Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
  • A61K31/565—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol
  • A61K31/566—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol having an oxo group in position 17, e.g. estrone
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
  • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
  • A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
  • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
  • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
  • A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
  • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
  • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
  • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
  • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
  • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
  • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
  • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
  • A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
  • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
  • A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
  • A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
  • A61K47/6951—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J41/00—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
  • C07J41/0033—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005
  • C07J41/005—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 the 17-beta position being substituted by an uninterrupted chain of only two carbon atoms, e.g. pregnane derivatives
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J41/00—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
  • C07J41/0033—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005
  • C07J41/0088—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 containing unsubstituted amino radicals
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J71/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton is condensed with a heterocyclic ring
  • C07J71/0005—Oxygen-containing hetero ring
  • C07J71/0026—Oxygen-containing hetero ring cyclic ketals
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J71/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton is condensed with a heterocyclic ring
  • C07J71/0005—Oxygen-containing hetero ring
  • C07J71/0026—Oxygen-containing hetero ring cyclic ketals
  • C07J71/0031—Oxygen-containing hetero ring cyclic ketals at positions 16, 17

Definitions

• novel steroids are novel steroids, protein conjugates thereof, and methods for treating diseases, disorders, and conditions comprising administering the steroids and conjugates.
• ADCs Antibody-drug conjugates
• ADCs are antibodies that are covalently linked to biologically active small molecule drugs, thus combining the targeting specificity of antibodies with the mode-of-action and potency of small molecule drugs.
• the therapeutic utility of ADC(s) has been validated in cancer treatment and is a major ongoing focus of study.
• ADCETPJS ® (bentruximab vedotin) and KADCYLA ® (ado-trastuzumab emtansine) are ADCs approved for the treatment of certain cancer types, and at least forty ADCs are currently in clinical development.
• Glucocorticoids are small molecule steroids that bind to glucocorticoid receptors (GRs) and are utilized in anti-inflammatory and immunosuppressive therapies.
• glucocorticoid treatments are compromised by toxicities to most organ systems.
• novel glucocorticoids as well as novel therapies that minimize the side effects arising from glucocorticoid administration, particularly those arising from activating glucocorticoid receptors in non-target cells.
• the instant disclosure provides solutions to the aforementioned needs as well as other unmet needs in the field to which the instant disclosure pertains. Included in the instant disclosure are antibody-drug conjugates comprising glucocorticoid payloads.
• the compounds have the structure of Formula (A):
• R 1 and R 2 are, inde endently, -H, alkyl, alkyl-C(0)-0- -OH, or halo; or R 1 and
• R 4 is alkyl, aryl, arylalkyl, or an N-containing heterocycloalkyl
• alkyl, aryl, arylalkyl, and N-containing heterocycloalkyl independently in each instance, optionally substituted with -NR a R b ;
• R 3 is -OH, R z -C(0)-X- heteroalkyl, piperidinyl, -NR a R b , -oxyaryl-NR a R b
• R 5 is, independently in each instance, -OH, halo, alkyl, or arylalkyl;
• R z is alkyl
• X is O or NR 3 ;
• Z is S, S(O), S(0) 2 , S0 2 NR a , O, C(0)NR a , C(O), or NR a ;
• A is aryl, arylalkyl, or heteroaryl
• R p is, independently in each instance, halo, optionally substituted alkyl, -OH,
• R a and R b are, independently in each instance, -H, optionally substituted alkyl, or optionally subtitued aryl;
• n is an integer from 0-19;
• t is an integer from 1-3; with the proviso that:
• R 3 is not -OH (a) when R 1 is -OH or (b) when R 1 and R 2 together form A,
• the compounds are protein-drug conjugates, e.g., antibody- drug conjugates, comprising an antigen-binding protein, e.g., antibody and a compound of Formula (A).
• the compounds are protein-drug conjugates, e.g., antibody- drug conjugates, comprising an antigen-binding protein, e.g., antibody, a compound of Formula (A), and a cyclodextrin moiety.
• FIG. 1. shows a sequence for synthesizing the certain steroids described herein.
• FIG. 2. shows a sequence for modifying the primary alcohol position of budesonide.
• FIG. 3. shows a sequence for modifying the primary alcohol position of
• FIG. 4. shows a sequence for modifying the primary alcohol position of dexamethasone.
• FIG. 5 shows a two-dimensional nuclear Overhauser effect (NOE) magnetic resonance spectrum (hereinafter “2D-NOESY”) for compound 7-1R in Table 1.
• NOE nuclear Overhauser effect
• FIG. 6 shows a 2D-NOESY for compound 7-lS in Table 1.
• FIG. 7 shows a 2D-NOESY spectrum for 1 1-5R in Table 1.
• FIG. 8 shows a 2D-NOESY spectrum for compound 1 1 -5S in Table 1.
• FIG. 9 shows a general approaches for synthesizing certain Linker-Payloads.
• FIG. 10 shows a sequence for synthesizing DIBAC-Suc-NHS (Compound
• FIG. 11 shows a sequence for synthesizing DIBAC-Suc-PEG 4 -acid/NHS
• FIG. 12 shows a sequence for synthesizing BCN-PEG4-Acid (Compound
• FIG. 13 shows a sequence for synthesizing DIBAC-Suc-PEG 4 -VC-pAB-P P
• FIG. 14 shows a sequence for synthesizing Linker-Payload 1 (LP1).
• FIG. 15 shows a sequence for synthesizing Linker-Payload 2 (LP2) and
• FIG. 16 shows a sequence for synthesizing Linker-Payloads 4-11 (LP4-LP11).
• FIG. 17 shows a sequence for synthesizing Linker-Payload 12 (LP12).
• FIG. 18 shows a synthesis sequence for making Linker-Payload 12 (LP13) and
• FIG. 19 shows a general synthetic process for an ADC conjugation via a [2+3] click reaction with LP4.
• FIG. 20 shows a Coomassie-stained SDS-PAGE Gel of an anti-PRLR antibody, azido-functionalized anti-PRLR antibody, and anti-PRLR antibody-LP4 conjugate as described in Example 59
• FIG. 21 shows size exclusion chromatography (SEC) of an anti-PRLR antibody, azido-functionalized antibody, and 4DAR anti-PRLR-LP4 Conjugate as described in Example 59.
• FIG. 22 shows an ESI-MS of anti-PRLR antibody, azido-functionalized anti-
• FIG. 23 shows selective GR activation in 293/PRLR/GRE-Luc cells (FIG. 23 A) and 293/MMTV-Luc cells (FIG. 23B) by steroid ADCs and budesonide control as described in Example 64.
• FIG. 24 shows the linker-payload contribution to GR activation by steroid ADC and budesonide control as tested in 293/PRLR/GRE-Luc cells as described in Example 65.
• FIG. 25 shows activation of glucocorticoid receptor in a HEK293/MMTV- luc/IL-2Ry/IL7R cell line by Budesonide, 11-5 in Table 1, and anti-IL2Ry ncADC at twenty- four (24), forty-eight (48), or seventy-two (72) hours as described in Example 66.
• FIG. 26 shows a sequence for synthesizing Linker-Payload (LP7).
• FIG. 27 shows a synthetic process for preparing compound (27b).
• FIG. 28 shows a sequence for synthesizing Linker-Payloads (LP15 and LP16)
• FIG. 29 shows a general synthetic process for an ADC conjugation via a [2+3] click reaction with Cyclodextrin-Linker-Payloads.
• FIG. 30 shows bioactivity of steroid ADCs with and without cyclodextrin linkers in a plot of relative light units (RLU) vs. Logio [M].
• FIG. 31 shows a sequence for synthesizing certain steroids (payloads 1-6) described herein.
• FIG. 32 shows a sequence for synthesizing certain linker-steroids (LP101 to
• FIG. 33 shows a general synthetic process for an ADC conjugation via [2+3] click reaction.
• FIG. 34 shows ESI-MS of anti-PRLR Ab, anti-PRLR Ab-N 3 , and anti-PRLR-
• FIG. 35 shows ESI-MS of anti-Fel dl Ab, anti- Fel dl Ab-PEG 3 -N 3 , and anti-
• FIG. 36 shows bioactivity of steroid ADCs in antigen positive cells
• FIG. 37A shows mean blood concentration-time provides for compounds 4b and 6-1.
• FIG. 37B shows T F-a level in blood samples of payloads 4b and 6-1 as described in Examples 120-121.
• alkyl refers to a monovalent and saturated hydrocarbon radical moiety. Alkyl is optionally substituted and can be linear, branched, or cyclic, i.e., cycloalkyl. Alkyl includes, but is not limited to, those having 1-20 carbon atoms, i.e., Ci-20 alkyl; 1-12 carbon atoms, i.e., Ci-12 alkyl; 1-8 carbon atoms, i.e., Ci-8 alkyl; 1-6 carbon atoms, i.e., Ci-6 alkyl; and 1-3 carbon atoms, i.e., C1-3 alkyl.
• alkyl moieties include, but are not limited to methyl, ethyl, ⁇ -propyl, /-propyl, «-butyl, s-butyl, t-butyl, i— butyl, a pentyl moiety, a hexyl moiety, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.
• Alkylene is divalent alkyl.
• haloalkyl refers to alkyl, as defined above, wherein the alkyl includes at least one substituent selected from a halogen, e.g., F, CI, Br, or I.
• alkenyl refers to a monovalent hydrocarbon radical moiety containing at least two carbon atoms and one or more non-aromatic carbon-carbon double bonds. Alkenyl is optionally substituted and can be linear, branched, or cyclic.
• Alkenyl includes, but is not limited to, those having 2-20 carbon atoms, i.e., C2-20 alkenyl; 2-12 carbon atoms, i.e., C2-12 alkenyl; 2-8 carbon atoms, i.e., C2-8 alkenyl; 2-6 carbon atoms, i.e., C2-6 alkenyl; and 2-4 carbon atoms, i.e., C2-4 alkenyl.
• alkenyl moieties include, but are not limited to vinyl, propenyl, butenyl, and cyclohexenyl.
• Alkenylene is divalent alkenyl.
• alkynyl refers to a monovalent hydrocarbon radical moiety containing at least two carbon atoms and one or more carbon-carbon triple bonds. Alkynyl is optionally substituted and can be linear, branched, or cyclic.
• Alkynyl includes, but is not limited to, those having 2-20 carbon atoms, i.e., C2-20 alkynyl; 2-12 carbon atoms, i.e., C2-12 alkynyl; 2-8 carbon atoms, i.e., C2-8 alkynyl; 2-6 carbon atoms, i.e., C2-6 alkynyl; and 2-4 carbon atoms, i.e., C2-4 alkynyl.
• alkynyl moieties include, but are not limited to ethynyl, propynyl, and butynyl.
• Alkynylene is divalent alkynyl.
• alkoxy refers to a monovalent and saturated hydrocarbon radical moiety wherein the hydrocarbon includes a single bond to an oxygen atom and wherein the radical is localized on the oxygen atom,e.g, CH3CH2-O for ethoxy.
• Alkoxy substituents bond to the compound which they substitute through this oxygen atom of the alkoxy substituent.
• Alkoxy is optionally substituted and can be linear, branched, or cyclic, i.e., cycloalkoxy.
• Alkoxy includes, but is not limited to, those having 1-20 carbon atoms, i.e., Ci- 20 alkoxy; 1-12 carbon atoms, i.e., Ci-12 alkoxy; 1-8 carbon atoms, i.e., Ci-8 alkoxy; 1-6 carbon atoms, i.e., Ci-6 alkoxy; and 1-3 carbon atoms, i.e., C1-3 alkoxy.
• alkoxy moieties include, but are not limited to methoxy, ethoxy, «-propoxy, /-propoxy, «-butoxy, s- butoxy, t-butoxy, z-butoxy, a pentoxy moiety, a hexoxy moiety, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexoxy.
• haloalkoxy refers to alkoxy, as defined above, wherein the alkoxy includes at least one substituent selected from a halogen, e.g., F, CI, Br, or I.
• aryl refers to a monovalent moiety that is a radical of an aromatic compound wherein the ring atoms are carbon atoms.
• Aryl is optionally substituted and can be monocyclic or polycyclic, e.g., bicyclic or tricyclic.
• aryl moieties include, but are not limited to those having 6 to 20 ring carbon atoms, i.e., C6-20 aryl; 6 to 15 ring carbon atoms, i.e., G5-15 aryl, and 6 to 10 ring carbon atoms, i.e., Ce-w aryl.
• aryl moieties include, but are limited to phenyl, naphthyl, fluorenyl, azulenyl, anthryl, phenanthryl, and pyrenyl.
• arylalkyl refers to an monovalent moiety that is a radical of an alkyl compound, wherein the alkyl compound is substituted with an aromatic substituent, i.e., the aromatic compound includes a single bond to an alkyl group and wherein the radical is localized on the alkyl group.
• An arylalkyl group bonds to the illustrated chemical structure via the alkyl group.
• An arylalkyl can be represented by the structure, e.g.,
• B is an aromatic moiety, e.g., phenyl.
• Arylalkyl is optionally substituted, i.e., the aryl group and/or the alkyl group, can be substituted as disclosed herein. Examples of arylalkyl include, but are not limited to, benzyl.
• aryloxy refers to a monovalent moiety that is a radical of an aromatic compound wherein the ring atoms are carbon atoms and wherein the ring is substituted with an oxygen radical, i.e. , the aromatic compound includes a single bond to an
• Aryloxy substituents bond to the compound which they substitute through this oxygen atom.
• Aryloxy is optionally substituted.
• Aryloxy includes, but is not limited to those having 6 to 20 ring carbon atoms, i.e., C6-20 aryloxy; 6 to 15 ring carbon atoms, i.e., C 6 -i5 aryloxy, and 6 to 10 ring carbon atoms, i.e., C 6 -io aryloxy.
• Examples of aryloxy moieties include, but are not limited to phenoxy, naphthoxy, and anthroxy.
• R a R b N-aryloxy refers to a monovalent moiety that is a radical of an aromatic compound wherein the ring atoms are carbon atoms and wherein the ring is substituted with an R a R b N- substituent and an oxygen radical, i.e., the aromatic compound includes a single bond to an R a R b N- substituent and a single bond to an oxygen atom and
• R a R b N-aryloxy substituents bond to the compound which they substitute through this oxygen atom.
• R a R b N- aryloxy is optionally substituted.
• R a R b N-aryloxy includes, but is not limited to those having 6 to 20 ring carbon atoms, 6 to 15 ring carbon atoms; and 6 to 10 ring carbon atoms.
• An example of n R a R b N-aryloxy moiety includes, but is not limited to 4-(dimethyl-amino)-
• arylene refers to a divalent moiety of an aromatic compound wherein the ring atoms are only carbon atoms.
• Arylene is optionally substituted and can be monocyclic or poly cyclic, e.g., bicyclic or tricyclic.
• Examples of arylene moieties include, but are not limited to those having 6 to 20 ring carbon atoms, i.e., C6-20 arylene; 6 to 15 ring carbon atoms, i.e., C 6 -i5 arylene, and 6 to 10 ring carbon atoms, i.e., C 6 -io arylene.
• heteroalkyl refers to an alkyl in which one or more carbon atoms are replaced by heteroatoms.
• heteroalkenyl refers to an alkenyl in which one or more carbon atoms are replaced by heteroatoms.
• heteroalkynyl refers to an alkynyl in which one or more carbon atoms are replaced by heteroatoms. Suitable heteroatoms include, but are not limited to, nitrogen, oxygen, and sulfur atoms. Heteroalkyl is optionally substituted.
• heteroalkyl moieties include, but are not limited to, aminoalkyl, sulfonylalkyl, sulfinylalkyl. Examples of heteroalkyl moieties also include, but are not limited to, methylamino, methylsulfonyl, and methyl sulfinyl.
• heteroaryl refers to a monovalent moiety that is a radical of an aromatic compound wherein the ring atoms contain carbon atoms and at least one oxygen, sulfur, nitrogen, or phosphorus atom. Examples of heteroaryl moieties include, but are not limited to those having 5 to 20 ring atoms; 5 to 15 ring atoms; and 5 to 10 ring atoms.
• Heteroaryl is optionally substituted.
• heteroarylene refers to an arylene in which one or more ring atoms of the aromatic ring are replaced with an oxygen, sulfur, nitrogen, or phosphorus atom. Heteroarylene is optionally substituted.
• heterocycloalkyl refers to a cycloalkyl in which one or more carbon atoms are replaced by heteroatoms. Suitable heteroatoms include, but are not limited to, nitrogen, oxygen, and sulfur atoms. Heterocycloalkyl is optionally substituted. Examples of heterocycloalkyl moieties include, but are not limited to, morpholinyl, piperidinyl, tetrahydropyranyl, pyrrolidinyl, imidazolidinyl, oxazolidinyl, thiazolidinyl, dioxolanyl, dithiolanyl, oxanyl, or thianyl.
• N-containing heterocycloalkyl refers to a cycloalkyl in which one or more carbon atoms are replaced by heteroatoms and wherein at least one heteroatom is a nitrogen atom. Suitable heteroatoms in addition to nitrogen, include, but are not limited to oxygen and sulfur atoms. N-containing heterocycloalkyl is optionally substituted. Examples of N-containing heterocycloalkyl moieties include, but are not limited to, morpholinyl, piperidinyl, pyrrolidinyl, imidazolidinyl, oxazolidinyl, or thiazolidinyl.
• optionally substituted when used to describe a radical moiety, e.g., optionally substituted alkyl, means that such moiety is optionally bonded to one or more substituents.
• substituents include, but are not limited to halo, cyano, nitro, haloalkyl, azido, epoxy, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, + «* , + ⁇ *> .
• R A , R B , and R c are, independently at each occurrence, a hydrogen atom, alkyl, alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heteroalkyl, heteroaryl, or heterocycloalkyl, or R A and R B , together with the atoms to which they are bonded, form a saturated or unsaturated carbocyclic ring, wherein the ring is optionally substituted and wherein one or more ring atoms is optionally replaced with a heteroatom.
• a radical moiety is optionally substituted with an optionally substituted heteroaryl, optionally substituted heterocycloalkyl, or optionally substituted saturated or unsaturated carbocyclic ring
• the substituents on the optionally substituted heteroaryl, optionally substituted heterocycloalkyl, or optionally substituted saturated or unsaturated carbocyclic ring, if they are substituted, are not substituted with substituents which are further optionally substituted with additional substituents.
• the substituent bonded to the group is unsubstituted unless otherwise specified.
• binding agent refers to any molecule capable of binding with specificity to a given binding partner.
• the binding agent is an antibody, or an antigen binding fragment thereof.
• linker refers to a divalent moiety that covalently links the binding agent to the steroid described herein.
• amide synthesis conditions refers to reaction conditions suitable facilitate the formation of an amide, e.g., by the reaction of a carboxylic acid, activated carboxylic acid, or acyl halide with an amine.
• amide synthesis conditions refers to reaction conditions suitable to facilitate the formation of an amide bond between a carboxylic acid and an amine.
• the carboxylic acid is first converted to an activated carboxylic acid before the activated carboxylic acid reacts with an amine to form an amide.
• Suitable conditions to effect the formation of an amide include, but are not limited to, those utilizing reagents to effect the reaction between a carboxylic acid an amine, including, but not limited to, dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), (benzotriazol-l-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), (benzotriazol-l-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP), (7- azabenzotriazol-l-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBrOP), 0-(benzotriazol-l-yl)- ⁇ , ⁇ , ⁇ ', ⁇ '-tetramethyluron
• HATU hexafluorophosphate
• EEDQ 2-Ethoxy-l-ethoxycarbonyl-l,2-dihydroquinoline
• EEDQ l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide
• CIP 2-Chloro-l ,3- dimethylimidazolidinium hexafluorophosphate
• CDMT 2-chloro-4,6-dimethoxy-l,3,5-triazine
• CDI carbonyldiimidazole
• a carboxylic acid is first converted to an activated carboxylic ester before reacting with an amine to form an amide bond.
• the carboxylic acid is reacted with a reagent.
• the reagent activates the carboxylic acid by deprotonating the carboxylic acid and then forming a product complex with the deprotonated carboxylic acid as a result of nucleophilic attack by the deprotonated carboxylic acid onto the protonated reagent.
• this activated ester is more susceptible subsequently to nucleophilic attack by an amine than the carboxylic acid is before it is converted. This results in amide bond formation.
• the carboxylic acid is described as activated.
• Exemplary reagents include DCC and DIC.
• terapéuticaally effective amount refers to an amount (of a compound) that is sufficient to provide a therapeutic benefit to a patient in the treatment or management of a disease or disorder, or to delay or minimize one or more symptoms associated with the disease or disorder.
• pharmaceutically acceptable derivative refers to any form, e.g., ester or prodrug of a compound, which provides said compound upon administration to a patient.
• salt refers to any salt suitable for administration to a patient. Suitable salts include, but are not limited to, those disclosed in. Berge et al., "Pharmaceutical Salts", J. Pharm. Sci., 1977, 66: 1, incorporated herein by reference.
• salts include, but are not limited to, acid-derived, base-derived, organic, inorganic, amine, and alkali or alkaline earth metal salts, including but not limited to calcium salts, magnesium salts, potassium salts, sodium salts, salts of hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, 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, p-toluenesulfonic acid, and salicylic acid, and the like.
• cyclic group may be substituted with that substituent at any ring position in the cyclic group or on any ring in the fused ring group, according to techniques set forth herein or which are known in the field to which the instant disclosure pertains.
• the cyclic group may be substituted with that substituent at any ring position in the cyclic group or on any ring in the fused ring group, according to techniques set forth herein or which are known in the field to which the instant disclosure pertains.
• n is an integer from 0 to 19 and in which the positions of substituent R 5 are described generically, i.e., depicted as not directly attached to any vertex of the bond line structure, includes the following, non-limiting examples of, groups in which
• the phrase "reactive linker,” or the abbreviation "RL” refers to a monovalent group that comprises a reactive group and linking group, depicted as 3 ⁇ 4 , wherein RG is the reactive group and L is the linking group.
• the linking group is any divalent moiety that bridges the reactive group to a payload.
• the reactive linkers (RL), together with the payloads to which they are bonded, comprise intermediates ("linker-payloads”) useful as synthetic precursors for the preparation of the antibody steroid conjugates described herein.
• the reactive linker contains a reactive group (“RG”), which is a functional group or moiety that reacts with a reactive portion of an antibody, modified antibody, or antigen binding fragment thereof.
• the "reactive group” is a functional group or moiety (e.g., maleimide or NHS ester) that reacts with a cysteine or lysine residue of an antibody or antigen-binding fragment thereof.
• the "reactive group” is a functional group or moiety that is capable of undergoing a click chemistry reaction.
• the reactive group is an alkyne that is capable of undergoing a 1,3 cycloaddition reaction with an azide.
• Such suitable reactive groups include, but are not limited to, strained alkynes, e.g., those suitable for strain-promoted alkyne-azide cycloadditions (SPAAC), cycloalkynes, e.g., cyclooctynes, benzannulated alkynes, and alkynes capable of undergoing 1,3 cycloaddition reactions with azides in the absence of copper catalysts.
• Suitable alkynes also include, but are not limited to, DIBAC, DIBO, BARAC, DIFO, substituted, e.g., fluorinated alkynes, aza-cycloalkynes, BCN, and derivatives thereof.
• Linker-payloads comprising such reactive groups are useful for conjugating antibodies that have been functionalized with azido groups.
• Such functionalized antibodies include antibodies functionalized with azido-polyethylene glycol groups.
• such functionalized antibody is derived by reacting an antibody comprising at least one glutamine residue, e.g., heavy chain Q295 (EU numbering), with a compound according to the formula H2N-LL-N3, wherein LL is a divalent polyethylene glycol group, in the presence of the enzyme transglutaminase.
• the reactiv which can react via click chemistry with an azide, e.g., , to form a click chemistry product
• the reactive group is an alkyne, e.g., which can react via click chemistry with an alkyne
• the reactive group is an alkyne, e.g., chemistry
• the reactive group is a functional group
• the reactive group is a functional group
• Ab refers to an antibody or antigen-binding fragment thereof and N refers to the N atom on a lysine residue through which the functional group bonds to the Ab.
• binding agent linker refers to any divalent group or moiety that links, connects, or bonds a binding agent (e.g., an antibody or an antigen- binding fragment thereof) with a payload compound set forth herein (e.g., steroid).
• a binding agent e.g., an antibody or an antigen- binding fragment thereof
• a payload compound set forth herein (e.g., steroid).
• suitable binding agent linkers for the antibody conjugates described herein are those that are sufficiently stable to exploit the circulating half-life of the antibody and, at the same time, capable of releasing its payload after antigen-mediated internalization of the conjugate. Linkers can be cleavable or non-cleavable.
• Cleavable linkers are linkers that are cleaved by intracellular metabolism following internalization, e.g., cleavage via hydrolysis, reduction, or enzymatic reaction.
• Non-cleavable linkers are linkers that release an attached payload via lysosomal degradation of the antibody following internalization.
• Suitable linkers include, but are not limited to, acid-labile linkers, hydrolysis-labile linkers, enzymatically cleavable linkers, reduction labile linkers, self-immolative linkers, and non-cleavable linkers.
• Suitable linkers also include, but are not limited to, those that are or comprise glucuronides, succinimide-thioethers, polyethylene glycol (PEG) units, hydrazones, mal-caproyl units, disulfide units (e.g., -S-S-, -S-QR 1 ] 2 ) -, wherein R 1 and R 2 are independently hydrogen or hydrocarbyl), carbamate units, para-amino-benzyl units (PAB), phosphate units, e.g., mono-, bis-, or tris- phosphate units, and peptide units, e.g., peptide units containing two, three four, five, six, seven, eight, or more amino acids, including but not limited to valine-citrulline and units.
• PEG polyethylene glycol
• hydrazones mal-caproyl units
• disulfide units e.g., -S-S-, -S-QR 1 ] 2
• the binding agent linker (BL) comprises a moiety that is formed by the reaction of the reactive group (RG) of a reactive linker (RL) and reactive portion of the binding agent, e.g., antibody, modified antibody, or antigen binding fragment thereof.
• the BL comprises the following moiety:
• ⁇ is the bond to the binding agent.
• the BL comprises the following moiety: ? its regioisomer, or
• the BL is the nd to the bindin agent.
• ⁇ is the nd to the bindin agent.
• the BL comprises the following
• the BL comprises the following moiety: , wherein * is the bond to the cysteine of the antibody or antigen-binding fragment thereof. In some examples, the BL comprises the following moiety: , wherein i 5 is the bond to the lysine of the antibody or antigen-binding fragment thereof.
• the bond to the binding agent is direct or via a linker. In particular embodiments, the binding agent is modified with an azide to facilitate linkage to BL. Examples are described below.
• R 1 and R 2 are, independently, -H, alkyl, alkylene-C(0)-0- -OH, or halo; or R 1 and
• R 4 is alkyl, aryl, arylalkyl, or an N-containing heterocycloalkyl
• alkyl, aryl, arylalkyl, and N-containing heterocycloalkyl are, independently in each instance, optionally substituted with
• R 3 is -OH, R z -C(0)-X- heteroalkyl, piperidinyl, - R a R b , -oxyaryl- R a R b , or -Z-A(R P ),;
• R 5 is, independently in each instance, -OH, halo, alkyl, or arylalkyl;
• R z is alkyl
• X is O or R 3 ;
• Z is S, S(O), S(0) 2 , S0 2 R a , O, C(0) R a , C(O), or R a ;
• A is aryl or heteroaryl
• R p is, independently in each instance, halo, optionally substituted alkyl, -OH,
• R a and R b are, independently in each instance, -H or optionally substituted alkyl; n is an integer from 0-19; and
• t is an integer from 1-3;
• R 3 is not -OH (a) when 1 is -OH or (b) when R 1 and R 2
• the compound of Formula (A) has the structure of
• R ⁇ R are as defined above and R 5A and R 5B are each, independently, halo or a hydrogen atom.
• R 5A and R 5B are hydrogen atoms. In some embodiments of the compound of Formula (A 1 ), R 5A and R 5B are fluoro. In some embodiments of the compound of Formula (A 1 ), R 5A is a hydrogen atom and R 5B is fluoro. [0077] In some embodiments of the compound of Formula (A 1 ), R 1 is alkylene-C(O)-
• R 2 is alkyl
• R 4 is -aryl- R a R b .
• R 4 is -phenyl- R a R b . 0079 In some embodiments of the compound of Formula (A 1 ), R 1 and R 2 together
• R' is -OH, -NR a R b ,
• R b is H or alkyl
• X is O or H
• R z is alkyl
• R 3 is -OH, -NH2
• R is aryl, arylalkyl, or alkyl, wherein the aryl, arylalkyl, and alkyl are optionally substituted with -NR a R b ;
• R 3 is -OH, -NR a R b , R z -C(0)-X-, or
• R p is halo
• t is an integer from 0 to 2
• R a is H
• R b is H or alkyl
• X is O or H
• R z is alkyl
• R 5 independently at each occurrence, is fluoro or a hydrogen atom.
• n is an integer from 0 to 4 and R 3 is -OH or R z -C(0)-0-; wherein R z is alkyl. In certain embodiments, n is 0 or 1.
• n is an integer from 1-4 and R 3 is -OH or R z -C(0)-0-; wherein R z is alkyl. In certain embodiments, n is 2.
• R 3 is - R a R b and R 4 is alkyl, wherein R a and R b are each, independently, a hydrogen atom or alkyl, or R a and R b , taken together form a 3-7 membered ring.
• R 4 is Ci-4 alkyl. In some embodiments, R 4 is propyl. In certain embodiments,
• R 4 is alkyl
• R P1 is halo or a hydrogen atom
• R P2 is -NR a R b or -OH, wherein R a and R b are each, independently, a hydrogen atom or alkyl.
• R 4 is Ci-4 alkyl and R P2 is - H2.
• X is O or aryl or heteroaryl
• R p is halo
• t is an integer from 0-2
• R a and R b are each, independently, a hydrogen atom or alkyl
• R z is alkyl
• R Q is alkoxy
• R 4 is alkyl.
• R 3 is
• R wherein X is O or R a , is aryl or heteroaryl
• R p is halo
• t is an integer from 0-2
• R a and R b are each, independently, a hydrogen atom or alkyl
• R 5A is a hydrogen atom or fluoro
• R 5B is fluoro.
• R 3 is
• R 1 and R 2 are, independently, -H, alkyl, alkyl-C(0)-0- -OH, or halo; or R 1 and R 2
• R 4 is alkyl, aryl, arylalkyl, or an N-containing heterocycloalkyl
• alkyl, aryl, arylalkyl, and N-containing heterocycloalkyl are, independently in each instance, optionally substituted with -NR a R b ;
• R 3 is -OH, alkyl-C(0)-0- heteroalkyl, -NR a R b , -NR a R b -aryloxy, or
• R a R b N-aryloxy- wherein the alkyl-C(0)-0- , heteroalkyl, -NR a R b , and
• R a R b N-aryloxy- are optionally substituted with halo
• R 5 is, independently in each instance, -OH, halo, alkyl, or arylalkyl;
• R a and R b are, independently in each instance, H or alkyl
• n is an integer from 0-19;
• R 3 is not -OH when either (a) or (b): (a) R 1 is -OH or (b) R 1 and
• R 1 and R 2 are, independently, selected from -H, alkyl, alkyl-C(0)-0- -OH, and halo. In some other examples, R 1 and R 2 together form
• R is -H. In certain other examples, R is alkyl. In some examples, R 1 is alkyl-C(0)-0- In some other examples, R 1 is -OH. In certain examples, R 1 is halo. In certain other examples, R 1 is -F. In some examples, R 1 is -CI. In some other examples, R 1 is -Br. In certain examples, R 1 is -I. In certain other examples, R 2 is -OH. In some examples, R 2 is halo. In some other examples, R 2 is -F. In certain examples, R 2 is -CI. In certain other examples, R 2 is -Br. In some examples, R 2 is -I.
• R 5 is -OH.
• R 5 is halo such as but not limited to -F, -CI, -Br, or -I.
• R 5 is -F.
• R 5 is -CI.
• R 5 is -Br.
• R 5 is -I.
• R 5 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl.
• R 5 is benzyl.
• R 3 is selected from -OH, alkyl-C(0)-0- and R a R b N-aryloxy. In some of these examples, alkyl-C(0)-0- or R a R b N-aryloxy is optionally substituted with halo. In some examples, R 3 is -OH. In some examples, R 3 is alkyl-C(0)-0- In some examples, R 3 is a R b N-aryloxy. In some examples, R 3 is some examples, R 3 is ome examples, R is
• R 3 is In some examples, R 3
• R 3 is -OH, alkyl-C(0)-0- heteroalkyl
• R a and R b are, independently in each instance, -H or alkyl.
• R 3 is R a R b N-aryloxy, wherein R a and R b are, independently in each instance, -H or alkyl.
• R is ⁇ / In some example:
• R 3 is
• R 3 is . In some examples, R 3 is
• R is . In some examples, R is In
• R 3 is
• R 3 is In some examples, R 3 is
• R 3 is R a R b N-aryloxy, wherein R a and R b are, independently in each instance, -H or alkyl.
• R 4 is selected from the group consisting of alkyl, aryl, arylalkyl, and an N-containing heterocycloalkyl.
• alkyl, aryl, arylalkyl, or N-containing heterocycloalkyl are optionally substituted with
• R 4 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl.
• R 4 is methyl.
• R 4 is ethyl.
• R 4 is n-propyl.
• R 4 is i-propyl.
• R 4 is n-butyl.
• R 4 is i-butyl.
• R 4 is t-butyl.
• R 4 is sec-butyl.
• R 4 is pentyl.
• R 4 is hexyl. In some examples, R 4 is heptyl. In some examples, R 4 is octyl, or nonyl. In some examples, R 4 is aryl such as but not limited to phenyl or naphthyl. In some examples, R 4 is phenyl. In some examples, R 4 is naphthyl. In some examples, R 4 is arylalkyl-such as but not limited to benzyl. In some examples, R 4 is N-containing heterocycloalkyl such as but not limited to piperidinyl. In some examples, R 4 is 4-amino-phenyl. In some examples, R 4 is 4- aminophenyl optionally substituted with halo.
• R 4 is , wherein R a and R b are, independently in each instance, H or alkyl.
• R 4 is
• R 4 is J
• R is
• R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is . In some examples, R 4 is examples, R 4 is In some examples, R is In some examples, R 4 is mples, examples, R 4 is . In some examples, R is In some examples, R 4 is . In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is . I Inn s soommee e exxaammnplleess., R R 4 i iss . In some examples, R is In some examples, R 4 is . In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is xamples, R 4
• R 4 is In some
• R 4 is 4
• R 4 is
• R 4 is alkyl substituted with amino such as, but not limited to, methyl-amino, ethyl-amino, propyl-amino, butyl-amino, pentyl-amino, hexyl-amino, heptyl- amino, octyl-amino, or nonyl-amino.
• R 4 is methyl-amino.
• R 4 is ethyl-amino.
• R 4 is n-propyl-amino.
• R 4 is i-propyl-amino.
• R 4 is n-butyl-amino.
• R 4 is i-butyl- amino. In some examples, R 4 is t-butyl-amino. In some examples, R 4 is pentyl-amino. In some examples, R 4 is hexyl-amino. In some examples, R 4 is heptyl-amino. In some examples, R 4 is octyl-amino. In some examples, R 4 is nonyl-amino. . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is
• R a and R b are, independently in each instance, selected from H and alkyl. In some examples, both R a and R b are H. In some examples, both R a and R b are methyl. In some examples, both R a and R b are ethyl. In some examples, both R a and R b are propyl. In some examples, one of R a or R b is -H and the other is alkyl. In some examples, one of R a or R b is -H and the other is methyl. In some examples, one of R a or R b is -H and the other is ethyl. In some examples, one of R a or R b is -H and the other is propyl.
• n is an integer from 0-19. In some examples, n is 0. In some other examples, n is 1. In certain examples, n is 2. In some other examples, n is 3. In certain examples, n is 4. In some examples, n is 5. In some other examples, n is 6. In certain examples, n is 7. In some other examples, n is 8. In certain examples, n is 9. In some examples, n is 10. In some other examples, n is 1 1. In certain examples, n is 12. In some other examples, n is 13. In certain examples, n is 14. In some examples, n is 15. In some other examples, n is 16. In certain examples, n is 17. In some other examples, n is 18. In certain examples, n is 19.
• R 3 is not -OH when R 1 is -OH.
• R 3 is not -OH when R 1 and R 2 together form wherein R 4 is a Ci-9alkyl or 4-(dimethyl-amino)-phenyl.
• R 4 and R 2 together form ⁇ " "7TM .
• R 4 is alkyl, aryl, arylalkyl, or a N- containing heterocycloalkyl.
• alkyl, aryl, heteroaryl, arylalkyl, or N- containing heterocycloalkyl are optionally substituted with -NR a R b .
• R 4 is alkyl.
• R 4 is aryl.
• R 4 is arylalkyl.
• R 4 is N-containing heterocycloalkyl.
• R 4 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl. In some examples, R 4 is methyl. In some examples, R 4 is ethyl. In some examples, R 4 is n-propyl. In some examples, R 4 is i-propyl. In some examples, R 4 is n- butyl. In some examples, R 4 is i-butyl. In some examples, R 4 is t-butyl. In some examples, R 4 is sec-butyl. In some examples, R 4 is pentyl.
• R 4 is hexyl. In some examples, R 4 is heptyl. In some examples, R 4 is octyl, or nonyl. In some examples, R 4 is aryl such as but not limited to phenyl or naphthyl. In some examples, R 4 is phenyl. In some examples, R 4 is naphthyl. In some examples, R 4 is heteroaryl-such as but not limited to thiophene or phenol. In some examples, R 4 is arylalkyl-such as but not limited to benzyl. In some examples, R 4 is N-containing heterocycloalkyl such as but not limited to piperidinyl. In some examples, R 4 is 4-amino-phenyl. In some examples, R 4 is 4-aminophenyl optionally substituted with halo.
• R 4 is selected from the group consisting of alkyl, aryl, arylalkyl, and a N-containing heterocycloalkyl; and wherein alkyl, aryl, arylalkyl, or N- containing heterocycloalkyl are optionally substituted with -NR a R b ; and wherein the stereochemistry of the carbon indicated by * is the R configuration.
• R 4 and R 2 together form T / wherein R 4 is selected from the group consisting of alkyl, aryl, arylalkyl, and a N-containing heterocycloalkyl; and wherein alkyl, aryl, arylalkyl, or N- containing heterocycloalkyl are optionally substituted with -NR a R b ; and wherein the stereochemistry of the carbon indicated by * is the S configuration.
• R 1 and R 2 are, independently, selected from -H, alkyl, alkyl-C(O)-
• R 1 and R 2 together form ⁇ .
• R 1 is -H.
• R 1 is alkyl.
• R 1 is alkyl- C(0)-0-
• R 1 is -OH.
• R 1 is halo.
• R 1 is -F.
• R 1 is -CI.
• R 1 is -Br.
• R 1 is -I.
• R 2 is -OH.
• R 2 is halo.
• R 2 is -F.
• R 2 is -CI.
• R 2 is -Br.
• R 2 is- 1.
• R 5 is -OH.
• R 5 is halo such as but not limited to -F, -CI, -Br, or -I.
• R 5 is -F.
• R 5 is -CI.
• R 5 is -Br.
• R 5 is -I.
• R 5 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl.
• R 3 is selected from -OH, alkyl-C(0)-0- and R a R b N-aryloxy. In some of these examples, alkyl-C(0)-0- or R a R b N-aryloxy is optionally substituted with halo. In some examples, R 3 is -OH. In some examples, R 3 is alkyl- C(0)-0- In some examples, R 3 is R a R b N-aryloxy- In some examples, R 3 is In some examples, R 3 i is ome examples, R is
• R 3 is In some examples, R 3
• R 3 is R a R b N-aryloxy-. In some examples,
• R 3 is - R a R b -aryloxy.
• R 4 is selected from the group consisting of alkyl, aryl, arylalkyl, and an N-containing heterocycloalkyl. In some of these examples, alkyl, aryl, arylalkyl, or N-containing heterocycloalkyl are optionally substituted with - R a R b .
• R 4 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl. In some examples, R 4 is methyl. In some examples, R 4 is ethyl.
• R 4 is n-propyl. In some examples, R 4 is i-propyl. In some examples, R 4 is n-butyl. In some examples, R 4 is i-butyl. In some examples, R 4 is t-butyl. In some examples, R 4 is pentyl. In some examples, R 4 is hexyl. In some examples, R 4 is heptyl. In some examples, R 4 is octyl, or nonyl. In some examples, R 4 is aryl such as but not limited to phenyl or naphthyl. In some examples, R 4 is phenyl. In some examples, R 4 is naphthyl.
• R 4 is arylalkyl-such as but not limited to benzyl. In some examples, R 4 is N- containing heterocycloalkyl such as but not limited to pipendinyl. In some examples, R 4 is 4- amino-phenyl. In some examples, R 4 is 4-aminophenyl optionally substituted with halo. 4 is In some examples, R 4 is
• R is . in some examples, R is
• R 4 is . In some examples, R 4 is ample e e xamples, R .4 is . In some examples, R 4 is . In
• R s examples, R 4 ome examples, R is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is .
• R 4 In some examples, R 4 In some examples, R is some examples, R is In some examples, R 4 is . in some examples, R 4
• R is . In some examples, R is ples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is
• R is
• R is In some examples, R 4 is
• R 4 is alkyl substituted with amino such as, but not limited to, methyl-amino, ethyl-amino, propyl-amino, butyl-amino, pentyl-amino, hexyl-amino, heptyl- amino, octyl-amino, or nonyl-amino.
• R 4 is methyl-amino.
• R 4 is ethyl-amino.
• R 4 is n-propyl-amino.
• R 4 is i-propyl-amino.
• R 4 is n-butyl-amino.
• R 4 is i-butyl- amino. In some examples, R 4 is t-butyl-amino. In some examples, R 4 is pentyl-amino. In some examples, R 4 is hexyl-amino. In some examples, R 4 is heptyl-amino. In some examples, R 4 is octyl-amino. In some examples, R 4 is nonyl-amino.
• R 4 is
• R 4 is
• R a and R b are, independently in each instance, selected from -H or alkyl. In some examples, both R a and R b are -H. In some examples, both R a and R b are methyl. In some examples, both R a and R b are ethyl. In some examples, both R a and R b are propyl. In some examples, one of R a or R b is -H and the other is alkyl. In some examples, one of R a or R b is -H and the other is methyl. In some examples, one of R a or R b is -H and the other is ethyl. In some examples, one of R a or R b is -H and the other is propyl.
• n is an integer from 0-19. In some examples, n is 0. In some other examples, n is 1. In certain examples, n is 2. In some other examples, n is 3. In certain examples, n is 4. In some examples, n is 5. In some other examples, n is 6. In certain examples, n is 7. In some other examples, n is 8. In certain examples, n is 9. In some examples, n is 10. In some other examples, n is 1 1. In certain examples, n is 12. In some other examples, n is 13. In certain examples, n is 14. In some examples, n is 15. In some other examples, n is 16. In certain examples, n is 17. In some other examples, n is 18. In certain examples, n is 19.
• R 3 is not -OH when R 1 is -OH. [0128 In some examples, in Formula (Pla), R 3 is not -OH when R 1 and R 2 together
• R 4 is a Ci-9alkyl or 4-(dimethyl-amino)-phenyl. In some examples, R 4
• n is 0. In some examples, n is 1. In some examples, n is 2.
• R 3 is selected from alkyl-C(0)-0- or R a R b N-aryloxy-; wherein alkyl-C(0)-0- or R a R b N-aryloxy- are optionally substituted with halo.
• set forth herein is a compound of Formula (Pla), (PIb-1), -2), (PIc-1), (PIc-2), (PId-1), (PId-2), (PIe-1), or (PIe-2), wherein R 3 is [0138] In some examples, set forth herein is a compound of Formula (Pla), (PIb-1), -2), (PIc-1), (PIc-2), (PId-1), (PId-2), (PIe-1), or (PIe-2), wherein R 3 is
• R 3 is selected from -OH, alkyl-C(0)-0- and R a R b N-aryloxy- In some of these examples, alkyl-C(0)-0- or R a R b N-aryloxy- is optionally substituted with halo. In some examples, R 3 is -OH. In some examples, R 3 is alkyl-C(0)-0- In some examples, R 3 is a R b N-aryloxy- In some
• R 3 is In some examples, R 3 is ome
• R 3 is In some examples, R 3 is In
• R 3 is In some examples, R 3 is R a R b N-aryloxy In some examples, R 3 is - R a R b -aryloxy.
• R 3 is R a R b N- aryloxy- wherein R a and R b are, independently in each instance, H or alkyl.
• R 4 is selected from the group consisting of alkyl, aryl, arylalkyl, and an N-containing heterocycloalkyl.
• alkyl, aryl, arylalkyl, or N-containing heterocycloalkyl are optionally substituted with -NR a R b .
• R 4 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl.
• R 4 is methyl. In some examples, R 4 is ethyl. In some examples, R 4 is n-propyl. In some examples, R 4 is i- propyl. In some examples, R 4 is n-butyl. In some examples, R 4 is i-butyl. In some examples, R 4 is t-butyl. In some examples, R 4 is pentyl. In some examples, R 4 is hexyl. In some examples, R 4 is heptyl. In some examples, R 4 is octyl, or nonyl. In some examples, R 4 is aryl such as but not limited to phenyl or naphthyl. In some examples, R 4 is phenyl.
• R 4 is naphthyl. In some examples, R 4 is arylalkyl-such as but not limited to benzyl. In some examples, R 4 is N-containing heterocycloalkyl such as but not limited to piperidinyl. In some examples, R 4 is 4-amino-phenyl. In some examples, R 4 is 4-aminophenyl optionally substituted with halo.
• R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is In some examples, R is . In some examples, R 4 is
• R 4 is . In some examples, R 4 is
• R is
• R is . In some examples, R is
• R 4 is In some examples, R 4 is . In some examples, R 4 is 3 ⁇ 4— " . In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is
• R is . In some examples, R 4 is
• R 4 is 3 ⁇ 4— V . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is In some examples, R 4 is . In some examples, R 4 is In some examples, R is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is in some examples, R 4 is in some examples, R 4 is 3 ⁇ 4— V . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is . In some examples, R 4 is .
• R 4 is In some examples, R is In some exam les R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is
• R 4 is In some examples, R is ome examples, R 4 is In some examples, [0146] In some examples, set forth herein is a compound of Formula (Pla), (PIb-1),
• R 4 is alkyl substituted with amino such as, but not limited to, methyl-amino, ethyl-amino, propyl-amino, butyl-amino, pentyl-amino, hexyl-amino, heptyl-amino, octyl-amino, or nonyl-amino.
• R 4 is methyl-amino.
• R 4 is ethyl-amino.
• R 4 is n-propyl-amino.
• R 4 is i-propyl-amino. In some examples, R 4 is n-butyl-amino. In some examples, R 4 is i-butyl-amino. In some examples, R 4 is t- butyl-amino. In some examples, R 4 is pentyl-amino. In some examples, R 4 is hexyl-amino. In some examples, R 4 is heptyl-amino. In some examples, R 4 is octyl-amino. In some examples, R 4 is nonyl-amino.
• R 4 is
• R 4 is
• the compound of Formula (I) is not one of the following compounds:
• R 3 is selected from -OH, alkyl-C(0)-0- or R a R b N-aryloxy.
• alkyl-C(0)-0- or R a R b N-aryloxy is optionally substituted with halo.
• R 3 is -OH. In some examples, R 3 is
• R 3 is a R b N-aryloxy- In some examples, R 3 is In some examples, R 3 is ome examples, R is
• R 3 is In some examples, R 3
• R 3 is R a R b N-aryloxy-.
• R 4 is selected from the group consisting of alkyl, aryl, arylalkyl, and an N-containing heterocycloalkyl.
• alkyl, aryl, arylalkyl, or N-containing heterocycloalkyl are optionally substituted with
• R 4 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl.
• R 4 is methyl.
• R 4 is ethyl.
• R 4 is n-propyl.
• R 4 is i-propyl.
• R 4 is n-butyl.
• R 4 is i-butyl.
• R 4 is t-butyl.
• R 4 is sec-butyl.
• R 4 is pentyl.
• R 4 is hexyl. In some examples, R 4 is heptyl. In some examples, R 4 is octyl, or nonyl. In some examples, R 4 is aryl such as but not limited to phenyl or naphthyl. In some examples, R 4 is phenyl. In some examples, R 4 is naphthyl. In some examples, R 4 is aryl alky 1-such as but not limited to benzyl. In some examples, R 4 is N-containing heterocycloalkyl such as but not limited to piperidinyl. In some examples, R 4 is 4-amino-phenyl. In some examples, R 4 is 4- aminophenyl optionally substituted with halo.
• R 4 is In some examples, R 4 is In some examples, R is In some examples, R is In some examples, R is In some examples, R 4 is . In some examples, R 4 is
• n some examples, n some examples, R 4 is
• R 4 is some examples, R 4 is some examples, R 4 is In some examples,
• R 4 is In some examples, R 4 is In some examples, R 4 is . In some examples, R 4 is In some examples, R is In some examples, R is In some examples, R is . In some examples, R 4 is . I Inn s soommee e exxaammnplleess., R R 4 i iss . In some examples, R is In some examples, R 4 is . In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is In some examples, R 4 is xamples, R 4 is In some examples, R 4 is
• R 4 is
• R 4 is In some examples, R is F
• R 4 is In some examples, R 4 is
• R 4 is alkyl substituted with amino such as, but not limited to, methyl-amino, ethyl-amino, propyl-amino, butyl-amino, pentyl-amino, hexyl-amino, heptyl- amino, octyl-amino, or nonyl-amino.
• R 4 is methyl-amino.
• R 4 is ethyl-amino.
• R 4 is n-propyl-amino.
• R 4 is i-propyl-amino.
• R 4 is n-butyl-amino.
• R 4 is i-butyl- amino. In some examples, R 4 is t-butyl-amino. In some examples, R 4 is pentyl-amino. In some examples, R 4 is hexyl-amino. In some examples, R 4 is heptyl-amino. In some examples, R 4 is octyl-amino. In some examples, R 4 is nonyl-amino. [0 In some examples, R 4 i is In some examples, R 4 i IS
• R a and R b are, independently in each instance, selected from H or alkyl. In some examples, both R a and R b are -H. In some examples, both R a and R b are methyl. In some examples, both R a and R b are ethyl. In some examples, both R a and R b are propyl. In some examples, one of R a or R b is -H and the other is alkyl. In some examples, one of R a or R b is -H and the other is methyl. In some examples, one of R a or R b is -H and the other is ethyl. In some examples, one of R a or R b is -H and the other is propyl.
• n is an integer from 0-19. In some examples, n is 0. In some other examples, n is 1. In certain examples, n is 2. In some other examples, n is 3. In certain examples, n is 4. In some examples, n is 5. In some other examples, n is 6. In certain examples, n is 7. In some other examples, n is 8. In certain examples, n is 9. In some examples, n is 10. In some other examples, n is 11. In certain examples, n is 12. In some other examples, n is 13. In certain examples, n is 14. In some examples, n is 15. In some other examples, n is 16. In certain examples, n is 17. In some other examples, n is 18. In certain examples, n is 19.
• R 4 is selected from 4-amino-phenyl, 4-amino-l-methyl-phenyl, 2-amino-ethyl, piperidinyl, or propyl.
• R 4 is 4-amino-phenyl.
• R 4 is 4- amino-l-methyl-phenyl.
• R 4 is 2-amino-ethyl.
• R 4 is piperidinyl.
• R 4 is propyl.
• R 4 is n-propyl.
• R 4 is i-propyl.
• set forth herein is a compound of Formula (Plla) or (Pllb), wherein R 3 is selected from alkyl-C(0)-0- or R a R b N-aryloxy; wherein alkyl-C(0)-0-, or
• R a R b N-aryloxy a re optionally substituted with halo.
• R 9 is selected from H or -NR a R b . In some examples, R 9 is
• R 9 is -NR a R b , R 4 , R 4 , and subscript n are defined as in Formula I and noted above.
• R 10 and R 11 are each, independently in each instance, selected from H, F, or -NR a R b .
• R 4 , R 4 , and subscript n are defined as in Formula I and noted above.
• R 3 is selected from alkyl-C(0)-0- or R a R b N-aryloxy, wherein alkyl-C(O)- 0-, or - R a R b -aryloxy are optionally substituted with halo.
• R 4 is selected from -H, -OH, halo, or alkyl.
• R 4 is halo such as but not limited to -F, -CI, -Br, or -I.
• R 4 is -F.
• R 4 is- CI.
• R 4 is -Br.
• R 4 is -I.
• R 4 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl.
• Subscript n is an integer from 0-19.
• n is 0. In some other examples, n is 1. In certain examples, n is 2. In some other examples, n is 3. In certain examples, n is 4. In some examples, n is 5. In some other examples, n is 6. In certain examples, n is 7. In some other examples, n is 8. In certain examples, n is 9. In some examples, n is 10. In some other examples, n is 11. In certain examples, n is 12. In some other examples, n is 13. In certain examples, n is 14. In some examples, n is 15. In some other examples, n is 16. In certain examples, n is 17. In some other examples, n is 18. In certain examples, n is 19.
• R 3 is selected from -OH, alkyl-C(0)-0-
• alkyl-C(0)-0- or R a R b N-aryloxy is optionally substituted with halo.
• R 3 is -OH.
• R 3 is alkyl- C(0)-0- In some examples, R 3 is R a R b N-aryloxy.
• R 3 is -NR a R b . In some examples, R 3 is -NH2. In some examples, R 3 is - H(CH 3 ).
• R 3 is R a R b N-aryloxy. examples, R is In some examples, R 3 is In some examples, R 3 is In some examples, R . In some examples, R is
• R 3 is
• R 3 is In some examples, R 3 is
• R 3 is In some examples, R 3 is
• R 3 is selected from alkyl-C(0)-0- or R a R b N-aryloxy, wherein alkyl- C(0)-0- or R a R b N-aryloxy are optionally substituted with halo.
• R 4 is selected from -H, -OH, halo, or alkyl.
• R 4 is halo such as but not limited to -F, -CI, -Br, or -I.
• R 4 is -F.
• R 4 is -CI.
• R 4 is -Br.
• R 4 is -I.
• R 4 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl.
• Subscript n is an integer from 0-19.
• n is 0. In some other examples, n is 1. In certain examples, n is 2. In some other examples, n is 3. In certain examples, n is 4. In some examples, n is 5. In some other examples, n is 6. In certain examples, n is 7. In some other examples, n is 8. In certain examples, n is 9. In some examples, n is 10. In some other examples, n is 11. In certain examples, n is 12. In some other examples, n is 13. In certain examples, n is 14. In some examples, n is 15. In some other examples, n is 16. In certain examples, n is 17. In some other examples, n is 18. In certain examples, n is 19. In some examples, in Formula (PVII), R 3 is selected from -OH, alkyl-C(0)-0-
• alkyl-C(0)-0- or R a R b -aryloxy is optionally substituted with halo.
• R 3 is -OH.
• R 3 is alkyl-C(0)-0-
• R 3 is R a R b N-aryloxy.
• R 3 is -NR a R b . In some examples, R 3 is -NH2. In some examples, R 3 is - H(CH 3 ).
• R 3 is R a R b N-aryloxy-
• R is . In some examples, R is
• R . 3 is . In some examples, R 3 is . In some examples, R 3 is In
• R 3 is
• R 3 is F F . i n some examples, R is F'
• R 3 is
• R is
• set forth herein is a compound of Formula (PVII), (PVIIa), or (PVIIb), wherein R 3 is [0188] In some examples, set forth herein is a compound of Formula (PVII), (PVIIa), or (PVIIb), wherein R 3 is
• R 1 and R 2 are,
• R is -H.
• R 1 is alkyl.
• R 1 is alkyl-C(0)-0-
• R 1 is -OH.
• R 1 is halo.
• R 1 is -F.
• R S -CI.
• R 1 is -Br.
• R 1 is -I.
• R 2 is -OH.
• R 2 is halo.
• R 2 is -F.
• R 2 is -CI.
• R 2 is -Br.
• R 2 is -I.
• R 5 is, independently in each instance, selected from -OH, halo, alkyl, or arylalkyl.
• R 5 is -OH.
• R 5 is halo such as but not limited to -F, -CI, -Br, or -I.
• R 5 is -F.
• R 5 is -CI.
• R 5 is -Br.
• R 5 is -I.
• R 5 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl. In some examples, R 5 is benzyl.
• R 4 is selected from the group consisting of alkyl, aryl, arylalkyl, and an N-containing heterocycloalkyl.
• alkyl, aryl, arylalkyl, or N-containing heterocycloalkyl are optionally substituted with
• R 4 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl.
• R 4 is methyl.
• R 4 is ethyl.
• R 4 is n-propyl.
• R 4 is i-propyl.
• R 4 is n-butyl.
• R 4 is i-butyl.
• R 4 is t-butyl.
• R 4 is sec-butyl.
• R 4 is pentyl.
• R 4 is hexyl. In some examples, R 4 is heptyl. In some examples, R 4 is octyl, or nonyl. In some examples, R 4 is aryl such as but not limited to phenyl or naphthyl. In some examples, R 4 is phenyl. In some examples, R 4 is naphthyl. In some examples, R 4 is arylalkyl-such as but not limited to benzyl. In some examples, R 4 is N-containing heterocycloalkyl such as but not limited to piperidinyl. In some examples, R 4 is 4-amino-phenyl. In some examples, R 4 is 4- aminophenyl optionally substituted with halo. » exa m m consentpilTMes, ⁇ R> 4 is In some examples, R is In some examples, R is n some examples, R 4 is
• R 4 is
• R is
• R 4 is
• R 4 is In some examples, R 4 is
• R 4 is In some examples,
• R 4 is alkyl substituted with amino such as, but not limited to, methyl-amino, ethyl-amino, propyl-amino, butyl-amino, pentyl-amino, hexyl-amino, heptyl- amino, octyl-amino, or nonyl-amino.
• R 4 is methyl-amino.
• R 4 is ethyl-amino.
• R 4 is n-propyl-amino.
• R 4 is i-propyl-amino.
• R 4 is n-butyl-amino.
• R 4 is i-butyl- amino. In some examples, R 4 is t-butyl-amino. In some examples, R 4 is sec-butyl. In some examples, R 4 is pentyl-amino. In some examples, R 4 is hexyl-amino. In some examples, R 4 is heptyl-amino. In some examples, R 4 is octyl-amino. In some examples, R 4 is nonyl-amino.
• R 4 is
• R 4 is [0200]
• R a and R b are, independently in each instance, selected from H or alkyl. In some examples, both R a and R b are H. In some examples, both R a and R b are methyl. In some examples, both R a and R b are ethyl. In some examples, both R a and R b are propyl. In some examples, one of R a or R b is H and the other is alkyl. In some examples, one of R a or R b is H and the other is methyl. In some examples, one of R a or R b is H and the other is ethyl. In some examples, one of R a or R b is H and the other is propyl.
• n is an integer from 0-19. In some examples, n is 0. In some other examples, n is 1. In certain examples, n is 2. In some other examples, n is 3. In certain examples, n is 4. In some examples, n is 5. In some other examples, n is 6. In certain examples, n is 7. In some other examples, n is 8. In certain examples, n is 9. In some examples, n is 10. In some other examples, n is 11. In certain examples, n is 12. In some other examples, n is 13. In certain examples, n is 14. In some examples, n is 15. In some other examples, n is 16. In certain examples, n is 17. In some other examples, n is 18. In certain examples, n is 19.
• R 3 is not -OH when R 1 is -OH.
• R 3 is not -OH when R 1 and R 2 together form
• R 4 is a Ci-9alkyl or 4-(dimethyl-amino)-phenyl.
• R 4 is alkyl, aryl, arylalkyl, or a N- containing heterocycloalkyl. In certain examples, alkyl, aryl, arylalkyl, or N-containing heterocycloalkyl are optionally substituted with -NR a R b . In some of these examples, R 4 is alkyl. In some of these examples, R 4 is aryl. In some of these examples, R 4 is arylalkyl. In some of these examples, R 4 is N-containing heterocycloalkyl.
• R 4 is alkyl such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, or nonyl. In some examples, R 4 is methyl. In some examples, R 4 is ethyl. In some examples, R 4 is n- propyl. In some examples, R 4 is i-propyl. In some examples, R 4 is n-butyl. In some examples, R 4 is i-butyl. In some examples, R 4 is t-butyl. In some examples, R 4 is sec-butyl. In some examples, R 4 is pentyl.
• R 4 is hexyl. In some examples, R 4 is heptyl. In some examples, R 4 is octyl. In some examples, R 4 is nonyl. In some examples, R 4 is aryl such as but not limited to phenyl or naphthyl. In some examples, R 4 is phenyl. In some examples, R 4 is naphthyl. In some examples, R 4 is arylalkyl-such as but not limited to benzyl. In some examples, R 4 is N-containing heterocycloalkyl such as but not limited to piperidinyl. In some examples, R 4 is 4-amino-phenyl. In some examples, R 4 is 4-aminophenyl optionally substituted with halo.
• R 4 and R 2 together form ⁇ " ⁇ TM wherein R 4 is selected from the group consisting of alkyl, aryl, arylalkyl, and a N-containing heterocycloalkyl; and wherein alkyl, aryl, arylalkyl, or N- containing heterocycloalkyl are optionally substituted with -NR a R b ; and wherein the stereochemistry of the carbon indicated by * is R.
• R 4 and R 2 together form "" " ⁇ f" wherein R 4 is selected from the group consisting of alkyl, aryl, arylalkyl, and a N-containing heterocycloalkyl; and wherein alkyl, aryl, arylalkyl, or N- containing heterocycloalkyl are optionally substituted with -NR a R b ; and wherein the stereochemistry of the carbon indicated by * is S.
• R 3 is not -OH when R 1 is -OH or when R 1 and R 2 together form
• R 4 is a Ci-9alkyl or 4-(dimethyl-amino)-phenyl.
• the payload set forth herein is a derivative or analog of budesonide or diflorasone.
• the derivative is an amine or aniline containing molecule which is related in structure to budesonide or diflorasone.
• the payloads set forth herein as well as other steroids can be conjugated to an antibody or an antigen-binding fragment thereof based on the methods set forth herein.
• the payloads set forth herein as well as other steroids can be conjugated to an antibody, or an antigen-binding fragment thereof, and also to a cyclodextrin moiety based on the methods set forth herein.
• stable linker-payloads can be use with these methods of conjugation to produce antibody-steroid-conjugates.
• the antibody-steroid conjugates also include a cyclodextrin moiety.
• R 1 and R 2 are, independently, -H, alkyl, alkyl-C(0)-0- -OH, or halo; or R 1 and R 2 together form ,
• R 4 is alkyl, aryl, arylalkyl, or an N-containing heterocycloalkyl, wherein the alkyl, aryl, arylalkyl, and N-containing heterocycloalkyl are, independently in each instance, optionally substituted with -NR a R b ;
• R 5 is, independently in each instance, -OH, halo, alkyl, or arylalkyl;
• R 3 is -OH, alkyl-C(0)-0- or -X-aryl-NR a R b , wherein X is selected from S, S(O), S(0) 2 , S0 2 NR a , CONR a , C(O), or NR a , wherein the alkyl-C(0)-0- and -X- aryl-NR a R b are optionally substituted with halo or prodrug.
• R a and R b are, independently in each instance, H or alkyl, aryl;
• R c is -H or alkyl
• n is an integer from 0-19;
• R 3 is not -OH when either (a) or (b): (a) R 1 is -OH or (b) R 1 and
• R 2 together form and R 4 is a Ci-9alkyl or
• alkyl-C(0)-0- alkyl-C(0)-0- or
• R 3 is -OH. In some examples, R 3 is alkyl-C(0)-0- In some examples, R 3 is R a R b N-aryloxy. In some examples, R 3 is In some examples, R 3 is
• R 3 is H 2
• R 3 is -X-aryl- R a R b
• R 3 is In some examples, R 3 is In some examples, R 3 is In some examples, R 3 is In some examples, R , is . In some examples, R , is In some examples, R 3 is n some examples, R 3 is . In some examples, R is . In some examples, R is
• R 3 is examples, R is . In some examples, R is some examples, R 3 is . In some examples, R is . In some examples, R is . In some examples, R is . In some examples, R is . In some examples, R is . In some examples, R is .
• R 3 is In some examples, R is F'
• R 3 is
• R is examples, R 3 is . In some examples, R 3 is . In some examples, R 3 is . In some examples, R 3 is . In some examples, R 3 is . In some examples, R 3 is
• R is
• R 3 is . In some examples, R 1 is . In some examples, R 3 is . In some examples, R 3 is . In some examples, R 3 is . In some examples, R 3 is some examples, R 3 is n some examples, R 3 is
• R 3 is examples, R is . In some examples, R is some examples, R 3 is
• R 3 is
• R 3 is F F . in some examples, R 3 is F'
• R 3 is
• R 3 has a structure selected from one of the following structures:
• R is . In some exam les, R is
• R 3 is .
• q is an integer from 0 to 5.
• R 3 has a structure selected from one of the following structures:
• R 3 is i n some examples, R 3 is
• I n some examples, .
• q is an integer from 0 to 5.
• R has a structure selected from one of the following structures:
• RR 3 some examp] In some examples, R 3 is
• R 3 is
• R 1 and R 2 are, independently, selected from the group consisting of -H, -OH, alkyl,
• R and R together form ⁇ " - ⁇ , wherein R is selected from the group consisting of alkyl, aryl, alkylaryl, arylalkyl, heteroaryl, -alkylene-NR a R b ,
• R 3 is selected from the group consisting of -OH, -0-C(0)-alkyl, -O-aiyl, - R a R b , -alkylene- R a R b , -X-arylene-Y- R a R b , -X-heteroarylene-Y- R a R b , and N-containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or -CH2-.
• R 5 is, independently in each instance, selected from a substituent in the group consisting of -OH, halo, and alkyl; n is an integer from 0-19; and each R 5 is positioned on any ring atom.
• R a and R b are, independently in each instance, selected from the group consisting of -H and alkyl; or R a and R b cyclize to form
• R a and R b are, independently in each instance, optionally substituted with at least one substituent selected from the group consisting of -OH, -PO4H, NH2,
• R 3 is selected from the group consisting of -alkylene-NR a R b ,
• R 4 is selected from the group consisting of alkyl, aryl, alkylaryl, arylalkyl, heteroaryl, -alkylene- R a R b ,
• R 3 is selected from the group consisting of -OH, -0-C(0)-alkyl, -O-aiyl, -NR a R b , -alkylene-NR a R b , -X-arylene-Y-NR a R b , -X-heteroarylene-Y-NR a R b , and N-containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or
• R 4 is selected from the group consisting of -alkylene-NR a R b , -X-arylene-Y-NR a R b , -X-heteroarylene-Y-NR a R b , and N-containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or -CH2-.
• R 3 is -NR a R b ; and R 4 is alkyl.
• R 3 is -NH2.
• R 4 is n-propyl.
• R 3 is -NH2 and R 4 is n-propyl.
• the compound of Formula 1000 is according to
• the compound of Formula 1000 is according to
• R 3 i is -OH or
• R 4 is -alkylene- R a R b , -X-arylene- R a R b , -X-heteroarylene- R a R b , or N-containing heterocycloalkyl; wherein X is absent or -CH2-.
• R 4 is -alkylene- H 2 , -CeHs-NHz or -CH2-C6H5- H2.
• R 3 is -O-aiyl
• R 4 is alkyl, aryl, alkylaryl, or arylalkyl.
• R 3 is
• R 3 is -0-phenyl-NR a R b , -0-heteroaiylene-NR a R b ; wherein phenyl or heteroaryl is optionally substituted with halogen or deuterium.
• R 4 is n-propyl.
• R 3 is -NR a R b ; and R 4 is alkyl.
• R 3 is -NH2.
• R 4 is n-propyl.
• R 3 is -NH2 and R 4 is n-propyl.
• R 3 can be any specific R 3 provided above.
• R 3 is -NH2, -N(H)CH 3 , -N(CH 3 ) 2 , or .
• R 3 is- or
• R is
• R 4 can be any specific R 4 provided above.
• R 4 is selected from -CH2-CH2-NH2,
• R 4 is n-propyl.
• compositions include pharmaceutically acceptable salts, solvates, crystalline forms, amorphous forms, polymorphic forms, regioisomers, stereoisomers, prodrugs, e.g., phosphatase-prodrugs, glucose-prodrugs, ester prodrugs, etc., metabolites, and physiological adducts of the steroid payloads described herein, including those of Formula (I), (I 1 ), and (A 1 )- ⁇ 7 ).
• prodrugs e.g., phosphatase-prodrugs, glucose-prodrugs, ester prodrugs, etc., metabolites, and physiological adducts of the steroid payloads described herein, including those of Formula (I), (I 1 ), and (A 1 )- ⁇ 7 ).
• conjugates of the steroids described herein include proteins, e.g., antibodies or antigen-binding fragments thereof, that are covalently linked, e.g., via the binding agent linkers described herein, to the compounds described in Section B above, e.g., the compounds of Formula (A), (A 1 ), (A 2 ), (A 3 ), (A 4 ), (A 5 ), (A 6 ), (A 7 ), (I), (I 1 ), (Pla), (PIb-1), (PIb-2), PIc-1), (PIc-2), (PId-1), (PId-2), (PIe-1), (PIe-2), (PII), (Plla), (Pllb), (PHI), (Pllla), (Plllb), (PIV), (PV), (PVa), (PVb), (PVI), (PVII), (PVIIa), (PVIIb), (PVIIb-1), (PVIIb-2), (PVIIb-2
• the binding agent linker can be linked to a steroid described herein at any suitable moiety or position of the steroid, including e.g., through an amide, ether, ester, carbamate, or amine.
• the binding agent linker can be attached to compounds through R 1 , R 3 , or R 4 or hydroxyl group depicted Formula (A 1 ):
• the steroids described herein are attached to the binding agent linker by reacting an amino or hydroxyl group of the steroid with a suitable reactive group present on the linker.
• the binding agent linker also includes a cyclodextrin moiety.
• the cyclodextrin moiety may be bonded to the chemical backbone structure of the binding agent linker.
• each PAY is a radical obtainable by removal of an atom, for example a hydrogen atom from a compound according to a Formula selected from the group consisting of
• R 3 is -BL- ,-BL-X- or ;
• R 1 and R 2 are each, independently, -H, alkyl, alkyl-C(0)-0- -OH, or
• R 4 halo; or R 1 and R 2 together form " ⁇ " , wherein R 4 is alkyl, aryl, arylalkyl, or an N-containing heterocycloalkyl; wherein the alkyl, aryl, arylalkyl, and
• N-containing heterocycloalkyl are optionally substituted with -NR a R b ;
• R 3 is -OH, alkyl-C(0)-0-, heteroalkyl, -NR a R b or aryloxy, wherein the alkyl-C(0)-0- heteroalkyl, or aryloxy is optionally substituted with
• R 4 is -BL- - BL - X - ⁇ -(CH 2 )o. 2 - _ BL -x-(CH 2 ) 1 -4 — or -BL-Y, wherein Y is an N-containing divalent heterocycle;
• -BL- is a divalent binding agent linker
• R 5 is, independently in each instance, -OH, halo, alkyl, or arylalkyl;
• R a and R b are, independently in each instance, -H or alkyl
• R p independently in each instance, is halo
• BA is a binding agent bonded to -BL-;
• ependently in each instance is NR a or O; is aryl or heteroaryl;
• t is an integer from 0-2;
• x is an integer from 1-30;
• n is an integer from 0-19.
• subscript x is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15,, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30. In some examples, subscript x is 0. In some examples, subscript x is 1. In some examples, subscript x is 2. In some examples, subscript x is 3. In some examples, subscript x is 4. In some examples, subscript x is 5. In some examples, subscript x is 6. In some examples, subscript x is 7. In some examples, subscript x is 8. In some examples, subscript x is 9. In some examples, subscript x is 10. In some examples, subscript x is 11. In some examples, subscript x is 12. In some examples, subscript x is 13.
• subscript x is 14. In some examples, subscript x is 15. In some examples, subscript x is 16. In some examples, subscript x is 17. In some examples, subscript x is 18. In some examples, subscript x is 19. In some examples, subscript x is 20. In some examples, subscript x is 21. In some examples, subscript x is 22. In some examples, subscript x is 23. In some examples, subscript x is 24. In some examples, subscript x is 25. In some examples, subscript x is 26. In some examples, subscript x is 27. In some examples, subscript x is 28. In some examples, subscript x is 29. In some examples, subscript x is 30.
• R 1 and R 2 are, each, independently, -H, alkyl, or -OH. In some examples of Formula (III), one of R 1 or R 2 is -H, alkyl, or -OH. In some examples of Formula (III), both R 1 and R 2 are either -H, alkyl, or -OH.
• R 1 and R 2 together form " " " " ⁇ .
• R 4 is -RL.
• R 4 is RL-NR a -aryl.
• R 4 is alkyl.
• R 4 is arylalkyl,
• R 4 is aryl.
• R 4 is N-containing heterocycloalkyl.
• the alkyl, aryl, arylalkyl, or N-containing heterocycloalkyl is optionally substituted.
• R 5 is -H or halo.
• R 5 is -H or fluoro. In some examples of Formula (III), one of R 5 is -H or halo. In some examples of Formula (III), R 5 is -H or halo and n is 2. In some examples of Formula (III), R 5 is -F and n is 1. In some examples of Formula (II), R 5 is -F and n is 2.
• R 3 is BL.
• R 3 is RL-NR a -aryloxy- In some other examples of Formula (III), R 3 is -OH. In some other examples of Formula (III), R 3 is alkyl-C(0)-0- In some other examples of Formula (III), R 3 is heteroalkyl. In some other examples of Formula (III), R 3 is -N-R a R b . In some other examples of Formula (III), R 3 is aryl. In some other examples of Formula (III), R 3 is aryloxy. In some other examples of Formula (III), alkyl-C(0)-0-, heteroalkyl, or aryloxy is optionally substituted with -NR a R b or halo.
• R 3 is -OH. In some examples of Formula (III), R 3 is alkyl- C(0)-0- In so is heteroalkyl. In s of
• R 3 is -NR a R b -aryloxy. In some examples, R 3 is Ra . In some
• R 3 is examples, R 3 is Rb . In some examples R 3 is aryloxy In some exampl In some
• R 3 is [0244] In Formula (III), subscript n is an integer from 0-19. In some examples, n is 0.
• n is 1. In certain examples, n is 2. In some other examples, n is 3. In certain examples, n is 4. In some examples, n is 5. In some other examples, n is 6. In certain examples, n is 7. In some other examples, n is 8. In certain examples, n is 9. In some examples, n is 10. In some other examples, n is 11. In certain examples, n is 12. In some other examples, n is 13. In certain examples, n is 14. In some examples, n is 15. In some other examples, n is 16. In certain examples, n is 17. In some other examples, n is 18. In certain examples, n is 19.
• BA is a binding agent
• R 5 is, independently in each instance, -OH, halo, or alkyl
• R 3 is selected from -OH, alkyl-C(0)-0- heteroalkyl, - R a R b , -NR a R b - aryloxy,
• alkyl-C(0)-0- heteroalkyl, or aryloxy is optionally substituted with -NR a R b or halo;
• BL is a binding agent linker
• R a and R b are, independently in each instance, selected from H, alkyl, and alkyl-C(O);
• n is an integer from 0 to 19;
• x is an integer from 1 to 30.
• BA is a binding agent
• R 5 is, independently in each instance, -OH, halo, or alkyl
• R 3 is -OH, alkyl-C(0)-0- heteroalkyl, -NR a R b , - R a R b -aryloxy, or aryloxy, wherein the alkyl-C(0)-0- heteroalkyl, or aryloxy is optionally substituted with -NR a R b or halo;
• BL is a binding agent linker
• R a and R b are, independently in each instance, selected from H, alkyl, or alkyl-C(O);
• n is an integer from 0 to 19;
• x is an integer from 0 to 30.
• Formula (IIIa 3 is -OH. In some examples of Formula (IIIa2), R 3 is
• R 3 is heteroalkyl. In some examples R 3 is In some examples
• R 3 is - R a R b . In some is . In some examples R is aryloxy. In some examples R 3 ; , In some examples R is . In some examples R 3 is
• R 3 is
• the compound of Formula (IIIa2) has the following structure:
• BA is a binding agent
• R 3 is -OH or alkyl-C(0)-0-
• R 5a and R 5b are each, independently, -F or H;
• BL is a binding agent linker
• x is an integer from 1 to 30.
• BA is a binding agent
• R 5 is, independently in each instance, -OH, halo, or alkyl
• R 4 is selected from alkyl, aryl, arylalkyl, or an N-containing heterocycloalkyl, wherein the alkyl, aryl, arylalkyl, or N-containing heterocycloalkyl are optionally
• RL is a binding agent linker
• R a and R b are, independently in each instance, selected from H, alkyl, and alkyl-C(O);
• n is an integer from 0 to 19;
• x is an integer from 0 to 30.
• R 5 is -H or halo.
• R 5 is fluoro. In some examples of Formula (Illb), n is at least 2, and two of R 5 is halo. In some examples of Formula (Illb), R 5 is -F and n is 1. In some examples of Formula (Illb), R 5 is -F.
• R 4 is alkyl.
• R 4 is methyl, ethyl, ⁇ -propyl, /-propyl, «-butyl, s-butyl, t-butyl, /-butyl, a pentyl moiety, a hexyl moiety, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.
• R 4 is ⁇ -propyl.
• the compound of Formula (Illb) has the following structure:
• BA is a binding agent
• R 4 is alkyl
• R 5a and R 5b are each, independently, -F or H;
• BL is a binding agent linker
• x is an integer from 1 to 30.
• BA is a binding agent
• R 1 and R 2 are, independently,— H, alkyl, alkyl— C(0)—0—,—OH, or halo;
• R 5 is, independently in each instance, selected from -OH, halo, or alkyl
• BL is a binding agent linker
• n is an integer from 0 to 19;
• x is an integer from 1 to 30.
• R 5 is halo.
• R 5 is fluoro. In some examples of Formula (IIIc), one of R 5 is halo. In some examples of Formula (IIIc), two of R 5 is halo. In some examples of Formula (IIIc), R 5 is -F and n is 2.
• R 1 is CH .
• R 1 is OH.
• R 1 is H.
• R 2 is CH .
• R 2 is OH.
• R 2 is H.
• R 1 is CH3 and R 2 is CH3.
• R 1 is CH 3 and R 2 is OH.
• R 1 is CH 3 and R 2 is H.
• R 1 is OH and R 2 is CH 3 .
• R 1 is OH and R 2 is OH.
• R 1 is H and R 2 is H.
• R 1 is H and R 2 is OH.
• R 1 is H and R 2 is H.
• the compound of Formula (IIIc) has the following structure:
• BA is a binding agent
• R 2 is methyl
• R 5a and R 5b are each, independently, -F or H;
• BL is a binding agent linker
• x is an integer from 0 to 30.
• the compound of Formula (IIIc) has the following structure:
• BA is a binding agent
• RG is a reactive group residue
• CD is a cyclodextrin
• SP 1 is a spacer group
• AA 4 is an amino acid residue
• AA 5 is a dipeptide residue
• PEG is polyethylene glycol
• n is an integer from 0 to 4.
• x is an integer from 0 to 30;
• R 4 is alkyl, aryl, arylalkyl, or an N-containing heterocycloalkyl; wherein the alkyl, aryl, arylalkyl, and N-containing heterocycloalkyl are optionally substituted with -NR a R b ;
• R a and R b are, independently in each instance, -H or alkyl
• BA is a binding agent bonded to -BL-;
• SP 1 and SP 2 are each, independently in each instance, absent or a spacer group residue, and wherein SP 1 comprises a trivalent linker; AA 4 is a trivalent linker comprising an amino acid residue; AA 5 is a di-peptide residue; PEG is a polyethylene glycol residue; wherein the indicates the atom through which the indicated chemical group is bonded to the adjacent groups in the formula, CD is, independently in each instance, absent or a cyclodextrin residue, wherein at least one CD is present, subscript m is an integer from 0 to 5; In these examples, subscript m is 0, 1, 2, 3, 4, or 5. In some examples, subscript m is 0. In some examples, subscript m is 1. In some examples, subscript m is 2.
• any one of AA 4 or AA 5 comprises, independently in each instance, an amino acid selected from alanine, valine, leucine, isoleucine, methionine, tryptophan, phenylalanine, proline, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, histidine, or citrulline, a derivative thereof, or a combination thereof.
• AA 4 is an amino acid selected from alanine, valine, leucine, isoleucine, methionine, tryptophan, phenylalanine, proline, glycine, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, histidine, or citrulline, a derivative thereof, or a combination thereof.
• AA 4 is lysine.
• AA 4 is lysine or a derivative of lysine.
• the AA 5 is valine-citrulline.
• the AA 5 is citrulline-valine. In some embodiments, the AA 5 is valine-alanine. In some embodiments, the AA 5 is alanine-valine. In some embodiments, the AA 5 is valine-glycine. In some embodiments, the AA 5 is glycine-valine. In some embodiments, the AA 5 glutamate-valine-citrulline. In some embodiments, the AA 5 is glutamine-valine-citrulline. In some embodiments, the AA 5 is lysine-valine-alanine. In some embodiments, the AA 5 is lysine-valine-citrulline.
• the AA 5 is glutamate- valine-citrulline.
• SP 1 is independently in each instance, selected from the group consisting of Ci-e alkylene, - H-, -C(O)-, (-CH 2 -CH 2 -0)e, - H-CH 2 -CH 2 -(-0-CH 2 - CH 2 ) e -C(0)-, -C(0)-(CH 2 )u-C(0)-, -C(0)- H-(CH 2 )v-, and combinations thereof, wherein subscript e is an integer from 0 to 4, subscript u is an integer from 1 to 8, and subscript v is an integer from 1 to 8.
• SP 2 is independently in each instance, selected from the group consisting of Ci-e alkylene, - H-, -C(O)-, (-CH 2 -CH 2 -0) e , - H-CH 2 -CH 2 -(-0-CH 2 - CH 2 ) e -C(0)-, -C(0)-(CH 2 )u-C(0)-, -C(0)- H-(CH 2 )v-, and combinations thereof, wherein subscript e is an integer from 0 to 4, subscript u is an integer from 1 to 8, and subscript v is an integer from 1 to 8.
• n is an integer from 0 to 4, R 3 is -OH or R z -C(0)-0-; wherein R z is alkyl, BL is a binding agent linker, BA is a binding agent, and x is an integer from 1 to 30. In certain embodiments, n is 0 or 1 and x is an integer from 1-6. In certain embodiments, x is 4.
• n is an integer from 1-4, R 3 is -OH or R z -C(0)-0-; wherein R z is alkyl, BL is a binding agent linker, BA is a binding agent, and x is an integer from 1-30. In certain embodiments, n is 2 and x is an integer from 1-6. In certain embodiments, x is 4.
• R 4 is alkyl, wherein R a is a hydrogen atom or alkyl, BL is a binding agent linker, and BA is a binding agent.
• R 4 is Ci-4 alkyl.
• R 4 is propyl.
• R 3 is -NH 2 , - HCFL, or -N(CH 3 ) 2 .
• x is an integer from 1-6. In certain embodiments, x is 4.
• R 4 is alkyl, R P1 is halo or a hydrogen atom, and X is R a or O, wherein R a is a hydrogen atom or alkyl, BL is a binding agent linker, BA is a binding agent, and x is an integer from 1-30.
• R 4 is C1-4 alkyl, X is H, and x is an integer from 1-6. In certain embodiments, x is 4.
• X is O or NRa, ⁇ - ⁇ is aryl or heteroaryl, R p is halo, t is an integer from 0-2, R a is a hydrogen atom or alkyl, BL is a binding agent linker, BA is a binding agent, and x is an integer from 1-30, and R 4 is alkyl.
• X is O, R 4 is alkyl, and x is an integer from 1-6. In certain embodiments, x is 4.
• Ra is a hydrogen atom or alkyl
• BL is a binding agent linker
• BA is a binding agent
• x is an integer from 1 -30. In some embodiments, x is an integer from 1-6. In some embodiments, x is 4.
• binding agent linker refers to any divalent group or moiety that links, connects, or bonds a binding agent (e.g., an antibody or an antigen- binding fragment thereof) with a payload compound set forth herein (e.g., steroid).
• a binding agent e.g., an antibody or an antigen- binding fragment thereof
• a payload compound set forth herein (e.g., steroid).
• suitable binding agent linkers for the antibody conjugates described herein are those that are sufficiently stable to exploit the circulating half-life of the antibody and, at the same time, capable of releasing its payload after antigen-mediated internalization of the conjugate.
• Linkers can be cleavable or non-cleavable.
• Cleavable linkers are linkers that are cleaved by intracellular metabolism following internalization, e.g., cleavage via hydrolysis, reduction, or enzymatic reaction.
• Non-cleavable linkers are linkers that release an attached payload via lysosomal degradation of the antibody following internalization.
• Suitable linkers include, but are not limited to, acid-labile linkers, hydrolysis-labile linkers, enzymatically cleavable linkers, reduction labile linkers, self-immolative linkers, and non-cleavable linkers.
• Suitable linkers also include, but are not limited to, those that are or comprise glucuronides,
• succinimide-thioethers polyethylene glycol (PEG) units, carbamates, hydrazones, mal- caproyl units, disulfide units (e.g., -S-S-, -S-S- C(R 1 )(R 2 ) -, wherein R 1 and R 2 are independently hydrogen or hydrocarbyl), para-amino-benzyl (PAB) units, phosphate units, e.g., mono-, bis-, and tris- phosphate units, peptides, e.g., peptide units containing two, three, four, five, six, seven, eight, or more amino acid units, including but not limited to valine- citrulline units, valine-alanine units, valine-arginine units, valine-lysine units, -lysine-valine- citrulline units, and -lysine-valine-alanine units.
• PEG polyethylene glycol
• carbamates carbamates
• the binding agent linker group of the conjugates described herein are derived from the reaction of a "reactive linker” group of a linker-payload described herein with a reactive portion of an antibody.
• the reactive linker group refers to a monovalent group that comprises a reactive group and linking group, depicted as * , wherein RG is the reactive group, L is the linking group, and the wiggly line represents a bond to a payload.
• the linking group is any divalent moiety that bridges the reactive group to the payload.
• the linking group may also be any trivalent moiety that bridges the reactive group, the payload and a cyclodextrin moiety.
• the linking group is trivalent and includes a cyclodextrin moiety bonded to a trivalent group (e.g., a lysine residue) in the linking group.
• the reactive linkers (RL), together with the payloads to which they are bonded, comprise intermediates ("linker-payloads") useful as synthetic precursors for the preparation of the antibody steroid conjugates described herein.
• the reactive linker contains a reactive group (RG), which is a functional group or moiety that reacts with a reactive portion of an antibody, modified antibody, or antigen binding fragment thereof.
• BL binding agent linker
• RG is the moiety resulting from the reaction of a reactive group of a linker-payload with a reactive portion of a binding agent
• L is a linking group
• 5 is a bond to a payload.
• RG is derived from the reaction of RG with a cysteine or lysine residue of an antibody or antigen-binding fragment thereof.
• RG N is derived from a click chemistry reaction. In some embodiments of said click chemistry reaction, RG N is derived from a 1,3 cycloaddition reaction between an alkyne and an azide.
• Non-limiting examples of such RG N s include those derived from strained alkynes, e.g., those suitable for strain-promoted alkyne-azide cycloadditions (SPAAC), cycloalkynes, e.g., cyclooctynes, benzannulated alkynes, and alkynes capable of undergoing 1,3 cycloaddition reactions with azides in the absence of copper catalysts.
• Suitable RG ⁇ s also include, but are not limited to those derived from DIB AC, DIBO, BARAC, substituted, e.g., fluorinated alkynes, aza-cycloalkynes, BCN, and derivatives thereof.
• Conjugates containing such RG N groups can be derived from antibodies that have been functionalized with azido groups.
• Such functionalized antibodies include antibodies functionalized with azido-polyethylene glycol groups.
• such functionalized antibody is derived by reacting an antibody comprising at least one glutamine residue with a compound according to the formula H2N-LL-N3, wherein LL is a divalent polyethylene glycol group, in the presence of the enzyme transglutaminase, e.g., microbial transglutaminase.
• Suitable glutamine residues of an antibody include Q295, or those derived by insertion or mutation, e.g., N297Q mutation.
• BA of the conjugates described herein is an antibody or an antigen-binding fragment thereof. In some embodiments, the conjugates described herein are derived from azido-functionalized antibodies. In certain embodiments, BA of the conjugates described herein is: wherein Ab is an antibody or antigen-binding fragment thereof, n is an integer from 1 to 10, w is the number of linker payload moieties, and ⁇ is a bond to a single binding agent linker (BL), e.g., bond to a moiety derived from a 1,3-cycloaddition reaction between an alkyne and azide. In certain embodiments, w is 3. In certain embodiments, w is 2 or 4, i.e., the conjugate comprises 2 or 4 linker payload moieties.
• Ab is an antibody or antigen-binding fragment thereof
• n is an integer from 1 to 10
• w is the number of linker payload moieties
• ⁇ is a bond to a single binding agent linker (BL), e.
• BL is a divalent moiety of Formula (BL A );
• A is an amino acid or a peptide
• R a is H or alkyl
• B is aryl, heteroaryl, or heterocycloalkyl, wherein aryl, heteroaryl, or heterocycloalkyl is optionally substituted with alkyl, -OH, or -NR a R b ;
• SP 1 and SP 2 are, independently, a spacer groups; and q, z, s, t, u, v, and w are, independently in each instance, 0 or 1.
• BL is a trivalent moiety of Formula (BL B );
• RG N is as defined herein;
• A is tripeptide, wherein at least one of the amino acids in the tripeptide is bonded directly or indirectly to a cyclodextrin moiety;
• R a is H or alkyl
• B is aryl, heteroaryl, or heterocycloalkyl, wherein aryl, heteroaryl, or heterocycloalkyl is optionally substituted with alkyl, -OH, or -NR a R b ;
• SP 1 and SP 2 are, independently, a spacer groups; and q, z, s, t, u, v, and w are, independently in each instance, 0 or 1.
• the cyclodextrin (CD) is bonded directly to an amino acid residue, such as a lysine amino acid residue. This means that the CD is one bond position away from the lysine amino acid covalent linker.
• the covalent linker is also bonded directly to a payload moiety. This means that the covalent linker is one bond position away from a payload such as, but not limited to a steroid payload set forth herein.
• the covalent linker is also bonded directly to a CD moiety. This means that the covalent linker is one bond position away from a CD, such as the CD(s) set forth herein.
• the covalent linker is a lysine amino acid or a derivative thereof.
• the CD is bonded indirectly to a covalent linker in a linking group (e.g., a BL).
• a linking group e.g., a BL
• the CD is more than one bond position away from the covalent linker.
• the CD is bonded through another moiety to the covalent linker.
• the CD may be bonded to a maleimide group which is bonded to a polyethylene glycol group which is bonded to the covalent linker.
• the covalent linker is also bonded indirectly to a payload moiety. This means that the covalent linker is more than one bond position away from a payload such as, but not limited to a steroid payload set forth herein.
• the covalent linker is bonded through another moiety to the payload.
• the covalent linker may be bonded to a dipeptide, such as but not limited to Val-Ala or Val-Cit, which may be bonded to para-amino benzoyl which may be bonded to the payload.
• the covalent linker is also bonded indirectly to a cyclodextrin moiety. This means that the covalent linker is more than one bond position away from a cyclodextrin, such as the cyclodextrins set forth herein. This also means that the covalent linker is bonded through another moiety to the cyclodextrin.
• the covalent linker may be bonded to a polyethylene glycol group which may be bonded to reactive group which may be bonded to the cyclodextrin.
• the covalent linker is a lysine amino acid or a derivative thereof.
• BL is -RG ⁇ SP ⁇ q- ⁇ A) ⁇ . In some embodiments, BL is
• BL is a moiety of Formula (BL A1 )
• R AA1 and R AA2 are each, independently, amino acid side chains.
• SP 1 is a divalent polyethylene glycol group and RG 1 ⁇ is a 1,3-cycloaddition reaction adduct of the reaction between an alkyne and an azide.
• BL is -RG ⁇ SP ⁇ q- ⁇ A) ⁇ . In some embodiments, BL is
• BL is a moiety of Formula (BL B1 )
• R AA1 and R AA2 are each, independently, amino acid side chains.
• R AA3 is an amino acid side chain that is bonded directly or indirectly to a cyclodextrin moiety.
• SP 1 is a divalent polyethylene glycol group and RG N is a 1,3- cycloaddition reaction adduct of the reaction between an alkyne and an azide.
• BL has the following structure:
• RG N , SP 1 are as defined herein;
• q is 0 or 1
• Z 1 is a polyethylene glycol or caproyl group
• Z 2 is a dipeptide or tripeptide
• Z 3 is a PAB group.
• RG 1 ⁇ is derived from a click-chemistry reactive group and Z 1 is a polyethylene glycol group.
• RGN-(SPl)q-Zl- is:
• the dipeptide is valine-citrulline or valine alanine.
• the BL is attached to the payload via tertiary amine.
• BL is a binding agent linker as defined above;
• R a and R b are, independently in each instance, -H or alkyl.
• G ⁇ is derived from a click-chemistry reactive group.
• RG N is:
• RG is selected from a group which reacts with a cysteine or lysine residue on an antibody or an antigen-binding fragment thereof.
• RG N is ond to cysteine of a binding agent, e.g.,
• RG N is .
• SP is selected from
• SP 1 is . In some other examples, SP 1 is
• subscripts a, b, and c are independently, in each instance, an integer from 1 to 20.
• R AA3 is selected from , wherein CD is a
• R AA3 is selected from
• SP 1 is selected from: [0294] In some examples, SP 1 is . In some examples, SP 1 is
• SP 1 is In
• BL-SP 1 is:
• subscripts b, c, and d are independently, in each instance, an integer from 1 to 20.
• BL-SPHs selected from:
• A is a peptide selected from valine-citrulline, citrulline- valine, lysine-phenylalanine, phenylalanine-lysine, valine-asparagine, asparagine-valine, threonine-asparagine, asparagine-threonine, serine-asparagine, asparagine-serine, phenylalanine-asparagine, asparagine-phenylalanine, leucine-asparagine, asparagine-leucine, isoleucine-asparagine, asparagine-isoleucine, glycine-asparagine, asparagine-glycine, glutamic acid-asparagine, asparagine-glutamic acid, citrulline-asparagine, asparagine- citrulline, alanine-asparagine, or asparagine-a
• A is valine-citrulline or citrulline-valine. [0300] In some examples, A is valine-alanine or alanine-valine.
• A is lysine-valine-alanine or alanine-valine-lysine.
• A is lysine-valine-citrulline or citrulline-valine-lysine.
• A is valine
• A is alanine
• A is citrulline
• A is . In some of these examples,
• R AA1 is an amino acid side chain, and wherein R AA2 is an amino acid side chain.
• R AA1 is an amino acid side chain
• R ⁇ 2 is an amino acid side chain
• R AA3 is an amino acid side chain that is bonded directly or indirectly to a cyclodextrin moiety.
• A is N-(0,1] n-(0,1] n-(0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1]
• A is N-(0,1] n-(0,1] n-(0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1]
• A is represents a direct or indirect bond to a cyclodextrin moiety.
• CD is, independently in each instance, selected from
• A is .
• R a is H
• R a is alkyl
• R a is methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, i- butyl, or pentyl.
• B is aryl
• B is phenyl
• B is phenyl or pyridinyl.
• B is:
• R 10 is alkyl, alkenyl, alkynyl, alkoxy, aryl, alkylaiyl, aiylalkyl, halo, haloalkyl, haloalkoxy, heteroaryl, heterocycloalkyl, hydroxyl, cyano, nitro, " I "orA 3 -f-so 2 RA , NR a R b , or azido.
• subscripts p and m are independently, in each instance, selected from an inte er from 0 to 4.
• R 1 is, independently at each occurrence, alkyl, alkoxy, haloalkyl, or halo. In some examples, R 1 is alkyl. In some examples, R 1 is alkoxy. In some examples, R 1 is haloalkyl. In some examples, R 1 is halo.
• BA is a binding agent, and x is an integer from 1-30.
• BA is an antibody.
• x is an integer from 1 to 4.
• x is 4.
• x is 2.
• BA is a binding agent; each L is an optional linker; BA or L is covalently bonded to R 3 or R 4 ; and x is an integer from 1 to 30.
• L is bonded to R 3 or R 4 ; when L is not present, BA is bonded to R 3 or R 4 .
• the groups R 3 or R 4 are described in detail below.
• BA is an antibody.
• x is an integer from 1 to 4.
• x is 4.
• x is 2.
• R 1 and R 2 are, independently, selected from the group consisting of -H, -OH, alkyl, -0-C(0)-alkyl, and halo; or R 1 and R 2
• R is selected from the group consisting of -alkylene-NR a R b , -X-arylene-Y- R a R b , -X-heteroarylene-Y- R a R b , and N-containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or -CH2-; and R 4 is selected from the group consisting of alkyl, aryl, alkylaryl, arylalkyl, heteroaryl, -alkylene-NR a R b , -X-arylene-Y- R a R b , -X-heteroarylene-Y- R a R b , and N-containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or -CH2-.
• R 3 is selected from the group consisting of -OH, -0-C(0)-alkyl, -O-aiyl, - R a R b , -alkylene- R a R b , -X-arylene-Y- R a R b , -X-heteroarylene-Y- R a R b , and N-containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or -CH2-; and R 4 is selected from the group consisting of -alkylene-NR a R b , -X-arylene-Y- R a R b , -X-heteroarylene-Y- R a R b , and N-containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or
• R 3 is -NR a R b ; and R 4 is alkyl.
• BA or L is bonded to a functional group in R 3 or R 4 .
• R 3 or R 4 comprises a amino group
• BA or L can be bonded to the amino group, substituting for a hydrogen atom.
• R 5 is, independently in each instance, selected from a substituent in the group consisting of -OH, halo, and alkyl; n is an integer from 0-19; and each R 5 is positioned on any ring atom.
• R a and R b are, independently in each instance, selected from the group consisting of -H and alkyl; or R a and R b cyclize to form cycloheteroalkyl with three to six ring atoms, including one hetero atom, which is the N to which they are attached.
• BA is an antibody.
• x is an integer from 1 to 4. In some embodiments, x is 4. In some embodiments, x is 2.
• R 3 is covalently bonded to L or BA.
• R 1 and R 2 are, independently, selected from the group consisting of -H, -OH, alkyl, -0-C(0)-alkyl, and halo;
• R is selected from the group consisting of -alkylene- R a R b , -X-arylene-Y- R a R b , -X-heteroarylene-Y- R a R b , and N- containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or
• R 4 is selected from the group consisting of alkyl, aryl, alkylaryl, arylalkyl, heteroaryl, -alkylene- R a R b , -X-arylene-Y- R a R b , -X-heteroarylene-Y- R a R b , and N- containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or
• R 3 is - R a R b ; and R 4 is alkyl.
• BA or L is bonded to an amino group in R 3 , for instance, substituting for a hydrogen atom.
• R a and R b are, independently in each instance, selected from the group consisting of -H and alkyl; or R a and R b cyclize to form cycloheteroalkyl with three to six ring atoms, including one hetero atom, which is the N to which they are attached.
• BA is an antibody.
• x is an integer from 1 to 4. In some embodiments, x is 4. In some embodiments, x is 2.
• R 3 is selected from the group consisting of -OH, -0-C(0)-alkyl, -O-aiyl, - R a R b , -alkylene- R a R b , -X- arylene-Y- R a R b , -X-heteroarylene-Y-NR a R b , and N-containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or -0-; wherein Y is absent or -CH2-; and R 4 is selected from the group consisting of -alkylene- R a R b , -X-arylene-Y- R a R b , -X-heteroarylene-Y- R a R b , and N- containing heterocycloalkyl; wherein X is absent, -N-, -CH2-, or
• BA or L is bonded to an amino group in R 4 , for instance, substituting for a hydrogen atom.
• R a and R b are, independently in each instance, selected from the group consisting of -H and alkyl; or R a and R b cyclize to form cycloheteroalkyl with three to six ring atoms, including one hetero atom, which is the N to which they are attached.
• BA is an antibody.
• x is an integer from 1 to 4. In some embodiments, x is 4. In some embodiments, x is 2.
• Ab is an antibody and x is an integer from 1-30. In some embodiments, x is an integ from 1 to 4. In some embodiments, x is 4. In some embodiments, x is 2.
• Ab is an antibody and x is an integer from 1-30. In some embodiments, x is an integer from 1 to 4. In some embodiments, x is 4. In some embodiments, x is 2.
• Ab is an antibody and x is an integer from 1-30. In some embodiments, x is an integer from 1 to 4. In some embodiments, x is 4. In some
• x is 2.
• binding agent conjugates of budesonide or diflorasone are also provided herein.
• Suitable binding agents for any of the conjugates provided in the instant disclosure include, but are not limited to, antibodies, lymphokines, hormones, growth factors, viral receptors, interleukins, or any other cell binding or peptide binding molecules or substances.
• the binding agent is an antibody.
• antibody means any antigen-binding molecule or molecular complex comprising at least one complementarity determining region (CDR) that specifically binds to or interacts with a particular antigen.
• CDR complementarity determining region
• the term “antibody” includes immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, as well as multimers thereof (e.g., IgM).
• Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region.
• the heavy chain constant region comprises three domains, CHI , CH2 and CH3.
• Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region.
• the light chain constant region comprises one domain (CL I).
• the VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR).
• CDRs complementarity determining regions
• FR framework regions
• Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy -terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
• the FRs of the antibody may be identical to the human germline sequences, or may be naturally or artificially modified.
• An amino acid consensus sequence may be defined based on a side-by-side analysis of two or more CDRs.
• antibody also includes antigen-binding fragments of full antibody molecules.
• antigen-binding portion of an antibody, “antigen-binding fragment” of an antibody, and the like, as used herein, include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds an antigen to form a complex.
• Antigen-binding fragments of an antibody may be derived, e.g., from full antibody molecules using any suitable standard techniques such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding antibody variable and optionally constant domains.
• DNA is known and/or is readily available from, e.g., commercial sources, DNA libraries (including, e.g., phage-antibody libraries), or can be synthesized.
• the DNA may be sequenced and manipulated chemically or by using molecular biology techniques, for example, to arrange one or more variable and/or constant domains into a suitable configuration, or to introduce codons, create cysteine residues, modify, add or delete amino acids, etc.
• Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of the amino acid residues that mimic the hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR) such as a CDR3 peptide), or a constrained FR3- CDR3-FR4 peptide.
• CDR complementarity determining region
• Other engineered molecules such as domain-specific antibodies, single domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted
• antibodies diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g. monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the expression "antigen- binding fragment," as used herein.
• nanobodies e.g. monovalent nanobodies, bivalent nanobodies, etc.
• SMIPs small modular immunopharmaceuticals
• shark variable IgNAR domains are also encompassed within the expression "antigen- binding fragment," as used herein.
• An antigen-binding fragment of an antibody will typically comprise at least one variable domain.
• the variable domain may be of any size or amino acid composition and will generally comprise at least one CDR which is adjacent to or in frame with one or more framework sequences.
• the VH and VL domains may be situated relative to one another in any suitable arrangement.
• the variable region may be dimeric and contain VH-VH, VH-VL or VL-VL dimers.
• the antigen-binding fragment of an antibody may contain a monomeric VH or VL domain.
• an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain.
• variable and constant domains that may be found within an antigen-binding fragment of an antibody of the present disclosure include: (i) VH-CH1 ; (ii) VH-CH2; (iii) VH-CH3; (iv) VH-CH1-CH2; (V) VH-CH1-CH2-CH3; (vi) VH-CH2-CH3; (vii) VH-CL; (viii) VL-CH1 ; (ix) VL-CH2; (X) VL-CH3; (xi) VL-CH1-CH2; (xii) VL-CH1-CH2-CH3; (xiii) VL- CH2-CH3; and (xiv) VL-CL.
• variable and constant domains may be either directly linked to one another or may be linked by a full or partial hinge or linker region.
• a hinge region may consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids which result in a flexible or semi-flexible linkage between adjacent variable and/or constant domains in a single polypeptide molecule.
• an antigen-binding fragment of an antibody of the present disclosure may comprise a homo-dimer or hetero-dimer (or other mul timer) of any of the variable and constant domain configurations listed above in non-covalent association with one another and/or with one or more monomeric VH or VL domain (e.g., by disulfide bond(s)).
• antigen-binding fragments may be any antigen-binding fragments.
• a multispecific antigen-binding fragment of an antibody will typically comprise at least two different variable domains, wherein each variable domain is capable of specifically binding to a separate antigen or to a different epitope on the same antigen.
• Any multispecific antibody format, including the exemplary bispecific antibody formats disclosed herein, may be adapted for use in the context of an antigen-binding fragment of an antibody of the present disclosure using routine techniques available in the art.
• the antibodies of the present disclosure may function through complement- dependent cytotoxicity (CDC) or antibody-dependent cell-mediated cytotoxicity (ADCC).
• CDC complement-dependent cytotoxicity
• ADCC antibody-dependent cell-mediated cytotoxicity
• FcRs Fc receptors
• NK Natural Killer
• the constant region of an antibody is important in the ability of an antibody to fix complement and mediate cell-dependent cytotoxicity.
• the isotype of an antibody may be selected on the basis of whether it is desirable for the antibody to mediate cytotoxicity.
• the antibodies useful for the compounds herein include human antibodies.
• human antibody is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences.
• the human antibodies can include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo), for example in the CDRs and in particular CDR3.
• the term "human antibody”, as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.
• the term "human antibody” does not include naturally occurring molecules that normally exist without modification or human intervention/manipulation, in a naturally occurring, unmodified living organism.
• the antibodies can, in some embodiments, be recombinant human antibodies.
• recombinant human antibody is intended to include all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described further below), antibodies isolated from a recombinant, combinatorial human antibody library (described further below), antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes (see e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295) or antibodies prepared, expressed, created or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences.
• Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences.
• such recombinant human antibodies are subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline repertoire in vivo.
• an immunoglobulin molecule comprises a stable four chain construct of approximately 150-160 kDa in which the dimers are held together by an interchain heavy chain disulfide bond.
• the dimers are not linked via inter-chain disulfide bonds and a molecule of about 75-80 kDa is formed composed of a covalently coupled light and heavy chain (half-antibody).
• the frequency of appearance of the second form in various intact IgG isotypes is due to, but not limited to, structural differences associated with the hinge region isotype of the antibody.
• a single amino acid substitution in the hinge region of the human IgG4 hinge can significantly reduce the appearance of the second form (Angal et al. (1993) Molecular Immunology 30: 105) to levels typically observed using a human IgGl hinge.
• the instant disclosure encompasses antibodies having one or more mutations in the hinge, CH2 or CH3 region which may be desirable, for example, in production, to improve the yield of the desired antibody form.
• the antibodies useful for the compounds herein can be isolated antibodies.
• isolated antibody means an antibody that has been identified and separated and/or recovered from at least one component of its natural environment.
• an antibody that has been separated or removed from at least one component of an organism, or from a tissue or cell in which the antibody naturally exists or is naturally produced is an "isolated antibody” for purposes of the instant disclosure.
• An isolated antibody also includes an antibody in situ within a recombinant cell. Isolated antibodies are antibodies that have been subjected to at least one purification or isolation step. According to certain embodiments, an isolated antibody may be substantially free of other cellular material and/or chemicals.
• the antibodies useful for the compounds disclosed herein may comprise one or more amino acid substitutions, insertions and/or deletions in the framework and/or CDR regions of the heavy and light chain variable domains as compared to the corresponding germline sequences from which the antibodies were derived. Such mutations can be readily ascertained by comparing the amino acid sequences disclosed herein to germline sequences available from, for example, public antibody sequence databases.
• the present disclosure includes antibodies, and antigen-binding fragments thereof, which are derived from any of the amino acid sequences disclosed herein, wherein one or more amino acids within one or more framework and/or CDR regions are mutated to the corresponding residue(s) of the germline sequence from which the antibody was derived, or to the corresponding residue(s) of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are referred to herein collectively as "germline mutations").
• Germline mutations A person of ordinary skill in the art, starting with the heavy and light chain variable region sequences disclosed herein, can easily produce numerous antibodies and antigen-binding fragments which comprise one or more individual germline mutations or combinations thereof.
• all of the framework and/or CDR residues within the V H and/or V L domains are mutated back to the residues found in the original germline sequence from which the antibody was derived.
• only certain residues are mutated back to the original germline sequence, e.g., only the mutated residues found within the first 8 amino acids of FRl or within the last 8 amino acids of FR4, or only the mutated residues found within CDRl, CDR2 or CDR3.
• one or more of the framework and/or CDR residue(s) are mutated to the corresponding residue(s) of a different germline sequence ⁇ i.e., a germline sequence that is different from the germline sequence from which the antibody was originally derived).
• the antibodies of the present disclosure may contain any combination of two or more germline mutations within the framework and/or CDR regions, e.g., wherein certain individual residues are mutated to the corresponding residue of a particular germline sequence while certain other residues that differ from the original germline sequence are maintained or are mutated to the corresponding residue of a different germline sequence.
• antibodies and antigen-binding fragments that contain one or more germline mutations can be easily tested for one or more desired property such as, improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as the case may be), reduced immunogenicity, etc.
• the antibody is a monoclonal antibody, polyclonal antibody, antibody fragment (Fab, Fab', and F(ab)2, minibody, diabody, tribody, and the like), or bispecific antibody.
• Antibodies herein can be humanized using methods described in US Patent No. 6,596,541 and US Publication No. 2012/0096572, each incorporated by reference in their entirety.
• the binding agent binds to an antigen binding partner that is a polypeptide and may be a transmembrane molecule (e.g., receptor) or a growth factor that might be glycosylated or phosphorylated.
• an antigen binding partner that is a polypeptide and may be a transmembrane molecule (e.g., receptor) or a growth factor that might be glycosylated or phosphorylated.
• Suitable targets to which the binding agent binds include any target to which steroid delivery is desirable.
• the binding agent is an antibody, modified antibody, or antigen binding fragment there of that binds a target selected from: AXL, BAFFR, BCMA, BCR-list components, BDCA2, BDCA4, BTLA, BTNL2 BTNL3, BTNL8, BTNL9, C10orf54, CCR1, CCR3, CCR4, CCR5, CCR6, CCR7, CCR9, CCR10, CDl lc, CD137, CD 138, CD 14, CD 168, CD 177, CD 19, CD20, CD209, CD209L, CD22, CD226, CD248, CD25, CD27, CD274, CD276, CD28, CD30, CD300A, CD33, CD37, CD38, CD4, CD40, CD44, CD45, CD46, CD47, CD48, CD5, CD52, CD55, CD56, CD59, CD62E, CD68
• the binding agent linkers can be bonded to the binding agent, e.g., antibody or antigen-binding molecule, through an attachment at a particular amino acid within the antibody or antigen-binding molecule.
• exemplary amino acid attachments that can be used in the context of this aspect of the disclosure include, e.g., lysine (see, e.g., US 5,208,020;
• WO 2013/053872 WO 2011/130598; US 2013/0101546; and US 7,750,116
• selenocysteine see, e.g., WO 2008/122039; and Hofer et al, Proc. Natl. Acad. Sci., USA, 2008, 705: 12451- 12456
• formyl glycine see, e.g., Carrico et al., Nat. Chem. Biol, 2007, 3 :321-322; Agarwal et al, Proc. Natl. Acad. Sci., USA, 2013, 770:46-51, and Rabuka et al, Nat.
• Linkers can be conjugated via glutamine via transglutaminase-based chemo-enzymatic conjugation (see, e.g., Dennler et al,
• Linkers can also be conjugated to an antigen- binding protein via attachment to carbohydrates (see, e.g., US 2008/0305497,
• the binding agent is an antibody, and the antibody is bonded to the linker through a lysine residue. In some embodiments, the antibody is bonded to the linker through a cysteine residue.
• the conjugates described herein can be synthesized by coupling the linker- payloads described herein with a binding agent, e.g., antibody under standard conjugation conditions (see, e.g., DrugDeliv. 2016 Jun;23(5): 1662-6; AAPS Journal, Vol. 17, No. 2, March 2015; and Int. J. Mol. Sci. 2016, 17, 561, the entireties of which are incorporated herein by reference).
• Linker-payloads are synthetic intermediates comprising the payload of interest and linking moiety that ultimately serves as the moiety (or portion thereof) that connects the binding agent with the payload.
• Linker-payloads comprise a reactive group that reacts with the binding agent to form the conjugates described herein.
• the antibody can be coupled to a linker-payload via one or more cysteine, lysine, or other residue of the antibody.
• Linker payloads can be coupled to cysteine residues, for example, by subjecting the antibody to a reducing agent, e.g., dithiotheritol, to cleave the disulfide bonds of the antibody, purifying the reduced antibody, e.g., by gel filtration, and subsequently reacting the antibody with a linker-payload containing a reactive moiety, e.g., a maleimido group.
• Suitable solvents include, but are not limited to water, DMA, DMF, and DMSO.
• Linker- payloads containing a reactive group can be coupled to lysine residues.
• Suitable solvents include, but are not limited to water, DMA, DMF, and DMSO.
• Conjugates can be purified using known protein techniques, including, for example, size exclusion chromatography, dialysis, and ultrafiltration/diafiltration.
• Binding agents e.g., antibodies
• the linker-payload comprises a reactive group, e.g., alkyne that is capable of undergoing a 1,3 cycloaddition reaction with an azide.
• Such suitable reactive groups include, but are not limited to, strained alkynes, e.g., those suitable for strain-promoted alkyne-azide cycloadditions (SPAAC), cycloalkynes, e.g., cyclooctynes, benzannulated alkynes, and alkynes capable of undergoing 1,3 cycloaddition reactions with azides in the absence of copper catalysts.
• Suitable alkynes also include, but are not limited to, DIBAC, DIBO, BARAC, DIFO, substituted, e.g., fluorinated alkynes, aza- cycloalkynes, BCN, and derivatives thereof.
• Linker-payloads comprising such reactive groups are useful for conjugating antibodies that have been functionalized with azido groups.
• Such functionalized antibodies include antibodies functionalized with azido-polyethylene glycol groups.
• such functionalized antibody is derived by reacting an antibody comprising at least one glutamine residue, e.g., heavy chain Q295, with a compound according to the formula H2N-LL-N3, wherein LL is a divalent polyethylene glycol group, in the presence of the enzyme transglutaminase.
• the antibody Ab is a modified antibody with one or more covalently linked -LL-N3 groups, or residues thereof.
• each -LL-N3 is covalently bonded to an amino acid side chain of a glutamine residue of the antibody.
• the -LL-N3 is or can be reacted with a reactive group RG to form a covalent bond to a linker-payload.
• the -LL-N3 groups are expressly drawn. [0357] Set forth here are methods of synthesizing the conjugates described herein comprising contacting a binding agent, e.g., antibody, with a linker-payload described herein.
• the linker-payload includes a cyclodextrin moiety.
• the linker payload is a compound of Formula (II):
• R 1 and R 2 are each, independently, -H, alkyl, alkyl-C(0)-0- -OH, or
• arylalkyl or an N-containing heterocycloalkyl; wherein the alkyl, aryl, arylalkyl, and N-containing heterocycloalkyl are optionally substituted with -NR a R b ; or
• R 3 is -OH, alkyl-C(0)-0-, heteroalkyl, -NR a R b or aryloxy, wherein the alkyl-C(0)-0- heteroalkyl, or aryloxy is optionally substituted with
• R 1 and R 2 together form " ⁇ TM ⁇ , wherein R 4 is
• RL is a reactive linker
• R 5 is, independently in each instance, -OH, halo, alkyl, or arylalkyl;
• R a and R b are, independently in each instance, -H or alkyl

Landscapes

• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Engineering & Computer Science (AREA)
• Animal Behavior & Ethology (AREA)
• Public Health (AREA)
• Veterinary Medicine (AREA)
• Pharmacology & Pharmacy (AREA)
• Medicinal Chemistry (AREA)
• Organic Chemistry (AREA)
• Epidemiology (AREA)
• Immunology (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Chemical & Material Sciences (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Cell Biology (AREA)
• Pain & Pain Management (AREA)
• Rheumatology (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Peptides Or Proteins (AREA)
• Steroid Compounds (AREA)
• Preparation Of Compounds By Using Micro-Organisms (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
• Medicinal Preparation (AREA)
• Micro-Organisms Or Cultivation Processes Thereof (AREA)

Priority Applications (41)

Applications Claiming Priority (4)

Related Child Applications (3)

Publications (3)

Family

ID=60409441

Family Applications (1)

Country Status (17)

Cited By (25)

Families Citing this family (12)

Citations (30)

Family Cites Families (127)

• 2017
  • 2017-11-07 KR KR1020247004647A patent/KR102835535B1/ko active Active
  • 2017-11-07 KR KR1020197015823A patent/KR20190074310A/ko not_active Ceased
  • 2017-11-07 CN CN201780082517.1A patent/CN110291097A/zh active Pending
  • 2017-11-07 CN CN202211283749.2A patent/CN115651056A/zh active Pending
  • 2017-11-07 ES ES17801239T patent/ES3046537T3/es active Active
  • 2017-11-07 AU AU2017359043A patent/AU2017359043B2/en active Active
  • 2017-11-07 US US15/806,197 patent/US10711032B2/en active Active
  • 2017-11-07 JP JP2019523684A patent/JP7330101B2/ja active Active
  • 2017-11-07 EP EP17801239.9A patent/EP3538539B1/en active Active
  • 2017-11-07 IL IL266353A patent/IL266353B2/en unknown
  • 2017-11-07 CA CA3042428A patent/CA3042428A1/en active Pending
  • 2017-11-07 IL IL307357A patent/IL307357A/en unknown
  • 2017-11-07 SG SG10202104259RA patent/SG10202104259RA/en unknown
  • 2017-11-07 WO PCT/US2017/060434 patent/WO2018089373A2/en not_active Ceased
  • 2017-11-07 MX MX2019005330A patent/MX394528B/es unknown
  • 2017-11-07 EP EP25196812.9A patent/EP4659809A2/en active Pending
  • 2017-11-07 KR KR1020257023550A patent/KR20250114431A/ko active Pending
  • 2017-11-07 MY MYPI2019002581A patent/MY199937A/en unknown
• 2019
  • 2019-05-07 ZA ZA2019/02871A patent/ZA201902871B/en unknown
  • 2019-05-07 MX MX2021012927A patent/MX2021012927A/es unknown
  • 2019-05-07 CL CL2019001259A patent/CL2019001259A1/es unknown
  • 2019-05-08 PH PH12019501021A patent/PH12019501021A1/en unknown
  • 2019-06-07 CO CONC2019/0005966A patent/CO2019005966A2/es unknown
• 2020
  • 2020-04-24 US US16/858,458 patent/US11760775B2/en active Active
• 2023
  • 2023-07-28 US US18/361,275 patent/US12377159B2/en active Active
  • 2023-08-08 JP JP2023129158A patent/JP2023153988A/ja active Pending