Classifications

• • 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
  • A61K47/68037—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a camptothecin [CPT] or derivatives
• • 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
  • A61K47/6851—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 determinant of a tumour cell
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents

Definitions

• the present disclosure relates to antibody drug conjugates (ADCs) comprising an antibody comprising an Fc variant, and methods of preparing the same. Also provided herein are methods of treating cancer using the ADCs described herein.
• ADCs antibody drug conjugates
• ADCs Antibody-Drug Conjugates
• ADCs allow for the targeted delivery of a drug moiety to a tumor, and, in some embodiments intracellular accumulation therein, where systemic administration of unconjugated drugs may result in unacceptable levels of toxicity to normal cells
• ADCs are targeted chemotherapeutic molecules which combine properties of both antibodies and cytotoxic drugs by targeting potent cytotoxic drugs to antigen-expressing tumor cells (Teicher, B.A. (2009) Current Cancer Drug Targets 9:982-1004), thereby enhancing the therapeutic index by maximizing efficacy and minimizing off-target toxicity (Carter, P.J. and Senter P.D. (2008) The Cancer Jour. 14(3):154-169; Chari, R.V. (2008) Acc. Chem. Res. 41:98-107.
• ADCs comprising a monoclonal antibody, comprising an Fc variant, said Fc variant comprising two amino acid substitutions L234A and L235A (also named “LALA mutation” herein), which is conjugated to camptothecin derivative toxins or duostatin derivative toxins through linker moieties.
• the monoclonal antibody is an anti- CD25 antibody, anti-B7-H3 antibody, anti-RORl antibody, anti-Trop-2 antibody or anti-BCMA antibody.
• the anti-CD25 antibody binds to CD25 -expressing cancer cells and allows for selective uptake of the ADC into the cancer cells.
• the anti-B7-H3 antibody binds to B7-H3-expressing cancer cells and allows for selective uptake of the ADC into the cancer cells.
• the anti-RORl antibody binds to R0R1 -expressing cancer cells and allows for selective uptake of the ADC into the cancer cells.
• the anti- Trop-2 antibody binds to Trop-2-expressing cancer cells and allows for selective uptake of the ADC into the cancer cells.
• the anti-BCMA antibody binds to BCMA- expressing cancer cells and allows for selective uptake of the ADC into the cancer cells.
• the ADCs provided herein selectively deliver an effective amount of the camptothecin derivative toxin or the duostatin derivative toxin to tumor tissue and reduce the non-specific toxicity associated with related ADCs.
• the ADC compounds described herein include those with anticancer activity.
• Camptothecin is a cytotoxic quinoline alkaloid isolated from Camptotheca acuminta, a type of tree natively growing in China. CPT was discovered in the 1960s (Wall M.E. et al., 1966, J. Am. Chem. Soc. 88:3888-3890). The antitumor activity of Camptothecin depends on a highly specific inhibition of Topoisomerase-I (TOPO 1). The enzyme TOPO 1 cleaves one strand of double stranded DNA, partially unwinds the DNA, and then reanneals the strand to relieve tension.
• TOPO 1 Topoisomerase-I
• Camptothecin and its derivatives bind to the TOPO 1/DNA complex to prevent reannealing, which can cause cell death due to the accumulation of partially cleaved DNA (Hsiang Y. Happel et al, 1985, J. Biol. Chem. 260:14873-14878).
• camptothecin The clinical application of camptothecin is limited due to its low solubility as well as serious side-effects (Joerger M. et al., 2015, Br. J. Clin. Pharmacol. 80:128-138; Joerger M. et al., 2015, Invest. New Drugs 33:472-479).
• camptothecin derivatives have been developed to date, including topotecan (9-dimethyl amino- 10-hydroxy camptothecin; TPT) and irinotecan (7-ethyl-10-[4-(l-piperidino)-l-piperidino] carbonyloxycamptothecin; CPT-11) (Naumczuk B.
• camptothecin derivative is exatecan, which is a water soluble derivative of camptothecin (US patent Nos. 10,195,288, 8,575,188). Unlike irinotecan currently used in clinical settings, an activation by an enzyme is unnecessary. Dxd is another useful camptothecin derivative.
• camptothecin drugs are widely applied clinically, and the main indications are bone cancer, prostatic cancer, breast cancer, gastric cancer, pancreatic cancer, ovarian cancer, esophageal cancer, endometrial cancer and the like (Iqbal et al., 2014, Mol. Biol. Int. 2014).
• Camptothecin drugs have a short half-life in plasma and maintaining drug efficacy in clinical use requires an increased dose or increased frequency of administration, thus possibly causing tolerance problems to patients.
• Dolastatins such as natural product Dolastatin 10, and its synthetic derivatives Monomehtyl Auristatin E (MMAE) and Monomethyl Auristatin F (MMAF) are products that show potent antineoplastic and tubulin inhibitory property. Because of their high toxicity, the direct use of Dolastatins as therapeutic agents has not been effective. Instead, they were conjugated to an antibody for targeted delivery to kill cancer cells.
• ADCs antibody-drug conjugates comprising a monoclonal antibody, said monoclonal antibody comprising an Fc variant, said Fc variant comprising two amino acid substitutions L234A and L235A.
• provided herein are methods of preparing ADCs comprising a monoclonal antibody, said monoclonal antibody comprising an Fc variant, said Fc variant comprising two amino acid substitutions L234A and L235A.
• methods for treating cancers using the ADCs disclosed herein are provided herein.
• Ab is a monoclonal antibody, said monoclonal antibody comprising an Fc variant, said Fc variant comprising two amino acid substitutions L234A and L235A;
• m is an integer from 1 to 8;
• L 1 is a linker bound to the monoclonal antibody;
• L 2 is a bond, -C(O)-, -NH-, Amino Acid Unit, -(CH2CH2O)n-, -(CHijn-, - (4-aminobenzyloxycarbonyl)-, -(C(O)CH2CH2NH)-, -(C(O)N(R 2 )CH2CH2N(R 3 ))-, -O-, or any combination thereof;
• L is a substituted or unsubstituted heterocycloalkylene or a
• R* is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl;
• R 1 is H or — C 1 -C 8 alkyl
• R 3 is H, halogen, -CCh, -CBr 3 , -CF3, -CI 3 , -CHCh, -CHBr 2 , -CHF 2 , -CHI 2 , -CH 2 C1, -CH 2 Br, -CH 2 F, -CH 2 1, -CN, -OR 3A , -NR 3A R 3B , -(CH 2 ) V OR 6 , substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl;
• R 4 is H, halogen, -OR 4A , -NR 4A R 4B , substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl;
• V is N, O, or C
• Z 1 is a substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocycloalkyl;
• Z 2 is a substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted cycloalkylene, or substituted or unsubstituted heterocycloalkylene;
• R 6 is H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl,
• v is an integer from 1 to 24;
• w is an integer from 1 to 24;
• M is -NH 2 , -OH, -COOH, or -OCH 3 ;
• R 10 is -OH, -OCH 3 or -COOH; and each R 3A , R 3B , R 4A , and R 4B is independently H or substituted or unsubstituted alkyl.
• a method of treating a CD25 -expressing cancer, B7-H3- expressing cancer, ROR1 -expressing cancer, Trop-2-expressing cancer, or BCMA-expressing cancer, in a subject in need thereof including administering the ADC described herein (including in an aspect, embodiment, table, example, or claim), or a pharmaceutically acceptable salt thereof, to the subject.
• ADC of formula (I) m , formula ( formula (III) m , or a pharmaceutically acceptable salt there iof, said method including reacting a monoclonal antibody, or a modified antibody with a molecule of formula (P-II) , or formula (P-III) , p ically acceptable salt thereof, wherein B is a reactive moiety capable of forming a bond with the monoclonal antibody;
• L 2 is a bond, -C(O)-, -NH-, Amino Acid Unit, -(CH 2 CH 2 O) n -, -(CH 2 ) n -, -(4-aminobenzyloxycarbonyl)-, -O-, -(C(O)CH 2 CH 2 NH)-, -(C(O)N(R 2 )CH 2 CH 2 N(R 3 ))-, or any combination thereof; wherein n is an integer from 1 to 24; each R 2 and R is independently H or substituted
• R 1 is H or — C 1 -C 8 alkyl
• R 3 is H, halogen, -CCh, -CBr 3 , -CF3, -CI 3 , -CHCh, -CHBr 2 , -CHF 2 , -CHI 2 , -CH 2 C1, -CH 2 Br, -CH 2 F, -CH 2 I, -CN, -OR 3A , -NR 3A R 3B , -(CH 2 ) V OR 6 , substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl;
• R 4 is H, halogen, -OR 4A , -NR 4A R 4B , substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl;
• V is N, O, or C
• Z 1 is a substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocycloalkyl;
• Z 2 is a substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted cycloalkylene, or substituted or unsubstituted heterocycloalkylene;
• R 6 is H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl,
• v is an integer from 1 to 24;
• w is an integer from 1 to 24;
• M is -NH 2 , -OH, -COOH, or -OCH 3 ;
• R 10 is -OH, -OCH 3 or -COOH; and each R 3A , R 3B , R 4A , and R 4B is independently H or substituted or unsubstituted alkyl.
• composition comprising the ADCs described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
• FIG. 1 shows the chemical structures of linker-payload compounds that were used to make the ADCs used in in vitro and in vivo efficacy studies.
• FIG. 2 shows binding of wildtype (WT) and double mutated antibodies (LALA) in various human cancer cell lines.
• CD25-WT and CD25-LALA antibodies binding to SU-DHL-1 cell line (CD25+) (FIG. 2A).
• CD25-WT and CD25-LALA antibodies binding to Daudi cell line (CD25-) (FIG. 2B).
• BCMA-WT and BCMA-LALA antibodies binding to NCI-H929 cell line (BCMA+) (FIG. 2C).
• BCMA-WT and BCMA-LALA antibodies binding to K562 cell line (BCMA-) (FIG. 2D).
• B7-H3-WT and B7-H3-LALA antibodies binding to Panc-1 cell line (B7- H3+) (FIG. 2E).
• B7-H3-WT and B7-H3-LALA antibodies binding to A375 cell line (B7-H3+) (FIG. 2F).
• B7-H3-WT and B7-H3-LALA antibodies binding to A549 cell line (B7-H3+) FIG. 2G).
• FIG. 3 shows results of an in vitro efficacy study of ADCs comprising anti-B7-H3 antibody or anti-B7-H3-LALA antibody conjugated to L014-077 in: PA-1 (B7-H3 +) cells (FIG. 3A) and NCLH929 (B7-H3-) cells (FIG. 3B). Positive controls are anti-B7-H3 antibodies of Daiichi or Macrogenics conjugated to L014-077.
• FIG. 4 shows results of an in vitro efficacy study of ADCs comprising anti-B7-H3 antibody or anti-B7-H3-LALA antibody conjugated to L078-118 in: PA-1 (B7-H3 +) cells (FIG. 4A) and NCLH929 (B7-H3-) cells (FIG. 4B). Positive controls are anti-B7-H3 conjugated to SET0218 (Daiichi’s linker-payload) and anti-B7-H3-LALA conjugated to SET0218 linkerpayload.
• FIG. 5 shows results of in vitro binding study in leukocytes from two donors. Binding of wildtype (WT) and mutated antibodies (LALA) (anti-CD25, anti-B7-H3, and anti-BCMA) to monocytes, neutrophils, NK cells, T cells, and B cells is shown. Dotted line at 1000 bound antibodies indicates the limit of quantification.
• WT wildtype
• LALA mutated antibodies
• FIG. 6 shows results of in vitro binding study in leukocytes from two donors. Binding of anti-B7-H3 wildtype (WT B7-H3) or mutated antibody (LALA B7-H3) to monocytes, neutrophils, NK cells, T cells, and B cells is shown. Dotted line at 1000 bound antibodies indicates the limit of quantification.
• WT B7-H3 wildtype
• LALA B7-H3 mutated antibody
• FIG. 7A shows results of an in vivo efficacy study in PA-1 xenograft in Nu/Nu nude mice of anti-B7-H3 antibody or anti-B7-H3-LALA antibody conjugated to L014-077, where the mice were treated once intravenously with 5 mg/kg of ADC. Positive controls are anti-B7-H3 antibodies of Daiichi or Macrogenics conjugated to L014-077.
• FIG. 7B shows results of an in vivo efficacy study in PA-1 xenograft in Nu/Nu nude mice of anti-B7-H3 antibody or anti-B7-H3-LALA antibody conjugated to L078-118, where the mice were treated once intravenously with 10 mg/kg of ADC. Positive control is anti-B7-H3- LALA conjugated to SET0218 (Daiichi’s linker-payload).
• FIG. 7C shows results of mouse body weight changes during an in vivo study in Nu/Nu nude mice following treatments described in FIG. 7B.
• FIG. 8 shows results of an in vivo efficacy study in SU-DHL-1 xenograft in Nu/Nu nude mice.
• anti-CD25 antibody or anti-CD25-LALA antibody conjugated to L078-118 where the mice were treated once intravenously with 3 mg/kg of ADC
• anti-CD25 antibody or anti-CD25-LALA antibody conjugated to L014-077 where the mice were treated once intravenously with 3 mg/kg of ADC
• FIG. 8B anti-CD25-LALA antibody conjugated to L078- 118, L014-077, L078-182, or L078-120, where the mice were treated once intravenously with 3 mg/kg of ADC
• FIG. 8C DETAILED DESCRIPTION OF THE INVENTION
• Enzymatic reactions and enrichment/purification techniques are also well known and are performed according to manufacturer's specifications, as commonly accomplished in the art or as described herein.
• the terminology used in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are well known and commonly used in the art. Standard techniques can be used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.
• the term “and/or” used herein is to be taken mean specific disclosure of each of the specified features or components with or without the other.
• the term “and/or” as used in a phrase such as “A and/or B” herein is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone).
• the term “and/or” as used in a phrase such as “A, B, and/or C” is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).
• the term “about” refers to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in pail on how the value or composition is measured or determined, i.e., the limitations of the measurement system.
• “about” or “approximately” can mean within one or more than one standard deviation per the practice in the art.
• “about” or “approximately” can mean a range of up to 10% (i.e., ⁇ 10%) or more depending on the limitations of the measurement system.
• about 5 mg can include any number between 4.5 mg and 5.5 mg.
• the terms can mean up to an order of magnitude or up to 5-fold of a value.
• the meaning of “about” or “approximately” should be assumed to be within an acceptable error range for that particular value or composition.
• about includes the specified value.
• Numerical ranges include the endpoints of the range. For example, “between 4.5 mg and 5.5 mg” includes 4.5 mg, 5.5 mg, and all values greater than 4.5 mg and less than 5.5 mg.
• polypeptide refers to a polymer of amino acid residues, wherein the polymer may in embodiments be conjugated to a moiety that does not consist of amino acids.
• the terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers.
• a “fusion protein” refers to a chimeric protein encoding two or more separate protein sequences that arc recombinantly expressed as a single moiety. Polypeptides include mature molecules that have undergone cleavage.
• polypeptide complex can be dimeric, trimeric, tetrameric, or higher order complexes depending on the number of polypeptide chains that form the complex.
• cancer As used herein, the terms “cancer,” “neoplasm,” and “tumor” are used interchangeably and, in either the singular or plural form, refer to cells that have undergone a malignant transformation that makes them pathological to the host organism.
• Primary cancer cells can be readily distinguished from non-cancerous cells by well-established techniques, particularly histological examination.
• the definition of a cancer cell includes not only a primary cancer cell, but any cell derived from a cancer cell ancestor. This includes metastasized cancer cells, and in vitro cultures and cell lines derived from cancer cells.
• a “clinically detectable” tumor is one that is detectable on the basis of tumor mass; e.g., by procedures such as computed tomography (CT) scan, magnetic resonance imaging (MRI), X-ray, ultrasound or palpation on physical examination, and/or which is detectable because of the expression of one or more cancer- specific antigens in a sample obtainable from a patient.
• CT computed tomography
• MRI magnetic resonance imaging
• X-ray X-ray
• ultrasound or palpation e.g., ultrasound or palpation on physical examination
• Tumors may be a hematopoietic (or hematologic or hematological or blood-related) cancer, for example, cancers derived from blood ceils or immune ceils, which may be referred to as “liquid tumors.”
• liquid tumors Specific examples of clinical conditions based on hematologic tumors include leukemias such as chronic myelocytic leukemia, acute myelocytic leukemia, chronic lymphocytic leukemia and acute lymphocytic leukemia; plasma cell malignancies such as multiple myeloma, MGUS and Waldenstrom's macroglobulinemia; lymphomas such as non-Hodgkin's lymphoma, Hodgkin's lymphoma; and the like.
• leukemias such as chronic myelocytic leukemia, acute myelocytic leukemia, chronic lymphocytic leukemia and acute lymphocytic leukemia
• plasma cell malignancies such as multiple myeloma,
• leukemia refers broadly to progressive, malignant diseases of the blood- forming organs and is generally characterized by a distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is generally clinically classified on the basis of (1) the duration and character of the disease-acute or chronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid (lymphogenous), or monocytic; and (3) the increase or non-increase in the number abnormal cells in the blood-leukemic or aleukemic (subleukemic).
• Exemplary leukemias that may be treated with a compound or method provided herein include, for example, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, a leukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic leukemia, leukemia cutis, embryonal leukemia, eosinophilic leukemia, Gross' leukemia, hairy-cell leukemia, hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphogenous leukemia,
• lymphoma refers to a group of cancers affecting hematopoietic and lymphoid tissues. It begins in lymphocytes, the blood cells that are found primarily in lymph nodes, spleen, thymus, and bone marrow. Two main types of lymphoma are non-Hodgkin lymphoma and Hodgkin’s disease. Hodgkin’s disease represents approximately 15% of all diagnosed lymphomas. This is a cancer associated with Reed-Sternberg malignant B lymphocytes. Non-Hodgkin’ s lymphomas (NHL) can be classified based on the rate at which cancer grows and the type of cells involved.
• B-cell lymphomas that may be treated with a compound or method provided herein include, but are not limited to, small lymphocytic lymphoma, Mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, extranodal (MALT) lymphoma, nodal (monocytoid B- ccll) lymphoma, splenic lymphoma, diffuse large cell B-lymphoma, Burkitt’s lymphoma, lymphoblastic lymphoma, immunoblastic large cell lymphoma, or precursor B -lymphoblastic lymphoma.
• small lymphocytic lymphoma Mantle cell lymphoma
• follicular lymphoma marginal zone lymphoma
• MALT extranodal lymphoma
• nodal lymphoma nodal lymphocytoid B- ccll lymphoma
• splenic lymphoma diffuse large cell B-lymp
• T-cell lymphomas that may be treated with a compound or method provided herein include, but are not limited to, cunateous T-cell lymphoma, peripheral T-cell lymphoma, anaplastic large cell lymphoma, mycosis fungoides, and precursor T-lymphoblastic lymphoma.
• cancer refers to all types of cancer, neoplasm or malignant tumors found in mammals (e.g. humans), including leukemias, lymphomas, carcinomas and sarcomas.
• the ADCs and methods provided herein are useful for treating CD25 -expressing cancers, B7-H3-expressing cancers, ROR1 -expressing cancers, Trop-2-expressing cancers, or BCMA-expressing cancers.
• Exemplary cancers that may be treated with a compound or method provided herein include brain cancer, glioma, glioblastoma, neuroblastoma, prostate cancer, colorectal cancer, pancreatic cancer, Medulloblastoma, melanoma, cervical cancer, gastric cancer, ovarian cancer, lung cancer, cancer of the head, Hodgkin's Disease, and Non-Hodgkin's Lymphomas.
• Exemplary cancers that may be treated with a compound or method provided herein include cancer of the thyroid, endocrine system, brain, breast, cervix, colon, head & neck, liver, kidney, lung, ovary, pancreas, rectum, stomach, and uterus.
• Additional examples include, thyroid carcinoma, cholangiocarcinoma, pancreatic adenocarcinoma, skin cutaneous melanoma, colon adenocarcinoma, rectum adenocarcinoma, stomach adenocarcinoma, esophageal carcinoma, head and neck squamous cell carcinoma, breast invasive carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, non- small cell lung carcinoma, mesothelioma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, primary brain tumors, malignant pancreatic insulanoma, malignant carcinoid, urinary bladder cancer, premalignant skin lesions, testicular cancer, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract
• leukemia refers broadly to progressive, malignant diseases of the blood- forming organs and is generally characterized by a distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is generally clinically classified on the basis of (1) the duration and character of the disease-acute or chronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid (lymphogenous), or monocytic; and (3) the increase or non-increase in the number abnormal cells in the blood-leukemic or aleukemic (subleukemic).
• Exemplary leukemias that may be treated with a compound or method provided herein include, for example, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, a leukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic leukemia, leukemia cutis, embryonal leukemia, eosinophilic leukemia, Gross' leukemia, hairy-cell leukemia, hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphogenous leukemia,
• lymphoma refers to a group of cancers affecting hematopoietic and lymphoid tissues. It begins in lymphocytes, the blood cells that are found primarily in lymph nodes, spleen, thymus, and bone marrow. Two main types of lymphoma are non-Hodgkin lymphoma and Hodgkin’s disease. Hodgkin’s disease represents approximately 15% of all diagnosed lymphomas. This is a cancer associated with Reed-Sternberg malignant B lymphocytes. Non-Hodgkin’ s lymphomas (NHL) can be classified based on the rate at which cancer grows and the type of cells involved.
• B-cell lymphomas that may be treated with a compound or method provided herein include, but are not limited to, small lymphocytic lymphoma, Mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, extranodal (MALT) lymphoma, nodal (monocytoid B- ccll) lymphoma, splenic lymphoma, diffuse large cell B-lymphoma, Burkitt’s lymphoma, lymphoblastic lymphoma, immunoblastic large cell lymphoma, or precursor B -lymphoblastic lymphoma.
• small lymphocytic lymphoma Mantle cell lymphoma
• follicular lymphoma marginal zone lymphoma
• MALT extranodal lymphoma
• nodal lymphoma nodal lymphocytoid B- ccll lymphoma
• splenic lymphoma diffuse large cell B-lymp
• T-cell lymphomas that may be treated with a compound or method provided herein include, but are not limited to, cunateous T-cell lymphoma, peripheral T-cell lymphoma, anaplastic large cell lymphoma, mycosis fungoides, and precursor T-lymphoblastic lymphoma.
• sarcoma generally refers to a tumor which is made up of a substance like the embryonic connective tissue and is generally composed of closely packed cells embedded in a fibrillar or homogeneous substance.
• Sarcomas that may be treated with a compound or method provided herein include a chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Abernethy's sarcoma, adipose sarcoma, liposarcoma, alveolar soft part sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma, chorio carcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcom
• melanoma is taken to mean a tumor arising from the melanocytic system of the skin and other organs.
• Melanomas that may be treated with a compound or method provided herein include, for example, acral-lentiginous melanoma, amelanotic melanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma, Harding-Passey melanoma, juvenile melanoma, lentigo maligna melanoma, malignant melanoma, nodular melanoma, subungal melanoma, or superficial spreading melanoma.
• carcinoma refers to a malignant new growth made up of epithelial cells tending to infiltrate the surrounding tissues and give rise to metastases.
• exemplary carcinomas that may be treated with a compound or method provided herein include, for example, medullary thyroid carcinoma, familial medullary thyroid carcinoma, acinar carcinoma, acinous carcinoma, adenocystic carcinoma, adenoid cystic carcinoma, carcinoma adenomatosum, carcinoma of adrenal cortex, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, carcinoma basocellulare, basaloid carcinoma, basosquamous cell carcinoma, bronchioalveolar carcinoma, bronchiolar carcinoma, bronchogenic carcinoma, cerebriform carcinoma, cholangiocellular carcinoma, chorionic carcinoma, colloid carcinoma, comedo carcinoma, corpus carcinoma, cribriform carcinoma, carcinoma en cuirasse, carcinoma cutaneum, cylindrical carcinoma, cylindrical cell carcinoma, duct carcinoma, carcinoma durum, embryonal carcinoma, encephaloid
• the terms “metastasis,” “metastatic,” and “metastatic cancer” can be used interchangeably and refer to the spread of a proliferative disease or disorder, e.g., cancer, from one organ or another non-adjacent organ or body part. “Metastatic cancer” is also called “Stage IV cancer.” Cancer occurs at an originating site, e.g., breast, which site is referred to as a primary tumor, e.g., primary breast cancer. Some cancer cells in the primary tumor or originating site acquire the ability to penetrate and infiltrate surrounding normal tissue in the local area and/or the ability to penetrate the walls of the lymphatic system or vascular system circulating through the system to other sites and tissues in the body.
• a second clinically detectable tumor formed from cancer cells of a primary tumor is referred to as a metastatic or secondary tumor.
• the metastatic tumor and its cells are presumed to be similar to those of the original tumor.
• the secondary tumor in the breast is referred to a metastatic lung cancer.
• metastatic cancer refers to a disease in which a subject has or had a primary tumor and has one or more secondary tumors.
• non-metastatic cancer or subjects with cancer that is not metastatic refers to diseases in which subjects have a primary tumor but not one or more secondary tumors.
• metastatic lung cancer refers to a disease in a subject with or with a history of a primary lung tumor and with one or more secondary tumors at a second location or multiple locations, e.g., in the breast.
• cutaneous metastasis or “skin metastasis” refer to secondary malignant cell growths in the skin, wherein the malignant cells originate from a primary cancer site (e.g., breast).
• a primary cancer site e.g., breast
• cancerous cells from a primary cancer site may migrate to the skin where they divide and cause lesions. Cutaneous metastasis may result from the migration of cancer cells from breast cancer tumors to the skin.
• visceral metastasis refers to secondary malignant cell growths in the interal organs (e.g., heart, lungs, liver, pancreas, intestines) or body cavities (e.g., pleura, peritoneum), wherein the malignant cells originate from a primary cancer site (e.g., head and neck, liver, breast).
• a primary cancer site e.g., head and neck, liver, breast.
• a primary cancer site e.g., head and neck, liver, breast
• Visceral metastasis may result from the migration of cancer cells from liver cancer tumors or head and neck tumors to internal organs.
• the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.
• an "antibody” and “antibodies” and related terms used herein refers to an intact immunoglobulin or to an antigen binding portion thereof that binds specifically to an antigen.
• Antigen binding portions may be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies.
• Antigen binding portions include, inter alia, Fab, Fab', F(ab')2, Fv, domain antibodies (dAbs), and complementarity determining region (CDR) fragments, single-chain antibodies (scFv), chimeric antibodies, diabodies, triabodies, tetrabodies, and polypeptides that contain at least a portion of an immunoglobulin that is sufficient to confer specific antigen binding to the polypeptide.
• Antibodies include recombinantly produced antibodies and antigen binding portions.
• Antibodies include non-human, chimeric, humanized and fully human antibodies.
• Antibodies include monospecific, multispecific (e.g., bispecific, trispecific and higher order specificities).
• Antibodies include tetrameric antibodies, light chain monomers, heavy chain monomers, light chain dimers, heavy chain dimers.
• Antibodies include F(ab’)2 fragments, Fab’ fragments and Fab fragments.
• Antibodies include single domain antibodies, monovalent antibodies, single chain antibodies, single chain variable fragment (scFv), camelized antibodies, affibodies, disulfide- linked Fvs (sdFv), anti-idiotypic antibodies (anti-Id), minibodies.
• Antibodies include monoclonal and polyclonal populations. Anti-CD25 antibodies, anti-B7-H3 antibodies, anti-RORl antibodies, anti-Trop-2 antibodies, and anti-BCMA antibodies are described herein.
• the term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical and/or bind the same epitope, except for possible variant antibodies, e.g., containing naturally occurring mutations or arising during production of a monoclonal antibody preparation, such variants generally being present in minor amounts.
• polyclonal antibody preparations typically include different antibodies directed against different determinants (epitopes)
• each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen.
• the modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method.
• the monoclonal antibodies to be used in accordance with the present invention may be made by a variety of techniques, including but not limited to the hybridoma method, recombinant DNA methods, phage-display methods, and methods utilizing transgenic animals containing all or part of the human immunoglobulin loci, such methods and other exemplary methods for making monoclonal antibodies being described herein.
• An "epitope" and related terms as used herein refers to a portion of an antigen that is bound by an antigen binding protein (e.g., by an antibody or an antigen binding portion thereof).
• An epitope can comprise portions of two or more antigens that are bound by an antigen binding protein.
• An epitope can comprise non-contiguous portions of an antigen or of two or more antigens (e.g., amino acid residues that are not contiguous in an antigen’s primary sequence but that, in the context of the antigen’s tertiary and quaternary structure, are near enough to each other to be bound by an antigen binding protein).
• the variable regions, particularly the CDRs, of an antibody interact with the epitope.
• Anti-CD25 antibodies, and antigen binding proteins thereof, that bind an epitope of a CD25 polypeptide are described herein.
• Anti-B7-H3 antibodies, and antigen binding proteins thereof, that bind an epitope of a B7-H3 polypeptide are described herein.
• Anti-RORl antibodies, and antigen binding proteins thereof, that bind an epitope of a ROR1 polypeptide are described herein.
• Anti-Trop-2 antibodies, and antigen binding proteins thereof, that bind an epitope of a Trop-2 polypeptide are described herein.
• Anti-BCMA antibodies, and antigen binding proteins thereof, that bind an epitope of a BCMA polypeptide are described herein.
• an "antibody fragment”, “antibody portion”, “antigen-binding fragment of an antibody”, or “antigen-binding portion of an antibody” and other related terms used herein refer to a molecule other than an intact antibody that comprises a portion of an intact antibody that binds the antigen to which the intact antibody binds.
• antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')i; Fd; and Fv fragments, as well as dAb; diabodies; linear antibodies; single-chain antibody molecules (e.g. scFv); polypeptides that contain at least a portion of an antibody that is sufficient to confer specific antigen binding to the polypeptide.
• Antigen binding portions of an antibody may be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies.
• Antigen binding portions include, inter alia, Fab, Fab', F(ab')2, Fv, domain antibodies (dAbs), and complementarity determining region (CDR) fragments, chimeric antibodies, diabodies, triabodies, tetrabodies, and polypeptides that contain at least a portion of an immunoglobulin that is sufficient to confer antigen binding properties to the antibody fragment.
• Antigen-binding fragments of anti-CD25 antibodies are described herein.
• Antigen-binding fragments of anti-B7-H3 antibodies are described herein.
• Antigen-binding fragments of anti-RORl antibodies are described herein.
• Antigen-binding fragments of anti-Trop-2 antibodies arc described herein.
• Antigen-binding fragments of anti-BCMA antibodies are described herein.
• An antigen binding protein can have, for example, the structure of an immunoglobulin.
• an "immunoglobulin” refers to a tetrameric molecule. Each tetrameric molecule is composed of two identical pairs of polypeptide chains, each pair having one “light” (about 25 kDa) and one “heavy” chain (about 50-70 kDa). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The carboxy-terminal portion of each chain defines a constant region primarily responsible for effector function. Human light chains are classified as kappa or lambda light chains.
• Heavy chains are classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively.
• the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with the heavy chain also including a "D” region of about 10 more amino acids. See generally, Fundamental Immunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, N.Y. (1989)) (incorporated by reference in its entirety for all purposes).
• the variable regions of each light/heavy chain pair form the antibody binding site such that an intact immunoglobulin has two antigen binding sites.
• an antigen binding protein can be a synthetic molecule having a structure that differs from a tetrameric immunoglobulin molecule but still binds a target antigen or binds two or more target antigens.
• a synthetic antigen binding protein can comprise antibody fragments, 1-6 or more polypeptide chains, asymmetrical assemblies of polypeptides, or other synthetic molecules.
• variable heavy chain refers to the variable region of an immunoglobulin heavy chain, including an Fv, scFv , dsFv or Fab
• variable light chain refers to the variable region of an immunoglobulin light chain, including of an Fv, scFv , dsFv or Fab
• variant region or “variable domain” refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to antigen.
• variable domains of the heavy chain and light chain (VH and VL, respectively) of a native antibody generally have similar’ structures, with each domain comprising four conserved framework regions (FRs) and three hypervariable regions (HVRs).
• FRs conserved framework regions
• HVRs hypervariable regions
• a single VH or VL domain may be sufficient to confer antigen-binding specificity.
• antibodies that bind a particular antigen may be isolated using a VH or VL domain from an antibody that binds the antigen to screen a library of complementary VL or VH domains, respectively. Sec, e.g., Portolano ct al., J.
• antibody functional fragments include, but are not limited to, complete antibody molecules, antibody fragments, such as Fv, single chain Fv (scFv), complementarity determining regions (CDRs), VL (light chain variable region), VH (heavy chain variable region), Fab, F(ab)2' and any combination of those or any other functional portion of an immunoglobulin peptide capable of binding to target antigen (see, e.g., FUNDAMENTAL IMMUNOLOGY (Paul ed., 4th ed. 2001).
• various antibody fragments can be obtained by a variety of methods, for example, digestion of an intact antibody with an enzyme, such as pepsin; or de novo synthesis.
• Antibody fragments are often synthesized de novo either chemically or by using recombinant DNA methodology.
• the term antibody includes antibody fragments either produced by the modification of whole antibodies, or those synthesized de novo using recombinant DNA methodologies (e.g., single chain Fv) or those identified using phage display libraries (see, e.g., McCafferty et al., (1990) Nature 348:552).
• the term "antibody” also includes bivalent or bispecific molecules, diabodies, triabodies, and tetrabodies. Bivalent and bispecific molecules are described in, e.g., Kostelny et al. (1992) J. Immunol.
• antigen binding protein refers to a protein comprising a portion that binds to an antigen and, optionally, a scaffold or framework portion that allows the antigen binding portion to adopt a conformation that promotes binding of the antigen binding protein to the antigen.
• antigen binding proteins include antibodies, antibody fragments (e.g., an antigen binding portion of an antibody), antibody derivatives, and antibody analogs.
• the antigen binding protein can comprise, for example, an alternative protein scaffold or artificial scaffold with grafted CDRs or CDR derivatives.
• Such scaffolds include, but are not limited to, antibody-derived scaffolds comprising mutations introduced to, for example, stabilize the three-dimensional structure of the antigen binding protein as well as wholly synthetic scaffolds comprising, for example, a biocompatible polymer. See, for example, Korndorfer et al., 2003, Proteins: Structure, Function, and Bioinformatics, Volume 53, Issue 1:121-129; Roque et al., 2004, Biotechnol. Prog. 20:639-654.
• PAMs peptide antibody mimetics
• Antigen binding proteins that bind CD25, B7-H3, ROR1, Trop-2, or BCMA are described herein.
• a dissociation constant can be measured using a BIACORE surface plasmon resonance (SPR) assay.
• SPR surface plasmon resonance refers to an optical phenomenon that allows for the analysis of real-time interactions by detection of alterations in protein concentrations within a biosensor matrix, for example using the BIACORE system (Biacore Life Sciences division of GE Healthcare, Piscataway, NJ).
• “Specifically binds” as used throughout the present specification in relation to CD25, B7-H3, ROR1, Trop-2, or BCMA antigen binding proteins means that the antigen binding protein binds human CD25 (hCD25), B7-H3 (hB7-H3), ROR1 (hRORl), Trop-2 (hTrop-2), or BCMA (hBCMA), respectively, with no or insignificant binding to other human proteins.
• an antibody specifically binds to a target antigen if it binds to the antigen with a dissociation constant KD of 10’ 5 M or less, or 10’ 6 M or less, or 10’ 7 M or less, or 10’ 8 M or less, or 10’ 9 M or less, or 10’ 10 M or less.
• the term “Fc” or “Fc region” as used herein refers to the portion of an antibody heavy chain constant region beginning in or after the hinge region and ending at the C-terminus of the heavy chain.
• the Fc region comprises at least a portion of the CH and CH3 regions, and may or may not include a portion of the hinge region. Two polypeptide chains each carrying a half Fc region can dimerize to form an Fc region.
• An Fc region can bind Fc cell surface receptors and some proteins of the immune complement system.
• An Fc region exhibits effector function, including any one or any combination of two or more activities including complementdependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent phagocytosis (ADP), opsonization and/or cell binding.
• An Fc region can bind an Fc receptor, including FcyRI (c.g., CD64), FcyRII (c.g, CD32) and/or FcyRIII (c.g., CD16a).
• the Fc region can include a mutation that increases or decreases any one or any combination of these functions (e.g., an effector-negative Fc region).
• the antigen-binding protein is an IgG, IgA, IgD, IgE, or IgM antibody having one or more mutations in the Fc region, for example one or more mutations that decreases antibody dependent enhancement (ADE) and/or one or more mutations that increases antibody half-life. Mutations that reduce or eliminate interaction of the Fc region of antibody with its receptor (e.g., FcyRs) on such cells can reduce or eliminate ADE.
• FcyRs antibody dependent enhancement
• an antigen-binding protein as provided herein can be an IgGl or IgG4 antibody having an ADE- reducing mutation, such as the LALA mutation, in the Fc region, or can be single chain antibody (ScFv) that optionally includes an Fc region that can optionally include an ADE-reducing mutation, such as the LALA mutation (mutation of Leucine residues at positions 234 and 235, in the EU index set forth in Kabat, to Alanine residues).
• an antigenbinding protein as provided herein can be a Fab, Fab’, or F(ab’)2 antibody fragment.
• the present disclosure provides ADCs comprising a monoclonal antibody comprising a mutation in the Fc region selected from L234A and L235A (a LA mutation).
• the mutation in the Fc region can include both mutations L234A and L235A (LALA).
• the mutation in the Fc region is a LA mutation or two LALA mutations.
• the LALA mutations in the Fc region reduce the effector function of the monoclonal antibody relative to monoclonal antibody without LALA mutations in the Fc region.
• the term “CD25,” as used herein, refers to any native CD25 from any vertebrate source, including mammals such as primates (e.g.
• cynomolgus monkey cyno
• rodents e.g., mice and rats
• the term encompasses “full-length,” unprocessed CD25 as well as any form of CD25 that results from processing in the cell.
• the term also encompasses naturally occurring variants of CD25, e.g., splice variants, allelic variants, and isoforms.
• the amino acid sequence of an exemplary human CD25 protein is shown in SEQ ID NO: 70.
• B7-H3 refers to any native B7-H3 from any vertebrate source, including mammals such as primates (e.g. humans, cynomolgus monkey (cyno)) and rodents (e.g., mice and rats), unless otherwise indicated.
• the term encompasses “full-length,” unprocessed B7-H3 as well as any form of B7-H3 that results from processing in the cell.
• the term also encompasses naturally occurring variants of B7-H3, e.g., splice variants, allelic valiants, and isoforms.
• the amino acid sequence of an exemplary human B7-H3 protein is shown in SEQ ID NO: 71.
• ROR1 refers to any native ROR1 from any vertebrate source, including mammals such as primates (e.g. humans, cynomolgus monkey (cyno)) and rodents (e.g., mice and rats), unless otherwise indicated.
• the term encompasses “full-length,” unprocessed ROR1 as well as any form of ROR1 that results from processing in the cell.
• the term also encompasses naturally occurring variants of ROR1, e.g., splice variants, allelic variants, and isoforms.
• the amino acid sequence of an exemplary human ROR1 protein is shown in SEQ ID NO: 72.
• Trop-2 refers to any native Trop-2 from any vertebrate source, including mammals such as primates (e.g. humans, cynomolgus monkey (cyno)) and rodents (e.g., mice and rats), unless otherwise indicated.
• the term encompasses “full-length,” unprocessed Trop-2 as well as any form of Trop-2 that results from processing in the cell.
• the term also encompasses naturally occurring variants of Trop-2, e.g., splice variants, allelic variants, and isoforms.
• the amino acid sequence of an exemplary human Trop-2 protein is shown in SEQ ID NO: 73.
• BCMA refers to any native BCMA from any vertebrate source, including mammals such as primates (e.g. humans, cynomolgus monkey (cyno)) and rodents (e.g., mice and rats), unless otherwise indicated.
• the term encompasses “full-length,” unprocessed BCMA as well as any form of BCMA that results from processing in the cell.
• the term also encompasses naturally occurring variants of BCMA, e.g., splice variants, allelic variants, and isoforms.
• the amino acid sequence of an exemplary human BCMA protein is shown in SEQ ID NO: 74.
• CD25-expressing cancer refers to a cancer comprising cells that express CD25 on their surface.
• CD25-expressing cancer refers to a cancer comprising cells that internalize CD25 inside the cells.
• B7-H3-expressing cancer refers to a cancer comprising cells that express B7-H3 on their surface.
• B7-H3-expressing cancer refers to a cancer comprising cells that internalize B7-H3 inside the cells.
• R0R1 -expressing cancer refers to a cancer comprising cells that express R0R1 on their surface.
• the term “ROR 1 -expressing cancer” refers to a cancer comprising cells that internalize R0R1 inside the cells.
• the term “Trop-2-cxprcssing cancer” refers to a cancer comprising cells that express Trop-2 on their surface. In embodiments, the term “Trop-2-expressing cancer” refers to a cancer comprising cells that internalize Trop-2 inside the cells.
• the term “BCMA-expressing cancer” refers to a cancer comprising cells that express BCMA on their surface. In embodiments, the term “BCMA -expressing cancer” refers to a cancer comprising cells that internalize BCMA inside the cells.
• anti-CD25 antibody and “an antibody that binds to CD25” refer to an antibody that is capable of binding CD25 with sufficient affinity such that the antibody is useful as a therapeutic agent in targeting CD25.
• the extent of binding of an anti- CD25 antibody to an unrelated, non-CD25 protein is less than about 10% of the binding of the antibody to CD25 as measured, e.g., by a radioimmunoassay (RIA).
• RIA radioimmunoassay
• an antibody that binds to CD25 has a dissociation constant (Kd) of ⁇ I pM, ⁇ 100 nM, ⁇ 10 nM, , ⁇ 5 nM , ⁇ 4 nM, ⁇ 3 nM, ⁇ 2 nM, ⁇ 1 nM, ⁇ 0.1 nM, ⁇ 0.01 nM, or ⁇ 0.001 nM (e.g., 10‘ 8 M or less, e.g. from 10' 8 M to 10‘ 13 M, e.g., from 10' 9 M to 10‘ 13 M).
• an anti- CD25 antibody binds to an epitope of CD25 that is conserved among CD25 from different species.
• anti-B7-H3 antibody and “an antibody that binds to B7-H3” refer to an antibody that is capable of binding B7-H3 with sufficient affinity such that the antibody is useful as a therapeutic agent in targeting B7-H3.
• the extent of binding of an anti- B7-H3 antibody to an unrelated, non-B7-H3 protein is less than about 10% of the binding of the antibody to B7-H3 as measured, e.g., by a radioimmunoassay (RIA).
• RIA radioimmunoassay
• an antibody that binds to B7-H3 has a dissociation constant (Kd) of ⁇ IpM, ⁇ 100 nM, ⁇ 10 nM, , ⁇ 5 nM , ⁇ 4 nM, ⁇ 3 nM, ⁇ 2 nM, ⁇ 1 nM, ⁇ 0.1 nM, ⁇ 0.01 nM, or ⁇ 0.001 nM (e.g., 10‘ 8 M or less, e.g. from 10' 8 M to 10‘ 13 M, e.g., from 10' 9 M to 10‘ 13 M).
• an anti- B7-H3 antibody binds to an epitope of B7-H3 that is conserved among B7-H3 from different species.
• anti-RORl antibody and “an antibody that binds to ROR1” refer to an antibody that is capable of binding ROR1 with sufficient affinity such that the antibody is useful as a therapeutic agent in targeting ROR1.
• the extent of binding of an anti- RORl antibody to an unrelated, non-RORl protein is less than about 10% of the binding of the antibody to ROR 1 as measured, e.g., by a radioimmunoassay (RIA).
• RIA radioimmunoassay
• an antibody that binds to ROR1 has a dissociation constant (Kd) of ⁇ IpM, ⁇ 100 nM, ⁇ 10 nM, , ⁇ 5 nM , ⁇ 4 nM, ⁇ 3 nM, ⁇ 2 nM, ⁇ 1 nM, ⁇ 0.1 nM, ⁇ 0.01 nM, or ⁇ 0.001 nM (e.g., 10‘ 8 M or less, e.g. from 10' 8 M to 10‘ 13 M, e.g., from 10' 9 M to 10‘ 13 M).
• an anti- RORl antibody binds to an epitope of ROR1 that is conserved among ROR1 from different species.
• anti-Trop-2 antibody and “an antibody that binds to Trop-2” refer to an antibody that is capable of binding Trop-2 with sufficient affinity such that the antibody is useful as a therapeutic agent in targeting Trop-2.
• the extent of binding of an anti- Trop-2 antibody to an unrelated, non-Trop-2 protein is less than about 10% of the binding of the antibody to Trop-2 as measured, e.g., by a radioimmunoassay (RIA).
• RIA radioimmunoassay
• an antibody that binds to Trop-2 has a dissociation constant (Kd) of ⁇ I , ⁇ 100 nM, ⁇ 10 nM, , ⁇ 5 nM , ⁇ 4 nM, ⁇ 3 nM, ⁇ 2 nM, ⁇ 1 nM, ⁇ 0.1 nM, ⁇ 0.01 nM, or ⁇ 0.001 nM (e.g., 10’ 8 M or less, e.g. from 10' 8 M to 10’ 13 M, e.g., from 10' 9 M to 10’ 13 M).
• an anti- Trop-2 antibody binds to an epitope of Trop-2 that is conserved among Trop-2 from different species.
• anti-BCMA antibody and “an antibody that binds to BCMA” refer to an antibody that is capable of binding BCMA with sufficient affinity such that the antibody is useful as a therapeutic agent in targeting BCMA.
• the extent of binding of an anti- BCMA antibody to an unrelated, non-BCMA protein is less than about 10% of the binding of the antibody to BCMA as measured, e.g., by a radioimmunoassay (RIA).
• RIA radioimmunoassay
• an antibody that binds to BCMA has a dissociation constant (Kd) of ⁇ I M, ⁇ 100 nM, ⁇ 10 nM, , ⁇ 5 nM , ⁇ 4 nM, ⁇ 3 nM, ⁇ 2 nM, ⁇ 1 nM, ⁇ 0.1 nM, ⁇ 0.01 nM, or ⁇ 0.001 nM (e.g., IO’ 8 M or less, e.g. from 10' 8 M to 10‘ 13 M, e.g., from 10' 9 M to 10‘ 13 M).
• an anti- BCMA antibody binds to an epitope of BCMA that is conserved among BCMA from different species.
• chimeric antibody refers to an antibody that contains one or more regions from a first antibody and one or more regions from one or more other antibodies.
• one or more of the CDRs are derived from a human antibody.
• all of the CDRs are derived from a human antibody.
• the CDRs from more than one human antibody arc mixed and matched in a chimeric antibody.
• a chimeric antibody may comprise a CDR1 from the light chain of a first human antibody, a CDR2 and a CDR3 from the light chain of a second human antibody, and the CDRs from the heavy chain from a third antibody.
• the CDRs originate from different species such as human and mouse, or human and rabbit, or human and goat.
• the framework regions may be derived from one of the same antibodies, from one or more different antibodies, such as a human antibody, or from a humanized antibody.
• a portion of the heavy and/or light chain is identical with, homologous to, or derived from an antibody from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is/are identical with, homologous to, or derived from an antibody (-ies) from another species or belonging to another antibody class or subclass.
• fragments of such antibodies that exhibit the desired biological activity (i.e., the ability to specifically bind a target antigen).
• Chimeric antibodies can be prepared from portions of any of the anti-CD25, anti-B7-H3, anti-RORl, anti-Trop-2, or anti- BCMA antibodies described herein.
• Antibody effector functions refer to those biological activities attributable to the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: Clq binding and complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors (e.g., B cell receptor); and B cell activation.
• variant polypeptides and variants of polypeptides refers to a polypeptide comprising an amino acid sequence with one or more amino acid residues inserted into, deleted from and/or substituted into the amino acid sequence relative to a reference polypeptide sequence.
• Polypeptide variants include fusion proteins.
• a variant polynucleotide comprises a nucleotide sequence with one or more nucleotides inserted into, deleted from and/or substituted into the nucleotide sequence relative to another polynucleotide sequence.
• Polynucleotide variants include fusion polynucleotides.
• the term “derivative” of a polypeptide is a polypeptide (e.g., an antibody) that has been chemically modified, e.g., via conjugation to another chemical moiety such as, for example, polyethylene glycol, albumin (e.g., human serum albumin), phosphorylation, and glycosylation.
• another chemical moiety such as, for example, polyethylene glycol, albumin (e.g., human serum albumin), phosphorylation, and glycosylation.
• the term “antibody” includes, in addition to antibodies comprising two full-length heavy chains and two full-length light chains, derivatives, variants, fragments, and mutcins thereof, examples of which arc described below.
• a hinge region refers to an amino acid segment that is generally found between two domains of a protein and may allow for flexibility of the overall construct and movement of one or both of the domains relative to one another.
• a hinge region comprises from about 10 to about 100 amino acids, e.g., from about 15 to about 75 amino acids, from about 20 to about 50 amino acids, or from about 30 to about 60 amino acids.
• the hinge region is 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 amino acids in length.
• the hinge region can be derived from a hinge region of a naturally-occurring protein, such as a CD 8 hinge region or a fragment thereof, a CD8a hinge region, or a fragment thereof, a hinge region of an antibody e.g., IgG, IgA, IgM, IgE, or IgD antibodies), or a hinge region that joins the constant domains CHI and CH2 of an antibody.
• a hinge region of a naturally-occurring protein such as a CD 8 hinge region or a fragment thereof, a CD8a hinge region, or a fragment thereof, a hinge region of an antibody e.g., IgG, IgA, IgM, IgE, or IgD antibodies
• an antibody e.g., IgG, IgA, IgM, IgE, or IgD antibodies
• the hinge region can be derived from an antibody and may or may not comprise one or more constant regions of the antibody, or the hinge region comprises the hinge region of an antibody and the CH3 constant region of the antibody, or the hinge region comprises the hinge region of an antibody and the CH2 and CH3 constant regions of the antibody, or the hinge region is a non-naturally occurring peptide, or the hinge region is disposed between the C-terminus of the scFv and the N-terminus of the transmembrane domain.
• the hinge region comprises any one or any combination of two or more regions comprising an upper, core or lower hinge sequences from an IgGl , IgG2, IgG3 or IgG4 immunoglobulin molecule.
• the hinge region comprises an IgGl upper hinge sequence EPKSCDKTHT (SEQ ID NO: 108). In embodiments, the hinge region comprises an IgGl core hinge sequence CPXCP, wherein X is P, R or S (SEQ ID NO: 109). In one embodiment, the hinge region comprises a lower hinge sequence APELLGGP (SEQ ID NO:110). In embodiments, the hinge is joined to an Fc region (CH2) having the amino acid sequence SVFLFPPKPKDT (SEQ ID NO: 111). In one embodiment, the hinge region includes the amino acid sequence of an upper, core and lower hinge and comprises
• the hinge region comprises one, two, three or more cysteines that can form at least one, two, three or more interchain disulfide bonds.
• labeled antibody refers to antibodies and their antigen binding portions thereof that are labeled or joined to a detectable label or moiety for detection, wherein the detectable label or moiety is radioactive, colorimetric, antigenic, enzymatic, a detectable bead (such as a magnetic or electrodense (e.g., gold) bead), biotin, streptavidin or protein A.
• detectable label or moiety is radioactive, colorimetric, antigenic, enzymatic, a detectable bead (such as a magnetic or electrodense (e.g., gold) bead), biotin, streptavidin or protein A.
• a variety of labels can be employed, including, but not limited to, radionuclides, fluorescers, enzymes, enzyme substrates, enzyme cofactors, enzyme inhibitors and ligands (e.g., biotin, haptens).
• one or more amino acid residues in one or more CDR sequences of a non-human antibody are changed to reduce the likely immunogenicity of the non-human antibody when it is administered to a human subject, wherein the changed amino acid residues either are not critical for immunospecific binding of the antibody to its antigen, or the changes to the amino acid sequence that are made are conservative changes, such that the binding of the humanized antibody to the antigen is not significantly worse than the binding of the non-human antibody to the antigen. Examples of how to make humanized antibodies may be found in U.S. Pat. Nos. 6,054,297, 5,886,152 and 5,877,293.
• human antibody refers to antibodies that have one or more variable and constant regions derived from human immunoglobulin sequences. In one embodiment, all of the variable and constant domains are derived from human immunoglobulin sequences (e.g., a fully human antibody). These antibodies may be prepared in a variety of ways, examples of which are described below, including through recombinant methodologies or through immunization with an antigen of interest of a mouse that is genetically modified to express antibodies derived from human heavy and/or light chain-encoding genes. Fully human anti-CD25 antibodies, anti-B7-H3 antibodies, anti-RORl antibodies, anti-Trop-2 antibodies and anti-BCMA antibodies, and antigen binding proteins thereof are described herein.
• isolated means altered “by the hand of man” from its natural state, has been changed or removed from its original environment, or both.
• isolated means altered “by the hand of man” from its natural state, has been changed or removed from its original environment, or both.
• isolated denotes that the nucleic acid or protein is essentially free of other cellular components with which it is associated in the natural state. It can be, for example, in a homogeneous state and may be in either a dry or aqueous solution. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis, high-performance liquid chromatography or mass spectrophotometry.
• a protein that is the predominant species present in a preparation is substantially purified.
• a polynucleotide or a polypeptide naturally present in a living organism is not “isolated,” but the same polynucleotide or polypeptide separated from the coexisting materials of its natural state is “isolated”, including but not limited to when such polynucleotide or polypeptide is introduced back into a cell, even if the cell is of the same species or type as that from which the polynucleotide or polypeptide was separated.
• CDRs are defined as the complementarity determining region amino acid sequences of an antibody which are the hypervariable domains of immunoglobulin heavy and light chains. There are three heavy chain and three light chain CDRs (or CDR regions) in the variable portion of an immunoglobulin. Thus, “CDRs” as used herein may refer to all three heavy chain CDRs, or all three light chain CDRs (or both all heavy and all light chain CDRs, if appropriate). [0082] CDRs provide the majority of contact residues for the binding of the antibody to the antigen or epitope.
• CDRs of interest in this invention are derived from donor antibody variable heavy and light chain sequences, and include analogs of the naturally occurring CDRs, which analogs also share or retain the same antigen binding specificity and/or neutralizing ability as the donor antibody from which they were derived.
• the minimum overlapping region using at least two of the Kabat, Chothia, AbM and contact methods can be determined to provide the “minimum binding unit”.
• the minimum binding unit may be a subportion of a CDR.
• domain refers to a folded protein structure which has tertiary structure independent of the rest of the protein. Generally, domains are responsible for discrete functional properties of proteins and in many cases may be added, removed or transferred to other proteins without loss of function of the remainder of the protein and/or of the domain.
• An “antibody single variable domain” is a folded polypeptide domain comprising sequences characteristic of antibody variable domains.
• variable domains and modified variable domains, for example, in which one or more loops have been replaced by sequences which are not characteristic of antibody variable domains, or antibody variable domains which have been truncated or comprise N- or C-terminal extensions, as well as folded fragments of variable domains which retain at least the binding activity and specificity of the full-length domain.
• cytotoxic agent refers to a substance that inhibits or prevents a cellular function and/or causes cell death or destruction.
• Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., 211 At, 131 I, 125 I, 90 Y, 186 Re, 188 Re, 153 Sm, 212 Bi, 32 P, 212 Pb and radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, adriamicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitory agents; enzymes and fragments thereof such as nucleolytic enzymes; antibiotics; toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin
• a “chemotherapeutic agent” is a chemical compound useful in the treatment of a cancer.
• chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN®); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicines; betulinic acid; a camptothecin (including the synthetic analogue topotecan
• calicheamicin especially calicheamicin gammall and calicheamicin omegall
• dynemicin including dynemicin A; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antiobiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, dctorubicin, 6-diazo-5-oxo-L-norlcucinc, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-dox
• an “antibody-drug conjugate” or “ADC” is an antibody conjugated to one or more heterologous molecule(s), including but not limited to a cytotoxic agent.
• conjugated when referring to two moieties means the two moieties are bonded, wherein the bond or bonds connecting the two moieties may be covalent or non-covalent.
• the two moieties are covalently bonded to each other (e.g. directly or through a covalently bonded intermediary).
• the two moieties are non-covalently bonded (e.g. through ionic bond(s), van der waal’s bond(s)/interactions, hydrogen bond(s), polar bond(s), or combinations or mixtures thereof).
• an “individual” or “subject” is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and nonhuman primates such as monkeys), rabbits, and rodents (e.g., mice and rats).
• the individual or subject is a human.
• the subject is an adult, an adolescent, a child, or an infant.
• the terms “individual” or “patient” are used and are intended to be interchangeable with “subject”.
• Percent (%) amino acid sequence identity with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, with the aid of the local homology algorithm by Smith and Waterman, 1981, Ads App. Math. 2, 482, with the aid of the local homology algorithm by Needleman and Wunsch, 1970, J. Mol. Biol. 48, 443, with the aid of the similarity search algorithm by Pearson and Lipman, 1988, Proc. Natl Acad.
• the amino acid sequence in the comparison window may comprise additions or deletions (e.g., gaps or overhangs) as compared to the reference sequence for optimal alignment of the two sequences.
• Local alignment between two sequences only includes segments of each sequence that are deemed to be sufficiently similar according to a criterion that depends on the algorithm used to perform the alignment (e.g., EMBOSS Water), "identical” or percent “identity,” refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region, when compared and aligned for maximum correspondence over a comparison window or designated region).
• the percentage identity is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100.
• Optimal alignment of sequences for comparison may be conducted by the local homology algorithm of Smith and Waterman (Add. APL. Math. 2:482, 1981), by the global homology alignment algorithm of Needleman and Wunsch (J. Mol. Biol. 48:443, 1970), by the search for similarity method of Pearson and Lipman (Proc. Natl. Acad. Sci. USA 85: 2444, 1988), or by inspection.
• GAP and BESTFIT as additional examples, can be employed to determine the optimal alignment of two sequences that have been identified for comparison. Typically, the default values of 5.00 for gap weight and 0.30 for gap weight length are used.
• a comparison of the sequences and determination of the percent identity between two polypeptide sequences, or between two polynucleotide sequences may be accomplished using a mathematical algorithm.
• the "percent identity” or “percent homology" of two polypeptide or two polynucleotide sequences may be determined by comparing the sequences using the GAP computer program (a part of the GCG Wisconsin Package, version 10.3 (Accelrys, San Diego, Calif.)) using its default parameters.
• Expressions such as “comprises a sequence with at least X% identity to Y” with respect to a test sequence mean that, when aligned to sequence Y as described above, the test sequence comprises residues identical to at least X% of the residues of Y.
• the amino acid sequence of a test antibody may be similar but not identical to any of the amino acid sequences of the polypeptides that make up the multi- specific antigen binding protein complexes described herein.
• the similarities between the test antibody and the polypeptides can be at least 95%, or at or at least 96% identical, or at least 97% identical, or at least 98% identical, or at least 99% identical, to any of the polypeptides that make up the multi- specific antigen binding protein complexes described herein.
• similar polypeptides can contain amino acid substitutions within a heavy and/or light chain.
• the amino acid substitutions comprise one or more conservative amino acid substitutions.
• a “conservative amino acid substitution” is one in which an amino acid residue is substituted by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity).
• R group side chain
• a conservative amino acid substitution will not substantially change the functional properties of a protein.
• the percent sequence identity or degree of similarity may be adjusted upwards to correct for the conservative nature of the substitution. Means for making this adjustment are well-known to those of skill in the art.
• Examples of groups of amino acids that have side chains with similar chemical properties include (1) aliphatic side chains: glycine, alanine, valine, leucine and isoleucine; (2) aliphatic-hydroxyl side chains: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartate and glutamate, and (7) sulfur-containing side chains are cysteine and methionine.
• Antibodies can be obtained from sources such as serum or plasma that contain immunoglobulins having varied antigenic specificity. If such antibodies are subjected to affinity purification, they can be enriched for a particular antigenic specificity. Such enriched preparations of antibodies usually are made of less than about 10% antibody having specific binding activity for the particular antigen. Subjecting these preparations to several rounds of affinity purification can increase the proportion of antibody having specific binding activity for the antigen. Antibodies prepared in this manner arc often referred to as "monospecific.” Monospecific antibody preparations can be made up of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 99.9% antibody having specific binding activity for the particular antigen. Antibodies can be produced using recombinant nucleic acid technology as described below.
• vector refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked.
• the term includes the vector as a selfreplicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced.
• Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors arc referred to herein as “expression vectors.”
• host cell refers to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells.
• Host cells include “transformants” and “transformed cells,” which include the primary transformed cell and progeny derived therefrom without regal'd to the number of passages. Progeny may not be completely identical in nucleic acid content to a parent cell, but may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are included herein.
• salts are meant to include salts of the active compounds that are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein.
• base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent.
• pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt.
• acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent.
• Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogen sulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, oxalic, methanesulfonic, and the like.
• inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogen sulfuric, hydriodic, or phosphorous
• salts of amino acids such as arginate and the like
• salts of organic acids like glucuronic or galactunoric acids and the like see, for example, Berge el al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19).
• Certain specific compounds of the present disclosure contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.
• the compounds of the present disclosure may exist as salts, such as with pharmaceutically acceptable acids.
• the present disclosure includes such salts.
• Non-limiting examples of such salts include hydrochlorides, hydrobromides, phosphates, sulfates, methanesulfonates, nitrates, maleates, acetates, citrates, fumarates, proprionates, tartrates (e.g., (+)-tartrates, (-)-tartrates, or mixtures thereof including racemic mixtures), succinates, benzoates, and salts with amino acids such as glutamic acid, and quaternary ammonium salts (e.g. methyl iodide, ethyl iodide, and the like). These salts may be prepared by methods known to those skilled in the art.
• the neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner.
• the parent form of the compound may differ from the various salt forms in certain physical properties, such as solubility in polar solvents.
• the present disclosure provides compounds, which are in a prodrug form.
• Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present disclosure.
• Prodrugs of the compounds described herein may be converted in vivo after administration.
• prodrugs can be converted to the compounds of the present disclosure by chemical or biochemical methods in an ex vivo environment, such as, for example, when contacted with a suitable enzyme or chemical reagent.
• Certain compounds of the present disclosure can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present disclosure. Certain compounds of the present disclosure may exist in multiple crystalline or amorphous forms. In general, all physical forms arc equivalent for the uses contemplated by the present disclosure and are intended to be within the scope of the present disclosure.
• “Pharmaceutically acceptable excipient” and “pharmaceutically acceptable carrier” refer to a substance that aids the administration of an active agent to and absorption by a subject and can be included in the compositions of the present disclosure without causing a significant adverse toxicological effect on the patient.
• Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer’s, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, and the like.
• Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with the compounds of the disclosure.
• auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with the compounds of the disclosure.
• auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with the compounds of the disclosure.
• pharmaceutical formulation refers to a preparation which is in such form as to permit the biological activity of an active ingredient contained therein to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered.
• administering refers to the physical introduction of an agent to a subject, using any of the various methods and delivery systems known to those skilled in the art.
• exemplary routes of administration for the formulations disclosed herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes of administration, for example by injection or infusion.
• parenteral administration means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion, as well as in vivo electroporation.
• the formulation is administered via a non-parenteral route, e.g., orally.
• non -parenteral routes include a topical, epidermal or mucosal route of administration, for example, intranasally, vaginally, rectally, sublingually or topically.
• Administering can also be performed, for example, once, a plurality of times, and/or over one or more extended periods.
• an “effective amount” of an agent refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.
• saccharide means carbohydrate (or sugar). In embodiments, the saccharide is a monosaccharide. In embodiments, the saccharide is a polysaccharide. The most basic unit of saccharide is a monomer of carbohydrate. The general formula is CnthnOn.
• saccharide derivative means sugar molecules that have been modified with substituents other than hydroxyl groups. Examples include glycosylamines, sugar phosphates, and sugar esters. Other saccharide derivatives include for example beta-D-glucuronyl, D-galactosyl, and D-glucosyl.
• Charged Group means a chemical group bearing a positive or a negative charge, such as for example phosphate, phosphonate, sulfate, sulfonate, nitrate, carboxylate, carbonate, etc.
• a Charged Group is at least 50% ionized in aqueous solution at least one pH in the range of 5-9.
• a Charged Group is an anionic Charged Group.
• alkyl by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched carbon chain (or carbon), or combination thereof, which may be fully saturated, mono- or polyunsaturated and can include mono-, di- and multivalent radicals.
• the alkyl may include a designated number of carbons (e.g., Ci-Cio means one to ten carbons).
• Alkyl is an uncyclized chain.
• saturated hydrocarbon radicals include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec -butyl, methyl, homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.
• An unsaturated alkyl group is one having one or more double bonds or triple bonds.
• Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2- propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(l,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs and isomers.
• An alkoxy is an alkyl attached to the remainder of the molecule via an oxygen linker (-O-).
• An alkyl moiety may be an alkenyl moiety.
• An alkyl moiety may be an alkynyl moiety.
• An alkyl moiety may be fully saturated.
• An alkenyl may include more than one double bond and/or one or more triple bonds in addition to the one or more double bonds.
• An alkynyl may include more than one triple bond and/or one or more double bonds in addition to the one or more triple bonds.
• alkylene by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkyl, as exemplified, but not limited by, -CH2CH2CH2CH2-.
• an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being preferred herein.
• a “lower alkyl” or “lower alkylene” is a shorter chain alkyl or alkylene group, generally having eight or fewer carbon atoms.
• alkenylene by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkene.
• heteroalkyl by itself or in combination with another term, means, unless otherwise stated, a stable straight or branched chain, or combinations thereof, including at least one carbon atom and at least one heteroatom (e.g., O, N, P, Si, or S), and wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quatemized.
• the heteroatom(s) e.g., O, N, S, Si, or P
• Hctcroalkyl is an uncyclizcd chain.
• a heteroalkyl moiety may include one heteroatom (e.g., O, N, S, Si, or P).
• a heteroalkyl moiety may include two optionally different heteroatoms (e.g., O, N, S, Si, or P).
• a heteroalkyl moiety may include three optionally different heteroatoms (e.g., O, N, S, Si, or P).
• a heteroalkyl moiety may include four optionally different heteroatoms (e.g., O, N, S, Si, or P).
• a heteroalkyl moiety may include five optionally different heteroatoms (e.g., O, N, S, Si, or P).
• a heteroalkyl moiety may include up to
• heteroalkenyl by itself or in combination with another term, means, unless otherwise stated, a heteroalkyl including at least one double bond.
• a heteroalkenyl may optionally include more than one double bond and/or one or more triple bonds in addition to the one or more double bonds.
• heteroalkynyl by itself or in combination with another term, means, unless otherwise stated, a heteroalkyl including at least one triple bond.
• a heteroalkynyl may optionally include more than one triple bond and/or one or more double bonds in addition to the one or more triple bonds.
• heteroalkylene by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from heteroalkyl, as exemplified, but not limited by, -CH2-CH2-S-CH2-CH2- and -CH2-S-CH2-CH2-NH-CH2-.
• heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like).
• no orientation of the linking group is implied by the direction in which the formula of the linking group is written.
• heteroalkyl groups include those groups that are attached to the remainder of the molecule through a heteroatom, such as - C(O)R', -C(O)NR', -NR'R", -OR', -SR', and/or -SO2R'.
• heteroalkyl is recited, followed by recitations of specific heteroalkyl groups, such as -NR'R" or the like, it will be understood that the terms heteroalkyl and -NR'R" are not redundant or mutually exclusive. Rather, the specific heteroalkyl groups are recited to add clarity. Thus, the term “heteroalkyl” should not be interpreted herein as excluding specific heteroalkyl groups, such as -NR'R” or the like.
• heterocycloalkyl examples include, but are not limited to, 1-(1, 2,5,6- tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1- piperazinyl, 2-piperazinyl, and the like.
• a “cycloalkylene” and a “heterocycloalkylene,” alone or as part of another substituent, means a divalent radical derived from a cycloalkyl and heterocycloalkyl, respectively.
• cycloalkyl means a monocyclic, bicyclic, or a multicyclic cycloalkyl ring system.
• monocyclic ring systems are cyclic hydrocarbon groups containing from 3 to 8 carbon atoms, where such groups can be saturated or unsaturated, but not aromatic.
• cycloalkyl groups are fully saturated. Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.
• Bicyclic cycloalkyl ring systems are bridged monocyclic rings or fused bicyclic rings.
• bridged monocyclic rings contain a monocyclic cycloalkyl ring where two non adjacent carbon atoms of the monocyclic ring are linked by an alkylene bridge of between one and three additional carbon atoms (i.e., a bridging group of the form (CFEjw , where w is 1, 2, or 3).
• the fused bicyclic cycloalkyl is a 5 or 6 membered monocyclic cycloalkyl ring fused to either a phenyl ring, a 5 or 6 membered monocyclic cycloalkyl, a 5 or 6 membered monocyclic cycloalkcnyl, a 5 or 6 membered monocyclic heterocyclyl, or a 5 or 6 membered monocyclic heteroaryl, wherein the fused bicyclic cycloalkyl is optionally substituted by one or two groups which are independently oxo or thia.
• multicyclic cycloalkyl ring systems are a monocyclic cycloalkyl ring (base ring) fused to either (i) one ring system selected from the group consisting of a bicyclic aryl, a bicyclic heteroaryl, a bicyclic cycloalkyl, a bicyclic cycloalkenyl, and a bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of a phenyl, a bicyclic aryl, a monocyclic or bicyclic heteroaryl, a monocyclic or bicyclic cycloalkyl, a monocyclic or bicyclic cycloalkenyl, and a monocyclic or bicyclic heterocyclyl.
• a cycloalkyl is a cycloalkenyl.
• the term “cycloalkenyl” is used in accordance with its plain ordinary meaning.
• a cycloalkenyl is a monocyclic, bicyclic, or a multicyclic cycloalkenyl ring system.
• monocyclic cycloalkenyl ring systems are cyclic hydrocarbon groups containing from 3 to 8 carbon atoms, where such groups are unsaturated (i.e., containing at least one annular carbon carbon double bond), but not aromatic. Examples of monocyclic cycloalkenyl ring systems include cyclopentenyl and cyclohexenyl.
• bicyclic cycloalkenyl rings are bridged monocyclic rings or a fused bicyclic rings.
• bridged monocyclic rings contain a monocyclic cycloalkenyl ring where two non adjacent carbon atoms of the monocyclic ring are linked by an alkylene bridge of between one and three additional carbon atoms (i.e., a bridging group of the form (CHijw, where w is 1, 2, or 3).
• Representative examples of bicyclic cycloalkenyls include, but are not limited to, norbomenyl and bicyclo[2.2.2]oct 2 enyl.
• fused bicyclic cycloalkenyl ring systems contain a monocyclic cycloalkenyl ring fused to either a phenyl, a monocyclic cycloalkyl, a monocyclic cycloalkcnyl, a monocyclic hctcrocyclyl, or a monocyclic heteroaryl.
• the bridged or fused bicyclic cycloalkenyl is attached to the parent molecular moiety through any carbon atom contained within the monocyclic cycloalkenyl ring.
• cycloalkenyl groups are optionally substituted with one or two groups which are independently oxo or thia.
• multicyclic cycloalkenyl rings contain a monocyclic cycloalkenyl ring (base ring) fused to either (i) one ring system selected from the group consisting of a bicyclic aryl, a bicyclic heteroaryl, a bicyclic cycloalkyl, a bicyclic cycloalkenyl, and a bicyclic heterocyclyl; or (ii) two ring systems independently selected from the group consisting of a phenyl, a bicyclic aryl, a monocyclic or bicyclic heteroaryl, a monocyclic or bicyclic cycloalkyl, a monocyclic or bicyclic cycloalkenyl, and a monocyclic or bicyclic heterocyclyl.
• the 6 or 7 membered ring contains zero, one or two double bonds and one, two or three heteroatoms selected from the group consisting of O, N and S.
• the heterocyclyl monocyclic heterocycle is connected to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the heterocyclyl monocyclic heterocycle.
• aryl means, unless otherwise stated, a polyunsaturated, aromatic, hydrocarbon substituent, which can be a single ring or multiple rings (preferably from 1 to 3 rings) that are fused together (i.e., a fused ring aryl) or linked covalently.
• a fused ring aryl refers to multiple rings fused together wherein at least one of the fused rings is an aryl ring.
• heteroaryl refers to aryl groups (or rings) that contain at least one heteroatom such as N, O, or S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized.
• heteroaryl includes fused ring heteroaryl groups (i.e., multiple rings fused together wherein at least one of the fused rings is a heteroaromatic ring).
• a 5,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 5 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring.
• a 6,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring.
• Nonlimiting examples of aryl and heteroaryl groups include phenyl, naphthyl, pyrrolyl, pyrazolyl, pyridazinyl, triazinyl, pyrimidinyl, imidazolyl, pyrazinyl, purinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidyl, benzothiazolyl, benzoxazoyl benzimidazolyl, benzofuran, isobenzofuranyl, indolyl, isoindolyl, benzothiophenyl, isoquinolyl, quinoxalinyl, quinolyl, 1- naphthyl, 2-naphthyl, 4-biphcnyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4- imidazo
• arylene and heteroarylene independently or as part of another substituent, mean a divalent radical derived from an aryl and heteroaryl, respectively.
• a heteroaryl group substituent may be -O- bonded to a ring heteroatom nitrogen.
• a fused ring heterocyloalkyl-aryl is an aryl fused to a heterocycloalkyl.
• a fused ring heterocycloalkyl-heteroaryl is a heteroaryl fused to a heterocycloalkyl.
• a fused ring heterocycloalkyl-cycloalkyl is a heterocycloalkyl fused to a cycloalkyl.
• a fused ring heterocycloalkyl-heterocycloalkyl is a heterocycloalkyl fused to another heterocycloalkyl.
• Fused ring heterocycloalkyl-aryl, fused ring heterocycloalkyl-heteroaryl, fused ring heterocycloalkyl- cycloalkyl, or fused ring heterocycloalkyl-heterocycloalkyl may each independently be unsubstituted or substituted with one or more of the substitutents described herein.
• Spirocyclic rings are two or more rings wherein adjacent rings are attached through a single atom.
• the individual rings within spirocyclic rings may be identical or different.
• Individual rings in spirocyclic rings may be substituted or unsubstituted and may have different substituents from other individual rings within a set of spirocyclic rings. Possible substituents for individual rings within spirocyclic rings are the possible substituents for the same ring when not part of spirocyclic rings (e.g. substituents for cycloalkyl or heterocycloalkyl rings).
• Spirocylic rings may be substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heterocycloalkylene and individual rings within a spirocyclic ring group may be any of the immediately previous list, including having all rings of one type (e.g. all rings being substituted heterocycloalkylene wherein each ring may be the same or different substituted heterocycloalkylene).
• heterocyclic spirocyclic rings means a spirocyclic rings wherein at least one ring is a heterocyclic ring and wherein each ring may be a different ring.
• substituted spirocyclic rings means that at least one ring is substituted and each substituent may optionally be different.
• oxo means an oxygen that is double bonded to a carbon atom.
• alkylsulfonyl means a moiety having the formula -S(O2)-R', where R' is a substituted or unsubstituted alkyl group as defined above. R' may have a specified number of carbons (e.g., “C1-C4 alkylsulfonyl”).
• alkylarylene as an arylene moiety covalently bonded to an alkylene moiety (also referred to herein as an alkylene linker).
• alkylarylene group has the formula:
• An alkylarylene moiety may be substituted (e.g. with a substituent group) on the alkylene moiety or the arylene linker (e.g. at carbons 2, 3, 4, or 6) with halogen, oxo, -N3, -CF3, - CCh, -CBr 3 , -CI3, -CN, -CHO, -OH, -NH 2 , -COOH, -CONH 2 , -NO 2 , -SH, -SO2CH3 -SO3H, , - OSO3H, -SO2NH2, -NHNH 2 , -ONH2, -NHC(O)NHNH 2 , substituted or unsubstituted C1-C5 alkyl or substituted or unsubstituted 2 to 5 membered heteroalkyl).
• the alkylarylene is unsubstituted.
• Each of the above terms e.g., “alkyl,” “heteroalkyl,” “cycloalkyl,” “heterocycloalkyl,” “aryl,” and “heteroaryl” includes both substituted and unsubstituted forms of the indicated radical. Preferred substituents for each type of radical are provided below.
• R, R', R", R'", and R" each preferably independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1-3 halogens), substituted or unsubstituted heteroaryl, substituted or unsubstituted alkyl, alkoxy, or thioalkoxy groups, or arylalkyl groups.
• aryl e.g., aryl substituted with 1-3 halogens
• substituted or unsubstituted heteroaryl substituted or unsubstituted alkyl, alkoxy, or thioalkoxy groups, or arylalkyl groups.
• each of the R groups is independently selected as are each R', R", R'", and R"" group when more than one of these groups is present.
• R' and R" are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring.
• -NR'R includes, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl.
• alkyl is meant to include groups including carbon atoms bound to groups other than hydrogen groups, such as haloalkyl (e.g., -CF 3 and -CH 2 CF 3 ) and acyl (e.g., -C(O)CH 3 , -C(O)CF 3 , -C(O)CH 2 OCH 3 , and the like).
• haloalkyl e.g., -CF 3 and -CH 2 CF 3
• acyl e.g., -C(O)CH 3 , -C(O)CF 3 , -C(O)CH 2 OCH 3 , and the like.
• each of the R groups is independently selected as are each R', R", R'", and R"" groups when more than one of these groups is present.
• Substituents for rings e.g. cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene
• substituents on the ring may be depicted as substituents on the ring rather than on a specific atom of a ring (commonly referred to as a floating substituent).
• the substituent may be attached to any of the ring atoms (obeying the rules of chemical valency) and in the case of fused rings or spirocyclic rings, a substituent depicted as associated with one member of the fused rings or spirocyclic rings (a floating substituent on a single ring), may be a substituent on any of the fused rings or spirocyclic rings (a floating substituent on multiple rings).
• the multiple substituents may be on the same atom, same ring, different atoms, different fused rings, different spirocyclic rings, and each substituent may optionally be different.
• a point of attachment of a ring to the remainder of a molecule is not limited to a single atom (a floating substituent)
• the attachment point may be any atom of the ring and in the case of a fused ring or spirocyclic ring, any atom of any of the fused rings or spirocyclic rings while obeying the rules of chemical valency.
• a ring, fused rings, or spirocyclic rings contain one or more ring heteroatoms and the ring, fused rings, or spirocyclic rings are shown with one more floating substituents (including, but not limited to, points of attachment to the remainder of the molecule), the floating substituents may be bonded to the heteroatoms.
• the ring heteroatoms are shown bound to one or more hydrogens (e.g. a ring nitrogen with two bonds to ring atoms and a third bond to a hydrogen) in the structure or formula with the floating substituent, when the heteroatom is bonded to the floating substituent, the substituent will be understood to replace the hydrogen, while obeying the rules of chemical valency.
• Two or more substituents may optionally be joined to form aryl, heteroaryl, cycloalkyl, or heterocycloalkyl groups.
• Such so-called ring-forming substituents are typically, though not necessarily, found attached to a cyclic base structure.
• the ring-forming substituents are attached to adjacent members of the base structure.
• two ring-forming substituents attached to adjacent members of a cyclic base structure create a fused ring structure.
• the ring-forming substituents are attached to a single member of the base structure.
• two ring-forming substituents attached to a single member of a cyclic base structure create a spirocyclic structure.
• the ring-forming substituents are attached to non-adjacent members of the base structure.
• Two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally form a ring of the formula -T-C(O)-(CRR') P -U-, wherein T and U are independently - NR-, -O-, -CRR'-, or a single bond, and p is an integer of from 0 to 3.
• two of the substituents on adjacent atoms of the aryl or hctcroaryl ring may optionally be replaced with a substituent of the formula -A-(CH2) r -B-, wherein A and B are independently -CRR'-, -O-, -NR-, - S-, -S(O) -, -S(O) 2 -, -S(O)2NR'-, or a single bond, and r is an integer of from 1 to 4.
• One of the single bonds of the new ring so formed may optionally be replaced with a double bond.
• two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -(CRR')s-X'- (C"R"R"')d-, where s and d are independently integers of from 0 to 3, and X' is -O-, -NR'-, -S-, -S(O)-, -S(O) 2 -, or - S(O) 2 NR’-.
• R, R', R", and R' are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.
• heteroatom or “ring heteroatom” are meant to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).
• a “substituent group,” as used herein, means a group selected from the following moieties
• alkyl e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl
• heteroalkyl e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl
• cycloalkyl e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl
• heterocycloalkyl e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl
• aryl e.g., Ce-Cio aryl, C10 aryl, or phenyl
• heteroaryl e.g., 5 to 10 membered hctcroaryl, 5 to 9 membered hctcroary
• alkyl e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl
• heteroalkyl e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl
• cycloalkyl e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl
• heterocycloalkyl e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl
• aryl e.g., Ce-Cio aryl, C10 aryl, or phenyl
• heteroaryl e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membere
• alkyl e.g., C 1 -C 8 alkyl, C1-C6 alkyl, or C1-C4 alkyl
• heteroalkyl e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl
• cycloalkyl e.g., C 3 -Cs cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl
• heterocycloalkyl e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl
• aryl e.g., Ce-Cio aryl, C10 aryl, or phenyl
• heteroaryl e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membere
• unsubstituted alkyl e.g., Ci-C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl
• unsubstituted heteroalkyl e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl
• unsubstituted cycloalkyl e.g., C 3 -Cs cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl
• unsubstituted heterocycloalkyl e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl
• unsubstituted aryl e.g., Ci-C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl
• a “size-limited substituent” or “ size-limited substituent group,” as used herein, means a group selected from all of the substituents described above for a “substituent group,” wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C20 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C8 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 8 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted Ce-Cio aryl, and each substituted or unsubstituted heteroaryl is a group selected
• a “lower substituent” or “ lower substituent group,” as used herein, means a group selected from all of the substituents described above for a “substituent group,” wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C 1 -C 8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted phenyl, and each substituted or unsubstituted heteroaryl is a substituted or unsubstitute
• each substituted group described in the compounds herein is substituted with at least one substituent group. More specifically, in some embodiments, each substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene described in the compounds herein are substituted with at least one substituent group. In other embodiments, at least one or all of these groups are substituted with at least one size-limited substituent group. In other embodiments, at least one or all of these groups are substituted with at least one lower substituent group.
• each substituted or unsubstituted alkyl may be a substituted or unsubstituted C1-C20 alkyl
• each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20 membered heteroalkyl
• each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C8 cycloalkyl
• each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 8 membered heterocycloalkyl
• each substituted or unsubstituted aryl is a substituted or unsubstituted Ce-Cio aryl
• each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5 to 10 membered heteroaryl.
• each substituted or unsubstituted alkylene is a substituted or unsubstituted C1-C20 alkylene
• each substituted or unsubstituted heteroalkylene is a substituted or unsubstituted 2 to 20 membered heteroalkylene
• each substituted or unsubstituted cycloalkylene is a substituted or unsubstituted C3-C8 cycloalkylene
• each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3 to 8 membered heterocycloalkylene
• each substituted or unsubstituted arylene is a substituted or unsubstituted C6-C10 arylene
• each substituted or unsubstituted heteroarylene is a substituted or unsubstituted 5 to 10 membered heteroarylene.
• each substituted or unsubstituted alkyl is a substituted or unsubstituted C 1 -C 8 alkyl
• each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8 membered hctcroalkyl
• each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C7 cycloalkyl
• each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7 membered heterocycloalkyl
• each substituted or unsubstituted aryl is a substituted or unsubstituted Ce-Cio aryl
• each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5 to 9 membered heteroaryl.
• each substituted or unsubstituted alkylene is a substituted or unsubstituted C 1 -C 8 alkylene
• each substituted or unsubstituted heteroalkylene is a substituted or unsubstituted 2 to 8 membered heteroalkylene
• each substituted or unsubstituted cycloalkylene is a substituted or unsubstituted C3-C7 cycloalkylene
• each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3 to 7 membered heterocycloalkylene
• each substituted or unsubstituted arylene is a substituted or unsubstituted Ce-Cio arylene
• each substituted or unsubstituted heteroarylene is a substituted or unsubstituted 5 to 9 membered heteroarylene.
• the compound is a chemical species set forth in the Examples section, figures, or tables below.
• a substituted or unsubstituted moiety e.g., substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, and/or substituted or unsubstituted heteroarylene) is unsubstituted (e.g., is an unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted
• a substituted or unsubstituted moiety e.g., substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, and/or substituted or unsubstituted heteroarylene) is substituted (e.g., is a substituted alkyl, substituted hctcroalkyl, substituted cycloalkyl, substituted hctcrocycloalkyl, substituted aryl, substituted
• a substituted moiety e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene
• is substituted with at least one substituent group wherein if the substituted moiety is substituted with a plurality of substituent groups, each substituent group may optionally be different. In embodiments, if the substituted moiety is substituted with a plurality of substituent groups, each substituent group is different.
• a substituted moiety e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene
• is substituted with at least one size-limited substituent group wherein if the substituted moiety is substituted with a plurality of size-limited substituent groups, each size-limited substituent group may optionally be different. In embodiments, if the substituted moiety is substituted with a plurality of size-limited substituent groups, each size-limited substituent group is different.
• a substituted moiety e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene
• is substituted with at least one lower substituent group wherein if the substituted moiety is substituted with a plurality of lower substituent groups, each lower substituent group may optionally be different. In embodiments, if the substituted moiety is substituted with a plurality of lower substituent groups, each lower substituent group is different.
• a substituted moiety e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene
• each substituent group, size-limited substituent group, and/or lower substituent group is different.
• Certain compounds of the present disclosure possess asymmetric carbon atoms (optical or chiral centers) or double bonds; the enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisometric forms that may be defined, in terms of absolute stereochemistry, as (R)-or (S)- or, as (D)- or (L)- for amino acids, and individual isomers are encompassed within the scope of the present disclosure.
• the compounds of the present disclosure do not include those that are known in art to be too unstable to synthesize and/or isolate.
• the present disclosure is meant to include compounds in racemic and optically pure forms.
• Optically active (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.
• the compounds described herein contain olefinic bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers.
• the term “isomers” refers to compounds having the same number and kind of atoms, and hence the same molecular weight, but differing in respect to the structural arrangement or configuration of the atoms.
• the term “tautomer,” as used herein, refers to one of two or more structural isomers which exist in equilibrium and which are readily converted from one isomeric form to another.
• tautomer refers to one of two or more structural isomers which exist in equilibrium and which are readily converted from one isomeric form to another.
• structures depicted herein are also meant to include all stereochemical forms of the structure; i.e., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the disclosure.
• Linker refers to a chemical moiety comprising a covalent bond or a chain of atoms that covalently attaches an antibody to a drug moiety.
• linkers include a divalent radical.
• linkers can comprise one or more amino acid residues.
• the linker is a non-cleavable linker.
• the linker is an enzyme- cleavable linker (e.g., Val-Cit or Val-Cit-PAB linker).
• “Amino Acid Unit” has the formula hydrogen, methyl, isopropyl, isobutyl, sec -butyl, benzyl, p-hydroxybenzyl, — CH2OH, — CH(OH)CH3, — CH2CH2SCH , — CH2CONH2, — CH2COOH, — CH2CH2CONH2, — CH2CH2COOH,
• Amino Acid Unit includes not only naturally occurring amino acids but also minor amino acids, and non- naturally occurring amino acid analogs, such as citrulline, norleucine, selenomethionine, - alanine, N-dimethyl lysine etc.
• An amino acid unit may be referred to by its standard three-letter code for the amino acid (e.g., Ala, Cys, Asp, Glu, Vai, Phe, Lys, etc.).
• aldehyde or ketone groups such that subsequent derivatization is possible via formation of carbonyl derivatives such as, for example, imines, hydrazones, semicarbazones or oximes, or via such mechanisms as Grignard addition or alkyllithium addition;
• amine or thiol groups e.g., present in cysteine
• cysteine e.g., aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoeine)
• cysteine e.g., present in cysteine
• alkenes which can undergo, for example, cycloadditions, acylation, Michael addition, etc;
• biotin conjugate can react with avidin or strepavidin to form a avidin-biotin complex or streptavidin-biotin complex.
• bioconjugate reactive groups can be chosen such that they do not participate in, or interfere with, the chemical stability of the conjugate described herein.
• a reactive functional group can be protected from participating in the crosslinking reaction by the presence of a protecting group.
• the bioconjugate comprises a molecular entity derived from the reaction of an unsaturated bond, such as a maleimide, and a thiol group.
• an analog is used in accordance with its plain ordinary meaning within Chemistry and Biology and refers to a chemical compound that is structurally similar to another compound (i.c., a so-called “reference” compound) but differs in composition, c.g., in the replacement of one atom by an atom of a different element, or in the presence of a particular functional group, or the replacement of one functional group by another functional group, or the absolute stereochemistry of one or more chiral centers of the reference compound. Accordingly, an analog is a compound that is similar or comparable in function and appearance but not in structure or origin to a reference compound.
• R 4 is H, halogen, -OR 4A , -NR 4A R 4B , substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl;
• L 2 is a bond, -C(O)-, -NH-, -Vai-, -Phe-, -Lys-, -Gly-, -(4-aminobenzyloxycarbonyl)-, -(C(O)N(CH3)CH2CH2N(CH3))-, -Ser-, -Thr-, -Ala-, -f>-Ala-, -O-, -citrulline- (Cit), -(CH2) n - -(CH2CH2O) n -, N-dimethyl lysine, or any combination thereof.
• L 3 is substituted with one or more substituent groups. In embodiments, L 3 is substituted with one or more size-limited substituent groups. In embodiments, L 3 is substituted with one or more lower substituent groups.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocycloalkylene, or 5 to 6 membered heterocycloalkylene), or substituted (e.g.
• L 3 is substituted with one or more substituent groups. In embodiments, L 3 is substituted with one or more size-limited substituent groups. In embodiments, L 3 is substituted with one or more lower substituent groups.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocycloalkylene, or 5 to 6 membered heterocycloalkylene).
• L 3 is substituted with one or more substituent groups.
• L 3 is substituted with one or more size-limited substituent groups.
• L 3 is substituted with one or more lower substituent groups.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocycloalkylene (e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocyclo alkylene, or 5 to 6 membered heterocycloalkylene).
• L 3 is unsubstituted heterocycloalkylene (e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocycloalkylene, or 5 to 6 membered heterocycloalkylene).
• L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene).
• heteroarylene e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene.
• L 3 is unsubstituted heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -OCH2-(heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered hctcroarylcnc) ).
• L 3 is unsubstituted -OCH2-(heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene)).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -CH2NCH2-(heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl)).
• L 3 is unsubstituted -CH2NCH2-(heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl)).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 8 membered heterocycloalkylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) 3 to 8 membered heterocycloalkylene. In embodiments, L 3 is unsubstituted 3 to 8 membered heterocycloalkylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) or unsubstituted -CH2NCH2-(3 to 8 membered heterocycloalkyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -CH2NCH2-(3 to 8 membered heterocycloalkyl). In embodiments, L 3 is unsubstituted - CH2NCH2-(3 to 8 membered heterocycloalkyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) or unsubstituted -OCH2-(3 to 8 membered heterocycloalkylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -OCH2-(3 to 8 membered heterocycloalkylene). In embodiments, L 3 is unsubstituted -OCH2-(3 to 8 membered heterocycloalkylene). [00200] In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 8 membered heterocycloalkylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 6 membered heterocycloalkylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) 3 to 6 membered heterocycloalkylene. In embodiments, L 3 is unsubstituted 3 to 6 membered heterocycloalkylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) or unsubsitutcd -CH2NCH2-(3 to 6 membered hctcrocycloalkyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -CH2NCH2-(3 to 6 membered heterocycloalkyl). In embodiments, L 3 is unsubsituted - CH2NCH2-(3 to 6 membered heterocycloalkyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) or unsubsituted -OCH2-(3 to 6 membered heterocycloalkylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -OCH2-(3 to 6 membered heterocycloalkylene). In embodiments, L 3 is unsubsituted -OCH2-(3 to 6 membered heterocycloalkylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 6 membered heterocycloalkylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene.
• L is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclobutylene, heterocyclopentylene or heterocyclohexylene.
• L 3 is unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) -CH2NCH2-(heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl).
• L 3 is unsubstituted -CH2NCH2- (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene). In embodiments, L 3 is unsubstituted -OCH2-(heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted hctcrocyclobutylcnc, heterocyclopentylene or heterocyclohexylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclobutylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclobutylene. In embodiments, L 3 is unsubstituted heterocyclobutylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) or unsubstituted -CH2NCH2- (heterocyclobutyl).
• L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) -CH2NCH2-(heterocyclobutyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(heterocyclobutyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(heterocyclobutylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (heterocyclobutylene). In embodiments, L 3 is unsubstituted -OCH2-(heterocyclobutylene). [00206] In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclopentylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclopentylene.
• L 3 is unsubstituted heterocyclopentylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) or unsubstituted -CH2NCH2- (heterocyclopentyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CHoNCth- heterocyclopentyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(heterocyclopentyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(heterocyclopentylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (heterocyclopentylene). In embodiments, L 3 is unsubstituted -OCH2-(heterocyclopentylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclohexylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) hctcrocyclohcxylcnc.
• L 3 is unsubstituted heterocyclohexylene.
• L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) or unsubstituted -CH2NCH2- (heterocyclohexyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) -Ct NCth-lheterocyclohexyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(heterocyclohexyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(heterocyclohexylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (heterocyclohexylene). In embodiments, L 3 is unsubstituted -OCH2-(heterocyclohexylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(5 to 10 membered heteroaryl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2-(5 to 10 membered heteroaryl). In embodiments, L 3 is unsubstituted -CH2NCH2-(5 to 10 membered heteroaryl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 5 to 9 membered heteroarylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 9 membered heteroarylene. In embodiments, L 3 is unsubstituted 5 to 9 membered heteroarylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(5 to 9 membered heteroaryl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2-(5 to 9 membered heteroaryl). In embodiments, L 3 is unsubstituted -CH2NCH2-(5 to 9 membered heteroaryl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(5 to 9 membered heteroarylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(5 to 9 membered heteroarylene). In embodiments, L 3 is unsubstituted -OCH2-(5 to 9 membered heteroarylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 5 to 6 membered heteroarylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 6 membered heteroarylene. In embodiments, L 3 is unsubstituted 5 to 6 membered heteroarylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(5 to 6 membered heteroaryl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2-(5 to 6 membered heteroaryl). In embodiments, L 3 is unsubstituted -CH2NCH2-(5 to 6 membered heteroaryl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(5 to 6 membered heteroarylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(5 to 6 membered heteroarylene). In embodiments, L 3 is unsubstituted -OCH2-(5 to 6 membered heteroarylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene.
• L 3 is unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2- (furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl).
• L 3 is unsubstituted -CH2NCH2-(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene).
• L 3 is unsubstituted -OCH2-(furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted furanylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) furanylene. In embodiments, L 3 is unsubstituted furanylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(furanyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) - CH2NCH2- (furanyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(furanyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(furanylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (furanylene). In embodiments, L 3 is unsubstituted -OCH2-(furanylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyrrolylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyrrolylene. In embodiments, L 3 is unsubstituted pyrrolylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(pyrrolyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2- (pyrrolyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(pyrrolyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(pyrrolylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size- limited substituent group or a lower substituent group) -OCH2-(pyrrolylene). In embodiments, L 3 is unsubstituted -OCH2-(pyrrolylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyridylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyridylene. In embodiments, L 3 is unsubstituted pyridylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(pyridyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) - CH2NCH2-(pyridyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(pyridyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(pyridylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (pyridylene). In embodiments, L 3 is unsubstituted -OCH2-(pyridylene).
• each R 3A , R 3B , R 4A , and R 4B is independently H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, or pentyl.
• each R 3A , R B , R 4A , and R 4B is independently H.
• each R 3A , R 3B , R 4A , and R 4B is independently methyl.
• each R 3A , R 3B , R 4A , and R 4B is independently ethyl.
• R 6 is H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -CO(CH2CH2O) W CH2CH2M,
• w is an integer from 1 to 24; M is -NH2, -OH, -COOH, or -OCH3; R 10 is -OH, -OCH3 or -COOH.
• R 6 is H or substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., Ci- Cs alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl), substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) or unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6
• alkyl e
• R 6 is H or substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl).
• heterocycloalkyl e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl.
• R 6 is H.
• w is an integer from 1 to 24. In embodiments, w is 1 . In embodiments, w is 2. In embodiments, w is 3. In embodiments, w is 4. In embodiments, w is 5. In embodiments, w is 6. In embodiments, w is 7. In embodiments, w is 8. In embodiments, w is 9. In embodiments, w is 10. In embodiments, w is 11. In embodiments, w is 12. In embodiments, w is 13. In embodiments, w is 14. In embodiments, w is 15. In embodiments, w is 16. In embodiments, w is 17. In embodiments, w is 18. In embodiments, w is 19. In embodiments, w is 20. In embodiments, w is 21. In embodiments, w is 22. In embodiments, w is 23. In embodiments, w is 24.
• Z 2 is a substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted arylene (e.g., Ce-Cio arylene, C10 arylene, or phenylene).
• Z 2 is a substituted (e.g. with a substituent group, a sizelimited substituent group or a lower substituent group) arylene (e.g., Ce-Cio arylene, C10 arylene, or phenylene).
• Z 2 is an unsubstituted arylene (e.g., C6-C10 arylene, C10 arylene, or phenylene).
• Z 2 is a substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene).
• Z 2 is a substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene).
• Z 2 is an unsubstituted heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene).
• each G is independently Cl, Br, I, F, -CH 3 , -CH2CH3, -CH2CH2CH3, -OCH3, -OCH2CH3, -OH, or -NH2; and p is an integer from 0-4.
• ring A is a substituted or unsubstituted heterocycloalkylene or a substituted or unsubstituted heteroarylene, connected to L 2 through a heteroatom Y ;
• ring A’ is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl, connected to D’ through a heteroatom Y;
• each Y is independently N, P, or S;
• L 1 , L 2 , Ab, m, D, and D’ are each as defined herein including embodiments.
• ring A is a substituted with one or more (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 6 membered heterocycloalkylene, where ring A is connected to L 2 through a heteroatom Y.
• ring A’ is a substituted with one or more (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 6 membered heterocycloalkyl, where ring A' is connected to D’ through a heteroatom Y.
• each Y is N.
• each R 7 is independently H, oxo, halogen, -CCh, -CBn, -CF3, -CI3, -CH2CI, -CHiBr, -CH 2 F, -CH2I, -CHCI2, -CHBr 2 , -CHF 2 , -CHI2, -CN, -OR 7A , -NR 7A R 7B , -COOR 7A , -CONR 7A R 7B , -NO2, -SR 7A , -SOn?R 7A , -SOV7NR 7A R 7B , -PO(OH) 2 , -PO m7 R 7A , PO r7 NR 7A R 7B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or
• X is -Cl, -Br, -I or -F; each n7 is independently an integer from 0 to 4; each v7 is independently 1 or 2; each m7 is independently an integer from 0 to 3; each r7 is independently 1 or 2; and
• Y, m, D, D’, L 1 , L 2 and Ab are each as defined herein including embodiments.
• an ADC of formula (ID1) or formula (IID 1) or a pharmaceutically acceptable salt thereof; wherein R 7 , D, D’, m, Y, L 1 , L 2 , and Ab are each as defined herein including embodiments.
• ring W is a substituted or unsubstituted cycloalkylene or a substituted or unsubstituted arylene
• ring C is a substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl
• D, m, L 1 , L 2 , and Ab are each as defined herein including embodiments.
• -NH- and -D are connected to different carbon atoms on ring W. In embodiments, -NH- and -D are connected to the same carbon atom on ring W.
• ring W is a substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C8 cycloalkylene, C3-C6 cycloalkylene, or C5-C6 cycloalkylene) or substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted arylene (e.g., C5-C10 arylene, Cs-Cs arylene, or C5-C6 arylene).
• ring W is substituted with one or more substituent groups.
• ring W is substituted with one or more size-limited substituent groups.
• ring W is substituted with one or more lower substituent groups.
• ring W is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted C -C8 cycloalkylene.
• ring W is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) C3-C8 cycloalkylcnc.
• ring W is an unsubstituted C3-C8 cycloalkylene.
• ring W is a substituted with one or more (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) C3-C8 cycloalkylene.
• ring W is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted cyclobutylene.
• ring W is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted cyclopentylene.
• ring W is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted cyclohexylene.
• ring W is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted C5-C6 arylene.
• ring W is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) C5-C6 arylene.
• ring W is an unsubstituted C5-C6 arylene.
• ring W is a substituted with one or more (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) C5-C6 arylene.
• ring C is a substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl) or substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl).
• ring C is substituted with one or more substituent groups.
• ring C is substituted with one or more size-limited substituent groups.
• ring C is substituted with one or more lower substituent groups.
• ring C is a substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl) or substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl).
• ring C is substituted with one or more substituent groups.
• ring C is substituted with one or more size-limited substituent groups.
• ring C is substituted with one or more lower substituent groups.
• ring C is a substituted with one or more (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 9 membered heteroaryl. In embodiments, ring C is an unsubstituted 5 to 9 membered heteroaryl.
• ring C is a substituted with one or more (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 6 membered heteroaryl. In embodiments, ring C is an unsubstituted 5 to 6 membered heteroaryl.
• ring C is a substituted with one or more (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 3 to 8 membered heterocycloalkyl. In embodiments, ring C is a substituted with one or more (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 6 membered heterocycloalkyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl.
• ring C is unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted furanyl. In embodiments, ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) furanyl. In embodiments, ring C is unsubstituted furanyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyrrolyl. In embodiments, ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyrrolyl. In embodiments, ring C is unsubstituted pyrrolyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyridyl. In embodiments, ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyridyl. In embodiments, ring C is unsubstituted pyridyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyranyl. In embodiments, ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyranyl. In embodiments, ring C is unsubstituted pyranyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted imidazolyl. In embodiments, ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) imidazolyl. In embodiments, ring C is unsubstituted imidazolyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted thiazolyl. In embodiments, ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) thiazolyl. In embodiments, ring C is unsubstituted thiazolyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted thienyl. In embodiments, ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) thienyl. In embodiments, ring C is unsubstituted thienyl.
• ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted oxazolyl. In embodiments, ring C is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) oxazolyl. In embodiments, ring C is unsubstituted oxazolyl. [00307] In embodiments, ring C is a substituted (e.g.
• ring C is substituted with one or more substituent groups.
• ring C is substituted with one or more size-limited substituent groups.
• ring C is substituted with one or more lower substituent groups.
• ring C is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted C 3 -C 8 cycloalkyl.
• ring C is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) C 3 -C 8 cycloalkyl.
• ring C is an unsubstituted C 3 -C 8 cycloalkyl.
• ring C is a substituted with one or more (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) C 3 -C 8 cycloalkyl.
• ring C is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted cyclobutyl. In embodiments, ring C is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted cyclopentyl. In embodiments, ring C is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted cyclohexyl.
• ring C is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted C5-C6 aryl.
• ring C is a substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) C5-C6 aryl.
• ring C is an unsubstituted C5-C6 aryl.
• ring C is a substituted with one or more (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) C5-C6 aryl.
• Z is S, N, or O; and wherein R 7 , D, m, L 1 , L 2 , and Ab arc each as defined herein including embodiments.
• Z is N. In embodiments, Z is O. In embodiments, Z is S.
• an ADC having the structure: or a pharmaceutically acceptable salt thereof.
• an ADC having the structure: pharmaceutically acceptable salt thereof.
• an ADC having the structure
• the ADC comprises an antibody that binds to CD25.
• CD25 has been reported to be upregulated, for example, in leukemias and lymphomas independent of baseline levels of CD25 expression.
• the ADC compounds described herein comprise an anti-CD25 antibody.
• the ADC compounds described herein comprise an anti- CD25 antibody comprising one or more mutation(s) in the Fc region, wherein the mutation(s) can include the mutation L234A and/or L235A (the substitution of the residue at position 234 in the EU index set forth in Rabat and/or substitution of the residue at position 235 in the EL) index sei forth in Rabat).
• the ADC compounds described herein comprise an anti- CD25 antibody comprising a mutation in the Fc region, wherein the mutation is L234A (LA mutation). In embodiments, the ADC compounds described herein comprise an anti-CD25 antibody comprising a mutation in the Fc region, wherein the mutation is L235A (LA mutation). In embodiments, the ADC compounds described herein comprise an anti-CD25 antibody comprising the double mutation in the Fc region, wherein the mutations are L234A and L235A (LALA mutations). In embodiments, the anti-CD25 antibody (MAA-V clone) includes the mutations L234A and L235A in the Fc region (MAA-V LALA).
• the ADC provided herein comprises an anti-CD25 antibody (e.g., MAA-V or MAA-V LALA) comprising at least one, two, three, four, five, or six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 1; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDR1 comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• an anti-CD25 antibody e.g., MAA-V or MAA-V LALA
• the ADC comprises an anti-CD25 antibody comprising at least three CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 1; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDR1 comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the ADC comprises anti-CD25 antibody comprising at least four CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 1; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDR1 comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the ADC comprises an anti-CD25 antibody comprising at least five CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 1; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDR1 comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the ADC comprises an anti-CD25 antibody comprising at least six CDRs selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 1; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDRI comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the ADC comprises an anti-CD25 antibody (e.g., MAA-V or MAA- V LALA) comprising one CDR selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 1 ; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDRI comprising the sequence of SEQ ID NO: 4; (c) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• an anti-CD25 antibody e.g., MAA-V or MAA- V LALA
• the ADC comprises an anti-CD25 antibody comprising two CDRs selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 1; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDRI comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the ADC comprises an anti-CD25 antibody comprising three CDRs selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 1; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDRI comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the ADC comprises an anti-CD25 antibody comprising four CDRs selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 1; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDRI comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the ADC comprises an anti-CD25 antibody comprising five CDRs selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 1 ; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDRI comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the ADC comprises an anti-CD25 antibody comprising six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 1; (b) VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) VH CDR1 comprising the sequence of SEQ ID NO: 4; (e) VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the anti-CD25 antibody (e.g., MAA-V or MAA-V LALA) comprises a VL CDR1 comprising the sequence of SEQ ID NO: 1, a VL CDR2 comprising the sequence of SEQ ID NO: 2, a VL CDR3 comprising the sequence of SEQ ID NO: 3, a VH CDR1 comprising the sequence of SEQ ID NO: 4, a VH CDR2 comprising the sequence of SEQ ID NO: 5, and a VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the anti-CD25 antibody comprises a VL CDR1 comprising the sequence of SEQ ID NO: 1.
• the anti-CD25 antibody comprises a VL CDR2 comprising the sequence of SEQ ID NO: 2. In embodiments, the anti-CD25 antibody comprises a VL CDR3 comprising the sequence of SEQ ID NO: 3. In embodiments, the anti-CD25 antibody comprises a VH CDR1 comprising the sequence of SEQ ID NO: 4. In embodiments, the anti-CD25 antibody comprises a VH CDR2 comprising the sequence of SEQ ID NO: 5. In embodiments, the anti-CD25 antibody comprises and a VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the ADC comprises an anti-CD25 antibody (e.g., MAA-V or MAA- V LALA) comprising the light chain CDR1 has the amino acid sequence of SEQ ID NO:1, the light chain CDR2 has the amino acid sequence of SEQ ID NO:2, the light chain CDR3 has the amino acid sequence of SEQ ID NO:3, the heavy chain CDR1 has the amino acid sequence of SEQ ID NO:4, the heavy chain CDR2 has the amino acid sequence of SEQ ID NO:5, and the heavy chain CDR3 has the amino acid sequence of SEQ ID NO:6.
• an anti-CD25 antibody e.g., MAA-V or MAA- V LALA
• the anti-CD25 antibody (e.g., MAA-V or MAA-V LALA) comprises a VL having a sequence with at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 10.
• the anti-CD25 antibody (e.g., MAA-V or MAA-V LALA) comprises a VL having the sequence of SEQ ID NO: 10.
• a VL sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 10 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-CD25 antibody comprising that sequence retains the ability to bind to CD25.
• a total of 1 to 10 amino acids have been substituted, inserted and/or deleted in SEQ ID NO: 10.
• a total of 1 to 5 amino acids have been substituted, inserted and/or deleted in SEQ ID NO: 10.
• substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., in the FRs).
• the anti-CD25 antibody comprises the VL sequence of SEQ ID NO: 10, and includes post-translational modifications of that sequence.
• the anti-CD25 antibody (e.g., MAA-V or MAA-V LALA) comprises a VH having a sequence with at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11.
• the anti-CD25 antibody (e.g., MAA-V or MAA-V LALA) comprises a VH having the sequence of SEQ ID NO: 1 1 .
• a VH sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-CD25 antibody comprising that sequence retains the ability to bind to CD25.
• a total of 1 to 10 amino acids have been substituted, inserted and/or deleted in SEQ ID NO: 11.
• a total of 1 to 5 amino acids have been substituted, inserted and/or deleted in SEQ ID NO: 11.
• substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., in the FRs).
• the anti-CD25 antibody comprises the VH sequence of SEQ ID NO: 11, and includes post-translational modifications of that sequence.
• the anti-CD25 antibody is an IgG antibody. In embodiments, the anti-CD25 antibody is an IgGl, IgG2, IgG3 or IgG4 antibody. In embodiments, the anti-CD25 antibody is an IgGl or IgG4 antibody. In embodiments, the anti-CD25 antibody is an IgGl antibody.
• an anti-CD25 antibody binds a human CD25.
• the human CD25 has the amino acid sequence of SEQ ID NO: 70.
• an anti-CD25 antibody is humanized.
• an anti-CD25 antibody comprises CDRs as in any of the above embodiments, and further comprises a human acceptor framework, e.g., a human immunoglobulin framework or a human consensus framework.
• a humanized anti-CD25 antibody (e.g., MAA-V or MAA-V LALA) comprises (a) a VL CDR1 comprising the sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising the sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising the sequence of SEQ ID NO: 6.
• the anti-CD25 antibody is a monoclonal antibody, including a chimeric, humanized, or human antibody.
• an anti-CD25 antibody is an antibody fragment, e.g., a Fv, Fab, Fab’, scFv, diabody, or F(ab’)2 fragment.
• the antibody is a substantially full-length antibody, e.g., an IgGl antibody or other antibody class or isotype as defined herein.
• the ADC comprises an antibody that binds to B7-H3.
• B7-H3 has been reported to be upregulated, for example, in lung cancer independent of baseline levels of B7-H3 expression.
• the ADC compounds described herein comprise an anti-B7- H3 antibody.
• the B7-H3 compounds described herein comprise an anti-B7-H3 antibody comprising one or more mutation(s) in the Fc region, wherein the mutation(s) can include the mutation L234A and/or L235A (the substitution of the residue at position 234 in the EU index set forth in Kabat and/or substitution of the residue at position 235 in the EU index set forth in Kabat).
• the ADC compounds described herein comprise an anti-B7-H3 antibody comprising a mutation in the Fc region, wherein the mutation is L234A (LA mutation). In embodiments, the ADC compounds described herein comprise an anti-B7-H3 antibody comprising a mutation in the Fc region, wherein the mutation is L235A (LA mutation). In embodiments, the ADC compounds described herein comprise an anti-B7-H3 antibody comprising the double mutation in the Fc region, wherein the mutations are L234A and L235A (LALA mutations). In embodiments, the anti-B7-H3 antibody (VA clone) includes the mutations L234A and L235A in the Fc region (VA LALA).
• the anti-B7-H3 antibody provided herein comprises a cysteine.
• the anti-B7-H3 antibody is bound to a drug, via linker, through the sulfur of a cysteine residue.
• the anti-B7-H3 antibody is bound to a drug, via linker, through the sulfur of two cysteine residues.
• the anti-B7-H3 antibody provided herein comprises a lysine.
• the anti-B7-H3 antibody is bound to a drug, via linker, through the amine of a lysine residue.
• the anti-B7-H3 antibody is bound to a drug, via linker, through the amine of one or two lysine residues.
• the ADC provided herein comprises an anti-B7-H3 antibody comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a light chain complementarity determining region 1 (CDR1) a light chain CDR2 and a light chain CDR3, and the heavy chain variable region comprises a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3.
• the light chain variable region comprises a light chain complementarity determining region 1 (CDR1) a light chain CDR2 and a light chain CDR3
• the heavy chain variable region comprises a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3.
• the ADC provided herein comprises an anti-B7-H3 antibody (e.g., VA or VA LALA) comprising at least one, two, three, four, five, or six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• VL CDR1 comprising the sequence of SEQ ID NO: 12
• VL CDR2 comprising the sequence of SEQ ID NO: 13
• VL CDR3 comprising the sequence of SEQ ID NO: 14
• VH CDR1 comprising the sequence of SEQ ID NO: 15
• VH CDR2 comprising the sequence of S
• the ADC comprises an anti-B7-H3 antibody comprising at least one CDR selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody comprising at least two CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody comprising at least three CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises anti-B7-H3 antibody comprising at least four CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody comprising at least five CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody comprising at least six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody (e.g., VA or VA LALA) comprising one CDR selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• VA or VA LALA comprising one CDR selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2
• the ADC comprises an anti-B7-H3 antibody comprising two CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody comprising three CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody comprising four CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody comprising five CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (e) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody comprising six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 13; (c) VL CDR3 comprising the sequence of SEQ ID NO: 14; (d) VH CDR1 comprising the sequence of SEQ ID NO: 15; (c) VH CDR2 comprising the sequence of SEQ ID NO: 16; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the anti-B7-H3 antibody (e.g., VA or VA LALA) comprises a VL CDR1 comprising the sequence of SEQ ID NO: 12, a VL CDR2 comprising the sequence of SEQ ID NO: 13, a VL CDR3 comprising the sequence of SEQ ID NO: 14, a VH CDR1 comprising the sequence of SEQ ID NO: 15, a VH CDR2 comprising the sequence of SEQ ID NO: 16, and a VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the anti-B7-H3 antibody comprises a VL CDR1 comprising the sequence of SEQ ID NO: 12.
• the anti-B7-H3 antibody comprises a VL CDR2 comprising the sequence of SEQ ID NO: 13. In embodiments, the anti-B7-H3 antibody comprises a VL CDR3 comprising the sequence of SEQ ID NO: 14. In embodiments, the anti-B7-H3 antibody comprises a VH CDR1 comprising the sequence of SEQ ID NO: 15. In embodiments, the anti-B7-H3 antibody comprises a VH CDR2 comprising the sequence of SEQ ID NO: 16. In embodiments, the anti- B7-H3 antibody comprises and a VH CDR3 comprising the sequence of SEQ ID NO: 17.
• the ADC comprises an anti-B7-H3 antibody (e.g., VA or VA LALA) comprising the light chain CDR1 has the amino acid sequence of SEQ ID NO: 12, the light chain CDR2 has the amino acid sequence of SEQ ID NO: 13, the light chain CDR3 has the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR1 has the amino acid sequence of SEQ ID NO: 15, the heavy chain CDR2 has the amino acid sequence of SEQ ID NO: 16, and the heavy chain CDR3 has the amino acid sequence of SEQ ID NO: 17.
• VA or VA LALA anti-B7-H3 antibody
• the anti-B7-H3 antibody (e.g., VA or VA LALA) comprises a VL having a sequence with at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 20.
• the anti-B7-H3 antibody (e.g., VA or VA LALA) comprises a VL having the sequence of SEQ ID NO: 20.
• a VL sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 20 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-B7-H3 antibody comprising that sequence retains the ability to bind to B7-H3.
• a total of 1 to 10 amino acids have been substituted, inserted and/or deleted in SEQ ID NO: 20.
• a total of 1 to 5 amino acids have been substituted, inserted and/or deleted in SEQ ID NO: 20.
• substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., in the FRs).
• the anti-B7-H3 antibody comprises the VL sequence of SEQ ID NO:
• the anti-B7-H3 antibody (e.g., VA or VA LALA) comprises a VH having a sequence with at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 21.
• the anti-B7-H3 antibody (e.g., VA or VA LALA) comprises a VH having the sequence of SEQ ID NO: 21.
• a VH sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 21 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-B7-H3 antibody comprising that sequence retains the ability to bind to B7-H3.
• a total of 1 to 10 amino acids have been substituted, inserted and/or deleted in SEQ ID NO: 21.
• a total of 1 to 5 amino acids have been substituted, inserted and/or deleted in SEQ ID NO: 21.
• substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., in the FRs).
• the anti-B7-H3 antibody comprises the VH sequence of SEQ ID NO:
• the anti-B7-H3 antibody is an IgG antibody. In embodiments, the anti-B7-H3 antibody is an IgGl, IgG2, IgG3 or IgG4 antibody. In embodiments, the anti-B7-H3 antibody is an IgGl or IgG4 antibody. In embodiments, the anti-B7-H3 antibody is an IgGl antibody.
• the anti- RORl antibody (RO6A-a7gm) includes the mutations L234A and L235A in the Fc region (RO6A-a7gm LALA).
• the anti-RORl antibody provided herein comprises a cysteine.
• the anti-RORl antibody is bound to a drug, via linker, through the sulfur of a cysteine residue.
• the anti-RORl antibody is bound to a drug, via linker, through the sulfur of two cysteine residues.
• the ADC comprises an anti-RORl antibody comprising at least three CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 22; (b) VL CDR2 comprising the sequence of SEQ ID NO: 23; (c) VL CDR3 comprising the sequence of SEQ ID NO: 24; (d) VH CDR1 comprising the sequence of SEQ ID NO: 25; (e) VH CDR2 comprising the sequence of SEQ ID NO: 26; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 27.
• the ADC comprises an anti-RORl antibody comprising at least five CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 22; (b) VL CDR2 comprising the sequence of SEQ ID NO: 23; (c) VL CDR3 comprising the sequence of SEQ ID NO: 24; (d) VH CDR1 comprising the sequence of SEQ ID NO: 25; (e) VH CDR2 comprising the sequence of SEQ ID NO: 26; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 27.
• the ADC comprises an anti-RORl antibody comprising at least six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 22; (b) VL CDR2 comprising the sequence of SEQ ID NO: 23; (c) VL CDR3 comprising the sequence of SEQ ID NO: 24; (d) VH CDR1 comprising the sequence of SEQ ID NO: 25; (e) VH CDR2 comprising the sequence of SEQ ID NO: 26; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 27.
• the ADC provided herein comprises an anti-RORl antibody (e.g., RO6A-a7gm or RO6A-a7gm LALA) comprising at least one, two, three, four, five, or six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 32; (b) VL CDR2 comprising the sequence of SEQ ID NO: 33; (c) VL CDR3 comprising the sequence of SEQ ID NO: 34; (d) VH CDR1 comprising the sequence of SEQ ID NO: 35; (e) VH CDR2 comprising the sequence of SEQ ID NO: 36; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 37.
• an anti-RORl antibody e.g., RO6A-a7gm or RO6A-a7gm LALA
• the ADC comprises an anti-RORl antibody comprising at least one CDR selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 32; (b) VL CDR2 comprising the sequence of SEQ ID NO: 33; (c) VL CDR3 comprising the sequence of SEQ ID NO: 34; (d) VH CDR1 comprising the sequence of SEQ ID NO: 35; (e) VH CDR2 comprising the sequence of SEQ ID NO: 36; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 37.
• the ADC comprises an anti-RORl antibody comprising at least three CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 32; (b) VL CDR2 comprising the sequence of SEQ ID NO: 33; (c) VL CDR3 comprising the sequence of SEQ ID NO: 34; (d) VH CDR1 comprising the sequence of SEQ ID NO: 35; (e) VH CDR2 comprising the sequence of SEQ ID NO: 36; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 37.
• the ADC comprises an anti-RORl antibody comprising at least six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 32; (b) VL CDR2 comprising the sequence of SEQ ID NO: 33; (c) VL CDR3 comprising the sequence of SEQ ID NO: 34; (d) VH CDR1 comprising the sequence of SEQ ID NO: 35; (e) VH CDR2 comprising the sequence of SEQ ID NO: 36; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 37.
• the ADC comprises an anti-RORl antibody (e.g., R06D8-slO or RO6D8-S10 LALA) comprising one CDR selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 22; (b) VL CDR2 comprising the sequence of SEQ ID NO: 23; (c) VL CDR3 comprising the sequence of SEQ ID NO: 24; (d) VH CDR1 comprising the sequence of SEQ ID NO: 25; (e) VH CDR2 comprising the sequence of SEQ ID NO: 26; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 27.
• R06D8-slO or RO6D8-S10 LALA comprising one CDR selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 22; (b) VL CDR2 comprising the sequence of SEQ ID NO: 23; (c) VL CDR3 comprising the sequence of SEQ ID NO: 24; (d)
• the ADC comprises an anti- R0R1 antibody comprising six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 22; (b) VL CDR2 comprising the sequence of SEQ ID NO: 23; (c) VL CDR3 comprising the sequence of SEQ ID NO: 24; (d) VH CDR1 comprising the sequence of SEQ ID NO: 25; (e) VH CDR2 comprising the sequence of SEQ ID NO: 26; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 27.
• the ADC comprises an anti-RORl antibody (e.g., RO6A-a7gm or RO6A-a7gm LALA) comprising one CDR selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 32; (b) VL CDR2 comprising the sequence of SEQ ID NO: 33; (c) VL CDR3 comprising the sequence of SEQ ID NO: 34; (d) VH CDR1 comprising the sequence of SEQ ID NO: 35; (e) VH CDR2 comprising the sequence of SEQ ID NO: 36; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 37.
• an anti-RORl antibody e.g., RO6A-a7gm or RO6A-a7gm LALA
• the ADC comprises an anti- ROR1 antibody comprising three CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 32; (b) VL CDR2 comprising the sequence of SEQ ID NO: 33; (c) VL CDR3 comprising the sequence of SEQ ID NO: 34; (d) VH CDR1 comprising the sequence of SEQ ID NO: 35; (e) VH CDR2 comprising the sequence of SEQ ID NO: 36; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 37.
• the ADC comprises an anti- ROR1 antibody comprising six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 32; (b) VL CDR2 comprising the sequence of SEQ ID NO: 33; (c) VL CDR3 comprising the sequence of SEQ ID NO: 34; (d) VH CDR1 comprising the sequence of SEQ ID NO: 35; (e) VH CDR2 comprising the sequence of SEQ ID NO: 36; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 37.
• a humanized anti-RORl antibody (e.g., RO6D8-S10 or RO6D8- slO LALA) comprises (a) a VL CDR1 comprising the sequence of SEQ ID NO: 22; (b) a VL CDR2 comprising the sequence of SEQ ID NO: 23; (c) a VL CDR3 comprising the sequence of SEQ ID NO: 24; (d) a VH CDR1 comprising the sequence of SEQ ID NO: 25; (e) a VH CDR2 comprising the sequence of SEQ ID NO: 26; and (f) a VH CDR3 comprising the sequence of SEQ ID NO: 27.
• a VL CDR1 comprising the sequence of SEQ ID NO: 22
• a VL CDR2 comprising the sequence of SEQ ID NO: 23
• a VL CDR3 comprising the sequence of SEQ ID NO: 24
• a VH CDR1 comprising the sequence of SEQ ID NO: 25
• a humanized anti-RORl antibody (e.g., R06D8-slO or RO6D8-S10 LALA) comprises (a) a VL CDR1 comprising the sequence of SEQ ID NO: 32; (b) a VL CDR2 comprising the sequence of SEQ ID NO: 33; (c) a VL CDR3 comprising the sequence of SEQ ID NO: 34; (d) a VH CDR1 comprising the sequence of SEQ ID NO: 35; (e) a VH CDR2 comprising the sequence of SEQ ID NO: 36; and (f) a VH CDR3 comprising the sequence of SEQ ID NO: 37.
• R06D8-slO or RO6D8-S10 LALA comprises (a) a VL CDR1 comprising the sequence of SEQ ID NO: 32; (b) a VL CDR2 comprising the sequence of SEQ ID NO: 33; (c) a VL CDR3 comprising the sequence of SEQ ID NO:
• the ADC comprises an antibody that binds to Trop-2.
• Trop-2 has been reported to be upregulated, for example, in carcinomas independent of baseline levels of Trop-2 expression.
• the ADC compounds described herein comprise an anti- Trop-2 antibody.
• the Trop-2 compounds described herein comprise an anti- Trop-2 antibody comprising one or more mutation(s) in the Fc region, wherein the mutation(s) can include the mutation L234A and/or L235A (die substitution of the residue at position 234 in the EU index set forth in Kabat and/or substitution of the residue at position 235 in the EU index set forth in Kabat).
• the ADC compounds described herein comprise an anti- Trop-2 antibody comprising a mutation in the Fc region, wherein the mutation is L234A (LA mutation). In embodiments, the ADC compounds described herein comprise an anti-Trop-2 antibody comprising a mutation in the Fc region, wherein the mutation is L235A (LA mutation). In embodiments, the ADC compounds described herein comprise an anti-Trop-2 antibody comprising the double mutation in the Fc region, wherein the mutations are L234A and L235A (LALA mutations). In embodiments, the anti-Trop-2 antibody (s27 clone) includes the mutations L234A and L235A in the Fc region (s27 LALA).
• the anti-Trop-2 antibody provided herein comprises a cysteine.
• the anti-Trop-2 antibody is bound to a drug, via linker, through the sulfur of a cysteine residue.
• the anti-Trop-2 antibody is bound to a drug, via linker, through the sulfur of two cysteine residues.
• the anti-Trop-2 antibody provided herein comprises a lysine.
• the anti-Trop-2 antibody is bound to a drug, via linker, through the amine of a lysine residue.
• the anti-Trop-2 antibody is bound to a drug, via linker, through the amine of one or two lysine residues.
• the ADC provided herein comprises an anti-Trop-2 antibody comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a light chain complementarity determining region 1 (CDR1) a light chain CDR2 and a light chain CDR3, and the heavy chain variable region comprises a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3.
• the light chain variable region comprises a light chain complementarity determining region 1 (CDR1) a light chain CDR2 and a light chain CDR3
• the heavy chain variable region comprises a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3.
• the ADC provided herein comprises an anti-Trop-2 antibody (e.g., s27 or s27 LALA) comprising at least one, two, three, four, five, or six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDR1 comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• an anti-Trop-2 antibody e.g., s27 or s27 LALA
• the ADC comprises an anti-Trop-2 antibody comprising at least one CDR selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDR1 comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• VL CDR1 comprising the sequence of SEQ ID NO: 42
• VL CDR2 comprising the sequence of SEQ ID NO: 43
• VL CDR3 comprising the sequence of SEQ ID NO: 44
• VH CDR1 comprising the sequence of SEQ ID NO: 45
• VH CDR2 comprising the sequence of SEQ ID NO: 46
• VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises an anti-Trop-2 antibody comprising at least two CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDR1 comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises anti-Trop-2 antibody comprising at least four CDRs selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDRI comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises an anti-Trop-2 antibody comprising at least five CDRs selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDRI comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• VL CDRI comprising the sequence of SEQ ID NO: 42
• VL CDR2 comprising the sequence of SEQ ID NO: 43
• VL CDR3 comprising the sequence of SEQ ID NO: 44
• VH CDRI comprising the sequence of SEQ ID NO: 45
• VH CDR2 comprising the sequence of SEQ ID NO: 46
• VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises an anti-Trop-2 antibody comprising at least six CDRs selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDRI comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• VL CDRI comprising the sequence of SEQ ID NO: 42
• VL CDR2 comprising the sequence of SEQ ID NO: 43
• VL CDR3 comprising the sequence of SEQ ID NO: 44
• VH CDRI comprising the sequence of SEQ ID NO: 45
• VH CDR2 comprising the sequence of SEQ ID NO: 46
• VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises an anti-Trop-2 antibody (e.g., s27 or s27 LALA) comprising one CDR selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDRI comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• an anti-Trop-2 antibody e.g., s27 or s27 LALA
• the ADC comprises an anti-Trop-2 antibody comprising two CDRs selected from (a) VL CDRI comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDRI comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• VL CDRI comprising the sequence of SEQ ID NO: 42
• VL CDR2 comprising the sequence of SEQ ID NO: 43
• VL CDR3 comprising the sequence of SEQ ID NO: 44
• VH CDRI comprising the sequence of SEQ ID NO: 45
• VH CDR2 comprising the sequence of SEQ ID NO: 46
• VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises an anti-Trop-2 antibody comprising three CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDR1 comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises an anti-Trop-2 antibody comprising four CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDR1 comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises an anti-Trop-2 antibody comprising five CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 12; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDR1 comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises an anti-Trop-2 antibody comprising six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 42; (b) VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) VH CDR1 comprising the sequence of SEQ ID NO: 45; (e) VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the anti-Trop-2 antibody (e.g., s27 or s27 LALA) comprises a VL CDR1 comprising the sequence of SEQ ID NO: 42, a VL CDR2 comprising the sequence of SEQ ID NO: 43, a VL CDR3 comprising the sequence of SEQ ID NO: 44, a VH CDR1 comprising the sequence of SEQ ID NO: 45, a VH CDR2 comprising the sequence of SEQ ID NO: 46, and a VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the anti-Trop-2 antibody comprises a VL CDR1 comprising the sequence of SEQ ID NO: 42.
• the anti-Trop-2 antibody comprises a VL CDR2 comprising the sequence of SEQ ID NO: 43. In embodiments, the anti-Trop-2 antibody comprises a VL CDR3 comprising the sequence of SEQ ID NO: 44. In embodiments, the anti-Trop-2 antibody comprises a VH CDR1 comprising the sequence of SEQ ID NO: 45. In embodiments, the anti-Trop-2 antibody comprises a VH CDR2 comprising the sequence of SEQ ID NO: 46. In embodiments, the anti- Trop-2 antibody comprises and a VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the ADC comprises an anti-Trop-2 antibody (e.g., s27 or s27 LALA) comprising the light chain CDR1 has the amino acid sequence of SEQ ID NO:42, the light chain CDR2 has the amino acid sequence of SEQ ID NO:43, the light chain CDR3 has the amino acid sequence of SEQ ID NO:44, the heavy chain CDR1 has the amino acid sequence of SEQ ID NO:45, the heavy chain CDR2 has the amino acid sequence of SEQ ID NO:46, and the heavy chain CDR3 has the amino acid sequence of SEQ ID NO:47.
• an anti-Trop-2 antibody e.g., s27 or s27 LALA
• the anti-Trop-2 antibody (e.g., s27 or s27 LALA) comprises a VL having a sequence with at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 50.
• the anti-Trop-2 antibody (e.g., s27 or s27 LALA) comprises a VL having the sequence of SEQ ID NO: 50.
• a VL sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 50 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-Trop-2 antibody comprising that sequence retains the ability to bind to Trop-2.
• substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., in the FRs).
• the anti-Trop-2 antibody comprises the VL sequence of SEQ ID NO:
• the anti-Trop-2 antibody (e.g., s27 or s27 LALA) comprises a VH having a sequence with at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 51.
• the anti-Trop-2 antibody (e.g., s27 or s27 LALA) comprises a VH having the sequence of SEQ ID NO: 51.
• a VH sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 51 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-Trop-2 antibody comprising that sequence retains the ability to bind to Trop-2.
• substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., in the FRs).
• the anti-Trop-2 antibody comprises the VH sequence of SEQ ID NO:
• the anti-Trop-2 antibody is an IgG antibody. In embodiments, the anti-Trop-2 antibody is an IgGl, IgG2, IgG3 or IgG4 antibody. In embodiments, the anti-Trop-2 antibody is an IgGl or IgG4 antibody. In embodiments, the anti-Trop-2 antibody is an IgGl antibody.
• an anti-Trop-2 antibody binds a human Trop-2.
• the human Trop-2 has the amino acid sequence of SEQ ID NO: 73.
• an anti-Trop-2 antibody is humanized.
• an anti-Trop-2 antibody comprises CDRs as in any of the above embodiments, and further comprises a human acceptor framework, e.g., a human immunoglobulin framework or a human consensus framework.
• a humanized anti-Trop-2 antibody (e.g., s27 or s27 LALA) comprises (a) a VL CDR1 comprising the sequence of SEQ ID NO: 42; (b) a VL CDR2 comprising the sequence of SEQ ID NO: 43; (c) a VL CDR3 comprising the sequence of SEQ ID NO: 44; (d) a VH CDR1 comprising the sequence of SEQ ID NO: 45; (e) a VH CDR2 comprising the sequence of SEQ ID NO: 46; and (f) a VH CDR3 comprising the sequence of SEQ ID NO: 47.
• the anti-Trop-2 antibody is a monoclonal antibody, including a chimeric, humanized, or human antibody.
• an anti-Trop-2 antibody is an antibody fragment, e.g., a Fv, Fab, Fab’, scFv, diabody, or F(ab’)2 fragment.
• the antibody is a substantially full-length antibody, e.g., an IgGl antibody or other antibody class or isotype as defined herein.
• the ADC comprises an antibody that binds to BCMA.
• BCMA has been reported to be upregulated, for example, in multiple myeloma independent of baseline levels of BCMA expression.
• the ADC compounds described herein comprise an anti-BCMA antibody.
• the BCMA compounds described herein comprise an anti-BCMA antibody comprising one or more mutation(s) in the Fc region, wherein the mutation(s) can include the mutation L234A and/or L235A (the substitution of the residue at position 234 in the EU index set forth in Rabat and/or substitution of the residue at position 235 in the EU index set forth in Rabat).
• the ADC compounds described herein comprise an anti-BCMA antibody comprising a mutation in the Fc region, wherein the mutation is L234A (LA mutation). In embodiments, the ADC compounds described herein comprise an anti- antibody comprising a mutation in the Fc region, wherein the mutation is L235A (LA mutation). In embodiments, the ADC compounds described herein comprise an anti-BCMA antibody comprising the double mutation in the Fc region, wherein the mutations are L234A and L235A (LALA mutations). In embodiments, the anti-BCMA antibody (BCA7-2C5 clone) includes the mutations L234A and L235A in the Fc region (BCA7-2C5 LALA).
• the anti-BCMA antibody provided herein comprises a cysteine.
• the anti-BCMA antibody is bound to a drug, via linker, through the sulfur of a cysteine residue.
• the anti-BCMA antibody is bound to a drug, via linker, through the sulfur of two cysteine residues.
• the anti-BCMA antibody provided herein comprises a lysine.
• the anti-BCMA antibody is bound to a drug, via linker, through the amine of a lysine residue.
• the anti-BCMA antibody is bound to a drug, via linker, through the amine of one or two lysine residues.
• the ADC provided herein comprises an anti-BCMA antibody comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a light chain complementarity determining region 1 (CDR1) a light chain CDR2 and a light chain CDR3, and the heavy chain variable region comprises a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3.
• the light chain variable region comprises a light chain complementarity determining region 1 (CDR1) a light chain CDR2 and a light chain CDR3
• the heavy chain variable region comprises a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3.
• the ADC provided herein comprises an anti-BCMA antibody (e.g., BCA7-2C5 or BCA7-2C5 LALA) comprising at least one, two, three, four, five, or six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 52; (b) VL CDR2 comprising the sequence of SEQ ID NO: 53; (c) VL CDR3 comprising the sequence of SEQ ID NO: 54; (d) VH CDR1 comprising the sequence of SEQ ID NO: 55; (e) VH CDR2 comprising the sequence of SEQ ID NO: 56; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 57.
• an anti-BCMA antibody e.g., BCA7-2C5 or BCA7-2C5 LALA
• the ADC comprises anti-BCMA antibody comprising at least four CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 52; (b) VL CDR2 comprising the sequence of SEQ ID NO: 53; (c) VL CDR3 comprising the sequence of SEQ ID NO: 54; (d) VH CDR1 comprising the sequence of SEQ ID NO: 55; (e) VH CDR2 comprising the sequence of SEQ ID NO: 56; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 57.
• the ADC comprises an anti- BCMA antibody comprising six CDRs selected from (a) VL CDR1 comprising the sequence of SEQ ID NO: 52; (b) VL CDR2 comprising the sequence of SEQ ID NO: 53; (c) VL CDR3 comprising the sequence of SEQ ID NO: 54; (d) VH CDR1 comprising the sequence of SEQ ID NO: 55; (e) VH CDR2 comprising the sequence of SEQ ID NO: 56; and (f) VH CDR3 comprising the sequence of SEQ ID NO: 57.
• the anti-BCMA antibody (e.g., BCA7-2C5 or BCA7-2C5 LALA) comprises a VL CDR1 comprising the sequence of SEQ ID NO: 52, a VL CDR2 comprising the sequence of SEQ ID NO: 53, a VL CDR3 comprising the sequence of SEQ ID NO: 54, a VH CDR1 comprising the sequence of SEQ ID NO: 55, a VH CDR2 comprising the sequence of SEQ ID NO: 56, and a VH CDR3 comprising the sequence of SEQ ID NO: 57.
• the anti-BCMA antibody comprises a VL CDR1 comprising the sequence of SEQ ID NO: 52.
• the anti-BCMA antibody comprises a VL CDR2 comprising the sequence of SEQ ID NO: 53. In embodiments, the anti-BCMA antibody comprises a VL CDR3 comprising the sequence of SEQ ID NO: 54. In embodiments, the anti-BCMA antibody comprises a VH CDR1 comprising the sequence of SEQ ID NO: 55. In embodiments, the anti-BCMA antibody comprises a VH CDR2 comprising the sequence of SEQ ID NO: 56. In embodiments, the anti- BCMA antibody comprises and a VH CDR3 comprising the sequence of SEQ ID NO: 57.
• an anti-BCMA antibody binds a human BCMA.
• the human BCMA has the amino acid sequence of SEQ ID NO: 74.
• an anti-BCMA antibody “binds with an affinity of’ ⁇ 10 nM, or ⁇ 5 nM, or ⁇ 4 nM, or ⁇ 3 nM, or ⁇ 2 nM, can be determined using standard Scatchard analysis utilizing a non-linear curve fitting program (see, for example, Munson et al., Anal Biochem, 107: 220-239, 1980).
• Single-domain antibodies are antibody fragments comprising all or a portion of the heavy chain variable domain or all or a portion of the light chain variable domain of an antibody.
• a single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Patent No. 6,248,516 Bl).
• a chimeric antibody is a humanized antibody.
• a non- human antibody is humanized to reduce immunogenicity to humans, while retaining the specificity and affinity of the parental non-human antibody.
• a humanized antibody comprises one or more variable domains in which HVRs, e.g., CDRs, (or portions thereof) are derived from a non-human antibody, and FRs (or portions thereof) are derived from human antibody sequences.
• HVRs e.g., CDRs, (or portions thereof) are derived from a non-human antibody
• FRs or portions thereof
• a humanized antibody optionally will also comprise at least a portion of a human constant region.
• Human framework regions that may be used for humanization include but are not limited to: framework regions selected using the “best-fit” method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (1993)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci.
• the anti-CD25 antibody, anti-B7-H3 antibody, anti-RORl antibody, anti-Trop-2 antibody, and anti-BCMA antibody provided herein is a human antibody.
• Human antibodies can be produced using various techniques known in the art. Human antibodies are described generally in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20:450-459 (2008). [00421] Human antibodies may be prepared by administering an immunogen to a transgenic animal that has been modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigenic challenge.
• Such animals typically contain all or a portion of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci, or which are present extrachromosomally or integrated randomly into the animal’s chromosomes.
• the endogenous immunoglobulin loci have generally been inactivated.
• Human antibodies may also be generated by isolating Fv clone variable domain sequences selected from human-derived phage display libraries. Such variable domain sequences may then be combined with a desired human constant domain. Techniques for selecting human antibodies from antibody libraries are described below. vi. Multispecific Antibodies
• the anti-CD25 antibody, anti-B7-H3 antibody, anti-RORl antibody, anti-Trop-2 antibody, and anti-BCMA antibody provided herein is a multispecific antibody, e.g., a bispecific antibody.
• Multispecific antibodies are monoclonal antibodies that have binding specificities for at least two different sites. In embodiments, one of the binding specificities is for CD25, B7-H3, ROR1, Trop-2, or BCMA, and the other is for any other antigen.
• bispecific antibodies may bind to two different epitopes of CD25, B7-H3, ROR1, Trop-2, or BCMA, respectively. Bispecific antibodies may also be used to localize cytotoxic agents to cells which express CD25, B7-H3, ROR1 , Trop-2, or BCMA, respectively.
• Bispecific antibodies can be prepared as full-length antibodies or antibody fragments.
• Engineered antibodies with three or more functional antigen binding sites are also included herein (see, e.g., US 2006/0025576A1).
• Such modifications include, for example, deletions from, and/or insertions into and/or substitutions of residues within the amino acid sequences of the antibody. Any combination of deletion, insertion, and substitution can be made to arrive at the final construct, provided that the final construct possesses the desired characteristics, e.g., antigen-binding. a ) Substitution, Insertion, and Deletion Variants
• the anti-CD25 antibody, anti-B7-H3 antibody, anti-RORl antibody, anti-Trop-2 antibody, and anti-BCMA antibody provided herein has one or more amino acid substitutions.
• Sites of interest for substitutional mutagenesis include the HVRs and FRs.
• Conservative substitutions are shown in Table 1 under the heading of “preferred substitutions.” More substantial changes are provided in Table 1 under the heading of “exemplary substitutions,” and as further described below in reference to amino acid side chain classes.
• Amino acid substitutions may be introduced into an antibody of interest and the products screened for a desired activity, e.g., retained/improved antigen binding, decreased immunogenicity, or improved ADCC or CDC.
• substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (e.g. a humanized or human antibody).
• a parent antibody e.g. a humanized or human antibody
• the resulting variant(s) selected for further study will have modifications (e.g., improvements) in biological properties (e.g., increased affinity, reduced immunogenicity) relative to the parent antibody and/or will have substantially retained certain biological properties of the parent antibody.
• An exemplary substitutional variant is an affinity matured antibody, which may be conveniently generated, e.g., using phage display-based affinity maturation techniques such as those described herein. Briefly, one or more HVR residues are mutated and the variant antibodies displayed on phage and screened for a particular biological activity (e.g. binding affinity).
• Alterations may be made in HVRs, e.g., to improve antibody affinity. Such alterations may be made in HVR “hotspots,” i.e., residues encoded by codons that undergo mutation at high frequency during the somatic maturation process (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and/or SDRs (a-CDRs), with the resulting variant VH or VL being tested for binding affinity.
• HVR “hotspots” i.e., residues encoded by codons that undergo mutation at high frequency during the somatic maturation process (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and/or SDRs (a-CDRs), with the resulting variant VH or VL being tested for binding affinity.
• Affinity maturation by constructing and reselecting from secondary libraries has been described, e.g., in Hoogenboom
• affinity maturation diversity is introduced into the variable genes chosen for maturation by any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-directed mutagenesis).
• a secondary library is then created. The library is then screened to identify any antibody variants with the desired affinity.
• Another method to introduce diversity involves HVR-directed approaches, in which several HVR residues (e.g., 4-6 residues at a time) are randomized. HVR residues involved in antigen binding may be specifically identified, e.g., using alanine scanning mutagenesis or modeling. CDR-H3 and CDR-L3 in particular are often targeted.
• a useful method for identification of residues or regions of an antibody that may be targeted for mutagenesis is called “alanine scanning mutagenesis” as described by Cunningham and Wells (1989) Science, 244:1081-1085.
• a residue or group of target residues e.g., charged residues such as arg, asp, his, lys, and glu
• a neutral or negatively charged amino acid e.g., alanine or polyalanine
• a crystal structure of an antigen- antibody complex is used to identify contact points between the antibody and antigen. Such contact residues and neighboring residues may be targeted or eliminated as candidates for substitution. Variants may be screened to determine whether they contain the desired properties.
• an anti-CD25 antibody, anti-B7-H3 antibody, anti-RORl antibody, anti-Trop-2 antibody, and anti-BCMA antibody provided herein is altered to increase or decrease the extent to which the antibody is glycosylated.
• Addition or deletion of glycosylation sites to an antibody may be conveniently accomplished by altering the amino acid sequence such that one or more glycosylation sites is created or removed.
• the carbohydrate attached thereto may be altered.
• Native antibodies produced by mammalian cells typically comprise a branched, biantennary oligosaccharide that is generally attached by an N-linkage to Asn297 of the CH2 domain of the Fc region. See, e.g., Wright et al. T1BTECH 15:26-32 (1997).
• the oligosaccharide may include various carbohydrates, e.g., mannose, N-acetyl glucosamine (GlcNAc), galactose, and sialic acid, as well as a fucose attached to a GlcNAc in the “stem” of the biantennary oligo saccharide structure.
• modifications of the oligosaccharide in an antibody may be made in order to create antibody variants with certain improved properties.
• antibody variants having a carbohydrate structure that lacks fucose attached (directly or indirectly) to an Fc region.
• the amount of fucose in such antibody may be from 1% to 80%, from 1% to 65%, from 5% to 65% or from 20% to 40%.
• the amount of fucose is determined by calculating the average amount of fucose within the sugar chain at Asn297, relative to the sum of all glycostructures attached to Asn297 (e. g. complex, hybrid and high mannose structures) as measured by MALDI-TOF mass spectrometry, as described in WO 2008/077546, for example.
• Asn297 refers to the asparagine residue located at about position 297 in the Fc region (Eu numbering of Fc region residues); however, Asn297 may also be located about ⁇ 3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300, due to minor sequence variations in antibodies. Such fucosylation variants may have improved ADCC function. See, e.g., US Patent Publication Nos. US 2003/0157108 (Presta, L.); US 2004/0093621 (Kyowa Hakko Kogyo Co., Ltd).
• Examples of publications related to “defucosylated” or “fucose-deficient” antibody variants include: US 2003/0157108; WO 2000/61739; WO 2001/29246; US 2003/0115614; US 2002/0164328; US 2004/0093621; US 2004/0132140; US 2004/0110704; US 2004/0110282; US 2004/0109865; WO 2003/085119; WO 2003/084570; WO 2005/035586; WO 2005/035778; W02005/053742; W02002/031140; Okazaki et al. J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004).
• Examples of cell lines capable of producing defucosylated antibodies include Lee 13 CHO cells deficient in protein fucosylation (Ripka et al. Arch.
• Antibody variants are further provided with bisected oligosaccharides, e.g., in which a biantennary oligosaccharide attached to the Fc region of the antibody is bisected by GlcNAc. Such antibody variants may have reduced fucosylation and/or improved ADCC function. Examples of such antibody variants are described, e.g., in WO 2003/011878 (Jean-Mairet et al.); US Patent No. 6,602,684 (Umana et al.); and US 2005/0123546 (Umana et al.). Antibody variants with at least one galactose residue in the oligosaccharide attached to the Fc region are also provided.
• Such antibody variants may have improved CDC function. Such antibody variants are described, e.g., in WO 1997/30087 (Patel et al.); WO 1998/58964 (Raju, S.); and WO 1999/22764 (Raju, S.).
• c) Fc Region Variants [00439] In embodiments, one or more amino acid modifications may be introduced into the Fc region of an anti-CD25 antibody, anti-B7-H3 antibody, anti-RORl antibody, anti-Trop-2 antibody, or anti-BCMA antibody provided herein, thereby generating an Fc region variant.
• the Fc region variant may comprise a human Fc region sequence e.g., a human IgGl, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (e.g. a substitution) at one or more amino acid positions.
• a human Fc region sequence e.g., a human IgGl, IgG2, IgG3 or IgG4 Fc region
• an amino acid modification e.g. a substitution
• an antibody variant that possesses some but not all effector functions is contemplated, which make it a desirable candidate for applications in which the half life of the antibody in vivo is important yet certain effector functions (such as complement and ADCC) arc unnecessary or deleterious.
• In vitro and/or in vivo cytotoxicity assays can be conducted to confirm the reduction/depletion of CDC and/or ADCC activities.
• Fc receptor (FcR) binding assays can be conducted to ensure that the antibody lacks FcyR binding (hence likely lacking ADCC activity) but retains FcRn binding ability.
• NK cells express FcyRIII only, whereas monocytes express FcyRI, FcyRII and FcyRIII.
• FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991).
• Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest is described in U.S. Patent No.
• non-radioactive assays methods may be employed (see, for example, ACTITM non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc. Mountain View, CA; and CytoTox 96® nonradioactive cytotoxicity assay (Promega, Madison, WI).
• Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells.
• ADCC activity of the molecule of interest may be assessed in vivo, e.g., in a animal model such as that disclosed in Clynes et al. Proc. Nat’l Acad. Sci. USA 95:652- 656 (1998).
• Clq binding assays may also be carried out to confirm that the antibody is unable to bind Clq and hence lacks CDC activity. See, e.g., Clq and C3c binding ELISA in
• a CDC assay may be performed (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, M.S. et al., Blood 101:1045-1052 (2003); and Cragg, M.S. and M.J. Glennie, Blood 103:2738-2743 (2004)).
• FcRn binding and in vivo clearance/half life determinations can also be performed using methods known in the art (see, e.g., Petkova, S.B. et al., Int’l. Immunol. 18(12): 1759-1769 (2006)).
• Antibodies with reduced effector function include those with substitution of one or more of Fc region residues 238, 265, 269, 270, 297, 327 and 329 (U.S. Patent No. 6,737,056).
• Fc mutants include Fc mutants with substitutions at two or more of amino acid positions 265, 269, 270, 297 and 327, including the so-called “DANA” Fc mutant with substitution of residues 265 and 297 to alanine (US Patent No. 7,332,581).
• Antibodies with increased half-lives and improved binding to the neonatal Fc receptor (FcRn), which is responsible for the transfer of maternal IgGs to the fetus are described in US2005/0014934A1 (Hinton et al.). Those antibodies comprise an Fc region with one or more substitutions therein which improve binding of the Fc region to FcRn.
• Such Fc variants include those with substitutions at one or more of Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434, e.g., substitution of Fc region residue 434 (US Patent No. 7,371,826).
• the antibodies provided of the conjugates described herein comprise one or more amino acid modifications in the Fc region.
• the mutation(s) are at positions 234 and/or 235 in the EU index set forth in Kabat. In embodiments, the mutation is at position 234. In embodiments, the mutation is at position 235.
• the mutations are at positions 234 and 235.
• the Leucine at position 234 is replaced by Alanine (L234A; LA mutation).
• the Leucine at position 235 is replaced by Alanine (L235A; LA mutation).
• the Leucines at position 234 and position 235 are replaced by Alanines (L234AL235A; LALA double mutation). viii. Antibody Derivatives
• a monoclonal antibody such as an anti-CD25 antibody, anti-B7-H3 antibody, anti-RORl antibody, anti-Trop-2 antibody, or anti-BCMA antibody, provided herein may be further modified (e.g., derivatized) to contain one or more additional non-proteinaceous moieties that are known in the art and readily available.
• the moieties suitable for derivatization of the antibody include but are not limited to water soluble polymers.
• Non-limiting examples of water soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol/propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly-l ,3-dioxolane, poly-l ,3,6-trioxane, ethylene/maleic anhydride copolymer, polyaminoacids (cither homopolymers or random copolymers), and dextran or poly(n- vinyl pyrrolidone)poly ethylene glycol, propropylene glycol homopolymers, prolypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof.
• PEG polyethylene glycol
• copolymers of ethylene glycol/propylene glycol carboxymethylcellulose
• dextran polyvinyl alcohol
• Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water.
• the polymer may be of any molecular weight, and may be branched or unbranched.
• the number of polymers attached to the antibody may vary, and if more than one polymer are attached, they can be the same or different molecules. In general, the number and/or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular properties or functions of the antibody to be improved, whether the antibody derivative will be used in a therapy under defined conditions, etc. ix. Recombinant Methods and Compositions
• Antibodies may be produced using recombinant methods and compositions, e.g., as described in U.S. Patent No. 4,816,567.
• suitable host cells include eukaryotic cells, e.g. a Chinese Hamster Ovary (CHO) cell or lymphoid cell (e.g., Y0, NSO, Sp20 cell).
• eukaryotic cells e.g. a Chinese Hamster Ovary (CHO) cell or lymphoid cell (e.g., Y0, NSO, Sp20 cell).
• nucleic acid encoding an antibody is isolated and inserted into one or more vectors for further cloning and/or expression in a host cell.
• nucleic acid may be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the antibody).
• An ADC of formula (I) may be prepared by several routes employing organic chemistry reactions, conditions, and reagents known to those skilled in the art, including: (1) reaction of a nucleophilic group of an antibody with a bivalent linker reagent (L 1 ) to form Ab-L 1 via a covalent bond, followed by reaction with a drug-linker molecule D-L 3 or D-L 3 -L 2 and (2) reaction of a nucleophilic group of a drug moiety D with a bivalent linker reagent (L 3 -L 2 -L 1 or L’-L 1 ) to form D-L '-L 1 or D-L 3 -L 2 -L 1 via a covalent bond, followed by reaction with a nucleophilic group of an antibody or a reduced antibody.
• An ADC of formula (II) may be prepared by several routes employing organic chemistry reactions, conditions, and reagents known to those skilled in the art, including: (1) reaction of a nucleophilic group of an antibody with a bivalent linker reagent (L 1 ) to form Ab-L 1 via a covalent bond, followed by reaction with a drug-linker molecule R*-D’ or R*-D’-L 2 and (2) reaction of a nucleophilic group of a drug- linker molecule R -D’ with a bivalent linker reagent (L"-L or L ) to form R -D’-L or R -D’-L"- L 1 via a covalent bond, followed by reaction with a nucleophilic group of an antibody or a reduced antibody.
• An ADC of formula (III) may be prepared by several routes employing organic chemistry reactions, conditions, and reagents known to those skilled in the art, including: (1) reaction of a nucleophilic group of an antibody with a bivalent linker reagent (L 1 ) to form Ab-L 1 via a covalent bond, followed by reaction with a drug moiety D” or drug-linker molecule D”-L 2 ; and (2) reaction of a nucleophilic group of a drug moiety D” with a bivalent linker reagent (L 2 and/or L 1 ) to form D”-L 2 or D”-L 2 -L 1 via a covalent bond, followed by reaction with a nucleophilic group of an antibody or a reduced antibody.
• Several such methods are described by Agarwal et al., (2015), Bioconjugate Chem., 26: 176-192.
• an antibody may be reduced with a reducing agent such as dithiothreitol (DTT) or tricarboxyethylphosphine (TCEP), under partial or total reducing conditions, to generate reactive cysteine thiol groups.
• DTT dithiothreitol
• TCEP tricarboxyethylphosphine
• the inter-chain cysteine residues can then be alkylated for example using maleimide.
• the inter-chain cysteine residues can undergo bridging alkylation for example using bis sulfone linkers or propargyldibromomaleimide followed by Cu-click ligation.
• the antibody can be conjugated through lysine amino acid. Such conjugation can be a one-step conjugation or a two- step conjugation.
• the one-step conjugation entails conjugation of the e-amino group of lysine residue to the drug-linker molecule (D-L 3 -L 2 -L 1 or D-L 3 -L 1 ) containing an amine-reactive group via amide bonds.
• the one-step conjugation entails conjugation of the £-amino group of lysine residue to the drug-linker molecule (R*-D’-L 2 -L 1 or R*-D’-L 1 ) containing an amine-reactive group via amide bonds.
• the one-step conjugation entails conjugation of the £-amino group of lysine residue to the drug-linker molecule (D”-L 2 -L 1 or D”-L 1 ) containing an amine-reactive group via amide bonds.
• the amine-reactive group is an activated ester.
• the antibody can be conjugated via a two-step conjugation.
• the two-step conjugation entails a first step, where a bi-functional reagent containing both amine and thiol reactive functional groups is reacted with the lysine £-amino group(s).
• the drug-linker molecule (D-L 3 -L 2 -L 1 , D-L 3 -L 1 , R*-D’-L 2 -L 1 , R*-D’-L 1 , D”-L 2 -L 1 , or D”-L l ) is conjugated to the thiol reactive group of the bifunctional reagent.
• the drug-linker molecule (D-L 3 -L 2 -L 1 , D-L 3 -L 1 , R*-D’-L 2 -L 1 , R*-D’-L 1 , D”-L 2 -L 1 , or D”-L l ) is conjugated to the thiol reactive group of the bifunctional reagent.
• the first step may involve the functionalization of the antibody with azide followed by a click chemistry reaction with an alkyne modified linker or drug-linker molecule (D-L 3 -L 2 -L 1 , D-L 3 L 1 , R * -D’-L 2 -L 1 , R * -D’-L 1 , D”-L 2 -L 1 , or D”-L'y
• the first step may involve the functionalization of the antibody with an alkyne followed by a click chemistry reaction with an azide modified linker or drug-linker molecule (D-L 3 -L 2 -L 1 , D-L ’-L 1 , R * -D’-L 2 L 1 , R * -D’-L 1 , D”-L 2 -L 1 , or D”-L l ).
• the first step may involve the functionalization of the antibody with an aldehyde followed by a click chemistry reaction with an alkoxyamine or hydrazine modified linker or drug-linker molecule (DL 3 -L 2 - L 1 , D-L 3 -L 1 , R*-D’- lAL 1 , R*-D’-L 1 , D’’-JAL 1 , or D”-L l ).
• the first step may involve the functionalization of the antibody with a tetrazine followed by a click chemistry reaction with a trans-cyclooctene or cyclopropene modified linker or drag-linker molecule (D-L 3 -L 2 -L 1 , D-L 3 - L 1 , R * D’-L 2 -L 1 , R*-D’-L', D”-L 2 -L 1 , or D”-L 1 ).
• a trans-cyclooctene or cyclopropene modified linker or drag-linker molecule D-L 3 -L 2 -L 1 , D-L 3 - L 1 , R * D’-L 2 -L 1 , R*-D’-L', D”-L 2 -L 1 , or D”-L 1 ).
• the first step may involve the functionalization of the antibody with a trans-cyclooctene or cyclopropene followed by a click chemistry reaction with a tetrazine modified linker or drug-linker molecule (D-L 3 -L 2 -L 1 , D-L 3 - L 1 , R * D’-lAL 1 , R*-D’-L', D”-L 2 -L 1 or D”-L 1 ).
• a tetrazine modified linker or drug-linker molecule D-L 3 -L 2 -L 1 , D-L 3 - L 1 , R * D’-lAL 1 , R*-D’-L', D”-L 2 -L 1 or D”-L 1 .
• an ADC of formula (I), formula (II), or formula (III) can be prepared by reacting a monoclonal antibody (Ab), said monoclonal antibody comprising an Fc variant, said Fc variant comprising two amino acid substitutions L234A and L235A, with a molecule of formula (P-I), formula (P-II), or formula (P-III):
• B is a reactive moiety capable of forming a bond with the monoclonal antibody
• L 2 is a bond, -C(O)-, -NH-, Amino Acid Unit, -(CH2CH O) n -, -(CH2)n-, -O-, -(4-aminobenzyloxycarbonyl)-, -(C(O)CH2CH2NH)-, -(C(O)N(R 2 )CH2CH2N(R 5 ))-, or any combination thereof; wherein n is an integer from 1 to 24; each R 2 and R 5 is independently H or substituted or unsubstituted alkyl;
• L 3 is a substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted hctcroarylcnc; or L 3 is substituted or unsubstituted -OCH2-(hctcrocycloalkylcnc) or substituted or unsubstituted -OCH2-(heteroarylene), wherein L 3 is linked to D through oxygen; or L 3 is substituted or unsubstituted -CH2NCH2-(heteroaryl) or substituted or unsubstituted -CH2NCH2-(heterocycloalkyl), wherein L 3 is linked to D through -CH2-, and through nitrogen to L 2 ;
• R* is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl
• R 1 is H or — C 1 -C 8 alkyl
• R 3 is H, halogen, -CCh, -CBr 3 , -CF3, -CI 3 , -CHCk, -CHBr 2 , -CHF 2 , -CHI 2 , -CH 2 C1, -CH 2 Br, -CH 2 F, -CH 2 I, -CN, -OR 3A , -NR 3A R 3B , -(CH 2 ) V OR 6 , substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl;
• R 4 is H, halogen, -OR 4A , -NR 4A R 4B , substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl; V is N, O, or C;
• Z 1 is a substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocycloalkyl;
• Z 2 is a substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted cycloalkylene, or substituted or unsubstituted heterocycloalkylene;
• R 6 is H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl,
• the monoclonal antibody is modified with a reactive moiety such as an aldehyde, azide, alkyne, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene or cyclopropene.
• a reactive moiety such as an aldehyde, azide, alkyne, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene or cyclopropene.
• the monoclonal antibody is modified with an aldehyde.
• the monoclonal antibody is modified with an azide.
• the monoclonal antibody is modified with a tetrazine.
• the monoclonal antibody is modified with an alkoxyamine.
• the monoclonal antibody is modified with a hydrazine.
• the monoclonal antibody is modified with a trans-cyclooctene.
• the monoclonal antibody is modified with a cyclopropene.
• B is a reactive moiety capable of forming a bond with a monoclonal antibody. In embodiments, B is a reactive moiety capable of forming a bond with a modified monoclonal antibody.
• B is a reactive moiety capable of forming a bond with one or two thiol or amine groups of the monoclonal antibody, or of the modified monoclonal antibody.
• the monoclonal antibody is modified with an azide, aldehyde, alkyne, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene or cyclopropene.
• B is an alkyne, azide, aldehyde, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene, cyclopropene, activated ester, haloacetyl, cycloalkyne, maleimide, or bis- sulfone.
• B is dibromomaleimide.
• B is cyclooctyne.
• the activated ester may be for example pentafluorophenyl ester, tetrafluorophenyl ester, trifluorophenyl ester, difluorophenyl ester, monofluorophenyl or ester, N- hydroxysuccinimide ester.
• the monoclonal antibody is an anti-CD25 antibody, anti-B7-H3 antibody, anti-RORl antibody, anti-Trop-2 antibody, or anti-BCMA antibody.
• the monoclonal antibody is an anti-CD25 antibody. In embodiments, the monoclonal antibody is an anti-B7-H3 antibody. In embodiments, the monoclonal antibody is an anti-RORl antibody. In embodiments, the monoclonal antibody is an anti-Trop-2 antibody. In embodiments, the monoclonal antibody is an anti-BCMA antibody.
• a modified monoclonal antibody is a modified anti-CD25 antibody. In embodiments, a modified monoclonal antibody is a modified anti-B7-H3 antibody. In embodiments, a modified monoclonal antibody is a modified anti-RORl antibody. In embodiments, a modified monoclonal antibody is a modified anti-Trop-2 antibody. In embodiments, a modified monoclonal antibody is a modified anti-BCMA antibody.
• n is an integer from 1 to 24. In embodiments, n is an integer from 1 to 4. In embodiments, n is 1. In embodiments, n is 2. In embodiments, n is 3. In embodiments, n is 4. In embodiments, n is 5. In embodiments, n is 6. In embodiments, n is 7. In embodiments, n is 8. In embodiments, n is 9. In embodiments, n is 10. In embodiments, n is 11. In embodiments, n is 12. In embodiments, n is 13. In embodiments, n is 14. In embodiments, n is 15. In embodiments, n is 16. In embodiments, n is 17. In embodiments, n is 18. In embodiments, n is 19. In embodiments, n is 20. In embodiments, n is 21. In embodiments, n is 22. In embodiments, n is 23. In embodiments, n is 24. [00460] In embodiments, wherein:
• R 1 is H or — C 1 -C 8 alkyl
• R 3 is H, halogen, substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl;
• R 4 is H, halogen, or substituted or unsubstituted alkyl
• V is N
• Z 2 is a substituted or unsubstituted arylene.
• B is . In embodiments, B is . In embodiments, B is embodiments, B is . In embodiments, B is H . In embodiments,
• monoclonal antibodies In embodiments, monoclonal antibodies, modified monoclonal antibodies, or anti-
• CD25 unmodified or modified antibodies undergo conjugation reactions with the following reactive B moieties as follows:
• L 2 is a cleavable or a non-cleavable linker as described in US Patents Nos. US 9,884,127, US 9,981,046, US 9,801,951, US 10,117,944, US 10,590,165, and US 10,590,165, and US Patent publications Nos. US 2017/0340750, and US 2018/0360985, all of which are incorporated herein in their entireties.
• L 2 is a bond, -C(O)-, -NH-, -Vai-, -Phe-, -Lys-, -Gly-, -O- -(4-aminobcnzyloxycarbonyl)-, -(C(O)N(R 2 )CH2CH2N(R 5 ))-, -Ser-, -Thr-, -Ala-, - [3- Ala-, -citrulline- (Cit), -(Chhin-, -(CFhCF Oln-, N-dimethyl lysine, or any combination thereof.
• each R 2 and R 5 is independently H or substituted or unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 5 is independently H. In embodiments, each R 2 and R 5 is independently substituted or unsubstituted alkyl. In embodiments, each R 2 and R 5 is independently substituted or unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• each R 2 and R 5 is independently unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 5 is independently substituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• each R 2 and R 5 is independently H or substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• each R 2 and R 5 is independently substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• each R 2 and R 5 is independently unsubstituted alkyl (e.g., C 1 -C 8 alkyl, Ci-C 6 alkyl, or C1-C4 alkyl).
• each R 2 and R 5 is independently methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, or hexyl. In embodiments, each R 2 and R 5 is independently methyl. In embodiments, each R 2 and R 5 is independently ethyl. In embodiments, each R 2 and R 5 is independently propyl. In embodiments, each R 2 and R 5 is independently butyl. In embodiments, each R 2 and R 5 is independently isopropyl. In embodiments, each R 2 and R 5 is independently isobutyl.
• each R 2 and R 5 is independently tert-butyl. In embodiments, each R 2 and R 5 is independently isopentyl.
• L 2 is a bond, -C(O)-, -NH-, -Vai-, -Phe-, -Lys-, -Gly-, -(4-aminobenzyloxycarbonyl)-, -(C(O)N(CH3)CH2CH2N(CH3))-, -Ser-, -Thr-, -Ala-, -
• L 2 is -C(O)-, -NH-, -Vai-, -Gly-, -Cit-, -Ala-, -O-, -(4- aminobenzyloxycarbonyl)-, -(CH 2 )n- -(CH 2 CH 2 O)n- -(C(O)N(CH3)CH 2 CH2N(CH3))-, N- dimethyl lysine, or any combination thereof.
• L 2 is -C(O)-, -NH-, -Gly-, -(CH 2 ) n -, -(CH 2 CH 2 O)u-, or any combination thereof.
• L 2 is -C(O)-, -NH-, -Vai-, -Cit-, -(CEI 2 CEI 2 O)II- -(4- aminobenzyloxycarbonyl)-, -(CH2)n-, -(C(O)N(CH3)CH2CH2N(CH3))-, N-dimethyl lysine, or any combination thereof.
• L 2 is -C(O)-, -NH-, -Vai-, -(4-aminobenzyloxycarbonyl)-, -Gly-, -citrulline- (-Cit-), -(CH 2 )n-, -(CH2CH2O) n -, N-dimethyl lysine, or any combination thereof.
• L 2 is:
• L 2 is , embodiments, L 2 is
• L 2 is In embodiments, L 2 is
• L 2 is In embodiments, L 2 is 84
• L 2 is a bond. In embodiments, L 2 is -C(O)-. In embodiments, L 2 is -NH-. In embodiments, L 2 is -Vai-. In embodiments, L 2 is -Phe-. In embodiments, L 2 is -Lys-. In embodiments, L 2 is -(4-aminobenzyloxycarbonyl)-. In embodiments, L 2 is -(CH2 1-. In embodiments, L 2 is -(CthCthC ii-. In embodiments, L 2 is -Gly-. In embodiments, L 2 is -Ser-. In embodiments, L 2 is -Thr-.
• L 2 is -Ala-. In embodiments, L 2 is -
• heteroarylene e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene, substituted (e.g.
• heterocycloalkylene e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocycloalkylene, or 5 to 6 membered heterocycloalkylene
• substituted e.g., with a substituent group, a size-limited substituent group or a lower substituent group
• unsubstituted -OCH2-(heteroarylene e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene)
• substituted e.g.
• L 3 is substituted with one or more substituent groups. In embodiments, L 3 is substituted with one or more size-limited substituent groups. In embodiments, L 3 is substituted with one or more lower substituent groups.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocycloalkylene, or 5 to 6 membered heterocycloalkylene), or substituted (e.g.
• L 3 is substituted with one or more substituent groups. In embodiments, L 3 is substituted with one or more size-limited substituent groups. In embodiments, L 3 is substituted with one or more lower substituent groups.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocycloalkylene, or 5 to 6 membered heterocycloalkylene).
• L 3 is substituted with one or more substituent groups.
• L 3 is substituted with one or more size-limited substituent groups.
• L is substituted with one or more lower substituent groups.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocycloalkylene (e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocycloalkylene, or 5 to 6 membered heterocycloalkylene).
• L 3 is unsubstituted heterocycloalkylene (e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocycloalkylene, or 5 to 6 membered heterocycloalkylene).
• L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene).
• heteroarylene e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene.
• L 3 is unsubstituted heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) - OCH2-(heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl)).
• L 3 is unsubstituted -OCH2-(heterocycloalkylene (e.g., 3 to 8 membered heterocycloalkylene, 3 to 6 membered heterocycloalkylene, or 5 to 6 membered heterocycloalkylene)).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -OCH2-(hctcroarylcnc (e.g., 5 to 10 membered hctcroarylcnc, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene)).
• L 3 is unsubstituted -OCH2-(heteroarylene (e.g., 5 to 10 membered heteroarylene, 5 to 9 membered heteroarylene, or 5 to 6 membered heteroarylene)).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -CH2NCH2- (heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl)).
• L 3 is unsubstituted -CH2NCH2- (heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl)).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -CH2NCH2-(heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl)).
• L 3 is unsubstituted -CH2NCH2-(heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl)).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 8 membered heterocycloalkylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) 3 to 8 membered heterocycloalkylene. In embodiments, L 3 is unsubstituted 3 to 8 membered heterocycloalkylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) or unsubstituted -CH2NCH2-(3 to 8 membered heterocycloalkyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -CH2NCH2-(3 to 8 membered heterocycloalkyl). In embodiments, L 3 is unsubstituted - CH2NCH2-(3 to 8 membered heterocycloalkyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) or unsubstituted -OCH2-(3 to 8 membered heterocycloalkylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -OCH2-(3 to 8 membered heterocycloalkylene). In embodiments, L 3 is unsubstituted -OCH2-(3 to 8 membered heterocycloalkylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 8 membered heterocycloalkylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 6 membered heterocycloalkylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) 3 to 6 membered heterocycloalkylene. In embodiments, L 3 is unsubstituted 3 to 6 membered heterocycloalkylene.
• L 3 is substituted (e.g., with a substituent group, a size- limited substituent group, or a lower substituent group) or unsubsituted -CH2NCH2-(3 to 6 membered heterocycloalkyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size- limited substituent group, or a lower substituent group) -CH2NCH2-(3 to 6 membered heterocycloalkyl). In embodiments, L 3 is unsubsituted - CH2NCH2-(3 to 6 membered heterocycloalkyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) or unsubsituted -OCH2-(3 to 6 membered heterocycloalkylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group, or a lower substituent group) -OCH2-(3 to 6 membered heterocycloalkylene). In embodiments, L 3 is unsubsituted -OCH2-(3 to 6 membered heterocycloalkylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 6 membered heterocycloalkylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclobutylene, heterocyclopentylene or heterocyclohexylene.
• L 3 is unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl).
• L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) -Ct NCth-Cheterocyclobutyl, heterocyclopentyl, or heterocyclohexyl).
• L 3 is unsubstituted -CH2NCH2- (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(hctcrocyclobutylcnc, hctcrocyclopcntylcnc, or hctcrocyclohcxylcnc).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene). In embodiments, L 3 is unsubstituted -OCH2-(heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclobutylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclobutylene. In embodiments, L 3 is unsubstituted heterocyclobutylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) or unsubstituted -CH2NCH2- (heterocyclobutyl).
• L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) -CH2NCH2-(heterocyclobutyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(heterocyclobutyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(heterocyclobutylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (heterocyclobutylene). In embodiments, L 3 is unsubstituted -OCH2-(heterocyclobutylene). [00493] In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclopentylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclopentylene.
• L 3 is unsubstituted heterocyclopentylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) or unsubstituted -CH2NCH2- (heterocyclopentyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) -CH2NCH2-(heterocyclopentyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(heterocyclopentyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(heterocyclopentylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (heterocyclopentylene). In embodiments, L 3 is unsubstituted -OCH2-(heterocyclopentylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclohexylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclohexylene. In embodiments, L 3 is unsubstituted heterocyclohexylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) or unsubstituted -CH2NCH2- (heterocyclohexyl).
• L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) -CH2NCH2-(heterocyclohexyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(heterocyclohexyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(heterocyclohexylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (heterocyclohexylene). In embodiments, L 3 is unsubstituted -OCH2-(heterocyclohexylene). [00495] In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 5 to 10 membered heteroarylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 10 membered heteroarylene.
• L 3 is unsubstituted 5 to 10 membered heteroarylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(5 to 10 membered heteroaryl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2-(5 to 10 membered heteroaryl). In embodiments, L 3 is unsubstituted -CH2NCH2-(5 to 10 membered heteroaryl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(5 to 10 membered heteroarylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(5 to 10 membered heteroarylene). In embodiments, L 3 is unsubstituted -OCH2-(5 to 10 membered heteroarylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 5 to 9 membered heteroarylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 9 membered heteroarylene. In embodiments, L 3 is unsubstituted 5 to 9 membered heteroarylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(5 to 9 membered heteroaryl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2-(5 to 9 membered heteroaryl). In embodiments, L 3 is unsubstituted -CH2NCH2-(5 to 9 membered heteroaryl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(5 to 9 membered heteroarylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(5 to 9 membered heteroarylene). In embodiments, L 3 is unsubstituted -OCH2-(5 to 9 membered heteroarylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 5 to 6 membered heteroarylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 6 membered heteroarylene. In embodiments, L 3 is unsubstituted 5 to 6 membered heteroarylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(5 to 6 membered heteroaryl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2-(5 to 6 membered heteroaryl). In embodiments, L 3 is unsubstituted -CH2NCH2-(5 to 6 membered heteroaryl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(5 to 6 membered heteroarylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(5 to 6 membered heteroarylene). In embodiments, L 3 is unsubstituted -OCH2-(5 to 6 membered heteroarylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene.
• L 3 is unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2- (furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl).
• L 3 is unsubstituted -CH2NCH2-(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene).
• L 3 is unsubstituted -OCH2-(furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted furanylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) furanylene. In embodiments, L 3 is unsubstituted furanylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(furanyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) - CH2NCH2-(furanyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(furanyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(furanylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (furanylene). In embodiments, L 3 is unsubstituted -OCH2-(furanylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyrrolylcnc. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyrrolylene. In embodiments, L 3 is unsubstituted pyrrolylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(pyrrolyl).
• L 3 is substituted (e.g., with a substituent group, a size- limited substituent group or a lower substituent group) -CH2NCH2-(pyiTolyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(pyrrolyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(pyrrolylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size- limited substituent group or a lower substituent group) -OCH2-(pyrrolylene). In embodiments, L 3 is unsubstituted -OCH2-(pyrrolylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyridylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyridylene. In embodiments, L 3 is unsubstituted pyridylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(pyridyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) - CH2NCH2-(pyridyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(pyridyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(pyridylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (pyridylene). In embodiments, L 3 is unsubstituted -OCH2-(pyridylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyranylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyranylene. In embodiments, L 3 is unsubstituted pyranylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(pyranyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) - CH2NCH2-(pyranyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(pyranyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(pyranylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size- limited substituent group or a lower substituent group) -OCH2-(pyranylene). In embodiments, L 3 is unsubstituted -OCH2-(pyranylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted imidazolylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) imidazolylene. In embodiments, L 3 is unsubstituted imidazolylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(imidazolyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2-(imidazolyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(imidazolyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(imidazolylene). In embodiments, L is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(imidazolylene).
• L 3 is unsubstituted -OCH2-(imidazolylene). [00504] In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted thiazolylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) thiazolylene. In embodiments, L 3 is unsubstituted thiazolylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(thiazolyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -CH2NCH2-(thiazolyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(thiazolyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(thiazolylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(thiazolylene). In embodiments, L 3 is unsubstituted -OCH2-(thiazolylene). [00505] In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted thienylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) thienylene. In embodiments, L 3 is unsubstituted thienylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(thienyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) - CH2NCH2-(thienyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(thienyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(thienylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2- (thienylene). In embodiments, L 3 is unsubstituted -OCH2-(thienylene).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted oxazolylene. In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) oxazolylene. In embodiments, L 3 is unsubstituted oxazolylene. In embodiments, L is unsubstituted oxazolylene.
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -CH2NCH2-(oxazolyl). In embodiments, L 3 is substituted (e.g., with a substituent group, a sizelimited substituent group or a lower substituent group) -CH2NCH2-(oxazolyl). In embodiments, L 3 is unsubstituted -CH2NCH2-(oxazolyl).
• L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted -OCH2-(oxazolylene). In embodiments, L 3 is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) -OCH2-(oxazolylene). In embodiments, L 3 is unsubstituted -OCH2-(oxazolylene).
• R* is substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl) or substituted (e.g. with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl).
• heterocycloalkyl e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl
• substituted e.g. with a substituent group, a size-limited substituent group or a lower substituent group
• R* is substituted with one or more substituent groups. In embodiments, R* is substituted with one or more size-limited substituent groups. In embodiments, R* is substituted with one or more lower substituent groups. [00508] In embodiments, R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl).
• heterocycloalkyl e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl.
• R is unsubstituted heterocycloalkyl (e.g., 3 to 8 membered heterocycloalkyl, 3 to 6 membered heterocycloalkyl, or 5 to 6 membered heterocycloalkyl).
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl).
• R* is unsubstituted heteroaryl (e.g., 5 to 10 membered heteroaryl, 5 to 9 membered heteroaryl, or 5 to 6 membered heteroaryl).
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 8 membered heterocycloalkyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 3 to 8 membered heterocycloalkyl.
• R* is unsubstituted 3 to 8 membered heterocycloalkyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 3 to 6 membered heterocycloalkyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 3 to 6 membered heterocycloalkyl.
• R* is unsubstituted 3 to 6 membered heterocycloalkyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.
• R* is unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclobutyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclobutyl.
• R is unsubstituted heterocyclobutyl .
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclopentyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclopentyl.
• R* is unsubstituted heterocyclopentyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted heterocyclohexyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) heterocyclohexyl.
• R* is unsubstituted heterocyclohexyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 5 to 10 membered heteroaryl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 10 membered heteroaryl.
• R* is unsubstituted 5 to 10 membered heteroaryl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 5 to 9 membered heteroaryl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 9 membered heteroaryl.
• R is unsubstituted 5 to 9 membered heteroaryl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted 5 to 6 membered heteroaryl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) 5 to 6 membered heteroaryl.
• R* is unsubstituted 5 to 6 membered heteroaryl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl.
• R* is unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted furanyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) furanyl.
• R* is unsubstituted furanyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyrrolyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyrrolyl.
• R is unsubstituted pyrrolyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyridyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyridyl.
• R* is unsubstituted pyridyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted pyranyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) pyranyl.
• R is unsubstituted pyranyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted imidazolyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) imidazolyl.
• R* is unsubstituted imidazolyl.
• R* is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) or unsubstituted thiazolyl.
• R is substituted (e.g., with a substituent group, a size-limited substituent group or a lower substituent group) thiazolyl.
• R* is unsubstituted thiazolyl.
• R 1 is H. In embodiments, R 1 is — C 1 -C 8 alkyl.
• R 1 is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, or hexyl.
• R 1 is methyl.
• R 1 is ethyl.
• R 1 is propyl.
• R 1 is isopropyl.
• R 1 is butyl.
• R 1 is isobutyl.
• R 1 is tert-butyl.
• R 1 is pentyl.
• R 1 is hexyl.
• R 3 is H, halogen, -CCk, -CB , -CF3, -CI3, -CHCh,
• R 3 is H, -OR 3A , -(CH 2 )vOR 6 , substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl), or substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) or unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl).
• substituted e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group
• unsubstituted alkyl e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or
• v is an integer from 1 to 24. In embodiments, v is 1. In embodiments, v is 2. In embodiments, v is 3. In embodiments, v is 4. In embodiments, v is 5. In embodiments, v is 6. In embodiments, v is 7. In embodiments, v is 8. In embodiments, v is 9. In embodiments, v is 10. In embodiments, v is 11. In embodiments, v is 12. In embodiments, v is 13. In embodiments, v is 14. In embodiments, v is 15. In embodiments, v is 16. In embodiments, v is 17. In embodiments, v is 18. In embodiments, v is 19. In embodiments, v is 20. In embodiments, v is 21. In embodiments, v is 22. In embodiments, v is 23. In embodiments, v is 24.
• R 4 is H, or substituted or unsubstituted alkyl. In embodiments, R 4 is H, or substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• R 4 is H, or substituted or unsubstituted alkyl.
• R 4 is H, or substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• R 4 is a substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• R 4 is an unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• R 4 is a substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl).
• R 4 is an unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl).
• R 4 is H, -OH, methyl, ethyl, propyl or butyl. In embodiments, R 4 is H or -OH. In embodiments, R 4 is H or methyl. In embodiments, R 4 is methyl. In embodiments, R 4 is ethyl. In embodiments, R 4 is propyl. In embodiments, R 4 is butyl. In embodiments, R 4 is H. In embodiments, R 4 is -OH.
• each R 3A , R 3B , R 4A , and R 4B is independently H or substituted or unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C1-C6 alkyl, or C1-C4 alkyl).
• each R 3A , R 3B , R 4A , and R 4B is independently H or substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• each R 3A , R 3B , R 4A , and R 4B is independently H.
• each R 3A , R 3B , R 4A , and R 4B is independently substituted (e.g., substituted with at least one substituent group, size-limited substituent group, or lower substituent group) alkyl (e.g., C 1 -C 8 alkyl, C 1 -C 6 alkyl, or C1-C4 alkyl).
• each R 3A , R 3B , R 4A , and R 4B is independently unsubstituted alkyl (e.g., Ci-C 8 alkyl, Ci-C 6 alkyl, or C1-C4 alkyl).
• each R 3A , R 3B , R 4A , and R 4B is independently H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, or pentyl. In embodiments, each R 3A , R 3B , R 4A , and R 4B is independently H. In embodiments, each R 3A , R 3B , R 4A , and R 4B is independently methyl. In embodiments, each R 3A , R 3B , R 4A , and R 4B is independently ethyl.
• each R 3A , R 3B , R 4A , and R 4B is independently propyl. In embodiments, each R 3A , R 3B , R 4A , and R 4B is independently isopropyl. In embodiments, each R 3A , R 3B , R 4A , and R 4B is independently butyl. In embodiments, each R 3A , R 3B , R 4A , and R 4B is independently isobutyl. In embodiments, each R 3A , R 3B , R 4A , and R 4B is independently tcrt-butyl. In embodiments, each R 3A , R 3B , R 4A , and R 4B is independently pentyl.
• R 6 is H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -CO(CH2CH2O) W CH2CH2M,
• w is an integer from 1 to 24; M is -NH2, -OH, -COOH, or -OCH3; R 10 is -OH, -OCH3 or -COOH.

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