substituted 8- to 12- membered bicyclic carbocycle with one or more substituents independently selected from -OR10, -N(R10)2, and =0. In some embodiments, R5 is an optionally substituted indane or an optionally substituted
tetrahydronaphthalene. In some embodiments, R5 is selected from:
Figure imgf000073_0001
and
Figure imgf000073_0002
, either of which is optionally substituted. In some embodiments for a compound or salt of Formula (IIA), (IIB) or (IIC), R5 is selected from:
Figure imgf000073_0003
[0162] In some embodiments, for a compound or salt of Formula (IIA), (IIB) or (IIC), R5 is an optionally substituted unsaturated C4-8 carbocycle. In some embodiments, R5 is an optionally substituted unsaturated C4-6 carbocycle, e.g., cyclopentene, cyclopentadiene, cyclohexene, cyclobutene. In some embodiments, R5 is an optionally substituted unsaturated C4-6 carbocycle with one or more substituents independently selected from optionally substituted C3-12 carbocycle, and optionally substituted 3- to l2-membered heterocycle. In some embodiments, R5 is an optionally substituted unsaturated C4-6 carbocycle with one or more substituents
independently selected from optionally substituted phenyl, optionally substituted 3- to 12- heterocycle, optionally substituted Ci-io alkyl, optionally substituted C2-10 alkenyl, and halogen.
[0163] In certain embodiments, for a compound or salt of Formula (IIA), (IIB) or (IIC), R5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle. In some embodiments, R5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle with one or more substituents independently selected from -C(0)OR10, -N(R10)2, - OR10, and optionally substituted Ci-io alkyl. In some embodiments, R5 is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle substituted with -C(0)OR10. In some embodiments, R5 is an optionally substituted fused 6-6 bicyclic heterocycle. The fused 6-6 bicyclic heterocycle may be an optionally substituted pyridine-piperidine. In some embodiments, L10 is bound to a carbon atom of the pyridine of the fused pyridine-piperidine. In some embodiments, R5 is selected from tetrahydroquinoline, tetrahydroisoquinoline,
tetrahydronaphthyridine, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is optionally substituted. In some embodiments, R5 is an optionally substituted
tetrahydronaphthyridine. In some embodiments for a compound or salt of Formula (IIA), (IIB) or (IIC), R5 is selected from:
Figure imgf000074_0001
[0164] In some embodiments, for a compound or salt of Formula (IIA), (IIB) or (IIC), R5 is selected from tetrahydronaphthalene, tetrahydroquinoline, tetrahydroisoquinoline, indane, cyclopropylbenzene, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is optionally substituted. In some embodiments for a compound or salt of Formula (IIA), (IIB) or
Figure imgf000074_0002
Figure imgf000075_0001
any one of which is optionally substituted. In some embodiments for a compound or salt of Formula (IIA), (IIB) or (IIC), R5 is selected from:
Figure imgf000075_0002
[0165] In some embodiments, for a compound or salt of Formula (IIA), (IIB) or (IIC), when R5 is substituted, substituents on R5 are independently selected at each occurrence from: halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2 io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
[0166] In some embodiments, for a compound or salt of Formula (IIA), (IIB) or (IIC), when R5 is substituted, substituents on R5 are independently selected at each occurrence from: halogen, - OR10, -SR10, -C(O)N(R10)2, -N(RIO)C(0)RIO, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2- io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle.
[0167] In some embodiments, for a compound or salt of Formula (IIA), (IIB) or (IIC), when R5 is substituted, substituents on R5 are independently selected at each occurrence from: halogen, - OR10, -SR10, -C(O)N(R10)2, -N(RIO)C(0)RIO, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2- io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N02, =0, =S, =N(R10), and -CN.In certain embodiments, substituents on R5 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, and -CN; and Ci -lo alkyl optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(0)OR10, -N02, =0, and -CN. In certain embodiments, R5 is not substituted.
[0168] In some embodiments for a compound or salt of Formula (IIA), (IIB) or (IIC), R5 is selected from saturated and unsaturated C3-8 carbocycle, bicyclic carbocycle, and bicyclic heterocycle, any of which is optionally substituted and wherein substituents are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(Rio)C(0)Rio, - N(RIO)C(0)N(RIO)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; C1-10 alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-0 carbocycle, and 3- to l2-membered heterocycle in R5 is independently optionally substituted with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
[0169] In some embodiments, for a compound or salt of Formula (IIA), (IIB) or (IIC), R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -OH, -OR10, -N02, -CN, and Ci-10 alkyl. In some embodiments, R20, R21, R22, and R23 are each hydrogen. In some
embodiments, R21 is halogen, e.g. fluorine. In an exemplary embodiment, R21 is hydrogen. In other embodiments, R21 is -OR10. For example, R21 may be -OCH3.
[0170] In some embodiments, for a compound of Formula (IIA), (IIB) or (IIC), or a
pharmaceutically acceptable salt thereof: L10 is selected from -C(0)N(R10)-* and -C(O)-;
L2 is -C(O)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; and Ci -lo alkyl optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle;
R5 is selected from optionally substituted unsaturated C4-8 carbocycle, optionally substituted bicyclic carbocycle, and optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle wherein substituents on R5 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -
C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2- io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2,
-N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R5 is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10,
-N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl optionally substituted with one or more substituents independently selected from halogen, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, -NHC(0)0CH2C6H5, Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12.
[0171] In some embodiments, for a compound of Formula (IIA), (IIB) or (IIC), or a
pharmaceutically acceptable salt thereof:
L10 is selected from -C(0)N(R10)-* and -C(O)-;
L2 is -C(O)-;
R1 is hydrogen; R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10;
R5 is selected from optionally substituted optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle wherein substituents are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2,
-N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10, -N(R10)C(O)N(R10)2,
-N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R5 is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10,
-N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, -NHC(0)0CH2C6H5, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12.
[0172] In some embodiments, for a compound of Formula (IIB) or (IIC), or a pharmaceutically acceptable salt thereof:
L10 is selected from -C(0)N(R10)-*, such as -C(0)NH-*;
L2 is -C(O)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10, such as R4 is -N(R10)2;
R5 is selected from optionally substituted optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle, such as R5 is selected from a tetrahydroquinoline,
tetrahydroisoquinaline, dihydroindene, wherein substituents on R5 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -
N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), and -CN; and Ci-io alkyl optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle;
R10 is independently selected at each occurrence from hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, -NHC(0)0CH2C6H5, C3-i2 carbocycle, and 3- to l2-membered heterocycle;
R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -OR10, -SR10, - N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl, such as R20, R21, R22, and R23 are each hydrogen; and
R24, and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, -N(R10)2, - S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl, such as R24 and R25 are each hydrogen.
[0173] In certain embodiments for a compound or salt of Formula (IIA), (IIB) or (IIC), the compound may further comprise a linker (L3). The linker may be covalently bound to any position, valence permitting, on a compound or salt of Formula (IIA), (IIB) or (IIC). For example, the linker may be bound to R4, R5, X10 or X2. In certain embodiments, the linker may be bound to a nitrogen atom, or an oxygen atom of a compound or salt of Formula (IIA), (IIB) or (IIC). The linker may comprise a reactive moiety, e.g., an electrophile that can react to form a covalent bond with a moiety of an antibody, e.g., lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue of the antibody. In some embodiments, a compound or salt of Formula (IIA), (IIB) or (IIC), may be covalently bound through the linker to an antibody construct.
[0174] In some aspects, the present disclosure provides a compound represented by the structure of F ormul a (HI A) :
Figure imgf000079_0001
or a pharmaceutically acceptable salt thereof, wherein: represents an optional double bond;
L11 is -X11-;
L2 is selected from -X21-, -X21-CI-6 alkylene-X21-, -X21-C2-6 alkenylene-X21-, and - X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R12;
X11 is selected from -C(O)- and -C(0)N(R10)-*, wherein * represents where X11 is bound to R6;
X21 at each occurrence is independently selected from a bond, -O- , -S-, -N(R10)-, -C(O)-,
-C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-,
-C(O)N(R10)C(O)N(R10)-, -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(O)-, -0S(0)2-, -S(0)20- , -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-,
-N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10,
-SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R7 and R6 is further optionally substituted by one or more additional substituents independently selected from R12;
R7 is selected from -C(0)NHNH2, -C(0)NH-Ci-3 alkylene-NH(R10), -C(0)CH3, - Ci-3 alkyl ene-NHC(0)0R11 , -Ci-3alkylene-NHC(0)R10, -Ci-3alkylene-NHC(0)NHR10, -Ci- 3alkylene-NHC(0)-Ci-3alkylene-R10, and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R12; R10 is independently selected at each occurrence from hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, -Ci-10 alkyl, -Ci-io haloalkyl, -O-Ci-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to l2-membered heterocycle;
R11 is selected from C3-12 carbocycle and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from R12;
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
[0175] In certain embodiments, for a compound or salt of Formula (IIIA):
represents an optional double bond;
L11 is independently selected from -X11-;
L2 is independently selected from -X2-, -X2-CI-6 alkylene-X2-, -X2-Ci-(, alkenyl ene-X2-, and -X2-Ci-(, alkynylene-X2-, each of which is optionally substituted at each occurrence with one or more R10;
X11 at each occurrence is independently selected from -C(O)-, and -C(0)N(R10)-*, wherein * represents where X11 is bound to R6;
X2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R10)-, -C(O)-, -C(0)0-, -0C(0)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, - (O)N(R10)C(O)N(R10), -N(R10)C(O)-,
-N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-,
-C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -0S(0)-, -S(0)0-, -S(O)-, -0S(0)2- , -S(0)20,
-N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-,
and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, - OR10,
-SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2,
-N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10,
-N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R7;
R7 is selected from -C(0)NHNH2, -C(0)NH-Ci-3alkylene-NH(R10), -C(0)CH3, - Ci-3alkylene-NHC(0)OR10, -Ci-3alkylene-NHC(0)R10, -Ci-3alkylene-NHC(0)NHR10, and -Ci- 3alkylene-NHC(0) -Ci-3alkylene-(R10)2; and an optionally substituted 3- to l2-membered heterocycle;
R10 is independently selected at each occurrence from hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -CN, -N02, -NH2, =0, =S, - C(0)0CH2C6H5, -NHC(0)0CH2C6H5, CI-IO alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl; and wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)2, -S(0)R10, - S(0)2R10,
-C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -P(O)(OR10)2, -OP(O)(OR10)2, -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl, or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
[0176] In some embodiments for a compound or salt of Formula (IIIA), L2 can be attached at C2, C3, C4, or C5 of the benzazepine core, wherein the numbering of the benzazepine is as follows:
Figure imgf000083_0001
certain embodiments, for a compound or salt of Formula (IIIA), L2 is attached to the benzazepine core at C4. In certain embodiments for a compound or salt of Formula (IIIA), represents a double bond and L2 is attached to the benzazepine core at C4.
[0177] In some embodiments for a compound or salt of Formula (IIIA), L11 can be attached at C6, C7, C8 or C9. In certain embodiments, for a compound or salt of Formula (IIIA), L11 is attached to the benzazepine core at C8. In certain embodiments for a compound or salt of Formula (IIIA), represents a double bond, L2 is attached to the benzazepine core at C4 and L11 is attached to the benzazepine core at C8.
[0178] In some embodiments for a compound or salt of Formula (IIIA), the substitutable carbon on the benzazepine core is selected from C2, C3, C4, C5, C6, C7, C8, and C9. The benzazepine core for a compound or salt of Formula (IIIA), can be optionally substituted by a substituent selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)2, -S(0)R10, -S(0)2R10 - C(0)R10, -C(0)OR10, -OC(0)R10,
-N02, =0, =S, =N(R10), -P(O)(OR10)2, -OP(O)(OR10)2, -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl, or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle. In some embodiments for a compound or salt of Formula (IIIA), a moiety at any one of C2, C3, C4, C5, C6, C7, C8, and C9 of the benzazepine core is independently selected from hydrogen, halogen, -OR10, -SR10,
-N(R10)2, -S(0)R10, -S(0)2R10 -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl. [0179] In some embodiments, the compound of Formula (IIIA) is represented by Formula (IIIB):
Figure imgf000084_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen,
-OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10 -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; and
R24, and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, - N(R10)2, -S(0)R10, -S(0)2R10 -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
[0180] In one embodiment, the compound of Formula (IIIA) is represented by Formula (IIIC):
Figure imgf000084_0002
or a pharmaceutically acceptable salt thereof.
[0181] In some embodiments, for a compound or salt of Formula (IIIA), (IIIB) or (IIIC), L11 is selected from -C(O)-, and -C(0)N(R10)-*, wherein * represents where L11 is bound to R6. In certain embodiments, L11 is -C(O)-. In certain embodiments, L11 is selected from -C(0)N(R10)-*. In certain embodiments, R10 or L11 is selected from hydrogen and Ci-6 alkyl.
[0182] In certain embodiments, for a compound or salt of Formula (IIIA), (IIIB) or (IIIC), R6 is further optionally substituted by one or more additional substituents independently selected from R12.
In some embodiments, for a compound or salt of Formula (IIIA), (IIIB) or (IIIC), R7 is selected from -C(O)N(R10)N(R10)2, -C(O)N(R10)-CI-3 alkylene-N(R10)2, -C(0)CH3, -Ci-3 alkylene- N(R10)C(O)ORu, -Ci-3alkylene-N(R10)C(O)R10, -Ci-3alkylene-N(R10)C(O)N(R10)2, -Ci- 3alkylene-N(R10)C(O)-Ci-3alkylene-R10, and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R12. R11 may be selected from C3-12 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from R12. In certain embodiments, for a compound or salt of Formula (IIIA), (IIIB) or (IIIC), R7 is selected from -C(0)NHNH2, - C(0)NH-CI-3 alkylene-NH(R10), -C(0)CH3, -C1.3 alkylene-NHC
NHC(0)R10, -Ci-3alkylene-NHC(0)NHR10, -Ci-3alkylene-NHC(
Figure imgf000085_0001
l2-membered heterocycle optionally substituted with one or more substituents independently selected from R12. In certain embodiments, R7 is selected from -C(0)NHNH2, -C(0)NH-CI-3 alkyl ene-NH(R10), -C(0)CH3, -C1.3 alkylene-NHC(0)ORu, -Ci-3alkylene-NHC(0)NHR10, -Ci- 3alkylene-NHC(0)-Ci-3alkylene-R10, and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R12. In certain embodiment, R7 is a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R12.
[0183] In some embodiments, for a compound or salt of Formula (IIIA), (IIIB) or (IIIC), R6 is selected from an optionally substituted phenyl and an optionally substituted 5-6 membered heteroaryl, wherein R6 is substituted with one or more substituents independently selected from R7. In some embodiments, R6 is selected from an optionally substituted phenyl, wherein the phenyl is substituted with one or more substituents independently selected from R7. In some embodiments, R6 is selected from an optionally substituted phenyl, wherein the phenyl is substituted with one or more substituents independently selected from -C(0)NHNH2, -C(0)NH- Ci.3alkylene-NH(R10), and -C(0)CH3; and 3- to l2-membered heterocycle, which is optionally substituted with one or more substituents selected from -OH, -N(R10)2, -NHC(0)(R10), - NHC(0)0(R10), -NHC(O)N(R10)2, -C(O)R10,-C(O)2R10, and -Ci-3alkylene-(R10), and optionally further substituted with one or more additional substituents. In some embodiments, for a compound or salt of Formula (IIIA), (IIIB) or (IIIC), R6 is selected from:
Figure imgf000086_0001
[0184] In some embodiments for a compound or salt of Formula (IIIA), (IIIB) or (IIIC), R6 is selected from an optionally substituted 5-6 membered heteroaryl, wherein the heteroaryl is substituted with one or more substituents independently selected from R7. R6 may be selected from a 5- and 6-membered heteroaryl substituted with one or more substituents independently selected from R7, and R6 may be further optionally substituted with one or more additional substituents selected from R12. In some embodiments, R6 is selected from an optionally substituted 5-6 membered heteroaryl, wherein the heteroaryl is substituted with one or more substituents independently selected from
-C(0)CH3, -Ci-3alkylene-NHC(0)OR10, -Ci-3alkylene-NHC(0)R10, -Ci-salkylene- NHC(0)NHR10, and
-Ci-3alkylene-NHC(0) -Ci-3alkylene-(R10); and 3- to l2-membered heterocycle, which is optionally substituted with one or more substituents selected from -OH, -N(R10)2, -
NHC(0)(R10), -NHC(0)0(R10), -NHC(O)N(R10)2, -C(0)R10, -C(O)N(R10)2, -C(0)2R10, and -Ci- 3alkylene-(R10), and R6 is optionally further substituted with one or more additional substituents independently selected from R12. In some embodiments, R6 is selected from substituted pyridine, pyrazine, pyrimidine, pyridazine, furan, pyran, oxazole, thiazole, imidazole, pyrazole, oxadiazole, oxathiazole, and triazole, and R6 is optionally further substituted with one or more additional substituents independently selected from R12. R6 may be an optionally substituted pyridine and R6 is optionally further substituted with one or more additional substituents independently selected from R12. In certain embodiments, R6 may be represented as follows:
Figure imgf000086_0002
some embodiments, R is substituted pyridine, and wherein R7 is -Ci-3alkylene-NHC(0)-Ci-3alkylene-R1°. R7 may be -Cialkylene-NHC(0)-Cialkylene-R10. R7 may be -Cialkylene-NHC(0)-Cialkylene-NH2 . In some embodiments for a compound or salt
of Formula (IIIA), (IIIB) or (IIIC), R6 is selected from:
Figure imgf000087_0001
Figure imgf000087_0002
,
[0185] In certain embodiments for a compound or salt of Formula (IIIA), (IIIB) or (IIIC), the compound may further comprise a linker (L3). The linker may be covalently bound to any position, valence permitting, on a compound or salt of Formula (IIIA), (IIIB) or (IIIC). For example, the linker may be bound to R4, R6, X11 or X2. In certain embodiments, the linker is bound to a nitrogen or oxygen atom of a compound or salt of Formula (IIIA), (IIIB) or (IIIC).
The linker may comprise a reactive moiety, e.g., an electrophile, that can react to form a covalent bond with a moiety of an antibody construct, a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue of an antibody. In some embodiments, a compound or salt of Formula (IIIA), (IIIB) or (IIIC), may be covalently bound through the linker moiety to an antibody construct.
[0186] In certain embodiments, for a compound or salt of Formula (IIIA), (IIIB) or (IIIC):
L11 is -C(0)N(R10)- *, such as L11 is -C(0)NH-;
L2 is selected from -C(O)- and -C(0)NR10-;
R1 is hydrogen; R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; and Ci -lo alkyl optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)2, -C(0)R10, -N02, =0, =S,
=N(R10), -CN, C3 -i2 carbocycle, and 3- to l2-membered heterocycle;
R6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R7;
R7 is selected from -C(0)NHNH2, -C(0)NH-Ci-3alkylene-NH(R10), -C(0)CH3, -Ci- 3alkylene-NHC(0)ORu, -Ci-3alkylene-NHC(0)R10, -Ci-3alkylene-NHC(0)NHR10, and -Ci- 3alkylene-NHC(0) -Ci-3alkylene-(R10)2; and a 3 - to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R12;
R10 is independently selected at each occurrence from hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, CI-IO alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
R11 is selected from C3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from R12; and wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12.
[0187] In certain embodiments, for a compound or salt of Formula (IIIA), (IIIB) or (IIIC):
L11 is -C(0)N(R10)- *;
L2 is -C(O)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10;
R6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R7;
R7 is selected from -C(0)NHNH2, -C(0)NH-Ci-3alkylene-NH(R10), -C(0)CH3, -Ci- 3alkylene-NHC(0)ORu, -Ci-3alkylene-NHC(0)R10, -Ci-3alkylene-NHC(0)NHR10, and -Ci- 3alkylene-NHC(0) -Ci-3alkylene-(R10)2; R10 is independently selected at each occurrence from hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, CI-IO alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
R11 is selected from C3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from R12; and wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12.
[0188] In certain embodiments, for a compound or salt of Formula (IIIA), (IIIB) or (IIIC):
L11 is -C(0)N(R10)- *, such as L11 is -C(0)NH-;
L2 is -C(O)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10;
R6 is selected from phenyl and 5- or 6- membered heteroaryl, e.g., pyridine, any one of which is substituted with one or more substituents selected from R7;
R7 is selected from -C(0)NHNH2, -C(0)NH-Ci-3alkylene-NH(R10), -C(0)CH3, -Ci- 3alkylene-NHC(0)ORu, -Ci-3alkylene-NHC(0)NHR10, and -Ci-3alkylene-NHC(0) -Ci- 3alkylene-(R10)2; and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R12;
R10 is independently selected at each occurrence from hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, CI-IO alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
R11 is selected from C3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from R12; and wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12. [0189] In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (PIB), and (IIIC), R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -OH, -N02, -CN, and Ci-io alkyl. In some embodiments, R20, R21, R22, and R23 are each hydrogen.
[0190] In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R24 and R25 are independently selected from hydrogen, halogen, -OH,
-N02, -CN, and Ci-io alkyl, or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC),
(IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R24 and R25 are each hydrogen. In some
embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R24 and R25 taken together form an optionally substituted saturated C3-5 carbocycle, wherein substituents may be selected from halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0,
=S, =N(R10), and -CN; and Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is
independently optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -
N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-0 carbocycle, and 3- to l2-membered heterocycle.
[0191] In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA),
(IIB), (IIC), (IIIA), (IIIB), and (IIIC), R2 is an amine masking group selected from an acid-labile moiety or an enzymatically-labile moiety. In certain embodiments, R2 is selected from a moiety covalently bound to an amine that is selectively cleaved under intracellular conditions.
[0192] In certain embodiments, R2 together with the nitrogen to which it is attached forms a carbamate or an amide. In certain embodiments, R2 is represented by the formula:
Figure imgf000090_0001
wherein:
R101 is selected from an amino acid, a peptide, -0-(Ci-C6 alkyl) and -Ci-C6 alkyl, wherein alkyl of -0-(Ci-C6 alkyl) and -Ci-C6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N02, -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle and R10 is as previously defined; and
R100 is C(=0), wherein when R101 is selected from an amino acid or peptide R100 is the C- terminus of the amino acid or peptide. [0193] In certain embodiments, R101 is selected from -0-(Ci-C4 alkyl) and -Ci-C4 alkyl, wherein alkyl of -0-(Ci-C4 alkyl) and -Ci-C4 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N02, - CN, C3-13 carbocycle, and 3- to l2-membered heterocycle. In certain embodiments, R2 is selected from 9-fluorenylmethylcarbonyl-, /c/V-butoxy carbonyl-, benzyloxy carbonyl-, acetyl-, and trifluoroacetyl-.
[0194] In certain embodiments, the amino acid of R101 is selected from any natural or non natural amino acid. The amino acid may be selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. The amino acid may be selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In certain
embodiments, the amino acid is an L-amino acid.
[0195] In certain embodiments, the peptide of R101 includes amino acids each independently selected from any natural or non-natural amino acid. The first amino acid (including R100) may each be independently selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In certain
embodiments, the amino acids are each independently L-amino acids or D-amino acids. In certain embodiments, the peptide is a dipeptide, tripeptide or tetrapeptide. In certain
embodiments, each amino acid of a dipeptide, tripeptide or tetrapeptide, is independently selected from a D- and L-amino acid. In certain embodiments, the amino acid immediately attached to the amine is an L-amino acid, e.g., R101 is represented by the formula: -aal-aa2, or - aal-aa2-aa3, where aal is an L-amino acid and aa2 and aa3 are independently selected from D- and L-amino acids. In certain embodiments, the first amino acid (including R100) is an L-amino acid selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan and the remaining amino acids are D or L amino acids selected from arginine, histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. [0196] In certain embodiments, an amine masking group is selected from amine protecting groups described in Greene’s Protective Groups in Organic Synthesis (P. G. M. Wuts, John Wiley & Sons, Inc., NY, 2014).
In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (PIB), and (IIIC), L2 is independently selected from -X2-, -X2-CI-6
alkyl-X2-, -X2-C2-6 alkenyl-X2-, and -X2-C2-6 alkynyl-X2-, each of which is optionally substituted at each occurrence with one or more R10. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), L2 is selected from -C(O)- , and -C(0)NR10-. In some embodiments for a compound of any one of Formulas (IA), (IB),
(IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), L2 is selected from -C(O)-. In some
embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), L2 is selected from -C(0)NR10-. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R10 of -C(0)NR10- is selected from hydrogen and Ci-6 alkyl. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), L2 is -C(0)NH-.
[0197] In some embodiments, for a compound of any one of Formulas (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), X2 at each occurrence is independently selected from a bond, -0-,
-S-, -N(R10)-, -C(O)-, -C(0)0-, -0C(0)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-,
-C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -0S(0)-, -S(0)0-, -S(O), -OS(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-. In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), X2 at each occurrence is independently selected from a -0-,
-S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-,
-C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(O), -OS(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-.In certain embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), X2 is selected from -C(0)-, and
-C(0)NR10-. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), X2 is selected from -C(O)-. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), X2 is selected from -C(0)NR10-. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC),
(IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R10 of -C(0)NR10- is selected from hydrogen and Ci-6 alkyl. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA),
(IIB), (IIC), (IIIA), (IIIB), and (IIIC), X2 is -C(0)NH-.
[0198] In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, - C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
[0199] In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, - C(0)R10,
-C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2,
-N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle and 3- to l2-membered heterocycle each of which is optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -C(0)N(R10)-, -C(O)N(R10)2, -N(R10)C(O)R10,
-N(RIO)C(0)N(RIO)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
[0200] In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R4 is selected from: -OR10, and -N(R10)2; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10 -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci -lo alkyl, C2-io alkenyl, and C2-io alkynyl. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), and (IIIC), R4 is - N(R10)2.
[0201] In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R10 is independently selected at each occurrence from hydrogen, -MB, -C(0)0CH2C6H5; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to l2-membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OH, -CN, -NO2, -MB, =0, =S, -C(0)0CH2C6H5, -MIC(0)0CH2C6H5, CI-IO alkyl, -Ci-io haloalkyl, -O-Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, 3- to l2-membered heterocycle, and haloalkyl.
[0202] In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R4 is -N(R10)2, and each R10 is independently selected from optionally substituted Ci-6 alkyl. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), and (IIIC), R4 is -N(R10)2, and each R10 is independently selected from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted. In some embodiments for a compound of any one of Formulas (IA), (IB), (IC), (IIA),
Figure imgf000094_0001
[0203] In some embodiments, for a compound or salt of Formula (IA), (IIA), (IIIA), any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle. In some embodiments, R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), and -CN; C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -NO2, =0, =S, =N(R10), -CN, C3- 10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to lO-membered heterocycle in R12 is independently optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, - S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -NO2, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl. [0204] In some embodiments, for a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC) , (IIIA), (PIB) or (IIIC), R12 is independently selected at each occurrence from halogen, - OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, - S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle each of which is optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, - S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl.
[0205] In some embodiments, for a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC) , (IIIA), (IIIB) or (IIIC), R12 may be independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-10 alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle.
[0206] In some embodiments, for a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC) , (IIIA), (MB) or (IIIC), R12 may be independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and - CN; and C1-10 alkyl optionally substituted with one or more substituents selected from halogen, - OR10, -SR10, -N(R10)2, -C(0)R10, -N02, =0, =S, =N(R10), and -CN.
[0207] In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (MB), and (IIIC), R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-10 alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to lO-membered heterocycle in R12 is independently optionally substituted with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, - C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl.
[0208] In some embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (PIA), (PIB), and (IIIC), R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to lO-membered heterocycle in R12 is independently optionally substituted with one or more substituents selected from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -N02, =0, =S, =N(R10), -CN, and Ci-6 alkyl.
[0209] Any combination of the groups described above for the various variables is contemplated herein. Throughout the specification, groups and substituents thereof can be chosen to provide stable moieties and compounds.
[0210] In some aspects, the disclosure provides compounds that include a linker, L3. In certain embodiments, a compound bound to a linker is represented by the structure of Formula (IV A):
Figure imgf000096_0001
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond; L12 is independently selected from -X3-, -X3-CI-6 alkyl-X3-, -X3-C2-6 alkenyl-X3-, and
-X3-C2-6 alkynyl-X3-, each of which is optionally substituted at each occurrence with one or more
R10;
L22 is independently selected from -X4-, -X4-CI-6 alkyl-X4-, -X4-C2-6 alkenyl-X4-, and
-X4-C2-6 alkynyl-X4-, each of which is optionally substituted at each occurrence with one or more
R10;
X3 and X4 are independently selected at each occurrence from a bond, -O- , -S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10)-, -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -0S(0)-, -S(0)0-, -S(O), -0S(0)2-, -S(0)20-, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 and R8 are independently selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from L3, halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 and R8 is independently optionally substituted with one or more substituents selected from L3, halogen, - OR10, -SR10, -C(O)N(R10)2,
-N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from L3, hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -CN, -N02, -NH2, =0, =S, - C(0)0CH2C6H5, -NHC(0)0CH2C6H5, CI-IO alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl; L3 is a linker moiety, wherein at least one of R10 is L3 or at least one substituent on R4, R8, X3 and X4 is -L3; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent selected from halogen, -OR10, -SR10, -C(O)N(R10)2-, -N(R10)2, -S(0)R10, - S(0)2R10,
-C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -P(O)(OR10)2, -OP(O)(OR10)2, -CN,
Ci-io alkyl, C2-10 alkenyl, and C2-10 alkynyl, or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
[0211] In some embodiments, for a compound or salt of Formula (IV A), L22 can be attached at C2, C3, C4, or C5 of the benzazepine core, wherein the numbering of the benzazepine is as
follows:
Figure imgf000098_0001
certain embodiments, for a compound or salt of Formula (IV A), L22 is attached to the benzazepine core at C4. In certain embodiments for a compound or salt of Formula (IIA), represents a double bond and L22 is attached to the benzazepine core at
C4.
[0212] In some embodiments for a compound or salt of Formula (IV A), L12 can be attached at C6, C7, C8 or C9. In certain embodiments, for a compound or salt of Formula (IV A), L12 is attached to the benzazepine core at C8. In certain embodiments for a compound or salt of Formula (IV A), represents a double bond, L22 is attached to the benzazepine core at C4 and L12 is attached to the benzazepine core at C8.
[0213] In some embodiments for a compound or salt of Formula (IV A), the substitutable carbon on the benzazepine core is selected from C2, C3, C4, C5, C6, C7, C8, and C9. The benzazepine core for a compound or salt of Formula (IV A), can be optionally substituted by a substituent selected from halogen, -OR10, -SR10, -C(O)N(R10)2-, -N(R10)2, -S(0)R10, -S(0)2R10, - C(0)R10, -C(0)OR10, -OC(0)R10,
-NO¾ =0, =S, =N(R10), -P(O)(OR10)2, -OP(O)(OR10)2, -CN, C I-IO alkyl, C2-io alkenyl, and C2-io alkynyl, or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle. In some embodiments for a compound or salt of Formula (IV A), a moiety at any one of C2, C3, C4, C5, C6, C7, C8, and C9 of the benzazepine core is independently selected from hydrogen, halogen, -OR10, -SR10,
-N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-10 alkyl, C2-io alkenyl, and C2-io alkynyl.
[0214] In one embodiment, the compound of Formula (IV A) is represented by Formula (IVB):
Figure imgf000099_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen, - OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; and
R24 and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, - N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
[0215] In certain embodiments, the compound of Formula (IV A) is represented by Formula (IVC):
Figure imgf000099_0002
or a pharmaceutically acceptable salt thereof.
[0216] In certain embodiments, for a compound or salt of any one of Formulas (IV A), (IVB), and (IVC), L12 is selected from -X3-, -X3-CI-6 alkylene-X3-, -X3-C2-6 alkenylene-X3-, and -X3-C2-6 alkynyl ene-X3-, each of which is optionally substituted on the alkylene, alkenylene and alkyneylene with one or more R12.
[0217] In certain embodiments, for a compound or salt of any one of Formulas (IV A), (IVB), and (IVC),
L22 is selected from -X4-, -X4-CI-6 alkylene-X4-, -X4-C2-6 alkenylene-X4-, and -X4-C2-6 alkynylene-X4-, optionally substituted on the alkylene, alkenylene and alkynylene with one or more R12.
[0218] In some embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), L12 is independently selected from -X3-, -X3-CI-6 alkyl-X3-, -X3-C2-6 alkenyl-X3-, and -X3-C2-6 alkynyl-X3-, each of which is optionally substituted at each occurrence with one or more R10. In some embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), L12 is selected from -C(O)-, and -C(0)NR10-. In some embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), L12 is -C(0)N(R10)-. In some embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), R10 of -C(0)N(R10)- is selected from hydrogen, Ci-6 alkyl, and -L3. In some embodiments for a compound or salt of Formula (IV A), (IVB) or (IVC), L12 is -C(0)NH-. In some embodiments for a compound or salt of Formula (IV A), (IVB) or (IVC), L12 is - C(0)N(L3)-.
[0219] In certain embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), R4 and R8 are independently selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from L3, halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is independently optionally substituted with one or more substituents selected from L3, halogen, -OR10, -SR10, -C(O)N(R10)2,
-N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
[0220] In some embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), R8 is an optionally substituted 5-6 membered heteroaryl. In some embodiments, R8 is an optionally substituted 5-6 membered heteroaryl, substituted with -L3. R8 may be an optionally substituted pyridine, substituted with -L3.
[0221] In some embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), L22 is independently selected from -X4-, -X4-CI-6 alkyl-X4-, -X4-C2-6 alkenyl-X4-, and -X4-C2-6 alkynyl-X4-, each of which is optionally substituted at each occurrence with one or more R10. In some embodiments for a compound or salt of Formula (IV A), (IVB) or (IVC), L22 is selected from -C(O)-, and -C(0)NR10-. L22 may be -C(O)-. In some embodiments, L22 is -C(0)NR10-, wherein, R10 of -C(0)NR10- may be selected from hydrogen, Ci-6 alkyl, and -L3. In some embodiments, L22 is -C(0)NH-. In certain embodiments, L12 is -C(0)N(L3)-.
[0222] In some embodiments, for a compound of Formulas (IV A), (IVB) or (IVC), R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from -L3, halogen, -OR10, - SR10, -C(O)N(R10)2, -N(RIO)C(0)RIO,
-N(RIO)C(0)N(RIO)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to 12- membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is independently optionally substituted with one or more substituents selected from -L3, halogen, -OR10, -SR10,
-C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10,
-OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl. In some embodiments, R4 is selected from: -OR10, and -N(R10)2; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, aryl, and heteroaryl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from -L3, halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, -S(O)2R10,-C(O)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl. In some embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), R10 of -N(R10)2 is selected from -L3 and hydrogen, and wherein at least one R10 of -N(R10)2 is -L3.
[0223] In certain embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; and Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from L3, halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-12 carbocycle, and 3- to l2-membered heterocycle.
[0224] In certain embodiments, for a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC), (PIA), (PIB), (IIIC), (IV A), (IVB) or (IVC), L3 is a noncleavable linker. In some embodiments, for a compound or salt of Formula (IV A), (IVB) or (IVC), L3 is cleavable linker such as a linker cleavable by a lysosomal enzyme.
[0225] In certain embodiments, a moiety described herein includes the symbol
Figure imgf000101_0001
which indicates the point of attachment, e.g., the point of attachment of a chemical moiety to the remainder of the compound, the point of attachment of a linker to a compound of the disclosure, or the point of attachment of a linker to an antibody construct, as described herein.
[0226] Some examplary linkers (L3) are described in the following paragraphs and additional linkers are described in the subsequent section entitled“Linkers”. In some embodiments for a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), peptide
(IVB) or (IVC), -L3 is represented by the formula:
Figure imgf000102_0001
, wherein peptide is a group comprising from one to ten amino acids.
In some embodiments for a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (PIB), (IIIC), (IV A), (IVB) or (IVC), -L3 is represented by the formula:
peptide-RX
Figure imgf000102_0002
wherein peptide is a group comprising from one to ten
amino acids and RX is a reactive moiety,
Figure imgf000102_0003
which L4 represents the C-terminus of the peptide and L5 is selected from a bond, alkyl ene and heteroalkylene, wherein L5 is optionally substituted with one or more groups independently selected from R32, RX is a reactive moiety; and R32 is independently selected at each occurrence from halogen, -OH, -CN, -O-alkyl, -SH, =0, =S, -NH2, -N02; and Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OH, -CN, -O-alkyl, -SH, =0, =S, -NH2, and -N02.
[0227] In certain embodiments, for a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB) or (IVC), -L3 is represented by the formula:
Figure imgf000102_0004
, in which L4 represents the C-terminal of the peptide and L5 is selected from a bond, alkylene and heteroalkylene, wherein L5 is optionally substituted with one or more groups independently selected from R32 ; RX* is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein on RX* represents the point of attachment to the residue of the antibody construct; and,R32 is independently selected at each occurrence from halogen, -OH, -CN, -O- alkyl, -SH, =0, =S, -NH2, -N02; and Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OH, -CN, -O-alkyl, -SH, =0, =S, -NH2, and -N02. The reactive moiety may be selected from an electrophile, e.g., an a, ?-un saturated carbonyl, such as a maleimide, and a leaving group.
[0228] In some embodiments for a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB) or (IVC), -L3 is represented by the formula:
Figure imgf000103_0004
, in which L4 represents the C-terminal of the peptide and L5 is selected from a bond, alkyl ene and heteroalkylene, wherein L5 is optionally substituted with one or more groups independently selected from R32 ; RX* is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein Antibody is an antibody construct; and,R32 is independently selected at each occurrence from halogen, -OH, -CN, -O-alkyl, -SH, =0, =S, -NH2, -N02; and Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OH, -CN, -O-alkyl, -SH, =0, =S, -NH2, and -N02. L3 may be represented by the formula:
Figure imgf000103_0001
, wherein L4 and L5 are independently selected from a bond, an alkylene and a heteroalkylene, each of which is optionally substituted with one or more groups independently selected from R12; *¾· on the left represents the point of attachment to the remainder of the compound, RX* is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety attached at the
Figure imgf000103_0002
on the right to a residue of an antibody construct; peptide is a group comprising from one to 10 amino acids.
[0229] In some embodiments, the compound is:
Figure imgf000103_0003
salt thereof. [0230] In some embodiments for a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB) or (IVC), -L3 is represented by the formula:
Figure imgf000104_0003
, p y, .g., a leaving group, n = 0-9 and ^represents the point of attachment to the remainder of the compound.
[0232] In some embodiments, for a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (PIB), (IIIC), (IV A), (IVB) or (IVC), -L3 is represented by the formula:
Figure imgf000104_0001
bond, a succinimide moiety, or a hydrolyzed succinimide moiety attached at the
Figure imgf000104_0002
on the right to a residue of an antibody construct, n = 0-9 and ^on the left represents the point of attachment to the remainder of the compound.
[0233] Chemical entities having carbon-carbon double bonds or carbon-nitrogen double bonds may exist in Z- or E- form (or cis- or trans- form). Furthermore, some chemical entities may exist in various tautomeric forms. Unless otherwise specified, compounds described herein are intended to include all Z-, E- and tautomeric forms as well.
[0234] A "tautomer" refers to a molecule wherein a proton shift from one atom of a molecule to another atom of the same molecule is possible. The compounds presented herein, in certain embodiments, exist as tautomers. In circumstances where tautomerization is possible, a chemical equilibrium of the tautomers will exist. The exact ratio of the tautomers depends on several factors, including physical state, temperature, solvent, and pH. Some examples of tautomeric equilibrium include:
Figure imgf000105_0001
[0235] The compounds disclosed herein, in some embodiments, are used in different enriched isotopic forms, e.g., enriched in the content of 2H, ¾, UC, 13C and/or 14C. In one particular embodiment, the compound is deuterated in at least one position. Such deuterated forms can be made by the procedure described in U.S. Patent Nos. 5,846,514 and 6,334,997. As described in U.S. Patent Nos. 5,846,514 and 6,334,997, deuteration can improve the metabolic stability and or efficacy, thus increasing the duration of action of drugs.
[0236] Unless otherwise stated, compounds described herein are intended to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by 13C- or 14C-enriched carbon are within the scope of the present disclosure.
[0237] The compounds of the present disclosure optionally contain unnatural proportions of atomic isotopes at one or more atoms that constitute such compounds. For example, the compounds may be labeled with isotopes, such as for example, deuterium (2H), tritium (¾), iodine-l25 (125I) or carbon-l4 (14C). Isotopic substitution with 2H, UC, 13C, 14C, 15C, 12N, 13N,
Figure imgf000105_0002
contemplated. All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.
[0238] In certain embodiments, the compounds disclosed herein have some or all of the ¾ atoms replaced with 2H atoms. The methods of synthesis for deuterium-containing compounds are known in the art and include, by way of non-limiting example only, the following synthetic methods. [0239] Deuterium substituted compounds are synthesized using various methods such as described in: Dean, Dennis C.; Editor. Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [In: Curr., Pharm. Des., 2000; 6(10)] 2000, 110 pp; George W.; Varma, Rajender S. The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; and Evans, E. Anthony. Synthesis of radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32.
[0240] Deuterated starting materials are readily available and are subjected to the synthetic methods described herein to provide for the synthesis of deuterium-containing compounds. Large numbers of deuterium-containing reagents and building blocks are available commercially from chemical vendors, such as Aldrich Chemical Co.
[0241] Compounds of the present invention also include crystalline and amorphous forms of those compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof.
[0242] In some aspects, the present disclosure provides a method for treating cancer. In some embodiments, the present disclosure provides a method comprising administering a conjugate, compound or salt of Formula (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IV A),
(IVB), or (IVC) to a subject in need thereof.
[0243] In some aspects, the compounds of the disclosure exhibit selective binding or agonizing properties for one receptor over another receptor. In some embodiments, a compound described herein selectively binds or modulates the activity of one toll-like receptor over another, e.g., TLR8 and TLR7. For example, a compound or salt of Formula (IA), (IB), (IC), (IIA), (IIB),
(IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB), or (IVC) may have weaker binding to TLR7 as compared to TLR8, as measured by the Kd values, e.g., a compound’s Kd for TLR7 is two times or greater than two times the Kd for TLR8, or an order of magnitude or greater, or even two orders of magnitude or greater than the Kd for TLR8. In certain embodiments, a compound or salt of of Formula (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or
(IVC) has greater activity for one toll-like receptor over another, e.g., TLR8 and TLR7. In certain embodiments, a compound of the disclosure agonizes TLR8 with an EC so of 500 nM or less while the same compound agonizes TLR7 with an EDso of greater than 1 mM. In certain embodiments, a compound of the disclosure agonizes TLR8 with at least an ECso of an order of magnitude or even two orders of magnituge less than the amount of the same compound required to show an agonizing effect on TLR7. [0244] Included in the present disclosure are salts, particularly pharmaceutically acceptable salts, of the compounds described herein. The compounds of the present disclosure that possess a sufficiently acidic, a sufficiently basic, or both functional groups, can react with any of a number of inorganic bases, and inorganic and organic acids, to form a salt. Alternatively, compounds that are inherently charged, such as those with a quaternary nitrogen, can form a salt with an appropriate counterion, e.g., a halide such as bromide, chloride, or fluoride, particularly bromide.
[0245] The compounds described herein may in some cases exist as diastereomers, enantiomers, or other stereoisomeric forms. The compounds presented herein include all diastereomeric, enantiomeric, and epimeric forms as well as the appropriate mixtures thereof. Separation of stereoisomers may be performed by chromatography or by forming diastereomers and separating by recrystallization, or chromatography, or any combination thereof. (Jean Jacques, Andre Collet, Samuel H. Wilen,“Enantiomers, Racemates and Resolutions”, John Wiley And Sons,
Inc., 1981, herein incorporated by reference for this disclosure). Stereoisomers may also be obtained by stereoselective synthesis.
[0246] The methods and compositions described herein include the use of amorphous forms as well as crystalline forms (also known as polymorphs). The compounds described herein may be in the form of pharmaceutically acceptable salts. As well, active metabolites of these compounds having the same type of activity are included in the scope of the present disclosure. In addition, the compounds described herein can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. The solvated forms of the compounds presented herein are also considered to be disclosed herein.
[0247] In certain embodiments, compounds or salts of the compounds of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) may be prodrugs, e.g., wherein a hydroxyl in the parent compound is presented as an ester or a carbonate, or carboxylic acid present in the parent compound is presented as an ester. The term “prodrug” is intended to encompass compounds which, under physiologic conditions, are converted into pharmaceutical agents of the present disclosure. One method for making a prodrug is to include one or more selected moieties which are hydrolyzed under physiologic conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by an enzymatic activity of the host animal such as specific target cells in the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids and esters of phosphonic acids) are preferred prodrugs of the present disclosure.
[0248] Prodrug forms of the herein described compounds, wherein the prodrug is metabolized in vivo to produce a compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB), or (IVC) as set forth herein are included within the scope of the claims. In some cases, some of the herein-described compounds may be a prodrug for another derivative or active compound.
[0249] Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent is not. Prodrugs may help enhance the cell permeability of a compound relative to the parent drug. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. Prodrugs may be designed as reversible drug derivatives, for use as modifiers to enhance drug transport to site-specific tissues or to increase drug residence inside of a cell.
[0250] In some embodiments, the design of a prodrug increases the lipophilicity of the pharmaceutical agent. In some embodiments, the design of a prodrug increases the effective water solubility. See, e.g., Fedorak e/ a/., Am. J Physiol ., 269:G2l0-2l8 (1995); McLoed e/ a/., Gastroenterol , 106:405-413 (1994); Hochhaus et al., Biomed. Chrom., 6:283-286 (1992); J. Larsen and H. Bundgaard, Int. J. Pharmaceutics, 37, 87 (1987); J. Larsen et al., Int. J.
Pharmaceutics , 47, 103 (1988); Sinkula et al., J. Pharm. Sci., 64: 181-210 (1975); T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems , Vol. 14 of the A.C.S. Symposium Series; and Edward B. Roche, Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, all incorporated herein for such disclosure). According to another embodiment, the present disclosure provides methods of producing the above-defined compounds. The compounds may be synthesized using conventional techniques.
Advantageously, these compounds are conveniently synthesized from readily available starting materials.
[0251] Synthetic chemistry transformations and methodologies useful in synthesizing the compounds described herein are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations (1989); T. W. Greene and P. G. M.
Wuts, Protective Groups in Organic Synthesis, 2d. Ed. (1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis (1995).
Linkers
[0252] The compounds and salts described herein may be bound to a linker also referred to herein as L3, e.g., a peptide linker. In certain embodiments, the linker is also bound to an antibody construct and referred to as an antibody construct conjugate or conjugate. Linkers of the conjugates described herein may not affect the binding of active portions of a conjugate, e.g, the antigen binding domains, Fc domains, target binding domains, antibodies, agonists or the like, to a target, which can be a cognate binding partner such as an antigen. A conjugate can comprise multiple linkers, each having one or more compounds attached. These linkers can be the same linkers or different linkers. Some examplary linkers (L3) are described in the preceding section entitled“Compounds” and additional linkers are described in this section.
[0253] A linker can be short, flexible, rigid, cleavable, non-cleavable, hydrophilic, or hydrophobic. A linker can contain segments that have different characteristics, such as segments of flexibility or segments of rigidity. The linker can be chemically stable to extracellular environments, for example, chemically stable in the blood stream, or may include linkages that are not stable or selectively stable. The linker can include linkages that are designed to cleave and/or immolate or otherwise breakdown specifically or non- specifically inside cells. A cleavable linker can be sensitive to enzymes. A cleavable linker can be cleaved by enzymes such as proteases. A cleavable linker may comprise a valine-citrulline linker or a valine-alanine peptide. A valine-citrulline- or valine-alanine-containing linker can contain a pentafluorophenyl group. A valine-citrulline- or valine-alanine-containing linker can contain a maleimide or succinimide group. A valine-citrulline- or valine-alanine-containing linker can contain a para aminobenzyl alcohol (PABA) group or para-aminobenzyl carbamate (PABC).
[0254] A valine-citrulline- or valine-alanine-containing linker can contain a PABA group and a pentafluorophenyl group. A valine-citrulline- or valine-alanine-containing linker can contain a PABA group and a maleimide or succinimide group.
[0255] A non-cleavable linker can be protease insensitive. A non-cleavable linker can be maleimidocaproyl linker. A maleimidocaproyl linker can comprise N- maleimidomethylcyclohexane-l-carboxylate. A maleimidocaproyl linker can contain a succinimide group. A maleimidocaproyl linker can contain pentafluorophenyl group. A linker can be a combination of a maleimidocaproyl group and one or more polyethylene glycol molecules. A linker can be a maleimide-PEG4 linker. A linker can be a combination of a maleimidocaproyl linker containing a succinimide group and one or more polyethylene glycol molecules. A linker can be a combination of a maleimidocaproyl linker containing a
pentafluorophenyl group and one or more polyethylene glycol molecules. A linker can contain maleimides linked to polyethylene glycol molecules in which the polyethylene glycol can allow for more linker flexibility or can be used lengthen the linker. A linker can be a
(maleimidocaproyl)-(valine-citrulline)-(para-aminobenzyloxycarbonly) linker. A linker can be a linker suitable for attachment to an engineered cysteine (THIOMAB). A THIOMAB linker can be a (maleimidocaproyl)-(valine-citrulline)-(para-aminobenzyloxycarbonly)-linker. [0256] A linker can also comprise alkylene, alkenylene, alkynylene, polyether, polyester, polyamide group(s) and also, polyamino acids, polypeptides, cleavable peptides, or
aminobenzylcarbamates. A linker can contain a maleimide at one end and an N- hydroxysuccinimidyl ester at the other end. A linker can contain a lysine with an N-terminal amine acetylated, and a valine-citrulline cleavage site. A linker can be a link created by a microbial transglutaminase, wherein the link can be created between an amine-containing moiety and a moiety engineered to contain glutamine as a result of the enzyme catalyzing a bond formation between the acyl group of a glutamine side chain and the primary amine of a lysine chain. A linker can contain a reactive primary amine. A linker can be a Sortase A linker. A Sortase A linker can be created by a Sortase A enzyme fusing an LXPTG recognition motif to an N-terminal GGG motif to regenerate a native amide bond. The linker created can therefore link a moiety attached to the LXPTG recognition motif with a moiety attached to the N-terminal GGG motif.
[0257] In the conjugates described herein, a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB), or (IVC) is linked to the antibody construct by way of a linker(s), also referred to herein as L3. L3, as used herein, may be selected from any of the linker moieties discussed herein. The linker linking the compound or salt to the antibody construct of a conjugate may be short, long, hydrophobic, hydrophilic, flexible or rigid, or may be composed of segments that each independently have one or more of the above- mentioned properties such that the linker may include segments having different properties. The linkers may be polyvalent such that they covalently link more than one compound or salt to a single site on the antibody construct, or monovalent such that covalently they link a single compound or salt to a single site on the antibody construct.
[0258] Linkers of the disclosure (L3) may have from about 10 to about 500 atoms in a linker, such as from about 10 to about 400 atoms, such as about 10 to about 300 atoms in a linker. In certain embodiments, linkers of the disclosure have from about 30 to about 400 atoms, such as from about 30 to about 300 atoms in the linker.
[0259] As will be appreciated by skilled artisans, the linkers may link a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) to the antibody construct by a covalent linkages between the linker and the antibody construct and compound. As used herein, the expression "linker" is intended to include (i) unconjugated forms of the linker that include a functional group capable of covalently linking the linker to a benzazepine compound(s) and a functional group capable of covalently linking the linker to an antibody construct; (ii) partially conjugated forms of the linker that include a functional group capable of covalently linking the linker to an antibody construct and that is covalently linked to a compound(s) or salt(s) of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (MB), (IIIC), (IV A), (IVB), or (IVC), or vice versa; and (iii) fully conjugated forms of the linker that is covalently linked to both a compound(s) or salt(s) of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (MC), (IV A), (IVB), or (IVC) and an antibody construct. One embodiment pertains to a conjugate formed by contacting an antibody construct that binds a cell surface receptor or tumor associated antigen expressed on a tumor cell with a linker-compound described herein under conditions in which the linker-compound covalently links to the antibody construct. One embodiment pertains to a method of making a conjugate formed by contacting a linker-compound described herein under conditions in which the linker-compound covalently links to the antibody construct. One embodiment pertains to a method of stimulating immune activity in a cell that expresses CD40, comprising contacting the cell with a conjugate described herein that is capable of binding the cell, under conditions in which the conjugate binds the cell.
[0260] Exemplary polyvalent linkers that may be used to link many benzazepine compounds to an antibody construct are described. For example, Fleximer® linker technology has the potential to enable high-DAR conjugates with good physicochemical properties. As shown below, the Fleximer® linker technology is based on incorporating drug molecules into a solubilizing poly acetal backbone via a sequence of ester bonds. The methodology renders highly-loaded conjugates (DAR up to 20) whilst maintaining good physicochemical properties. This methodology could be utilized with a benzazepine compound as shown in the Scheme below.
Figure imgf000111_0001
Figure imgf000111_0002
[0261] To utilize the Fleximer® linker technology depicted in the scheme above, an aliphatic alcohol can be present or introduced into the benzazepine compound. The alcohol moiety is then conjugated to an alanine moiety, which is then synthetically incorporated into the Fleximer® linker. Liposomal processing of the conjugate in vitro releases the parent alcohol-containing drug.
[0262] By way of example and not limitation, some cleavable and noncleavable linkers that may be included in the conjugates described herein are described below.
[0263] Cleavable linkers can be cleavable in vitro and in vivo. Cleavable linkers can include chemically or enzymatically unstable or degradable linkages. Cleavable linkers can rely on processes inside the cell to liberate a benzazepine compound, such as reduction in the cytoplasm, exposure to acidic conditions in the lysosome, or cleavage by specific proteases or other enzymes within the cell. Cleavable linkers can incorporate one or more chemical bonds that are either chemically or enzymatically cleavable while the remainder of the linker can be non cleavable.
[0264] A linker can contain a chemically labile group such as hydrazone and/or disulfide groups. Linkers comprising chemically labile groups can exploit differential properties between the plasma and some cytoplasmic compartments. The intracellular conditions that can facilitate benzazepine compound release for hydrazone containing linkers can be the acidic environment of endosomes and lysosomes, while the disulfide containing linkers can be reduced in the cytosol, which can contain high thiol concentrations, e.g., glutathione. The plasma stability of a linker containing a chemically labile group can be increased by introducing steric hindrance using substituents near the chemically labile group.
[0265] Acid-labile groups, such as hydrazone, can remain intact during systemic circulation in the blood’s neutral pH environment (pH 7.3-7.5) and can undergo hydrolysis and can release the benzazepine compound once the antibody construct benzazepine compound conjugate is internalized into mildly acidic endosomal (pH 5.0-6.5) and lysosomal (pH 4.5-5.0) compartments of the cell. This pH dependent release mechanism can be associated with nonspecific release of the drug. To increase the stability of the hydrazone group of the linker, the linker can be varied by chemical modification, e.g., substitution, allowing tuning to achieve more efficient release in the lysosome with a minimized loss in circulation.
[0266] Hydrazone-containing linkers can contain additional cleavage sites, such as additional acid-labile cleavage sites and/or enzymatically labile cleavage sites. Antibody construct benzazepine compound conjugates including exemplary hydrazone-containing linkers can include, for example, the following structures:
Figure imgf000113_0001
wherein D is a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC) and Ab is an antibody construct, respectively, and n represents the number of compound-bound linkers (LP) bound to the antibody construct. In certain linkers, such as linker (la), the linker can comprise two cleavable groups, a disulfide and a hydrazone moiety.
For such linkers, effective release of the unmodified free benzazepine compound can require acidic pH or disulfide reduction and acidic pH. Linkers such as (lb) and (Ic) can be effective with a single hydrazone cleavage site.
[0267] Other acid-labile groups that can be included in linkers include c/.s-aconityl -containing linkers. c .s-Aconityl chemistry can use a carboxylic acid juxtaposed to an amide bond to accelerate amide hydrolysis under acidic conditions.
[0268] Cleavable linkers can also include a disulfide group. Disulfides can be
thermodynamically stable at physiological pH and can be designed to release the benzazepine compound upon internalization inside cells, wherein the cytosol can provide a significantly more reducing environment compared to the extracellular environment. Scission of disulfide bonds can require the presence of a cytoplasmic thiol cofactor, such as (reduced) glutathione (GSH), such that disulfide-containing linkers can be reasonably stable in circulation, selectively releasing the benzazepine compound in the cytosol. The intracellular enzyme protein disulfide isomerase, or similar enzymes capable of cleaving disulfide bonds, can also contribute to the preferential cleavage of disulfide bonds inside cells. GSH can be present in cells in the concentration range of 0.5-10 mM compared with a significantly lower concentration of GSH or cysteine, the most abundant low-molecular weight thiol, in circulation at approximately 5 mM. Tumor cells, where irregular blood flow can lead to a hypoxic state, can result in enhanced activity of reductive enzymes and therefore even higher glutathione concentrations. The in vivo stability of a disulfide-containing linker can be enhanced by chemical modification of the linker, e.g., use of steric hindrance adjacent to the disulfide bond.
[0269] Antibody construct benzazepine compound conjugates including exemplary disulfide- containing linkers can include the following structures:
Figure imgf000114_0001
wherein D is a compound or salt of any one of Formulas (IA), (IIA), (IB), (IIB), (IIIA), and (IIIB) and Ab is an antibody construct, respectively, n represents the number of compounds bound to linkers (L3) bound to the antibody construct and R is independently selected at each occurrence from hydrogen or alkyl, for example. Increasing steric hindrance adjacent to the disulfide bond can increase the stability of the linker. Structures such as (Ila) and (lie) can show increased in vivo stability when one or more R groups is selected from a lower alkyl such as methyl.
[0270] Another type of linker that can be used is a linker that is specifically cleaved by an enzyme. For example, the linker can be cleaved by a lysosomal enzyme. Such linkers can be peptide-based or can include peptidic regions that can act as substrates for enzymes. Peptide based linkers can be more stable in plasma and extracellular milieu than chemically labile linkers.
[0271] Peptide bonds can have good serum stability, as lysosomal proteolytic enzymes can have very low activity in blood due to endogenous inhibitors and the unfavorably high pH value of blood compared to lysosomes. Release of a benzazepine compound from an antibody construct can occur due to the action of lysosomal proteases, e.g., cathepsin and plasmin. These proteases can be present at elevated levels in certain tumor tissues. The linker can be cleavable by a lysosomal enzyme. The lysosomal enzyme can be, for example, cathepsin B, cathepsin S, b- glucuronidase, or b-galactosidase. [0272] The cleavable peptide can be selected from tetrapeptides such as Gly-Phe-Leu-Gly, Ala- Leu-Ala-Leu or dipeptides such as Val-Cit, Val-Ala, and Phe-Lys. Dipeptides can have lower hydrophobicity compared to longer peptides.
[0273] A variety of dipeptide-based cleavable linkers can be used in the antibody constructs benzazepine compound conjugates described herein.
[0274] Enzymatically cleavable linkers can include a self-immolative spacer to spatially separate the benzazepine compound from the site of enzymatic cleavage. The direct attachment of a benzazepine compound to a peptide linker can result in proteolytic release of the benzazepine compound or of an amino acid adduct of the benzazepine compound, thereby impairing its activity. The use of a self-immolative spacer can allow for the elimination of the fully active, chemically unmodified benzazepine compound upon amide bond hydrolysis.
[0275] One self-immolative spacer can be a bifunctional para- aminobenzyl alcohol group, which can link to the peptide through the amino group, forming an amide bond, while amine containing benzazepine compounds can be attached through carbamate functionalities to the benzylic hydroxyl group of the linker (to give a / amidobenzyl carbarn ate, PABC). The resulting pro-benzazepine compound can be activated upon protease- mediated cleavage, leading to a 1,6- elimination reaction releasing the unmodified benzazepine compound, carbon dioxide, and remnants of the linker. The following scheme depicts the fragmentation of p- amidobenzyl carbamate and release of the benzazepine compound:
Figure imgf000115_0001
X-D
wherein X-D represents the unmodified benzazepine compound.
Heterocyclic variants of this self-immolative group have also been described.
[0276] The enzymatically cleavable linker can be a B-glucuronic acid-based linker. Facile release of the benzazepine compound can be realized through cleavage of the B-glucuronide glycosidic bond by the lysosomal enzyme B-glucuronidase. This enzyme can be abundantly present within lysosomes and can be overexpressed in some tumor types, while the enzyme activity outside cells can be low. B- Glucuronic acid-based linkers can be used to circumvent the tendency of an antibody construct benzazepine compound conjugate to undergo aggregation due to the hydrophilic nature of B-glucuronides. In certain embodiments, B-glucuronic acid-based linkers can link an antibody construct to a hydrophobic benzazepine compound. The following scheme depicts the release of a benzazepine compound (D) from an antibody construct benzazepine compound conjugate containing a B-glucuronic acid-based linker:
Figure imgf000116_0001
wherein Ab indicates the antibody construct.
[0277] A variety of cleavable b-glucuronic acid-based linkers useful for linking drugs such as auristatins, camptothecin and doxorubicin analogues, CBI minor-groove binders, and psymberin to antibodies have been described. These b-glucuronic acid-based linkers may be used in the conjugates described herein. In certain embodiments, the enzymatically cleavable linker is a b- galactoside-based linker. b-Galactoside is present abundantly within lysosomes, while the enzyme activity outside cells is low.
[0278] Additionally, benzazepine compounds containing a phenol group can be covalently bonded to a linker through the phenolic oxygen. One such linker relies on a methodology in which a diamino-ethane "Space Link" is used in conjunction with traditional "PABO"-based self- immolative groups to deliver phenols.
[0279] Cleavable linkers can include non-cleavable portions or segments, and/or cleavable segments or portions can be included in an otherwise non-cleavable linker to render it cleavable. By way of example only, polyethylene glycol (PEG) and related polymers can include cleavable groups in the polymer backbone. For example, a polyethylene glycol or polymer linker can include one or more cleavable groups such as a disulfide, a hydrazone or a dipeptide.
[0280] Other degradable linkages that can be included in linkers can include ester linkages formed by the reaction of PEG carboxylic acids or activated PEG carboxylic acids with alcohol groups on a benzazepine compound, wherein such ester groups can hydrolyze under
physiological conditions to release the benzazepine compound. Hydrolytically degradable linkages can include, but are not limited to, carbonate linkages; imine linkages resulting from reaction of an amine and an aldehyde; phosphate ester linkages formed by reacting an alcohol with a phosphate group; acetal linkages that are the reaction product of an aldehyde and an alcohol; orthoester linkages that are the reaction product of a formate and an alcohol; and oligonucleotide linkages formed by a phosphoramidite group, including but not limited to, at the end of a polymer, and a 5' hydroxyl group of an oligonucleotide.
[0281] A linker can contain an enzymatically cleavable peptide moiety, for example, a linker comprising structural formula (Ilia), (Mb), (Me), or (Hid):
Figure imgf000117_0001
(
Figure imgf000117_0002
Figure imgf000117_0003
Figure imgf000117_0004
or a salt thereof, wherein:“peptide” represents a peptide (illustrated in N C orientation, wherein peptide includes the amino and carboxy“termini”) that is cleavable by a lysosomal enzyme; T represents a polymer comprising one or more ethylene glycol units or an alkylene chain, or combinations thereof; Ra is selected from hydrogen, alkyl, sulfonate and methyl sulfonate; Ry is hydrogen or Ci-4 alkyl-(0)r-(Ci-4 alkylene)s-G1 or Ci-4 alkyl-(N)-[(Ci-4 alkylene)- G¾ Rz is Ci-4 alkyl-(0)r-(Ci-4 alkylene)s-G2; G1 is SO3H, C02H, PEG 4-32, or a sugar moiety; G2 is SO3H, CO2H, or a PEG 4-32 moiety; r is 0 or 1; s is 0 or 1; p is an integer ranging from 0 to 5; q is 0 or 1; x is 0 or 1; y is 0 or 1; * represents the point of attachment of the linker to a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), or (IIIC); and * represents the point of attachment to the remainder of the linker.
[0282] In certain embodiments, the peptide can be selected from natural amino acids, unnatural amino acids or combinations thereof. In certain embodiments, the peptide can be selected from a tripeptide or a dipeptide. In particular embodiments, the dipeptide can comprise L-amino acids and be selected from: Val-Cit; Cit-Val; Ala-Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala; Phe- Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu- Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit, or salts thereof.
[0283] Exemplary embodiments of linkers according to structural formula (Ilia) are illustrated below (as illustrated, the linkers include a reactive group suitable for covalently linking the linker to an antibody construct):
Figure imgf000118_0001
Figure imgf000118_0002
Figure imgf000118_0003
Figure imgf000118_0004
Figure imgf000119_0001
Figure imgf000119_0002
Figure imgf000119_0003
Figure imgf000119_0004
wherein indicates an attachment site of a linker (L3 ) to a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
[0284] Exemplary embodiments of linkers according to structural formula (Illb), (IIIc), or (Hid) that can be included in the conjugates described herein can include the linkers illustrated below (as illustrated, the linkers can include a reactive group suitable for covalently linking the linker to an antibody construct):
Figure imgf000120_0001
Figure imgf000120_0002
Figure imgf000120_0003
Figure imgf000120_0004
Figure imgf000121_0001
Figure imgf000121_0002
Figure imgf000121_0003
Figure imgf000121_0004
Figure imgf000121_0005
Figure imgf000121_0006
Figure imgf000122_0001
Figure imgf000122_0002
Figure imgf000122_0003
Figure imgf000122_0004
Figure imgf000122_0005
Figure imgf000123_0001
Figure imgf000123_0002
Figure imgf000123_0003
Figure imgf000123_0004
Figure imgf000123_0005
Figure imgf000124_0001
Figure imgf000124_0002
Figure imgf000124_0003
Figure imgf000124_0004
Figure imgf000124_0005
Figure imgf000125_0001
wherein ^ indicates an attachment site to a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
[0285] The linker can contain an enzymatically cleavable sugar moiety, for example, a linker comprising structural formula (IVa), (IVb), (IVc), (IVd), or (IVe):
Figure imgf000125_0002
Figure imgf000126_0001
or a salt thereof, wherein: q is 0 or 1; r is 0 or 1; X is CH2, O or NH; ' represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC); and * represents the point of attachment to the remainder of the linker.
[0286] Exemplary embodiments of linkers according to structural formula (IVa) that may be included in the antibody construct benzazepine compound conjugates described herein can include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
Figure imgf000127_0001
Figure imgf000127_0002
Figure imgf000127_0003
Figure imgf000127_0004
Figure imgf000128_0001
Figure imgf000128_0002
Figure imgf000128_0003
Figure imgf000128_0004
Figure imgf000128_0005
Figure imgf000129_0001
Figure imgf000129_0002
Figure imgf000129_0003
wherein » represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
[0287] Exemplary embodiments of linkers according to structural formula (IVb) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
Figure imgf000129_0004
Figure imgf000130_0001
Figure imgf000130_0002
Figure imgf000130_0003
Figure imgf000130_0004
Figure imgf000130_0005
Figure imgf000131_0001
Figure imgf000131_0002
Figure imgf000131_0003
Figure imgf000131_0004
wherein represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
[0288] Exemplary embodiments of linkers according to structural formula (IVc) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
Figure imgf000132_0001
Figure imgf000132_0002
Figure imgf000132_0003
Figure imgf000133_0001
Figure imgf000133_0002
Figure imgf000133_0003
Figure imgf000133_0004
Figure imgf000134_0001
Figure imgf000134_0002
Figure imgf000134_0003
Figure imgf000134_0004
wherein represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
[0289] Exemplary embodiments of linkers according to structural formula (IVd) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
Figure imgf000135_0001
Figure imgf000135_0002
Figure imgf000135_0003
Figure imgf000135_0004
Figure imgf000135_0005
Figure imgf000135_0006
wherein represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
[0290] Exemplary embodiments of linkers according to structural formula (IVe) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
Figure imgf000136_0001
wherein » represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
[0291] Although cleavable linkers can provide certain advantages, the linkers comprising the conjugate described herein need not be cleavable. For non-cleavable linkers, the benzazepine compound release may not depend on the differential properties between the plasma and some cytoplasmic compartments. The release of the benzazepine compound can occur after
internalization of the antibody construct benzazepine compound conjugate via antigen-mediated endocytosis and delivery to lysosomal compartment, where the antibody construct can be degraded to the level of amino acids through intracellular proteolytic degradation. This process can release a benzazepine compound derivative, which is formed by the benzazepine compound, the linker, and the amino acid residue or residues to which the linker was covalently attached.
The benzazepine compound derivative from antibody construct benzazepine compound conjugates with non-cleavable linkers can be more hydrophilic and less membrane permeable, which can lead to less bystander effects and less nonspecific toxicities compared to antibody construct benzazepine compound conjugates with a cleavable linker. Antibody construct benzazepine compound conjugates with non-cleavable linkers can have greater stability in circulation than antibody construct benzazepine compound conjugates with cleavable linkers. Non-cleavable linkers can include alkylene chains, or can be polymeric, such as, for example, based upon polyalkylene glycol polymers, amide polymers, or can include segments of alkylene chains, polyalkylene glycols and/or amide polymers. The linker can contain a polyethylene glycol segment having from 1 to 6 ethylene glycol units.
[0292] The linker can be non-cleavable in vivo , for example, a linker according to the formulations below:
(Va)
Figure imgf000137_0001
(Vb)
Figure imgf000137_0002
(Vc)
Figure imgf000137_0003
(Vd)
Figure imgf000137_0004
Figure imgf000137_0007
(Vf)
Figure imgf000137_0005
or salts thereof, wherein: Ra is selected from hydrogen, alkyl, sulfonate and methyl sulfonate; Rx is a reactive moiety including a functional group capable of covalently linking the linker to an antibody construct; and
Figure imgf000137_0006
represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IV A), (IVB), or (IVC).
[0293] Exemplary embodiments of linkers according to structural formula (Va)-(Vf) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct, and represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC):
Figure imgf000138_0001
(Vc.2)
Figure imgf000138_0002
(Vd.l)
Figure imgf000138_0003
(Vd.2)
Figure imgf000138_0004
(Vd.3)
Figure imgf000138_0005
(Vd.4)
Figure imgf000138_0006
Figure imgf000138_0007
[0294] Attachment groups that are used to attach the linkers to an antibody construct can be electrophilic in nature and include, for example, maleimide groups, alkynes, alkynoates, allenes and allenoates, activated disulfides, active esters such as NHS esters and HOBt esters, haloformates, acid halides, alkyl, and benzyl halides such as haloacetamides. There are also emerging technologies related to "self-stabilizing" maleimides and "bridging disulfides" that can be used in accordance with the disclosure. [0295] Maleimide groups are frequently used in the preparation of conjugates because of their specificity for reacting with thiol groups of, for example, cysteine groups of the antibody of a conjugate. The reaction between a thiol group of an antibody and a drug with a linker including a maleimide group proceeds according to the following scheme:
.SH Antibody
Antibody +
Linking Group Linking Group
Figure imgf000139_0001
/
Figure imgf000139_0002
/
Drug Drug
[0296] The reverse reaction leading to maleimide elimination from a thio-substituted
succinimide may also take place. This reverse reaction is undesirable as the maleimide group may subsequently react with another available thiol group such as other proteins in the body having available cysteines. Accordingly, the reverse reaction can undermine the specificity of a conjugate. One method of preventing the reverse reaction is to incorporate a basic group into the linking group shown in the scheme above. Without wishing to be bound by theory, the presence of the basic group may increase the nucleophilicity of nearby water molecules to promote ring opening hydrolysis of the succinimide group. The hydrolyzed form of the attachment group is resistant to deconjugation in the presence of plasma proteins. So-called“self-stabilizing” linkers provide conjugates with improved stability. A representative schematic is shown below:
Figure imgf000139_0005
Antibody Antibody
Figure imgf000139_0003
Figure imgf000139_0004
[0297] The hydrolysis reaction schematically represented above may occur at either carbonyl group of the succinimide group. Accordingly, two possible isomers may result, as shown below:
Figure imgf000139_0006
[0298] The identity of the base as well as the distance between the base and the maleimide group can be modified to tune the rate of hydrolysis of the thio- substituted succinimide group and optimize the delivery of a conjugate to a target by, for example, improving the specificity and stability of the conjugate.
[0299] Bases suitable for inclusion in a linker described herein, e.g., any L3 described herein with a maleimide group prior to conjugating to an antibody construct, may facilitate hydrolysis of a nearby succinimide group formed after conjugation of the antibody construct to the linker. Bases may include, for example, amines (e.g., -N(R26)(R27), where R26 and R27 are independently selected from H and Ci-6 alkyl), nitrogen-containing heterocycles (e.g., a 3- to l2-membered heterocycle including one or more nitrogen atoms and optionally one or more double bonds), amidines, guanidines, and carbocycles or heterocycles substituted with one or more amine groups (e.g., a 3- to l2-membered aromatic or non-aromatic cycle optionally including a heteroatom such as a nitrogen atom and substituted with one or more amines of the type - N(R26)(R27), where R26 and R27 are independently selected from H or Ci-6 alkyl). A basic unit may be separated from a maleimide group by, for example, an alkylene chain of the form - (CH2)m-, where m is an integer from 0 to 10. An alkylene chain may be optionally substituted with other functional groups as described herein.
[0300] A linker (L3) described herein with a maleimide group may include an electron withdrawing group such as, but not limited to, -C(0)R, =0, -CN, -N02, -CX3, -X, -COOR, - CONR2, -COR, -COX, -S02R, -S02OR, -S02NHR, -S02NR2, P03R2, -P(0)(CH3)NHR, -NO, - NR3 +, -CR=CR2, and -CºCR, where each R is independently selected from H and Ci-6 alkyl and each X is independently selected from F, Br, Cl, and I. Self-stabilizing linkers may also include aryl, e.g., phenyl, or heteroaryl, e.g., pyridine, groups optionally substituted with electron withdrawing groups such as those described herein.
[0301] Examples of self-stabilizing linkers are provided in, e.g., U.S. Patent Publication Number 2013/0309256, the linkers of which are incorporated by reference herein. It will be understood that a self-stabilizing linker useful in conjunction with the compounds of the present invention may be equivalently described as unsubstituted maleimide-including linkers, thio-substituted succinimide-including linkers, or hydrolyzed, ring-opened thio-substituted succinimide- including linkers. [0302] In certain embodiments, a linker of the disclosure (L3) comprises a stabilizing linker
moiety selected from:
Figure imgf000141_0001
[0303] In the scheme provided above, the bottom structure may be referred to as (maleimido)- DPR-Val-Cit-PAB, where DPR refers to diaminopropinoic acid, Val refers to valine, Cit refers to citrulline, and PAB refers to para-aminobenzylcarbonyl. ^ represents the point of attachment to compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (MC), (IV A), (IVB), or (IVC).
[0304] A method for bridging a pair of sulfhydryl groups derived from reduction of a native hinge disulfide bond has been disclosed and is depicted in the schematic below. An advantage of this methodology is the ability to synthesize homogenous DAR4 conjugates by full reduction of IgGs (to give 4 pairs of sulfhydryls from interchain disulfides) followed by reaction with 4 equivalents of the alkylating agent. Conjugates containing "bridged disulfides" are also claimed to have increased stability.
Figure imgf000141_0002
[0305] Similarly, as depicted below, a maleimide derivative that is capable of bridging a pair of sulfhydryl groups has been developed.
Figure imgf000142_0001
[0306] A linker of the disclosure, L3, can contain the following structural formulas (Via), (VIb), or (Vic):
Figure imgf000142_0002
or salts thereof, wherein: Rq is H or-0-(CH2CH20)n-CH3; x is 0 or 1; y is 0 or 1; G2 is- CH2CH2CH2SO3H or-CH2CH20-(CH2CH20)ii-CH3; Rw is-0-CH2CH2S03H or-NH(CO)- CH2CH20-(CH2CH20)i2-CH3; and * represents the point of attachment to the remainder of the linker.
[0307] Exemplary embodiments of linkers according to structural formula (Via) and (VIb) that can be included in the conjugates described herein can include the linkers illustrated below (as illustrated, the linkers can include a group suitable for covalently linking the linker to an antibody construct):
Figure imgf000143_0001
Figure imgf000143_0002
Figure imgf000143_0003
Figure imgf000143_0004
Figure imgf000144_0001
Figure imgf000144_0002
Figure imgf000144_0003
Figure imgf000145_0001
Figure imgf000145_0002
Figure imgf000145_0003
Figure imgf000146_0001
wherein > represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
[0308] Exemplary embodiments of linkers according to structural formula (Vic) that can be included in the antibody construct benzazepine compound conjugates described herein can include the linkers illustrated below (as illustrated, the linkers can include a group suitable for covalently linking the linker to an antibody construct):
Figure imgf000146_0002
Figure imgf000146_0003
Figure imgf000146_0004
Figure imgf000147_0001
Figure imgf000147_0002
Figure imgf000147_0003
wherein » represents the point of attachment of the linker (L3) to the compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC).
[0309] As is known by skilled artisans, the linker selected for a particular conjugate may be influenced by a variety of factors, including but not limited to, the site of attachment to the antibody construct (e.g., lys, cys or other amino acid residues), structural constraints of the drug pharmacophore and the lipophilicity of the drug. The specific linker selected for a conjugate should seek to balance these different factors for the specific antibody construct/drug
combination.
[0310] For example, conjugates have been observed to effect killing of bystander antigen negative cells present in the vicinity of the antigen-positive tumor cells. The mechanism of bystander cell killing by conjugates has indicated that metabolic products formed during intracellular processing of the conjugates may play a role. Neutral cytotoxic metabolites generated by metabolism of the conjugates in antigen-positive cells appear to play a role in bystander cell killing while charged metabolites may be prevented from diffusing across the membrane into the medium, or from the medium across the membrane, and therefore cannot affect bystander killing. In certain embodiments, the linker is selected to attenuate the bystander killing effect caused by cellular metabolites of the conjugate. In certain embodiments, the linker is selected to increase the bystander killing effect.
[0311] The properties of the linker, or linker-compound, may also impact aggregation of the conjugate under conditions of use and/or storage. Typically, conjugates reported in the literature contain no more than 3-4 drug molecules per antibody molecule. Attempts to obtain higher drug- to-antibody ratios (“DAR”) often failed, particularly if both the drug and the linker were hydrophobic, due to aggregation of the conjugate. In many instances, DARs higher than 3-4 could be beneficial as a means of increasing potency. In instances where the benzazepine compound is more hydrophobic in nature, it may be desirable to select linkers that are relatively hydrophilic as a means of reducing conjugate aggregation, especially in instances where DARs greater than 3-4 are desired. Thus, in certain embodiments, the linker incorporates chemical moieties that reduce aggregation of the conjugates during storage and/or use. A linker may incorporate polar or hydrophilic groups such as charged groups or groups that become charged under physiological pH to reduce the aggregation of the conjugates. For example, a linker may incorporate charged groups such as salts or groups that deprotonate, e.g., carboxylates, or protonate, e.g., amines, at physiological pH.
[0312] In particular embodiments, the aggregation of the conjugates during storage or use is less than about 40% as determined by size-exclusion chromatography (SEC). In particular embodiments, the aggregation of the conjugates during storage or use is less than 35%, such as less than about 30%, such as less than about 25%, such as less than about 20%, such as less than about 15%, such as less than about 10%, such as less than about 5%, such as less than about 4%, or even less, as determined by size-exclusion chromatography (SEC).
Attachment of Linkers to Antibody Construct
[0313] The conjugates described herein may comprise a linker, e.g., a cleavable linker such as a peptide linker or a noncleavable linker. Linkers of the conjugates and methods described herein may not affect the binding of active portions of a conjugate (e.g., active portions include antigen binding domains, Fc domains, target binding domains, antibodies, compounds or salts of the disclosed, e.g., Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (MB) or (IIIC), or linker- compounds of Formulas (IV A), (IVB), or (IVC) to a target, which can be a cognate binding partner such as an antigen. A linker sequence can form a linkage between different parts of a conjugate, e.g., between an antibody construct and a compound or salt of the disclosure. In certain embodiments, a conjugate comprises multiple linkers. In certain embodiments, wherein a conjugate comprises multiple linkers, the linkers may be the same linkers or different linkers.
[0314] A linker may be bound to an antibody construct by a bond between the antibody construct and the linker. A linker may be bound to an anti-tumor antigen antibody construct by a bond between the anti-tumor antigen antibody construct and the linker. A linker may be bound to a terminus of an amino acid sequence of an antibody construct, or could be bound to a side chain modification to the antibody construct, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue. A linker may be bound to a terminus of an amino acid sequence of an Fc domain or Fc region of an antibody construct, or may be bound to a side chain modification of an Fc domain or Fc region of an antibody construct, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue. A linker may be bound to a terminus of an amino acid sequence of an Fc domain of an antibody construct, or may be bound to a side chain modification of an Fc domain of an antibody construct, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, a non-natural amino acid residue, or glutamic acid residue.
[0315] A linker may be bound to an antibody construct at a hinge cysteine. A linker may be bound to an antibody construct at a light chain constant domain lysine. A linker may be bound to an antibody construct at a heavy chain constant domain lysine. A linker may be bound to an antibody construct at an engineered cysteine in the light chain. A linker may be bound to an antibody construct at an Fc region lysine. A linker may be bound to an antibody construct at an Fc domain lysine. A linker may be bound to an antibody construct at an Fc region cysteine. A linker may be bound to an antibody construct at an Fc domain cysteine. A linker may be bound to an antibody construct at a light chain glutamine, such as an engineered glutamine. A linker may be bound to an antibody construct at a heavy chain glutamine, such as an engineered glutamine. A linker may be bound to an antibody construct at an unnatural amino acid engineered into the light chain. A linker may be bound to an antibody construct at an unnatural amino acid engineered into the heavy chain. Amino acids can be engineered into an amino acid sequence of an antibody construct, for example, a linker of a conjugate. Engineered amino acids may be added to a sequence of existing amino acids. Engineered amino acids may be substituted for one or more existing amino acids of a sequence of amino acids.
[0316] A linker may be conjugated to an antibody construct via a sulfhydryl group on the antibody construct. A linker may be conjugated to an antibody construct via a primary amine on the antibody construct. A linker may be conjugated to an antibody construct via a residue of an unnatural amino acid on an antibody construct, e.g., a ketone moiety.
[0317] When one or more linkers are bound to an antibody construct at the sites described herein, an Fc domain of the antibody construct can bind to Fc receptors. In certain embodiments, an antibody construct bound through a linker or an antibody construct bound to a linker bound to a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB) or (IIIC), or bound through a compound-linker of Formulas (IV A), (IVB) or (IVC), retains the ability of the Fc domain of the antibody to bind to Fc receptors. In certain embodiments, when a linker is connected to an antibody construct at the sites described herein, the antigen binding domain of an antibody construct bound to a linker or an antibody construct bound through a linker bound to a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB) or (IIIC), or bound through a compound-linker of Formulas (IV A), (IVB), or (IVC) can bind its antigen. In certain embodiments, when a linker is connected to an antibody construct at the sites described herein, a target binding domain of an antibody construct bound to a linker or an antibody construct bound to a linker bound to a compound or salt of any one of Formulas
(IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) can bind its antigen.
[0318] In certain embodiments, a compound or a compound-linker of any one of Formulas (IA),
(IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) disclosed herein is attached to an antibody Fc region or domain at an engineered cysteine residue. In some embodiments, the engineered cysteine residue is at one or more of positions HC S239C, LC V205C, LC A114C, HC A140C, LC K149C, LC S168C, LC S153C, LC A127C, HC T116C, and HC Sl 15C, where HC refers to heavy chain, LC refers t light chain and the numbering of amino acid residues in the Fc region is according to the EU index as in Kabat. In certain embodiments, a compound or a compound-linker of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) disclosed herein may not be attached to an amino acid residue of an IgG Fc domain disclosed herein selected from: 221, 222, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 246, 247, 249,
250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 278, 280,
281, 283, 285, 286, 288, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 302, 305, 313,
317, 318, 320, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335 336, 396,
428, or any subset thereof wherein numbering of amino acid residues in the Fc domain is according to the EU index as in Kabat.
Lysine-based Bioconjugation [0319] An antibody construct can be conjugated to a linker via lysine-based bioconjugation. An antibody construct can be exchanged into an appropriate buffer, for example, phosphate, borate, PBS, Tris-Acetate, Tris-Glycine, HEPES, MOPS, MES, EPS, HEPPS, Histidine, or HEPBS at a concentration of about 2 mg/mL to about 10 mg/mL. An appropriate number of equivalents of a compound-linker, e.g., a compound or salt of Formula (IV A), (IVB), or (IVC), described herein can be added as a solution with stirring. Dependent on the physical properties of the
benzazepine-linker construct, a co-solvent can be introduced prior to the addition of the benzazepine-linker construct to facilitate solubility. The reaction can be stirred at room temperature for 2 hours to about 12 hours depending on the observed reactivity. The progression of the reaction can be monitored by LC-MS. Once the reaction is deemed complete, the remaining benzazepine-linker constructs can be removed by applicable methods and the antibody construct- benzazepine conjugate can be exchanged into the desired formulation buffer. Lysine- linked conjugates can be synthesized starting with antibody (mAb) or bispecific antibody (bsAb) and benzazepine-linker construct, e.g., 10 equivalents, following Scheme A below (Conjugate = antibody construct-benzazepine conjugate). Monomer content and benzazepine-antibody construct ratios (molar ratios) can be determined by methods described herein.
Scheme A:
10 equivs of compound- linker
sodium phosphate
Conjugate
Figure imgf000151_0001
Cysteine-based Bioconjugation
[0320] An antibody construct can be conjugated to a linker via cysteine-based bioconjugation.
An antibody construct can be exchanged into an appropriate buffer, for example, phosphate, borate, PBS, Tris-Acetate, Tris-Glycine, HEPES, MOPS, MES, EPS, HEPPS, Histidine, or HEPBS at a concentration of about 2 mg/mL to about 10 mg/mL with an appropriate number of equivalents of a reducing agent, for example, dithiothreitol or tris(2-carboxyethyl)phosphine.
The resultant solution can be stirred for an appropriate amount of time and temperature to effect the desired reduction. A compound-linker, e.g., a compound or salt of Formula (IV A), (IVB), or (IVC), described herein can be added as a solution with stirring. Dependent on the physical properties of the benzazepine-linker construct, a co-solvent can be introduced prior to the addition of the benzazepine-linker construct to facilitate solubility. The reaction can be stirred at room temperature for about 1 hour to about 12 hours depending on the observed reactivity. The progression of the reaction can be monitored by liquid chromatography-mass spectrometry (LC- MS). Once the reaction is deemed complete, the remaining free benzazepine-linker construct can be removed by applicable methods and the antibody construct-benzazepine conjugate can be exchanged into the desired formulation buffer. Such cysteine-based conjugates can be synthesized starting with antibody (mAb) and benzazepine-linker construct, e.g., 7 equivalents, using the conditions described in Scheme B below (Conjugate = antibody construct-benzazepine compound conjugate). Monomer content and drug-antibody ratios can be determined by methods described herein.
Scheme B:
1. reducing agent
mAb - ► Conjugate
2. 7 equivs of compound-linker
sodium phosphate
pH = 8
20% v/v DMSO
Pharmaceutical Formulations
[0321] The compositions, conjugates and methods described herein can be considered useful as pharmaceutical compositions for administration to a subject in need thereof. Pharmaceutical compositions can comprise at least the compounds, salts or conjugates described herein and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersing agents, suspending agents, and/or thickening agents. The composition can comprise the conjugate having an antibody construct and a compound or salt of any one of Formulas (IA), (IB), (IC),
(IIA), (PB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) connected via a linker, as described herein. The composition can comprise the conjugate having an antibody construct, a target binding domain, and a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA),
(IIB), (IIC), (IIIA), (IIIB), (IIIC), (IVA), (IVB), or (IVC) connected via a linker. The
composition can comprise any conjugate described herein. The antibody construct can be an anti- CD40 antibody. A conjugate can comprise an anti-CD40 antibody and a benzazepine. A conjugate can comprise an anti-HER2 antibody and a benzazepine. A conjugate can comprise an anti-TROP2 antibody and a benzazepine. A pharmaceutical composition can comprise at least the compounds, salts or conjugates described herein and one or more of buffers, antibiotics, steroids, carbohydrates, drugs (e.g., chemotherapy drugs), radiation, polypeptides, chelators, adjuvants and/or preservatives.
[0322] Pharmaceutical compositions can be formulated using one or more physiologically- acceptable carriers comprising excipients and auxiliaries. Formulation can be modified depending upon the route of administration chosen. Pharmaceutical compositions comprising a compound, salt orconjugate as described herein can be manufactured, for example, by
lyophilizing the compound, salt or conjugate, mixing, dissolving, emulsifying, encapsulating or entrapping the conjugate. The pharmaceutical compositions can also include the compounds, salts or conjugates described herein in a free-base form or pharmaceutically-acceptable salt form.
[0323] Methods for formulation of the conjugates described herein can include formulating any of the compounds, salts or conjugates described herein with one or more inert, pharmaceutically- acceptable excipients or carriers to form a solid, semi-solid, or liquid composition. Solid compositions can include, for example, powders, tablets, dispersible granules and capsules, and in some aspects, the solid compositions further contain nontoxic, auxiliary substances, for example wetting or emulsifying agents, pH buffering agents, and other pharmaceutically- acceptable additives. Alternatively, the compounds, salts or conjugates described herein can be lyophilized or in powder form for re-constitution with a suitable vehicle, e.g., sterile pyrogen- free water, before use.
[0324] Pharmaceutical compositions described herein can comprise at least one active ingredient (e.g., a compound, salt or conjugate). The active ingredients can be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin microcapsules and poly-(methylmethacylate) microcapsules, respectively), in colloidal drug-delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions.
[0325] Pharmaceutical compositions as described herein often further can comprise more than one active compound (e.g., a compound, salt or conjugate and other agents) as necessary for the particular indication being treated. The active compounds can have complementary activities that do not adversely affect each other. For example, the composition can also comprise a
chemotherapeutic agent, cytotoxic agent, cytokine, growth-inhibitory agent, anti-hormonal agent, anti -angiogenic agent, and/or cardioprotectant. Such molecules can be present in combination in amounts that are effective for the purpose intended.
[0326] The compositions and formulations can be sterilized. Sterilization can be accomplished by filtration through sterile filtration.
[0327] The compositions described herein can be formulated for administration as an injection. Non-limiting examples of formulations for injection can include a sterile suspension, solution or emulsion in oily or aqueous vehicles. Suitable oily vehicles can include, but are not limited to, lipophilic solvents or vehicles such as fatty oils or synthetic fatty acid esters, or liposomes.
Aqueous injection suspensions can contain substances which increase the viscosity of the suspension. The suspension can also contain suitable stabilizers. Injections can be formulated for bolus injection or continuous infusion. Alternatively, the compositions described herein can be lyophilized or in powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
[0328] For parenteral administration, the compounds, salts or conjugates can be formulated in a unit dosage injectable form (e.g., use letter solution, suspension, emulsion) in association with a pharmaceutically acceptable parenteral vehicle. Such vehicles can be inherently non-toxic, and non-therapeutic. Vehicles can be water, saline, Ringer’s solution, dextrose solution, and 5% human serum albumin. Non-aqueous vehicles such as fixed oils and ethyl oleate can also be used. Liposomes can be used as carriers. The vehicle can contain minor amounts of additives such as substances that enhance isotonicity and chemical stability (e.g., buffers and
preservatives).
[0329] Sustained-release preparations can also be prepared. Examples of sustained-release preparations can include semipermeable matrices of solid hydrophobic polymers that can contain the compound, salt or conjugate, and these matrices can be in the form of shaped articles (e.g., films or microcapsules). Examples of sustained-release matrices can include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate), or poly (vinyl alcohol)), polylactides, copolymers of L-glutamic acid and g ethyl -L-glutamate, non-degradable ethylene- vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LLIPRON DEPO™ (i.e., injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly- D-( -)-3-hydroxybutyric acid.
[0330] Pharmaceutical formulations described herein can be prepared for storage by mixing a compound, salt or conjugate with a pharmaceutically acceptable carrier, excipient, and/or a stabilizer. This formulation can be a lyophilized formulation or an aqueous solution. Acceptable carriers, excipients, and/or stabilizers can be nontoxic to recipients at the dosages and
concentrations used. Acceptable carriers, excipients, and/or stabilizers can include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives, polypeptides; proteins, such as serum albumin or gelatin; hydrophilic polymers; amino acids; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes; and/or non ionic surfactants or polyethylene glycol.
[0331] Pharmaceutical formulations of the conjugates described herein may have an average drug- antibody construct ratio (“DAR”) selected from about 1 to about 10, wherein the drug is a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), (IIC), (IIIA), (MB),
(IIIC), (IV A), (IVB), or (IVC). In certain embodiments, the average DAR of the formulation is from about 2 to about 8, such as from about 3 to about 8, such as from about 3 to about 7. In certain embodiments, a pharmaceutical formulation has an average DAR of about 3, about 3.5, about 4, about 4.5, about 5, about 5.5, about 6, or about 6.6.
Therapeutic Applications
[0332] The compositions, conjugates and methods of the present disclosure can be useful for a plurality of different subjects including, but are not limited to, a mammal, human, non-human mammal, a domesticated animal (e.g., laboratory animals, household pets, or livestock), non- domesticated animal (e.g., wildlife), dog, cat, rodent, mouse, hamster, cow, bird, chicken, fish, pig, horse, goat, sheep, rabbit, and any combination thereof.
[0333] The compositions, conjugates and methods described herein can be useful as a therapeutic, for example, a treatment that can be administered to a subject in need thereof. A therapeutic effect of the present disclosure can be obtained in a subject by reduction,
suppression, remission, or eradication of a disease state, including, but not limited to, a symptom thereof. A therapeutic effect in a subject having a disease or condition, or pre-disposed to have or is beginning to have the disease or condition, can be obtained by a reduction, a suppression, a prevention, a remission, or an eradication of the condition or disease, or pre-condition or pre- disease state.
[0334] In practicing the methods described herein, therapeutically-effective amounts of the compositions, and conjugates described herein can be administered to a subject in need thereof, often for treating and/or preventing a condition or progression thereof. A pharmaceutical composition can affect the physiology of the subject, such as the immune system, an
inflammatory response, or other physiologic affect. A therapeutically-effective amount can vary depending on the severity of the disease, the age and relative health of the subject, the potency of the compounds used, and other factors.
[0335] Treat and/or treating can refer to any indicia of success in the treatment or amelioration of the disease or condition. Treating can include, for example, reducing, delaying or alleviating the severity of one or more symptoms of the disease or condition, or it can include reducing the frequency with which symptoms of a disease, defect, disorder, or adverse condition, and the like, are experienced by a patient. Treat can be used herein to refer to a method that results in some level of treatment or amelioration of the disease or condition, and can contemplate a range of results directed to that end, including but not restricted to prevention of the condition entirely.
[0336] Prevent, preventing and the like can refer to the prevention of the disease or condition, e.g., tumor formation, in the patient. For example, if an individual at risk of developing a tumor or other form of cancer is treated with the methods of the present disclosure and does not later develop the tumor or other form of cancer, then the disease has been prevented, at least over a period of time, in that individual. Preventing can also refer to preventing re-occurrence of a disease or condition in a patient that has previously been treated for the disease or condition, e.g., by preventing relapse.
[0337] A therapeutically effective amount can be the amount of a composition or an active component thereof sufficient to provide a beneficial effect or to otherwise reduce a detrimental non-beneficial event to the individual to whom the composition is administered. A
therapeutically effective dose can be a dose that produces one or more desired or desirable (e.g., beneficial) effects for which it is administered, such administration occurring one or more times over a given period of time. An exact dose can depend on the purpose of the treatment, and can be ascertainable by one skilled in the art using known techniques.
[0338] The conjugates described herein that can be used in therapy can be formulated and dosages established in a fashion consistent with good medical practice taking into account the disease or condition to be treated, the condition of the individual patient, the site of delivery of the composition, the method of administration and other factors known to practitioners. The compositions described herein can be prepared according to the description of preparation described herein.
[0339] Pharmaceutical compositions can be used in the methods described herein and can be administered to a subject in need thereof using a technique known to one of ordinary skill in the art which can be suitable as a therapy for the disease or condition affecting the subject. One of ordinary skill in the art would understand that the amount, duration and frequency of
administration of a pharmaceutical composition described herein to a subject in need thereof depends on several factors including, for example but not limited to, the health of the subject, the specific disease or condition of the patient, the grade or level of a specific disease or condition of the patient, the additional therapeutics the subject is being or has been administered, and the like.
[0340] The methods and compositions described herein can be for administration to a subject in need thereof. Often, administration of the compositions described herein can include routes of administration, non-limiting examples of administration routes include intravenous, intraarterial, subcutaneous, subdural, intramuscular, intracranial, intrastemal, intratumoral, or
intraperitoneally. Additionally, a pharmaceutical composition can be administered to a subject by additional routes of administration, for example, by inhalation, oral, dermal, intranasal, or intrathecal administration.
[0341] Compositions and conjugates of the present disclosure can be administered to a subject in need thereof in a first administration, and in one or more additional administrations. The one or more additional administrations can be administered to the subject in need thereof minutes, hours, days, weeks or months following the first administration. Any one of the additional administrations can be administered to the subject in need thereof less than 21 days, or less than 14 days, less than 10 days, less than 7 days, less than 4 days or less than 1 day after the first administration. The one or more administrations can occur more than once per day, more than once per week or more than once per month. The administrations can be weekly, biweekly (every two weeks), every three weeks, monthly or bimonthly.
[0342] The compositions, conjugates and methods provided herein can be useful for the treatment of a plurality of diseases, conditions, preventing a disease or a condition in a subject or other therapeutic applications for subjects in need thereof. Often the compositions, conjugates and methods provided herein can be useful for treatment of hyperplastic conditions, including but not limited to, neoplasms, cancers, tumors and the like. The compositions, conjugates and methods provided herein can be useful for specifically targeting TLR8. In one embodiment, the compounds of the present disclosure serve as TLR8 agonists and activate an immune response.
In another embodiment, the conjugates of the present disclosure serve as TLR8 agonists and activate an immune response. A condition, such as a cancer, can be associated with expression of a molecule on the cancer cells. Often, the molecule expressed by the cancer cells can comprise an extracellular portion capable of recognition by the antibody construct of the conjugate. A molecule expressed by the cancer cells can be a tumor antigen. An antibody construct portion of the conjugate can recognize a tumor antigen. A tumor antigen can include CD5, CD 19, CD20, CD25, CD37, CD30, CD33, CD40, CD45, CAMPATH-l, BCMA, CS-l, PD-L1, B7-H3, B7- DC, HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, folate-binding protein, A33, G250, prostate-specific membrane antigen (PMSA), ferritin, GD2, GD3, GM2,
Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein, tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, avB3, WT1, LMP2, HPV E6, HPV E7, Her-2/neu, MAGE A3, p53 nonmutant, NY-ESO-l, MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyronsinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin Bl, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, sLe(animal), CYP1B1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-l, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, VEGFR2, MAD-CT-l, PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MUC16, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, MUC15, CA6, NAPI2B, TROP2, CLDN6, CLDN16, CLDN18.2, CLorfl86, RON, LY6E, FRA, DLL3, PTK7, STRA6,
TMPRSS3, TMPRSS4, TMEM238, UPK1B, VTCN1, LIV1, ROR1, or Fos-related antigen 1.
[0343] As described herein, an antigen binding domain portion of the conjugate may be configured to recognize a molecule expressed by a cancer cell, such as for example, a disease antigen, tumor antigen or a cancer antigen. Often such antigens are known to those of ordinary skill in the art, or newly found to be associated with such a condition, to be commonly associated with, and/or, specific to, such conditions. For example, a disease antigen, tumor antigen or a cancer antigen is, but is not limited to, CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD40, CD45, CAMPATH-l, BCMA, CS-l, PD-L1, B7-H3, B7-DC, HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, folate-binding protein, A33, G250, prostate-specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRVIII (de2-7 EGFR), fibroblast activation protein, tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, avB3, WT1, LMP2, HPV E6, HPV E7, Her-2/neu, MAGE A3, p53 nonmutant, NY-ESO-l, Mel an A/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyronsinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin Bl, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, sLe(animal), CYP1B1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-l, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, VEGFR2, MAD- CT-l, PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MUC16, MAGE A4, MAGE C2, GAGE,
EGFR, CMET, HER3, MUC15, CA6, NAPI2B, TROP2, CLDN6, CLDN16, CLDN18.2, CLorfl86, RON, LY6E, FRA, DLL3, PTK7, STRA6, TMPRSS3, TMPRSS4, TMEM238, EIPK1B, VTCN1, LIV1, ROR1 , or Fos-related antigen 1. Additionally, such tumor antigens can be derived from the following specific conditions and/or families of conditions, including but not limited to, cancers such as brain cancers, skin cancers, lymphomas, sarcomas, lung cancer, liver cancer, leukemias, uterine cancer, breast cancer, ovarian cancer, cervical cancer, bladder cancer, kidney cancer, hemangiosarcomas, bone cancers, blood cancers, testicular cancer, prostate cancer, stomach cancer, intestinal cancers, pancreatic cancer, and other types of cancers as well as pre-cancerous conditions such as hyperplasia or the like.
[0344] Non-limiting examples of cancers can include Acute lymphoblastic leukemia (ALL); Acute myeloid leukemia; Adrenocortical carcinoma; Astrocytoma, childhood cerebellar or cerebral; Basal-cell carcinoma; Bladder cancer; Bone tumor, osteosarcoma/malignant fibrous histiocytoma; Brain cancer; Brain tumors, such as, cerebellar astrocytoma, malignant glioma, ependymoma, medulloblastoma, visual pathway and hypothalamic glioma; Brainstem glioma; Breast cancer; Bronchial adenomas/carcinoids; Burkitt's lymphoma; Cerebellar astrocytoma; Cervical cancer; Cholangiocarcinoma; Chondrosarcoma; Chronic lymphocytic leukemia;
Chronic myelogenous leukemia; Chronic myeloproliferative disorders; Colon cancer; Cutaneous T-cell lymphoma; Endometrial cancer; Ependymoma; Esophageal cancer; Eye cancers, such as, intraocular melanoma and retinoblastoma; Gallbladder cancer; Glioma; Hairy cell leukemia; Head and neck cancer; Heart cancer; Hepatocellular (liver) cancer; Hodgkin lymphoma;
Hypopharyngeal cancer; Islet cell carcinoma (endocrine pancreas); Kaposi sarcoma; Kidney cancer (renal cell cancer); Laryngeal cancer; Leukemia, such as, acute lymphoblastic, acute myeloid, chronic lymphocytic, chronic myelogenous and, hairy cell; Lip and oral cavity cancer; Liposarcoma; Lung cancer, such as, non-small cell and small cell; Lymphoma, such as, AIDS- related, Burkitt; Lymphoma, cutaneous T-Cell, Hodgkin and Non-Hodgkin, Macroglobulinemia, Malignant fibrous histiocytoma of bone/osteosarcoma; Melanoma; Merkel cell cancer;
Mesothelioma; Multiple myeloma/plasma cell neoplasm; Mycosis fungoides; Myelodysplastic syndromes; Myelodysplasti c/myeloproliferative diseases; Myeloproliferative disorders, chronic; Nasal cavity and paranasal sinus cancer; Nasopharyngeal carcinoma; Neuroblastoma;
Oligodendroglioma; Oropharyngeal cancer; Osteosarcoma/malignant fibrous histiocytoma of bone; Ovarian cancer; Pancreatic cancer; Parathyroid cancer; Pharyngeal cancer;
Pheochromocytoma; Pituitary adenoma; Plasma cell neoplasia; Pleuropulmonary blastoma; Prostate cancer; Rectal cancer; Renal cell carcinoma (kidney cancer); Renal pelvis and ureter, transitional cell cancer; Rhabdomyosarcoma; Salivary gland cancer; Sarcoma, Ewing family of tumors; Sarcoma, Kaposi; Sarcoma, soft tissue; Sarcoma, uterine; Sezary syndrome; Skin cancer (non-melanoma); Skin carcinoma; Small intestine cancer; Soft tissue sarcoma; Squamous cell carcinoma; Squamous neck cancer with occult primary, metastatic; Stomach cancer; Testicular cancer; Throat cancer; Thymoma and thymic carcinoma; Thymoma; Thyroid cancer; Thyroid cancer, childhood; Uterine cancer; Vaginal cancer; Waldenstrom macroglobulinemia; Wilms tumor and any combination thereof.
[0345] The invention provides any therapeutic compound or conjugate disclosed herein for use in a method of treatment of the human or animal body by therapy. Therapy may be by any mechanism disclosed herein, such as by stimulation of the immune system. The invention provides any therapeutic compound or conjugate disclosed herein for use in stimulation of the immune system, vaccination or immunotherapy, including for example enhancing an immune response. The invention further provides any therapeutic compound or conjugate disclosed herein for prevention or treatment of any condition disclosed herein, for example cancer, autoimmune disease, inflammation, sepsis, allergy, asthma, graft rejection, graft-versus-host disease, immunodeficiency or infectious disease (typically caused by an infectious pathogen).
The invention also provides any therapeutic compound or conjugate disclosed herein for obtaining any clinical outcome disclosed herein for any condition disclosed herein, such as reducing tumour cells in vivo. The invention also provides use of any therapeutic compound or conjugate disclosed herein in the manufacture of a medicament for preventing or treating any condition disclosed herein.
General Synthetic Schemes and Examples
[0346] The following synthetic schemes are provided for purposes of illustration, not limitation. The following examples illustrate the various methods of making compounds described herein.
It is understood that one skilled in the art may be able to make these compounds by similar methods or by combining other methods known to one skilled in the art. It is also understood that one skilled in the art would be able to make, in a similar manner as described below by using the appropriate starting materials and modifying the synthetic route as needed. In general, starting materials and reagents can be obtained from commercial vendors or synthesized according to sources known to those skilled in the art or prepared as described herein.
Scheme 1
Synthesis of C-8 Carboxamide
Figure imgf000160_0001
[0347] React an aldehyde (i) with an appropriately Wittig reagent, such as tert-butyl 3-cyano-2- (triphenylphosphorylidene)propanoate, at elevated temperatures to afford an olefin (ii), which undergoes reductive cyclization by treating the olefin (ii) with a reducing agent, such as iron powder in hot acetic acid, to afford azepines (iii). Deprotect the C-4 ester group by using a strong acid such as HC1 to give compounds (iv), which is in turn coupled with a substituted amine using a coupling agent, such as BOP reagent. Protect the 2-amino substituent of compounds (v) with a tert-butoxy carbonyl group. Hydrolyze the resulting compounds (vi) with reagents such as LiOH in a mixture of THF and methanol to afford compounds (vii). Convert the C-8 carboxylic acid of (vii) to the amide group using known reagents such as HBTU and a tertiary amine base. Acid-mediated deprotection of compounds (viii) using a reagent such as TFA in dichloromethane provides the target compounds (ix).
Scheme 2
Alternative Synthesis of C-8 Carboxamides
Figure imgf000161_0001
[0348] React (i) under standard conditions used for the carbonylation of aryl halides such as carbon monoxide, a palladium catalyst such as Pd(OAc)2 and a ligand such as 4,5- bis(diphenylphosphino)-9,9-dimethylxanthene (XantPhos) and a base such as potassium phosphate in a mixture of THF and water to provide carboxylic acids (ii). Conversion to final products can then be carried out in a manner similar to that described in Scheme 1 (vii ix).
Scheme 3
Synthesis of C-8 Amine Analogs
Figure imgf000162_0001
[0349] React an aldehyde (i) with an appropriate Wittig reagent, such as ethyl 3-cyano-2- (triphenylphosphorylidene)propanoate, at ambient temperature to afford an olefin (ii), which undergoes reductive cyclization by treating the olefin (ii) with a reducing agent, such as iron powder in hot acetic acid, to afford azepines (iii). Protect the C-2 amine group by using Boc anhydride to give compounds (iii), which is in turn saponified with an alkaline metal hydroxide such as LiOH to afford the carboxylic acid which is coupled with a substituted amine using a coupling agent, such as BOP reagent to provide compounds (iv). Convert the C-8 carboxylic acid of (v) to the amide group using known reagents such as EDCI / HOBT and a tertiary amine base. Halogen-amine exchange can be effected using standard methodology such as copper- mediated or palladium-catalyzed couplings (benzophenone imine / Pd(II)) to provide C-8 anilines (vi). Functionalization of amines (vi) by acylation or sulfonylation provides anilides (X=C) or sulfonamides (X=SO) compounds (vii). Alternatively, compounds (vii) can be prepared directly through a palladium -mediated coupling of bromide (v) and an appropriately substituted amide or sulfonamide. Acid-mediated deprotection of compounds (vii) using a reagent such as TFA in dichloromethane provides the target compounds (viii).
Scheme 4
Synthesis of C-8 Sulfur Analogs
Figure imgf000163_0001
[0350] React (i) with benzyl thiol in the presence of a palladium catalyst such as Pd2(dba)3 and a ligand such as XantPhos at elevated temperatures to provide C-8 sulfides (ii). Oxidative chlorination of sulfides (ii) with a reagent such as l,3-dichloro-5,5-dimethylhydantoin (DCDMH) affords intermediate sulfonyl chlorides which can be reacted with an appropriately substituted amine of structure R”’NH2 to provides sulfonamides (iii). Acid-mediated deprotection of compounds (iii) using a reagent such as TFA in dichloromethane provides the target compounds (iv).
Scheme 5
Synthesis of Linker-Payloads
[0351] A linker-payload (LP) can be synthesized by various methods. For example, LP compounds can be synthesized as shown in Scheme 5-1.
Scheme 5-1 :
Figure imgf000163_0002
2 DIC / ROH
ii
R = NHS, pentafluorophenyl
ISC: immune-stimulatory compound
[0352] A PEGylated carboxylic acid (i) that has been activated for amide bond formation can be reacted with an appropriately substituted amine containing immune-stimulatory compound to afford an intermediate amide. Formation of an activated ester (ii) can be achieved by reaction the intermediate amide-containing carboxylic using a reagent such as N-hydroxysuccinimide or pentafluorophenol in the presence of a coupling agent such as diisopropyl carbodiimide (DIC) to provide compounds (ii).
[0353] An LP can be ssynthesized as shown in Scheme 5-2.
Scheme 5-2:
Figure imgf000164_0001
[0354] An activated carbonate such as (i) can be reacted with an appropriately substituted amine containing immune-stimulatory compound to afford carbamates (ii) which can be deprotected using standard methods based on the nature of the R3 ester group. The resulting carboxylic acid (iii) can then by coupled with an activating agent such as N-hydroxysuccinimide or
pentafluorophenol to provide compounds (iv).
[0355] An LP compound can be synthesized as shown in Scheme 5-3.
Scheme 5-3:
Figure imgf000164_0002
i-a; X = NHS
i-b; X = H
ISC: immune-stimulatory compound
[0356] An activated carboxylic ester such as (i-a) can be reacted with an appropriately
substituted amine containing immune-modstimulatory compound to afford amides (ii).
Alternatively, carboxylic acids of type (i-b) can be coupled to an appropriately substituted amine containing immune-stimulatory compound in the presence of an amide bond forming agent such as dicyclohexycarbodiimde (DCC) to provide the desired LP. [0357] An LP compound can be synthesized by various methods such as that shown in Scheme 5-4.
Scheme 5-4:
Figure imgf000165_0001
ISC: immune-stimulatory compound
[0358] An activated carbonate such as (i) can be reacted with an appropriately substituted amine containing immune-modstimulatory compound to afford carbamates (ii) as the target ISC.
[0359] An LP compound can also be synthesized as shown in Scheme 5-5.
Scheme 5-5:
Figure imgf000165_0002
[0360] An activated carboxylic acid such as (i-a, i-b, i-c) can be reacted with an appropriately substituted amine containing immune-stimulatory compound to afford amides (ii-a, ii-b, ii-c) as the target linkered payloads (LPs).
EXAMPLE 1
Synthesis of antibody benzazepine conjugates
[0361] Protocol for the preparation of Antibody Benzazepine Conjugate
[0362] The mAb (155 mg, 35.57 mL, 4.38 mg/mL in PBS) is exchanged into HEPES (100 mM, pH 7.0, 1 mM DTP A) via molecular weight cut-off centrifugal filtration (Millipore, 30 kDa). The resultant mAb solution is transferred to a 50 mL conical tube (31.873 mL by weight). The mAb concentration is determined to be 4.39 mg/mL by A2xo (total mAb content was 140 mg, 3 mg was removed for analytical applications). To the mAb solution is added TCEP (2.0 eq, 1.85 mL, 1 mM stock) at room temperature and the resultant mixture was incubated at 37 °C for 1 hr, with gentle shaking. Upon being cooled to room temperature, a stir bar is added to the reaction tube. With stirring, DMA (10% v/v, 3.0 mL) is added dropwise to the reaction mixture. A compound- linker is added dropwise and the resultant reaction mixture and allowed to stir at ambient temperature for 30 minutes, at which point N-ethyl maleimide (3.0 eq, 100 mM DMA, 28 pL) is added. After an additional 15 minutes of stirring, cysteine (6.0 equiv., 50 mM HEPES, 111 pL) is added. The crude conjugate is then exchanged into PBS and purified by preparative SEC (HiLoad 26/600, Superdex 200pg) using PBS as the mobile phase. The pure fractions are concentrated via molecular weight cut-off centrifugal filtration (Millipore, 30 kDa), sterile filtered and transferred to 15 mL conical tubes. Drug-antibody construct ratios (molar ratios) are determined by methods described in Example 2.
EXAMPLE 2
General Procedure for the Determination of the Drug- Antibody-Ratios Hydrophobic Interaction Chromatography
[0363] 10 pL of a 6 mg/mL solution of the conjugate is injected into an HPLC system set-up with a TOSOH TSKgel Butyl-NPR TM hydrophobic interaction chromatography (HIC) column (2.5 pM particle size, 4.6 mm x 35 mm) attached. Then, over the course of 18 minutes, a method is run in which the mobile phase gradient ran from 100% mobile phase A to 100% mobile phase B over the course of 12 minutes, followed by a six-minute re-equilibration at 100% mobile phase A. The flow rate is 0.8 mL/min and the detector is set at 280 nM. Mobile phase A is 1.5 M ammonium sulfate, 25 mM sodium phosphate (pH 7). Mobile phase B is 25% isopropanol in 25 mM sodium phosphate (pH 7). Post-run, the chromatogram was integrated and the molar ratio is determined by summing the weighted peak area.
Mass Spectrometry
[0364] One microgram of conjugate is injected into an LC/MS such as an Agilent 6550 iFunnel Q-TOF equipped with an Agilent Dual Jet Stream ESI source coupled with Agilent 1290 Infinity UHPLC system. Raw data is obtained and is deconvoluted with software such as Agilent MassHunter Qualitative Analysis Software with BioConfirm using the Maximum Entropy deconvolution algorithm. The average mass of intact antibody construct immune- stimulatorystimulatorystimulatory compound conjugates is calculated by the software, which used top peak height at 25% for the calculation. This data is then imported into another program to calculate the molar ratio of the conjugate such as Agilent molar ratio calculator.
EXAMPLE 3
TLR7/TLR8 Reporter Assays
Materials and general procedures
[0365] Reporter cells lines (namely HEK-Blue Nulll, HEK-Blue hTLR7 and HEK-Blue hTLR8) are obtained from InvivoGen. Cells are passed / expanded / stored in liquid nitrogen per supplier’s instruction. The cells are typically split twice per week in the growth media of DMEM supplemented with 10% fetal bovine serum, IX NEAA, lmM Pyruvate, 2mM glutamine, 50 pg/mL penicillin, 50 U/mL streptomycin (each from Gibco) in the presence of the following antibiotics, as shown in Table 1:
Table 1: Reporter cell lines treated with antibiotics
Figure imgf000167_0001
General procedure for in vitro small molecule screening
[0366] Test samples (at desired concentrations diluted in DMEM) are added to the 96-well assay plate, 20 pL per well. Reporter cells are harvested from the tissue culture flasks by incubation in PBS at 37 °C for two minutes after the media in the flask is removed and cells rinsed with PBS. Cells are counted and diluted in the HEK-Blue Detection media at 0.22 x 106 cells/mL. Then 180 pL of cells are added to the assay plate containing the 20 pL of test samples, and incubated for 17 hours at 37 °C in a 5% C02 humidified incubator. Optical density at 640 nm is analyzed using an Envision (Perkin Elmer) plate reader.
EXAMPLE 4
PBMC Screening Assay
Materials and general procedures
[0367] Human peripheral blood mononuclear cells (PBMC) are obtained from BenTek, frozen at 25 x 106 cell/mL in 10% DMSO (Sigma) prepared in fetal bovine serum (Gibco) and stored in liquid nitrogen. For culture PBMC are thawed quickly in a 37 °C water bath and diluted into pre- warmed RPMI 1640 (Lonza) supplemented with 10 % fetal bovine serum, 2 mM glutamine, 50 pg/mL penicillin, 50 U/mL streptomycin (all from Gibco) and centrifuged for 5 minutes at 500 x g. PBMC are suspended into the growth media described above and cultured at a concentration of 1 x 106 cells per mL at 37 °C in a 5% C02 incubator.
General procedure for in vitro small molecule screening
[0368] PBMC are thawed, suspended at a concentration of 1 x 106 cell/mL in growth media and 200 pL is aliquoted into each well of a 96-well plate for a total of 0.2 x 106 cells per well. PBMC are incubated for 16-18 hours at 37 °C in a 5% C02 humidified incubator. The following day PBMC plates are centrifuged at 500 x g for 5 minutes and the growth media is removed. 150 pL of twelve concentrations ranging from 1000 to 0.000238 nM of small molecules prepared in growth media are added to PBMC in duplicate and incubated for 24 hours at 37 °C in a 5% C02 incubator. Prior to supernatant harvest, cells are spun at 500 x g for 5 minutes to remove cell debris. TNF-a activity is assessed in the supernatant by ELISA (eBioscience) or HTRF (CisBio) per the manufacturer’s instructions. Optical density at 450 nm and 570 nm (ELISA) or luminescence (HTRF) is analyzed using an Envision (Perkin Elmer) plate reader.
EXAMPLE 5
Measuring TNFa Production by PBMCs
[0369] PBMCs are isolated from human blood as described above. Briefly, PBMCs are isolated by Ficoll gradient centrifugation, resuspended in RPMI, and plated in 96-well flat bottom microtiter plates (125, 000/well). Antigen-expressing tumor cells are then added (25,000/well) along with titrating concentrations of conjugates or unconjugated parental antibodies as controls. After overnight culture, supernatants are harvested, and TNFa levels are determined by
AlphaLISA. EXAMPLE 6
Measuring TNFa Production by Monocytes
[0370] Monocytes are prepared as follows: PBMCs are isolated from normal human blood using Ficoll purification, and monocytes are enriched from the PBMCs using Stem Cell Technologies Human Monocyte without CD 16 Depletion Negative Selection Kits according to the manufacturer’s instructions. Monocytes are then slowly frozen and stored in liquid nitrogen. Prior to assay monocytes are thawed and rested overnight at 37°C in 5% C02 in assay media (RPMI 1640 media supplemented with 10% FBS, 50 pg/mL Penicillin, 50 U/mL Streptomycin, lmM HEPES, IX non-essential amino acids, O.lmM sodium pyruvate). The following day, monocytes are plated at 4 x 104 cell/well with or without tumor cells at 4 x 104 cell/well in 96- well flat bottom plates in assay media described above. After overnight culture, supernatants are harvested, and TNFa levels are determined by AlphaLISA.
EXAMPLE 7
Measuring TNFa Production by Macrophages
[0371] Macrophages are generated as follows: Monocytes are isolated from human peripheral blood mononuclear cells (PBMCs) using magnetic bead-based negative selection and cultured 7 days in the presence of GM-CSF to generate macrophages. Macrophages are plated in 96-well flat bottom microtiter plates (40,000/well). Antigen-expressing tumor cells are then added (40,000/well) along with titrating concentrations of conjugates or unconjugated parental antibodies as controls. After overnight culture, supernatants are harvested, and TNFa levels are determined by AlphaLISA.
EXAMPLE 8
Antigen Dependent Monocyte Activation by a Conjugates in a Mouse Model
[0372] Human monocytes are prepared as described above. Trop2+ MDA-MB-453 cells or Trop2 MiaPaca2 are cultured by standard methods. TROP2-TLR8 benzazepine conjugates are prepared by conjugation of a compound-linker to a TROP2 antibody (sacituzumab) to form a conjugate with an average DAR of about 4.
[0373] On Day 1 NOD SCID mice are injected with 5xl05 Trop2+ MDA-MB-453 cells or Trop2 MiaPaca2 cells. Seven to ten days later when the tumor size reached about 300 mm3, the mice are injected intraveneously with 5-20 mg/kg of the TROP2-TLR8 benzazepine conjugate or TROP2 antibody alone. Sixteen hours later, 106 human monocytes are injected intratum orally. Four to six hours later, the tumors are harvested and digested. Monocytes are identified by staining for cell surface markers and monocyte activation is determined by intracellular staining for IL-6.
EXAMPLE 9
Murine TNFa Production by Murine Macrophages was Induced by TLR7 Small Molecules
[0374] Murine macrophages were derived from bone marrow cells from a Balb/c mouse.
Briefly, murine bone marrow cells were isolated and resuspended in cDMEM media (high glucose DMEM (HyClone) supplemented with 10 % fetal bovine serum (HyClone), 1 mM Sodium Pyruvate (HyClone), 2 mM L-glutamine (Gibco), 10 mM HEPES (HyClone), IX NEAA (Gibco), 50 U/mL penicillin, 50 U/mL streptomycin (HyClone)) and seeded at 5e5 cells/mL in the presence of 20ng/mL murine M-CSF (Peprotech) and incubated at 37 °C, 5% C02 for 7 days to differentiate into murine macrophages.
General procedure for in vitro small molecule screening
[0375] This example shows that TLR7 small molecules administered with and without amine masking groups can increase production of a pro-inflammatory cytokine, murine TNFa, from bone marrow-derived murine macrophages.
[0376] After differentiation, bone marrow-derived murine macrophages were plated in 96-well flat bottom microtiter plates (80,000/well) along with titrating concentrations of TLR7 small molecules (Table 2) ranging from 1000 to 0.06 nM in cRPMI media (RPMI-1640 (HyClone) supplemented with 10 % fetal bovine serum (HyClone), 1 mM Sodium Pyruvate (HyClone), 2 mM L-glutamine (Gibco), 10 mM HEPES (HyClone), IX NEAA (Gibco), 50 U/ L penicillin,
50 U/mL streptomycin (HyClone)). After overnight culture, supernatants were harvested and murine TNFa levels were determined by ELISA (BioLegend). Table 3. Data was analyzed using GraphPad Prism 7.01 software (GraphPad Software) and ECso values calculated using non-linear regression (FIG. 1). The TLR7 small molecules were active, stimulating production of murine TNFa in a dose-dependent manner from the murine macrophages. Compound A is represented by
Figure imgf000170_0001
the fomula: Table 2.
Figure imgf000171_0001
Figure imgf000172_0001
Figure imgf000173_0001
Figure imgf000174_0002
Table 3.
Figure imgf000174_0001

Claims

WHAT IS CLAIMED IS:
1. A compound represented by the structure of Formula (IIA):
Figure imgf000175_0001
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L10 is -X10-;
L2 is selected from -X2-, -X2-CI-6 alkylene-X2-, -X2-C2-6 alkenyl ene-X2-, and - X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R12;
X10 is selected from -C(O)-, and -C(0)N(R10)-*, wherein * represents where X10 is bound to R5;
X2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R10)-, -C(O)-,
-C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)
-C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(O), -0S(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-,
and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R5 is selected from unsaturated C4-8 carbocycle; bicyclic carbocycle; and fused 5- 5, fused 5-6, and fused 6-6 bicyclic heterocycle, wherein R5 is optionally substituted and wherein substituents are independently selected at each occurrence from: halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R5 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from hydrogen, -NH2, - C(0)0CH2C6H5; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, C MO alkyl, -Ci-io haloalkyl, -O-Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl; and wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
2. The compound or salt of claim 1, wherein the compound of Formula (IIA) is represented by Formula (IIB):
Figure imgf000177_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen, - OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-10 alkyl, C2-io alkenyl, and C2-io alkynyl; and
R24, and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, - N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-10 alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
3. The compound or salt of claim 2, wherein R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -OH, -OR10, -N02, -CN, and Ci-10 alkyl.
4. The compound or salt of claim 3, wherein R20, R21, R22, and R23 are each hydrogen.
5. The compound or salt of claim 3, wherein R21 is halogen.
6. The compound or salt of claim 3, wherein R21 is hydrogen.
7. The compound or salt of claim 3, wherein R21 is -OR10.
8. The compound or salt of claim 7, wherein R21 is -OCH3.
9. The compound or salt of any one of claims 2 to 8, wherein R24 and R25 are independently selected from hydrogen, halogen, -OH, -NO2, -CN, and C1-10 alkyl, or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
10. The compound or salt of claim 9, wherein R24 and R25 are each hydrogen.
11. The compound or salt of claim 9, wherein R24 and R25 taken together form an optionally substituted saturated C3-5 carbocycle, wherein substituents are selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle.
12. The compound or salt of any one of claims 1 to 11, wherein R2 is an
enzymatically-cleavable group.
13. The compound or salt of any one of claims 1 to 12, wherein R2 is represented by the formula:
Figure imgf000178_0001
wherein:
R101 is selected from an amino acid, a peptide, -0-(Ci-C6 alkyl) and -Ci-C6 alkyl, wherein alkyl of-0-(Ci-C6 alkyl) and -Ci-C6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N02, -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle; and
R100 is C(=0), wherein when R101 is selected from an amino acid or peptide R100 is the C- terminus of the amino acid or peptide.
14. The compound or salt of claim 13, wherein R101 is a peptide selected from a dipeptide, tripeptide and tetrapeptide.
15. The compound or salt of any one of claims 1 to 14, wherein L10 is selected from - C(0)N(R10)-*.
16. The compound or salt of claim 15, wherein R10 of -C(0)N(R10)-* is selected from hydrogen and Ci-6 alkyl.
17. The compound or salt of claim 16, wherein L10 is -C(0)NH-*.
18. The compound or salt of any one of claims 11 to 17, wherein R5 is an optionally substituted bicyclic carbocycle.
19. The compound or salt of claim 18, wherein R5 is an optionally substituted 8- to 12- membered bicyclic carbocycle.
20. The compound or salt of claim 19, wherein R5 is an optionally substituted 8- to 12- membered bicyclic carbocycle substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0,
=S, -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
21. The compound or salt of claim 20, wherein R5 is an optionally substituted 8- to 12- membered bicyclic carbocycle substituted with one or more substituents independently selected from -OR10, -N(R10)2, and =0.
22. The compound or salt of claim 21, wherein R5 is an optionally substituted indane, and optionally substituted tetrahydronaphthalene.
23. The compound or salt of claim 22, wherein R5 is selected from:
Figure imgf000179_0001
and
Figure imgf000179_0002
, any one of which is optionally substituted.
24. The compound or salt of claim 23, wherein R5 is selected from:
Figure imgf000179_0003
25. The compound or salt of any one of claims 1 to 17, wherein R5 is an optionally substituted unsaturated C4-8 carbocycle.
26. The compound or salt of claim 25, wherein R5 is an optionally substituted unsaturated C4-6 carbocycle.
27. The compound or salt of claim 26, wherein R5 is an optionally substituted unsaturated C4-6 carbocycle with one or more substituents independently selected from optionally substituted C3-12 carbocycle, and optionally substituted 3- to l2-membered heterocycle.
28. The compound or salt of claim 27, wherein R5 is an optionally substituted unsaturated C4-6 carbocycle with one or more substituents independently selected from optionally substituted phenyl, optionally substituted 3- to 12- heterocycle, optionally substituted Ci-io alkyl, optionally substituted C2-10 alkenyl, and halogen.
29. The compound or salt of any one of claims 1 to 17, wherein R5 is selected from an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle.
30. The compound or salt of claim 29, wherein R5 is an optionally substituted fused 5- 5, fused 5-6, and fused 6-6 bicyclic heterocycle with one or more substituents independently selected from -C(0)OR10, -N(R10)2, -OR10, and optionally substituted Ci-io alkyl.
31. The compound or salt of claim 30, wherein R5 is an optionally substituted fused 5- 5, fused 5-6, and fused 6-6 bicyclic heterocycle substituted with -C(0)OR10.
32. The compound or salt of claim 31, wherein R5 is an optionally substituted fused 6- 6 bicyclic heterocycle.
33. The compound or salt of claim 32, wherein the fused 6-6 bicyclic heterocycle is an optionally substituted pyridine-piperidine.
34. The compound or salt of claim 33, wherein L10 is bound to a carbon atom of the pyridine of the fused pyridine-piperidine.
35. The compound or salt of claim 29, wherein R5 is selected from
tetrahydroquinoline, tetrahydroisoquinoline, tetrahydronaphthyridine, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is optionally substituted.
36. The compound or salt of claim 35, wherein R5 is an optionally substituted tetrahy dronaphthyri dine .
37. The compound or salt of claim 29, wherein R5 is selected from:
Figure imgf000180_0001
The compound or salt of any one of claims 1 to 37, wherein when R5 is substituted, substituents on R5 are independently selected at each occurrence from: halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2- io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
39. The compound or salt of claim 38, wherein substituents on R5 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3 -i2 carbocycle, and 3- to l2-membered heterocycle.
40. The compound or salt of claim 39, wherein substituents on R5 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, and -CN; and Ci-io alkyl optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(0)OR10, -N02, =0, and -CN.
41. The compound or salt of any one of claims 1 to 40, wherein R5 is not substituted.
42. The compound or salt of any one of claims 1 to 41, wherein L2 is selected from -
C(O)-, and -C(0)NR10-.
43. The compound or salt of claim 42, wherein L2 is -C(O)-.
44. The compound or salt of claim 42, wherein L2 is selected from -C(0)NR10-.
45. The compound or salt of claim 44, wherein R10 of -C(0)NR10- is selected from hydrogen and Ci-6 alkyl.
46. The compound or salt of claim 45, wherein L2 is -C(0)NH-.
47. The compound or salt of any one of claims 1 to 46, wherein R4 is selected from: - OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN,
C3-12 carbocycle, and 3- to l2-membered heterocycle; and C3-12 carbocycle and 3- to 12- membered, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -
N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
48. The compound or salt of claim 47, wherein R4 is selected from: -OR10, and - N(R10)2; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, - OC(0)R10, -NO2, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-10 alkenyl, and C2-10 alkynyl.
49. The compound or salt of claim 48, wherein R4 is -N(R10)2.
50. The compound or salt of claim 49, wherein R10 of -N(R10)2 is independently selected at each occurrence from optionally substituted Ci-6 alkyl.
51. The compound or salt of claim 50, wherein R10 of -N(R10)2 is independently selected at each occurrence from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted.
52. The compound or salt of claim 51, wherein
Figure imgf000182_0001
53. The compound or salt of claim 52, wherein -
Figure imgf000182_0002
54. The compound or salt of any one of claims 1 to 53, wherein R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10,
-SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -NO2, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, Ci- 6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
55. The compound or salt of claims 54, wherein R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), and -CN; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle.
56. The compound or salt of claim 1, wherein the compound is selected from:
Figure imgf000183_0001
Figure imgf000184_0001
Figure imgf000185_0001
Figure imgf000185_0002
and a salt of any one thereof.
57 A compound represented by the structure of Formula (IIIA):
Figure imgf000185_0003
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L11 is -X11-;
L2 is selected from -X2-, -X2-CI-6 alkylene-X2-, -X2-C2-6 alkenyl ene-X2-, and - X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkyl ene, alkenylene or alkynylene with one or more R12;
X11 is selected from -C(O)- and -C(0)N(R10)-*, wherein * represents where X11 is bound to R6;
X2 at each occurrence is independently selected from a bond, -O- , -S-, -N(R10)-, -C(O)-,
-C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-,
-C(O)N(R10)C(O)N(R10)-, -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(O)-, -0S(0)2-, -S(0)20- , -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-,
-N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10,
-SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R6 is selected from phenyl and 5- or 6- membered heteroaryl, any one of which is substituted with one or more substituents selected from R7 and R6 is further optionally substituted by one or more additional substituents independently selected from R12;
R7 is selected from -C(0)NHNH2, -C(0)NH-Ci-3 alkylene-NH(R10), -C(0)CH3, - Ci-3 alkyl ene-NHC(0)0R11 , -Ci-3alkylene-NHC(0)R10, -Ci-3alkylene-NHC(0)NHR10, -Ci- 3alkylene-NHC(0)-Ci-3alkylene-R10, and a 3- to l2-membered heterocycle optionally substituted with one or more substituents independently selected from R12;
R10 is independently selected at each occurrence from hydrogen, -Nth, - C(0)0CH2C6H5; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, -CI-IO alkyl, -Ci-io haloalkyl, -O-Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to l2-membered heterocycle;
R11 is selected from C3-12 carbocycle and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from R12;
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(0)N(R10)2, -N(R10)C(0)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
58. The compound or salt of claim 57, wherein the compound of Formula (IIIA) is represented by Formula (IIIB):
Figure imgf000187_0001
or a pharmaceutically acceptable salt thereof, wherein: R20, R21, R22, and R23 are independently selected from hydrogen, halogen,
-OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; and
R24, and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, - N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
59. The compound or salt of claim 58, wherein R20, R21, R22, and R23 are
independently selected from hydrogen, halogen, -OH, -N02, -CN, and C1-10 alkyl.
60. The compound or salt of claim 59, wherein R20, R21, R22, and R23 are each hydrogen.
61. The compound or salt of any one of claims 58 to 60, wherein R24 and R25 are independently selected from hydrogen, halogen, -OH, -N02, -CN, and C1-10 alkyl, or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
62. The compound or salt of claim 61, wherein R24 and R25 are each hydrogen.
63. The compound or salt of claim 61, wherein R24 and R25 taken together form an optionally substituted saturated C3-5 carbocycle.
64. The compound or salt of any one of claims 57 to 63, wherein R2 is represented by the formula:
Figure imgf000188_0001
wherein:
R101 is selected from an amino acid, a peptide, -0-(Ci-C6 alkyl) and -Ci-C6 alkyl, wherein alkyl of -0-(Ci-C6 alkyl) and -Ci-C6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N02, -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle; and
R100 is C(=0), wherein when R101 is selected from an amino acid or peptide R100 is the C- terminus of the amino acid or peptide.
65. The compound or salt of claim 64, wherein R101 is a peptide selected from a dipeptide, tripeptide and tetrapeptide.
66. The compound or salt of any one of claims 57 to 65, wherein L11 is selected from -C(0)N(R10)- *.
67. The compound or salt of claim 66, wherein R10 of -C(0)N(R10)-* is selected from hydrogen and Ci-6 alkyl.
68. The compound or salt of claim 67, wherein L11 is -C(0)NH-*.
69. The compound or salt of claim of any one of claims 57 to 68, wherein R6 is phenyl substituted with R7 and R6 is further optionally substituted with one or more additional substituents independently selected from R12.
70. The compound or salt of claim 69, wherein R6 is selected from phenyl substituted with one or more substituents independently selected from -C(0)NHNH2, -C(0)NH-Ci- 3alkylene-NH(R10), -Ci-3alkylene-NHC(0)R10, and -C(0)CH3; and 3- to l2-membered
heterocycle, which is optionally substituted with one or more substituents selected from -OH, - N(R10)2, -NHC(0)(R10), -NHC(0)0(R10), -NHC(O)N(R10)2, -C(0)R10, -C(O)N(R10)2, - C(0)2R10, and -Ci-3alkylene-(R10) and R6 is further optionally substituted with one or more additional substituents independently selected from R12.
71. The compound or salt of claim 70, wherein R6 is selected from:
Figure imgf000189_0001
72. The compound or salt of any one of claims 57 to 68, wherein R6 is selected from a 5- and 6-membered heteroaryl substituted with one or more substituents independently selected from R7, and R6 is further optionally substituted with one or more additional substituents selected from R12.
73. The compound or salt of claim 72, wherein R6 is selected from 5- and 6- membered heteroaryl substituted with one or more substituents independently selected from - C(0)CH3, -Ci-3alkylene-NHC(0)OR10, -Ci-3 alkyl ene-NHC(0)R10, -Ci-3alkylene-NHC(0)NHR10, and -Ci-3alkylene-NHC(0) -Ci-3alkylene-(R10); and 3- to l2-membered heterocycle, which is optionally substituted with one or more substituents selected from -OH, -N(R10)2, -NHC(0)(R10), -NHC(0)0(R10), -NHC(O)N(R10)2, -C(0)R10, -C(O)N(R10)2, -C(0)2R10, and -Ci-3alkylene-(R10), and R6 is optionally further substituted with one or more additional substituents independently selected from R12.
74. The compound or salt of claim 73, wherein R6 is selected from substituted pyridine, pyrazine, pyrimidine, pyridazine, furan, pyran, oxazole, thiazole, imidazole, pyrazole, oxadiazole, oxathiazole, and triazole, and R6 is optionally further substituted with one or more additional substituents independently selected from R12.
75. The compound or salt of claim 74, wherein R6 is substituted pyridine and R6 is optionally further substituted with one or more additional substituents independently selected from R12.
76. The compound or salt of claim 75, wherein R6 is represented as follows:
Figure imgf000190_0001
77. The compound or salt of claim 75, wherein R6 is substituted pyridine, and wherein R7 is -Ci-3alkylene-NHC(0)-Ci-3alkylene-R1°.
78. The compound or salt of any one of claims 73 to 77, wherein R7 is -Cialkylene- NHC(0)-C ialkylene-R10.
79. The compound or salt of claim 78, wherein R7 is -Cialkylene-NHC(O)- Cialkylene-NTh.
80. The compound or salt of claim 72, wherein R6 is selected from:
Figure imgf000190_0002
Figure imgf000191_0001
81. The compound or salt of claim 80, wherein
Figure imgf000191_0002
82. The compound or salt of any one of claims 57 to 81, wherein L2 is selected from - C(O)-, and -C(0)NR10-.
83. The compound or salt of claim 82, wherein L2 is selected from -C(0)NR10-.
84. The compound or salt of claim 83, wherein R10 of -C(0)NR10- is selected from hydrogen and Ci-6 alkyl.
85. The compound or salt of claim 84, wherein L2 is -C(0)NH-.
86. The compound or salt of claim 82, wherein L2 is -C(O)-.
87. The compound or salt of any one of claims 57 to 86, wherein R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle and 3- to 12- membered, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
88. The compound or salt of claim 87, wherein R4 is selected from: -OR10 and - N(R10)2; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle and 3- to l2-membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10 - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2- io alkynyl.
89. The compound or salt of claim 88, wherein R4 is -N(R10)2.
90. The compound or salt of claim 89, wherein R10 of -N(R10)2 is independently selected at each occurrence from optionally substituted Ci-6 alkyl.
91. The compound or salt of claim 90, wherein R10 of -N(R10)2 is independently selected at each occurrence from methyl, ethyl, propyl, and butyl, any of which are optionally substituted.
92. The compound or salt of claim 91, wherein
Figure imgf000192_0001
93. The compound or salt of claim 92, wherein -
Figure imgf000192_0002
94. The compound or salt of any one of claims 57 to 93, wherein R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3. io carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
95. The compound or salt of claims 94, wherein R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), and -CN; C1-10 alkyl, C2-io alkenyl, C2-io alkynyl, each of which is independently optionally substituted at each occurrence with one or more substituents selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, - OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3. 10 carbocycle and 3- to lO-membered heterocycle.
96. The compound or salt of claim 57, wherein the compound is selected from:
Figure imgf000193_0001
Figure imgf000194_0001
Figure imgf000195_0001
Figure imgf000196_0001
salt of any one thereof.
97. A compound represented by the structure of Formula (IA):
Figure imgf000196_0002
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L1 is selected from -X1-, -X2-CI-6 alkyl ene-X2-Ci-6 alkylene-, -X2-C2-6 alkenylene- X2-, and -X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkylene, alkenylene or alkynylene with one or more R12;
L2 is selected from -X2-, -X2-CI-6 alkylene-X2-, -X2-C2-6 alkenyl ene-X2-, and -X2-C2-6 alkynylene-X2-, each of which is optionally substituted on alkylene, alkenylene or alkynylene with one or more R12;
X1 is selected from -S-*, -N(R10)-*, -C(0)0-*, -OC(O)-*, -0C(0)0-*, - C(O)N(R10)C(O)- *, -C(O)N(R10)C(O)N(R10)*, -N(R10)C(O)-*, -CR10 2N(R10)C
(O)-*, -N(R10)C(O)N(R10)-*, -N(R10)C(O)O-*, -OC(0)N(R10)-*, -C(NR10)-*, -N(R10)C(NR10)-*, -C(NR10)N(R10)-*, -N(R10)C(NR10)N(R10)-*, -S(0)2-*, -0S(0)-*, -S(0)0-*, -S(O), -0S(0)2- *, -S(0)20*, -N(R10)S(O)2-*, -S(0)2N(R10)-*, -N(R10)S(O)-*, -S(0)N(R10)-*, -N(R10)S(0)2N(R1 °)-*, and -N(R10)S(O)N(R10)-*, wherein * represents where X1 is bound to R3;
X2 is independently selected at each occurrence from -O- , -S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-, -C(O)N(R10)C(O)N(R10), -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -OS(O)-, -S(0)0-, -S(0), -0S(0)2-, -S(0)20, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R3 is selected from optionally substituted C3-0 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents on R3 are independently selected at each occurrence from: halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), and -CN; C1-10 alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, - N(R10)C(O)R10,
-N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to 12- membered heterocycle, wherein each C3-0 carbocycle, and 3- to l2-membered heterocycle in R3 is optionally substituted with one or more substituents independently selected from halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R4 is selected from: -OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, - S(0)R10, and -S(0)2R10; Ci -lo alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10,
-OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-0 carbocycle, and 3- to l2-membered heterocycle; and C3-0 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-0 carbocycle, and 3- to l2-membered heterocycle in R4 is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl;
R10 is independently selected at each occurrence from: hydrogen, -NH2, - C(0)0CH2C6H5; and C1-10 alkyl, C2-io alkenyl, C2-io alkynyl, C3-0 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -N02, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
98. The compound or salt of claim 97, wherein the compound of Formula (IA) is represented by Formula (IB):
Figure imgf000198_0001
or a pharmaceutically acceptable salt thereof, wherein:
R20, R21, R22, and R23 are independently selected from hydrogen, halogen, -
OR10, -SR10,
-N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, Ci-10 alkyl, C2-10 alkenyl, and C2-10 alkynyl; and
R24 and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, -
N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -NO2, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
99. The compound or salt of claim 98, wherein R20, R21, R22, and R23 are
independently selected from hydrogen, halogen, -OH, -N02, -CN, and C1-10 alkyl.
100. The compound or salt of claim 99, wherein R20, R21, R22, and R23 are each hydrogen.
101. The compound or salt of any one of claims 98 to 100, wherein R24 and R25 are independently selected from hydrogen, halogen, -OH, -N02, -CN, and C1-10 alkyl, or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
102. The compound or salt of claim 101, wherein R24 and R25 are each hydrogen.
103. The compound or salt of claim 101, wherein R24 and R25 taken together form an optionally substituted saturated C3-5 carbocycle.
104. The compound or salt of any one of claims 97 to 103, wherein R2 is represented by the formula:
Figure imgf000199_0001
wherein:
R101 is selected from an amino acid, a peptide, -0-(Ci-C6 alkyl) and -Ci-C6 alkyl, wherein alkyl of -0-(Ci-C6 alkyl) and -Ci-C6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N02, -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle; and
R100 is C(=0), wherein when R101 is selected from an amino acid or peptide R100 is the C- terminus of the amino acid or peptide.
105. The compound or salt of claim 104, wherein R101 is a peptide selected from a dipeptide, tripeptide and tetrapeptide.
106. The compound or salt of any one of claims 97 to 105, wherein L1 is selected from -N(R10)C(O)-*, -S(0)2N(R10)-*, -CR10 2N(R10)C (0)-*and -X2-Ci.6 alkylene-X2-Ci-6 alkylene-.
107. The compound or salt of claim 106, wherein L1 is selected from -N(R10)C(O)-*.
108. The compound or salt of claim 107, wherein R10 of -N(R10)C(O)-* is selected from hydrogen and Ci-6 alkyl.
109. The compound or salt of claim 108, wherein L1 is -NHC(O)-*.
110. The compound or salt of claim 106, wherein L1 is selected from -S(0)2N(R10)-*.
111. The compound or salt of claim 110, wherein R10 of -S(0)2N(R10)-* is selected from hydrogen and Ci-6 alkyl. 112 The compound or salt of claim 111, wherein L1 is -S(0)2NH-*.
113. The compound or salt of claim 106, wherein L1 is -CR10 2N(R10)C(O)-*.
114. The compound or salt of claim 113, wherein L1 is selected from -CH2N(H)C(0)-* and -CH(CH3)N(H)C(0)-*.
115. The compound or salt of any one of claims 97 to 114, wherein R3 is selected from optionally substituted C3-i2 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents on R3 are independently selected at each occurrence from: halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -
N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
116. The compound or salt of claim 115, wherein R3 is selected from optionally substituted C3-i2 carbocycle, and optionally substituted 3- to l2-membered heterocycle, wherein substituents on R3 are independently selected at each occurrence from: halogen, -OR10, - SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -
N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle.
117. The compound or salt of any one of claims 97 to 116, wherein R3 is selected from an optionally substituted aryl and an optionally substituted heteroaryl.
118. The compound or salt of claim 117, wherein R3 is an optionally substituted heteroaryl.
119. The compound or salt of claim 118, wherein R3 is an optionally substituted heteroaryl substituted with one or more substituents independently selected from halogen, - OR10, -SR10, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
120. The compound or salt of claim 118 or 119, wherein R3 is selected from an optionally substituted 6-membered heteroaryl.
121. The compound or salt of claim 120, wherein R3 is optionally substituted pyridine.
122. The compound or salt of claim 117, wherein R3 is an optionally substituted aryl.
123. The compound or salt of claim 122, wherein R3 is an optionally substituted aryl substituted with one or more substituents independently selected from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
124. The compound or salt of claim 122 or 123, wherein R3 is an optionally substituted phenyl.
125. The compound or salt of claim 117, wherein R3 is selected from pyridine, phenyl, tetrahydronaphthalene, tetrahydroquinoline, tetrahydroisoquinoline, indane, cyclopropylbenzene, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is optionally substituted.
126. The compound or salt of claim any one of claims 97 to 125, wherein R3 is
Figure imgf000201_0001
Figure imgf000201_0003
,
Figure imgf000201_0002
Figure imgf000202_0001
128. The compound or salt of any one of claims 97 to 127, wherein L2 is selected from -C(O)-, and -C(0)NR10-.
129. The compound or salt of claim 128, wherein L2 is -C(O)-.
130. The compound or salt of claim 128, wherein L2 is selected from -C(0)NR10-.
131. The compound or salt of claim 130, wherein R10 of -C(0)NR10- is selected from hydrogen and Ci-6 alkyl.
132. The compound or salt of claim 131, wherein L2 is -C(0)NH-.
133. The compound or salt of any one of claims 97 to 132, wherein R4 is selected from:
-OR10, -N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, - N(R10)C(O)N(R10)2, -N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl.
134. The compound or salt of claim 133, wherein R4 is selected from: -OR10, - N(R10)2, -C(O)N(R10)2, -C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle.
135. The compound or salt of claim 133, wherein R4 is selected from: -OR10, and - N(R10)2; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, and 3- to l2-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, - OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl.
136. The compound or salt of claim 135, wherein R4 is -N(R10)2.
137. The compound or salt of claim 136, wherein R10 of -N(R10)2 is independently selected at each occurrence from optionally substituted Ci-6 alkyl.
138. The compound or salt of claim 137, wherein R10 of -N(R10)2 is independently selected at each occurrence from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted.
139. The compound or salt of claim 138, wherein
Figure imgf000204_0001
140. The compound or salt of claim 139, wherein
Figure imgf000204_0002
141. The compound or salt of any one of claims 97 to 140, wherein R12 is
independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -
S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -
S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl.
142. The compound or salt of claim 141, wherein R12 is independently selected at each occurrence from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - N02, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -N02, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle.
143. The compound or salt of claim 97, wherein the compound is selected from:
Figure imgf000204_0003
salt of any one thereof.
144. A compound represented by the structure of Formula (IV A):
Figure imgf000205_0001
or a pharmaceutically acceptable salt thereof, wherein:
represents an optional double bond;
L12 is selected from -X3-, -X3-CI-6 alkylene-X3-, -X3-C2-6 alkenylene-X3-, and - X3-C2-6 alkynylene-X3-, each of which is optionally substituted on alkyl ene, alkenylene, or alkynylene with one or more substituents independently selected from R12;
L22 is independently selected from -X4-, -X4-CI-6 alkylene-X4-, -X4-C2-6 alkenylene-X4-, and -X4-C2-6 alkynylene-X4-, each of which is optionally substituted on alkyl ene, alkenylene, or alkynylene with one or more substituents independently selected from R10;
X3 and X4 are independently selected at each occurrence from a bond, -0-, -S-, -N(R10)-, -C(O)-, -C(0)0-, -OC(O)-, -0C(0)0-, -C(0)N(R10)-, -C(O)N(R10)C(O)-,
-C(O)N(R10)C(O)N(R10)-, -N(R10)C(O)-, -N(R10)C(O)N(R10)-, -N(R10)C(O)O-, -OC(0)N(R10)-, -C(NR10)-, -N(R10)C(NR10)-, -C(NR10)N(R10)-, -N(R10)C(NR10)N(R10)-, -S(0)2-, -0S(0)-, -S(O) 0-, -S(O)-, -0S(0)2-, -S(0)20-, -N(R10)S(O)2-, -S(0)2N(R10)-, -N(R10)S(O)-, -S(0)N(R10)-, -N(R10)S(O)2N(R10)-, and -N(R10)S(O)N(R10)-;
R1 is hydrogen;
R2 is an amine masking group;
R4 and R8 are independently selected from: -OR10, -N(R10)2, -C(O)N(R10)2, - C(0)R10, -C(0)OR10, -S(0)R10, and -S(0)2R10; Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from L3, halogen, -OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, - N(R10)2, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, C3-i2 carbocycle, and 3- to l2-membered heterocycle; and C3-i2 carbocycle, and 3- to l2-membered heterocycle, wherein each C3-i2 carbocycle, and 3- to l2-membered heterocycle in R4 and R8 is optionally substituted with one or more substituents independently selected from L3, halogen, - OR10, -SR10, -C(O)N(R10)2, -N(R10)C(O)R10, -N(R10)C(O)N(R10)2, -N(R10)2, - C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl; R10 is independently selected at each occurrence from L3, hydrogen, -NH2, -C(0)0CH2C6H5; and Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, and 3- to 12- membered heterocycle, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -NO2, -NH2, =0, =S, -C(0)0CH2C6H5, - NHC(0)0CH2C6H5, Ci-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-12 carbocycle, 3- to l2-membered heterocycle, and haloalkyl;
L3 is a linker moiety, wherein at least one R10 is L3 or at least one substituent on a group selected from R4, R8, X3 and X4 is L3; and
R12 is independently selected at each occurrence from halogen, -OR10, -SR10, - N(R10)2, -C(0)R10, -C(O)N(R10)2, -N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, - S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, -NO2, =0, =S, =N(R10), and -CN; Ci-io alkyl, C2-10 alkenyl, C2-10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C3-10 carbocycle and 3- to lO-membered heterocycle; and C3-10 carbocycle and 3- to lO-membered heterocycle, wherein each C3-10 carbocycle and 3- to 10- membered heterocycle in R12 is optionally substituted with one or more substituents
independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, -C(O)N(R10)2, - N(R10)C(O)R10 -C(0)OR10, -OC(0)R10, -S(0)R10, -S(0)2R10, -P(O)(OR10)2, -OP(O)(OR10)2, - NO2, =0, =S, =N(R10), -CN, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl; and
wherein any substitutable carbon on the benzazepine core is optionally substituted by a substituent independently selected from R12 or two substituents on a single carbon atom combine to form a 3- to 7- membered carbocycle.
145. The compound or salt of claim 144, wherein the compound of Formula (IV A) is represented by Formula (IVB):
Figure imgf000206_0001
or a pharmaceutically acceptable salt thereof, wherein: R20, R21, R22, and R23 are independently selected from hydrogen, halogen, - OR10, -SR10, -N(R10)2, -S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-io alkynyl; and
R24, and R25 are independently selected from hydrogen, halogen, -OR10, -SR10, -
N(R10)2,
-S(0)R10, -S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci- io alkyl, C2-io alkenyl, and C2-io alkynyl; or R24 and R25 taken together form an optionally substituted saturated C3-7 carbocycle.
146. The compound or salt of claims 144 or 145, wherein R2 is represented by the formula:
Figure imgf000207_0001
wherein:
R101 is selected from an amino acid, a peptide, -0-(Ci-C6 alkyl) and -Ci-C6 alkyl, wherein alkyl of -0-(Ci-C6 alkyl) and -Ci-C6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OR10, -SR10, -N(R10)2, -C(0)R10, - C(O)N(R10)2, -N02, -CN, C3-13 carbocycle, and 3- to l2-membered heterocycle; and
R100 is C(=0), wherein when R101 is selected from an amino acid or peptide R100 is the C- terminus of the amino acid or peptide.
147. The compound or salt of claim 146, wherein R101 is a peptide selected from a dipeptide, tripeptide and tetrapeptide.
148. The compound or salt of any one of claims 144 to 147, wherein L12 is - C(0)N(R10)-.
149. The compound or salt of claim 148, wherein R10 of -C(0)N(R10)- is selected from hydrogen, Ci-6 alkyl, and L3.
150. The compound or salt of claim 149, wherein L12 is -C(0)NH-.
151. The compound or salt of any one of claims 144 to 150, wherein R8 is an optionally substituted 5- or 6-membered heteroaryl.
152. The compound or salt of claim 151, wherein R8 is an optionally substituted 5- or 6- membered heteroaryl, substituted with L3.
153. The compound or salt of claim 152, wherein R8 is an optionally substituted pyridine, substituted with L3.
154. The compound or salt of any one of claims 144 to 153, wherein L22 is selected from -C(O)-, and -C(0)NR10-.
155. The compound or salt of claim 154, wherein L22 is -C(O)-.
156. The compound or salt of claim 154, wherein L22 is -C(0)NR10-.
157. The compound or salt of claim 156, wherein R10 of -C(0)NR10- is selected from hydrogen, Ci-6 alkyl, and -L3.
158. The compound or salt of claim 157, wherein L22 is -C(0)NH-.
159. The compound or salt of any one of claims 144 to 158, wherein R4 is selected from: -OR10, and -N(R10)2; and C1-10 alkyl, C2-io alkenyl, C2-io alkynyl, C3-i2 carbocycle, 3- to 12- membered heterocycle, aryl, and heteroaryl, each of which is optionally substituted with one or more substituents independently selected from L3, halogen, -OR10, -SR10, -N(R10)2, -S(0)R10, -
S(0)2R10, -C(0)R10, -C(0)OR10, -OC(0)R10, -N02, =0, =S, =N(R10), -CN, Ci-io alkyl, C2-io alkenyl, and C2-10 alkynyl.
160. The compound or salt of claim 159, wherein R4 is -N(R10)2 andR10 of -N(R10)2 is selected from L3 and hydrogen, and wherein at least one R10 of -N(R10)2 is L3.
161. The compound or salt of any one of claims 1 to 143, wherein the compound is further covalently bound to a linker, L3.
162. The compound or salt of any one of claims 144 to 160 or 161, wherein L3 is a noncleavable linker.
163. The compound or salt of any one of claims 144 to 160 or 161, wherein L3 is a cleavable linker.
164. The compound or salt of claim 163, wherein L3 is cleavable by a lysosomal enzyme.
165. The compound or salt of any one of claims 1 to 143, wherein the compound is covalently attached to an antibody construct.
166. The compound or salt of any of claims 144 to 160 or 161, wherein the compound is covalently attached to a targeting moiety, optionally through the linker.
167. The compound or salt of claim 165 or 166, wherein the targeting moiety or antibody construct specifically binds to a tumor antigen.
168. The compound or salt of any one of claims 165 to 167, wherein the antibody construct or targeting moiety further comprises a target binding domain.
169. The compound or salt of any one of claims 144 to 160 or 161, wherein L3 is
represented by the formula:
Figure imgf000208_0001
, wherein: L4 represents the C-terminus of the peptide and L5 is selected from a bond, alkylene and heteroalkylene, wherein L5 is optionally substituted with one or more groups independently selected from R32, and RX is a reactive moiety; and
R32 is independently selected at each occurrence from halogen, -OH, -CN, -O- alkyl, -SH, =0, =S, -NH2, -N02; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -
CN, -O-alkyl, -SH, =0, =S, -NH2, -N02.
170. The compound or salt of claim 169, wherein RX comprises a leaving group.
171. The compound or salt of claim 169, wherein RX comprises a maleimide.
172. The compound or salt of any one of claims 144 to 160 or 162, wherein L3 is further covalently bound to an antibody construct.
173. The compound or salt of claim 172, wherein the antibody construct is directed against a tumor antigen.
174. The compound or salt of claim 173, wherein the antibody construct further comprises target binding domain.
175. The compound or salt of claim 172, 173, or 174, wherein L3 is represented by the
formula:
Figure imgf000209_0001
, wherein L4 represents the C-terminal of the peptide and L5 is selected from a bond, alkyl ene and heteroalkylene, wherein L5 is optionally substituted with one or more groups independently selected from R32 ; RX* comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein ^ on RX* represents the point of attachment to the residue of the antibody construct; and,
R32 is independently selected at each occurrence from halogen, -OH, -CN, -O-alkyl, -SH, =0, =S, -NH2, -N02; and Ci-io alkyl, C2-io alkenyl, C2-io alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -O- alkyl, -SH, =0, =S, -NH2, -N02.
176. The compound or salt of any one of claims 169 to 171 or 175, wherein the peptide of L3 comprises Val— Cit or Val— Ala.
177. A compound or salt selected from:
Figure imgf000210_0001
Figure imgf000211_0001
210
Figure imgf000212_0001
211
Figure imgf000213_0001
and a salt of any one thereof.
178. A compound or salt selected from:
Figure imgf000213_0002
Figure imgf000214_0001
213
Figure imgf000215_0001
Figure imgf000216_0001
and a salt of any one thereof, wherein the RX* is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein * on RX* represents the point of attachment to the residue of the antibody construct.
179. The compound or salt of any one of claims 144 to 160 or 161, wherein L3 is
represented by the formula:
Figure imgf000217_0001
, wherein RX comprises a reactive moiety, and n = 0-9.
180. The compound or salt of claim 179, wherein RX comprises a leaving group.
181. The compound or salt of claim 179, wherein RX comprises a maleimide.
182. The compound or salt of any one of claims 172, 173, or 174, wherein L3 is
represented as follows:
Figure imgf000217_0002
, wherein RX comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein ^ on RX* represents the point of attachment to the residue of the antibody construct, and n = 0-9.
183. A compound or salt selected from:
Figure imgf000217_0003
Figure imgf000218_0001
salt of any one thereof.
184. A compound or salt selected from:
Figure imgf000218_0002
Figure imgf000219_0001
salt of any one thereof, wherein the RX* comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of an antibody construct, wherein
Figure imgf000219_0002
on RX* represents the point of attachment to the residue of the antibody construct.
185. The compound or salt of claim 175, 178, 182, or 184, wherein RX* comprises a succinamide moiety and is bound to a cysteine residue of an antibody construct.
186. The compound or salt of claim 175, 178, 182, or 184, wherein RX* comprises a hydrolyzed succinamide moiety and is bound to a cysteine residue of an antibody construct.
187. The compound or salt of any one of claims 172 to 175, 178, 182 or 184 to 186, wherein the antibody construct comprises a HER2 antigen binding domain.
188. The compound or salt of any one of claims 172 to 175, 178, 182 or 184 to 186, wherein the antibody construct comprises a TROP2 antigen binding domain.
189. The compound or salt of claim 187, wherein the antibody construct is pertuzumab or an antigen binding fragment thereof.
190. The compound or salt of claim 187, wherein the antibody construct is trastuzumab or an antigen binding fragment thereof.
191. The compound or salt of claim 188, wherein the antibody construct is
sacituzumab or an antigen binding fragment thereof.
192. A conjugate represented by the formula:
Antibody
Figure imgf000219_0003
, wherein Antibody is an antibody construct, D is a compound or salt selected from any one of claims 1 to 143, and L3 is a linker moiety.
193. A conjugate represented by the formula:
Antibody
Figure imgf000219_0004
, wherein Antibody is an antibody construct and D-L3 is a compound or salt selected from any of claims 144 to 160 or 161 to 164.
194. The conjugate of claim 192 or 193, wherein the antibody construct comprises a HER2 antigen binding domain.
195. The conjugate of claim 192 or 193, wherein the antibody construct comprises a TROP2 antigen binding domain.
196. The conjugate of claim 194, wherein the antibody construct is pertuzumab or an antigen binding fragment thereof.
197. The conjugate of claim 194, wherein the antibody construct is trastuzumab or an antigen binding fragment thereof.
198. The conjugate of claim 195, wherein the antibody construct is sacituzumab or an antigen binding fragment thereof.
199. A pharmaceutical composition comprising the compound or salt of any one of claims 1 to 143 and at least one pharmaceutically acceptable excipient.
200. A pharmaceutical composition, comprising the conjugate of any one of claims 192 or 198 and at least one pharmaceutically acceptable excipient.
201. The pharmaceutical composition of claim 200, wherein the average DAR of the conjugate is from about 2 to about 8, or about 1 to about 3, or about 3 to about 5.
202. A method of killing tumor cells in vivo, comprising contacting a tumor cell population with a conjugate of any one of claims 192 or 198.
203. A method for treatment, comprising administering to a subject a conjugate of any one of claims 192 or 198.
204. A method for treatment, comprising administering to a subject in need thereof a compound or salt of any one of claims 1 to 143.
205. A method for treating cancer, comprising administering to a subject in need thereof a conjugate of any one of claims 192 or 198.
206. A method for treating cancer, comprising administering to a subject in need thereof a compound or salt of any one of claims 1 to 143.
207. A compound or salt of any one of claims 1 to 143 for use in a method of treatment of a subject’s body by therapy.
208. A compound or salt of any one of claims 1 to 143 for use in a method of treating cancer.
209. A conjugate of any one of claims 192 or 198 for use in a method of treatment of a subject’s body by therapy.
210. A conjugate of any one of claims 192 or 198 for use in a method of treating cancer.
211. A method of delivering a benzazepine compound to a cancer cell in a subject comprising administering a conjugate of any one of claims 192 or 198 to a subject in need thereof.
Antibody
Figure imgf000221_0001
212 A method of preparing an antibody conjugate of the formula
wherein Antibody is an antibody construct, and L3-D is selected from a compound or salt of any one of claims 144 to 160, 161 to 164, 169 to 171, 177, 179 to 181, or 183 comprising contacting L3-D with an antibody construct.
213. The method of claim 212, wherein the antibody construct comprises a HER2 antigen binding domain.
214. The method of claim 212, wherein the antibody construct comprises a TROP2 antigen binding domain.
215. The method of claim 213, wherein the antibody construct is pertuzumab or an antigen binding fragment thereof.
216. The method of claim 213, wherein the antibody construct is trastuzumab or an antigen binding fragment thereof.
217. The method of claim 214, wherein the antibody construct is sacituzumab or an antigen binding fragment thereof.
218. The method of any one of claims 212 to 217, further comprising purifying the antibody conjugate.
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