WO2026002995A1 - Conjugués inhibiteurs de topoisomérase activés par fap spécifiques à une cible et leurs utilisations - Google Patents

Conjugués inhibiteurs de topoisomérase activés par fap spécifiques à une cible et leurs utilisations

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Publication number
WO2026002995A1
WO2026002995A1 PCT/EP2025/067754 EP2025067754W WO2026002995A1 WO 2026002995 A1 WO2026002995 A1 WO 2026002995A1 EP 2025067754 W EP2025067754 W EP 2025067754W WO 2026002995 A1 WO2026002995 A1 WO 2026002995A1
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WIPO (PCT)
Prior art keywords
pharmaceutically acceptable
acceptable salt
construct
optionally substituted
compound
Prior art date
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Pending
Application number
PCT/EP2025/067754
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English (en)
Inventor
Francis Wilson
Ellen WATTS
Thomas CLOUGH
David Jones
Graham Robert Pye-Smith SPENCE
Victoria JUSKAITE
Manuel Pinto
Douglas SAMMON
Ruairidh EDWARDS
Hanna BUIST
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Avacta Life Sciences Ltd
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Avacta Life Sciences Ltd
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Application filed by Avacta Life Sciences Ltd filed Critical Avacta Life Sciences Ltd
Publication of WO2026002995A1 publication Critical patent/WO2026002995A1/fr
Pending legal-status Critical Current
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    • 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
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    • 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
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    • A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
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Definitions

  • prodrugs remain inert in the body until they encounter the unique physiological conditions of the tumor, such as acidic pH, high enzymatic activity, or specific reductive environments.
  • This targeted activation ensures that the therapeutic agent is released directly at the tumor site, minimizing damage to healthy tissues and reducing systemic toxicity.
  • tumor-specific activation can improve drug efficacy, as the active form of the drug is concentrated where it is most needed, potentially allowing for higher doses to be used safely. This strategy enhances treatment precision and can overcome some of the challenges associated with traditional chemotherapy.
  • SUMMARY [4] Provided herein are compounds and constructs comprising a fibroblast activating protein (FAP)- cleavable moiety that are capable of delivering a camptothecin to FAP-expressing tissues (e.g., cancers). Also provided herein are pharmaceutical compositions comprising the constructs provided herein, and kits comprising the same.
  • FAP fibroblast activating protein
  • compounds of Formula (I′) and pharmaceutically acceptable salts thereof, wherein R′, L 1 , X, R 2A , R 3A , n, R 2 , R 3 , R 4 , m, L 2 , and CAM are as described herein.
  • compounds of Formula (I): and pharmaceutically acceptable salts thereof wherein R A , L A , L 1 , X, R 2A , R 3A , n, R 2 , R 3 , R 4 , m, L 2 , and CAM are as described herein.
  • a compound described herein e.g., a compound of Formula (I′) or (I)
  • a compound described herein is selected from those recited in Table 4 (infra), and pharmaceutically acceptable salts thereof.
  • constructs of Formula (II): and pharmaceutically acceptable salts thereof, wherein Z, R B , L A , L 1 , X, R 2A , R 3A , n, R 2 , R 3 , R 4 , m, L 2 , and CAM are as described herein.
  • a construct described herein e.g., a construct of Formula (II′) or (II)
  • a construct described herein is selected from those recited in Table 5A (infra), and pharmaceutically acceptable salts thereof.
  • compounds selected from those recited in Table 5B (infra) are compounds selected from those recited in Table 5B (infra), and pharmaceutically acceptable salts thereof.
  • pharmaceutical compositions comprising a construct described herein (e.g., a construct of Formula (II′) or (II)) or a pharmaceutically acceptable salt thereof,
  • a pharmaceutical composition provided herein comprises an effective amount (e.g., therapeutically effective amount) of a construct described herein (e.g., a construct of Formula (II′) or (II)), or a pharmaceutically acceptable salt thereof.
  • constructs and pharmaceutical compositions provided herein can deliver a camptothecin to FAP-expressing tissues (e.g., cancers) and are therefore useful for treating diseases characterized by fibroblast activation protein upregulation in a subject.
  • kits and uses of the constructs and pharmaceutical compositions provided herein including, but not limited to, the following: (a) Methods of treating a disease characterized by fibroblast activation protein upregulation (e.g., cancer, fibrosis, or inflammation) in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a construct described herein (e.g., a construct of Formula(II′) or (II)), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • a construct described herein e.g., a construct of Formula(II′) or (II)
  • FIG.1 shows size-exclusion chromatography (SEC) profiles of purified Affimer® conjugates containing either linker-payload 2 (LP2) or 1 (LP1) (see Table 13), as indicated.
  • SEC size-exclusion chromatography
  • the SEC profiles of the unconjugated Affimer® precursor (bottom peak) are also shown for FAP-2-TEV-CTCys-LP2 and FAP- 2-FAP-1-H-CTCys-LP2.
  • FIG.2 shows liquid chromatograph-mass spectrometry (LC-MS) analysis for purified Affimer® conjugates.
  • LC-MS liquid chromatograph-mass spectrometry
  • FIG.3 shows single-cycle kinetics SPR analysis of the binding of single-domain Affimer® conjugates to human FAPalpha.
  • the KD affinities are shown in the embedded table, showing equivalent FAP-binding affinities to the parent Affimer® protein.
  • FIG.4 shows single-cycle kinetics SPR analysis of the binding of two-domain Affimer® conjugates to human FAPalpha.
  • the KD affinities are shown in the embedded table, showing equivalent FAP-binding affinities to the parent Affimer® protein.
  • FIG.5 shows cellular cytotoxicity assays using HEK293T cells (FAP-negative) or cells overexpressing FAP (HEK-FAP).
  • Cells were incubated with the different LP2 Affimer® conjugates or unconjugated linker-payload, either alone or in the presence of hFAP or FAPi, as indicated, for 4 days in the presence of FBS substitute Panexin (which contains low levels of FAP).
  • the warhead was also included as a positive control.
  • the antiproliferative/cytotoxic effect of the compounds was measured as % inhibition relative to vehicle control.
  • FIG.6 shows cellular cytotoxicity assays using HEK293T cells (FAP-negative) or cells overexpressing FAP (HEK-FAP).
  • Cells were incubated with the different LP1 Affimer® conjugates or unconjugated linker-payload, either alone or in the presence of hFAP or FAPi, as indicated, for 4 days in the presence of FBS substitute Panexin (which contains low levels of FAP).
  • the warhead was also included as a positive control.
  • the antiproliferative/cytotoxic effect of the compounds was measured as % inhibition relative to vehicle control.
  • FIG.7 shows co-culture assays using LS174T-GFP colorectal adenocarcinoma cells (FAP- negative) in the presence (co-culture) or absence (mono-culture) of human colonic fibroblast (HCoF) cells.
  • FAP- negative human colonic fibroblast
  • HoF human colonic fibroblast
  • FIG.8 shows co-culture assay using MiaPaCa2-GFP pancreatic cancer cells (FAP-negative) in the presence (co-culture) or absence (mono-culture) of human pancreatic stellate (hPSC) stromal cells.
  • Cells were incubated with 10nM Affimer® conjugates (FAP-2-FAP-1-H-L3Cys-LP2 or 2FAP-2-FAP-1- H-L3Cys-LP1) for 5 days in the presence of FBS substitute Panexin (which contains low levels of FAP).
  • the warhead was also included as a positive control.
  • the antiproliferative/cytotoxic effect of the compounds was measured as % GFP positive cells using Incucyte.
  • FIG.9 shows size-exclusion chromatography (SEC) profiles of Affimer® constructs conjugated to 5 kDa PEG within Loop3, Loop7, the N-terminus, and/or the C-terminus.
  • the unconjugated parent proteins are also shown at the bottom of the figures.
  • Single-domain FAP-2 (left figure) and two-domain FAP-2-FAP-1-H (right figure) exemplars are shown.
  • a key to the right of each figure shows the location of the PEG conjugation sites: L3 – loop3, L7 – loop7, CT – C-terminus, NT – N-terminus.1x, 2x, and 3xPEG conjugated species are shown.
  • FIG.10 shows single-cycle kinetics SPR analysis of the binding of two-domain Affimer® PEG conjugates (1x, 2x, or 3x PEG conjugates) to human FAPalpha. The similar kinetic profiles demonstrate equivalent FAP-binding to the parent Affimer® protein.
  • FIG.11 shows single-cycle kinetics SPR analysis of the binding of single-domain Affimer® PEG conjugates (1x or 2x PEG conjugates) to human FAPalpha. The similar kinetic profiles demonstrate equivalent FAP-binding to the parent Affimer® protein.
  • FIG.12A shows the internalization of FAP-binding AFFIMER® proteins and monoclonal antibodies as measured by the percent downregulation of cell surface AFFIMER® protein. Internalizing monoclonal antibodies (three right-hand bars) were used as positive internalizing controls.
  • FIG.12B shows the internalization of FAP-binding AFFIMER® proteins in FAP-expressing cells as measured by area under the curve (AUC) over 24 hours. Dye-conjugated Sibrotuzumab antibody and human IgG1 were used as negative and positive controls, respectively.
  • FIG.12C shows the percent inhibition of FAP enzyme activity from a panel of 17 FAP-binding AFFIMER® proteins.
  • FIG.12D shows AFFIMER® protein stability under stress temperature conditions. FAP-2 single- domain (left) and FAP-2-FAP-1 two-domain (right) AFFIMER® proteins were incubated at 4°C, 37°C, or 45°C for 1 week. Protein aggregation in the samples was assessed by SEC. SEC profiles for the stressed samples are shown with the percent purity for the monomeric species indicated.
  • FIG.13 shows that FAP expression correlates with SLFN11 expression across multiple tumor types.
  • Scatter plots show the relationship between FAP and SLFN11 mRNA expression (log2(TPM+1)) in small cell lung cancer, pancreatic cancer, cervical cancer and gastric cancer. Each dot represents a tumor sample. Light grey (far left) indicates FAP negative patients, dark gray (bottom) indicates FAP positive, low SLFN11 expressing patients, and medium gray (top right) indicates FAP positive high SLFN11 patients. Linear regression analysis with correlation coefficients (R) and p-values are overlaid.
  • FIG.14 shows size-exclusion chromatography (SEC) profiles of anti-CEACAM5 conjugates containing different linker payloads (LP2, LP3, LP4 and LP10) following incubation of antibody conjugates at 37, or 45°C for 1 week at 1mg/ml or at baseline (time 0).
  • Table shows the percent aggregate increase at 37 and 45°C compared to time 0, and corresponds to the % of protein present in the SEC profile which is of a higher order than the monomeric peak.
  • FIG.15 shows binding of anti-EDB-F, anti-LRRC15 and anti-CEACAM5 antibody conjugates containing different linker payloads to target expressing cells, determined by flow cytometry. The respective naked antibodies are also shown.
  • FIG.16 shows cytotoxicity in FAP-negative (HEK293T) and FAP-overexpressing (HEK293T- FAP) cells after 4-day incubation with LP2 antibody conjugates or unconjugated linker-payload, with or without recombinant FAP or FAP inhibitor, in FBS substitute Panexin-containing medium (which contains low levels of FAP). Warhead was included as a positive control. Cytotoxicity was measured as % inhibition relative to vehicle control.
  • FIG.17 shows cytotoxicity in FAP-negative (HEK293T) and FAP-overexpressing (HEK293T- FAP) cells after 4-day incubation with LP3 antibody conjugates or unconjugated linker-payload, with or without recombinant FAP or FAP inhibitor, in FBS substitute Panexin-containing medium (which contains low levels of FAP). Warhead was included as a positive control. Cytotoxicity was measured as % inhibition relative to vehicle control.
  • FIG.18 shows cytotoxicity in FAP-negative (HEK293T) and FAP-overexpressing (HEK293T- FAP) cells after 4-day incubation with LP4 antibody conjugates or unconjugated linker-payload, with or without recombinant FAP or FAP inhibitor, in FBS substitute Panexin-containing medium (which contains low levels of FAP). Warhead was included as a positive control. Cytotoxicity was measured as % inhibition relative to vehicle control.
  • FIG.19 shows cytotoxicity in FAP-negative (HEK293T) and FAP-overexpressing (HEK293T- FAP) cells after 4-day incubation with LP5 antibody conjugates or unconjugated linker-payload, with or without recombinant FAP or FAP inhibitor, in FBS substitute Panexin-containing medium (which contains low levels of FAP). Warhead was included as a positive control. Cytotoxicity was measured as % inhibition relative to vehicle control.
  • FIGs.20A-20B show cytotoxicity of FAP-negative LS174T-GFP cells as mono-culture (FIG.
  • FIGs.21A-21B show cytotoxicity of FAP-negative MDA-MB-231-GFP cells as 3D spheroid mono-culture (FIG.21A) or co-culture with FAP-expressing human mammary fibroblast (HMF) cells (FIG.21B) after 7-day incubation with anti-CEACAM5-LP2 antibody conjugate, with or without recombinant FAP or FAP inhibition, in complete Mammocult medium (chemically defined with no exogenous FAP). Warhead was included as positive control. Tumour spheroid specific cytotoxicity was measured as % inhibition relative to vehicle control using GFP fluorescence. [43] FIGs.22A-22B show change in tumour volume (FIG.22A) or body weight (FIG.22B) following treatment with vehicle, anti-CEACAM5 antibody or anti-CEACAM5 conjugates containing
  • FIG.23 shows levels of released exatecan warhead in tumor and plasma, measured by LC/MS, after treatment with anti-CEACAM5-LP2 in a cell-line derived xenograft (CDX) model of LS174T cells engineered to express FAP. Data are plotted as mean + standard error of mean (SEM). D ETAILED D ESCRIPTION [45] FAP is a post-prolyl cleaving serine protease that can cleave on the C-terminal side of an internal proline residue.
  • camptothecin refers to a compound (or a radical thereof) belonging to the class of compounds considered to be camptothecins, camptothecin analogs, camptothecin derivatives or camptothecin conjugates.
  • camptothecin refers to a compound (or a radical thereof) derived from the camptothecin five-ring backbone: , optionally with one or more modifications or substituents. Camptothecins may exist in the lactone or carboxylate forms, and the term “camptothecin” refers to either or both alternatives.
  • camptothecins include, but are not limited to, irinotecan (7-ethyl-10-[4-(1-piperidino)-1-piperidino]- carbonyloxycamptothecin), belotecan, Dxd (N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13- dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-2- hydroxyacetamide), SN-38 ((4S)-4,11-Diethyl-4,9-dihydroxy-1,4-dihydro-3H,14H- pyrano[3’,4’:6,7]indolizino[1,2-b]quinoline-3,14-dione), topotecan ((S)-9-N,N-
  • self-immolative linker or “self-eliminating linker” refers to a temporary extender, spacer, or placeholder unit attaching two or more molecules together by chemical bonds that are cleaved under defined conditions to release the two molecules.
  • a self-immolative or self-eliminating linker may be linear or branched, and may link two or more of the same molecules together, or may link two or more different molecules together.
  • the self-immolative or self-eliminating linker may degrade, decompose, or fragment under, for example, physiological conditions, acidic conditions, basic conditions, or in the presence of specific chemical agents.
  • self-eliminating linkers include, but are not limited to, p-aminobenzyloxycarbonyl (PABC) and 2,4-bis(hydroxymethyl)aniline.
  • PABC p-aminobenzyloxycarbonyl
  • the self-immolative linker includes one or more moieties that are hydrolyzed under physiological conditions to reveal the desired molecule (e.g., a camptothecin).
  • the self-immolative linker is cleaved by an enzymatic activity of the host animal.
  • salt refers to any and all salts and encompasses pharmaceutically acceptable salts. Salts include ionic compounds that result from the neutralization reaction of an acid and a base. A salt is composed of one or more cations (positively charged ions) and one or more anions (negative ions) so that the salt is electrically neutral (without a net charge).
  • pharmaceutically acceptable salt refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al.
  • Pharmaceutically acceptable salts of the compounds of the present disclosure include those derived from suitable inorganic and organic acids and bases.
  • suitable inorganic and organic acids and bases include those derived from suitable inorganic and organic acids and bases.
  • pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid or with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
  • Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N + (C1-4 alkyl)4- salts.
  • Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
  • Further pharmaceutically acceptable salts include, when appropriate, nontoxic
  • references to “the compound” and “a compound” provided herein are intended to encompass the compound or group of compounds, and also pharmaceutically acceptable salts, stereoisomers, tautomers, solvates (e.g., hydrates), and isotopically labeled derivatives thereof.
  • amino acid or “amino acid residue” encompasses all compounds, whether natural or synthetic, which include both an amino functionality and an acid functionality, including amino acid analogues and derivatives.
  • the amino acids contemplated in the present invention are those naturally occurring amino acids found in proteins, or the naturally occurring anabolic or catabolic products of such amino acids, which contain amino and carboxyl groups.
  • Naturally occurring amino acids are identified throughout by the conventional three- letter and/or one-letter abbreviations, corresponding to the trivial name of the amino acid, in accordance with the following list. The abbreviations are accepted in the peptide art and are recommended by the IUPAC-IUB commission in biochemical nomenclature.
  • amino acid residue further includes analogues, derivatives, and congeners of any specific amino acid referred to herein, as well as C-terminal or N-terminal protected amino acid derivatives (e.g., modified with an N-terminal or C-terminal protecting group).
  • composition and “formulation” are used interchangeably.
  • a “subject” to which administration is contemplated refers to a human (i.e., male or female of any age group, e.g., pediatric subject (e.g., infant, child, or adolescent) or adult subject (e.g., young adult, middle-aged adult, or senior adult)) or non-human animal.
  • the non-human animal is a mammal (e.g., primate (e.g., cynomolgus monkey or rhesus monkey), commercially relevant mammal (e.g., cattle, pig, horse, sheep, goat, cat, or dog), or bird (e.g., commercially relevant bird, such
  • primate e.g., cynomolgus monkey or rhesus monkey
  • commercially relevant mammal e.g., cattle, pig, horse, sheep, goat, cat, or dog
  • bird e.g., commercially relevant bird, such
  • the non-human animal is a fish, reptile, or amphibian.
  • the non-human animal may be a male or female at any stage of development.
  • the non- human animal may be a transgenic animal or genetically engineered animal.
  • patient refers to a human subject in need of treatment of a disease or condition.
  • administer refers to injecting, implanting, providing or otherwise introducing a compound described herein, or a composition thereof, in, to or on a subject.
  • treatment refers to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease described herein.
  • treatment may be administered after one or more signs or symptoms of the disease have developed or have been observed.
  • treatment may be administered in the absence of signs or symptoms of the disease.
  • treatment may be administered to a susceptible subject prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of exposure to a pathogen). Treatment may also be continued after symptoms have resolved, for example, to delay or prevent recurrence.
  • condition “disease,” and “disorder” are used interchangeably.
  • an “effective amount” of a compound described herein refers to an amount sufficient to elicit the desired biological response.
  • An effective amount of a compound described herein may vary depending on such factors as the desired biological endpoint, severity of side effects, disease, or disorder, the identity, pharmacokinetics, and pharmacodynamics of the particular compound, the condition being treated, the mode, route, and desired or required frequency of administration, the species, age and health or general condition of the subject.
  • an effective amount is a therapeutically effective amount.
  • an effective amount is the amount of a compound described herein in a single dose.
  • an effective amount is the combined amounts of a compound described herein in multiple doses.
  • an effective amount is an amount sufficient for delivering a camptothecin to a FAP-expressing tissue. In certain embodiments, an effective amount is an amount sufficient for delivering a camptothecin to the site of a cancer.
  • a “therapeutically effective amount” of a compound described herein is an amount sufficient to provide a therapeutic benefit in the treatment of a condition or to delay or minimize one or more symptoms associated with the condition, alone or in combination with other therapies.
  • the term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms, signs, or causes of the condition, and/or enhances the therapeutic efficacy of another therapeutic agent.
  • a therapeutically effective amount is an amount sufficient for treating a disease characterized by FAP upregulation. In certain embodiments, a therapeutically effective amount is an amount sufficient for treating cancer, fibrosis, or inflammation.
  • fibroblast activating protein or “FAP” refers to fibroblast activation protein alpha (FAPa, or simply FAP; EC 3.4.21.-), also known as seprase or 170 kDa melanoma membrane-bound gelatinase.
  • FAP is a homodimeric integral membrane protein belonging to the serine protease family and to the dipeptidyl peptidase (DPP-IV)-like subfamily.
  • target tissue refers to any biological tissue of a subject (including a group of cells, a body part, or an organ) or a part thereof, including blood and/or lymph vessels, which is the object to which a compound, particle, and/or composition of the present disclosure is delivered.
  • a target tissue may be an abnormal or unhealthy tissue, which may need to be treated.
  • a target tissue may also be a normal or healthy tissue that is under a higher than normal risk of becoming abnormal or unhealthy, which may need to be prevented.
  • the target tissue is the liver.
  • the target tissue is the lung.
  • a “non-target tissue” is any biological tissue of a subject (including a group of cells, a body part, or an organ) or a part thereof, including blood and/or lymph vessels, which is not a target tissue.
  • a target tissue is a tissue that expresses FAP.
  • An “binding moiety” includes a specific part or component of a molecule that is responsible for its ability to bind or interact with another molecule (e.g., a target molecule), often with high specificity. In some embodiments, a binding moiety specifically binds to a target molecule.
  • Specifically binds refers to the ability of a binding moiety to bind to a target molecule binding partner with a degree of affinity or avidity that enables the molecule to be used to distinguish the target molecule from an appropriate control in a binding assay or other binding context.
  • “specifically binds” refers to the ability of the antibody or AFFIMER® to bind to a specific target molecule with a degree of affinity or avidity, compared with an appropriate reference molecule or molecules, that enables the antibody or AFFIMER® to be used to distinguish the specific target molecule from others, as described herein.
  • an antibody or AFFIMER® specifically binds to a target molecule if the antibody or AFFIMER® has a K D for binding the target molecule of at least about 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10- 12 M, 10 -13 M, or less.
  • An “antigen-binding moiety” includes a specific part or component of a molecule that binds to a target antigen (e.g., tumor, e.g., cancer antigen), and encompasses monoclonal antibodies, polyclonal antibodies, monospecific and multispecific antibodies (e.g., bispecific antibodies, trispecific antibodies etc.), and antibody fragments, provided they bind to the relevant target molecule(s).
  • a target antigen e.g., tumor, e.g., cancer antigen
  • a “cell-binding moiety” is a type of antigen-binding moiety that binds to a target antigen on the surface of a cell, such as a cancer cell.
  • an “antibody” includes a polypeptide that comprises at least one immunoglobulin variable domain or at least one site, e.g., paratope, that specifically binds to an antigen.
  • a typical antibody molecule comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), which are usually involved in antigen binding.
  • VH and VL regions can be further subdivided into regions of hypervariability, also known as “complementarity determining regions” (“CDR”), interspersed with regions that are more conserved, which are known as “framework regions” (“FR”).
  • CDR complementarity determining regions
  • Each VH and VL is typically composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.
  • the term “antibody” also includes antigen binding fragments thereof (e.g., Fab fragment, a F(ab')2 fragment, a Fv fragment or a scFv fragment).
  • An “AFFIMER ® polypeptide” refers to a polypeptide comprising a modified version of human Stefin A as well as one or two, preferably two, heterologous peptides.
  • AFFIMER® polypeptides include small, engineered proteins designed to bind specifically to target molecules. They are derived from human Stefin A scaffold proteins and have distinct loop structures that confer their binding properties. The different loop structures of an AFFIMER® polypeptide, based on their design and origin, typically include two variable “loop” structures.
  • a Variable Loop 2 (VL2) is one of the primary loops responsible for binding specificity. It is engineered to interact with the target molecule through various amino acid substitutions, providing a high degree of diversity and specificity.
  • VL4 also contributes significantly to the binding interaction.
  • the combination of VL2 and VL4 provides the structural diversity often needed for high-affinity binding to a wide range of targets.
  • the framework regions (collectively the scaffold) support the variable loops structurally. These regions are typically more conserved and provide a stable scaffold to present the variable loops in the correct orientation for target binding.
  • Chemical Definitions [65] Definitions of specific functional groups and chemical terms are described in more detail below. The chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th Ed., inside cover, and specific functional groups are generally defined as described therein.
  • the compounds described herein can be in the form of an individual enantiomer, diastereomer, or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer.
  • Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses.
  • range When a range of values (“range”) is listed, it encompasses each value and sub-range within the range.
  • a range is inclusive of the values at the two ends of the range unless otherwise provided.
  • C1-6 alkyl encompasses, C1, C2, C3, C4, C5, C6, C1-6, C1-5, C1-4, C1-3, C1-2, C2-6, C2-5, C2-4, C2-3, C3-6, C3-5, C3-4, C4-6, C4-5, and C5-6 alkyl.
  • At least one instance refers to 1, 2, 3, 4, or more instances, but also encompasses a range, e.g., for example, from 1 to 4, from 1 to 3, from 1 to 2, from 2 to 4, from 2 to 3, or from 3 to 4 instances, inclusive.
  • alkylene is the divalent moiety of alkyl
  • alkenylene is the divalent moiety of alkenyl
  • alkynylene is the divalent moiety of alkynyl
  • heteroalkylene is the divalent moiety of heteroalkyl
  • heteroalkenylene is the divalent moiety of heteroalkenyl
  • heteroalkynylene is the divalent moiety of heteroalkynyl
  • carbocyclylene is the divalent moiety of carbocyclyl
  • heterocyclylene is the divalent moiety of heterocyclyl
  • arylene is the divalent moiety of aryl
  • heteroarylene is the divalent moiety of heteroaryl.
  • halo refers to fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), or iodine (iodo, -I).
  • alkyl refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 20 carbon atoms (“C 1-20 alkyl”). In some embodiments, an alkyl group has 1 to 12 carbon atoms (“C1-12 alkyl”). In some embodiments, an alkyl group has 1 to 10 carbon atoms (“C1-10 alkyl”).
  • an alkyl group has 1 to 9 carbon atoms (“C1-9 alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C1-8 alkyl”). In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C1-7 alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C1-6 alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C1-5 alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C1-4 alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C1-3 alkyl”).
  • an alkyl group has 1 to 2 carbon atoms (“C1-2 alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C1 alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C2-6 alkyl”).
  • C1-6 alkyl groups include methyl (C1), ethyl (C2), propyl (C3) (e.g., n-propyl, isopropyl), butyl (C4) (e.g., n-butyl, tert-butyl, sec-butyl, isobutyl), pentyl (C5) (e.g., n-pentyl, 3-pentanyl, amyl, neopentyl, 3-methyl-2-butanyl, tert- amyl), and hexyl (C6) (e.g., n-hexyl).
  • alkyl groups include n-heptyl (C7), n-octyl (C8), n-dodecyl (C12), and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an “unsubstituted alkyl”) or substituted (a “substituted alkyl”) with one or more substituents (e.g., halogen, such as F).
  • substituents e.g., halogen, such as F
  • the alkyl group is an unsubstituted C1-12 alkyl (such as unsubstituted C1-6 alkyl, e.g., -CH3 (Me), unsubstituted ethyl (Et), unsubstituted propyl (Pr, e.g., unsubstituted n-propyl (n-Pr), unsubstituted isopropyl (i-Pr)), unsubstituted butyl (Bu, e.g., unsubstituted n-butyl (n-Bu), unsubstituted tert-butyl (tert-Bu or t-Bu), unsubstituted sec-butyl (sec- Bu or s-Bu), unsubstituted isobutyl (i-Bu)).
  • unsubstituted C1-6 alkyl e.g., -CH3 (Me), unsubstituted ethyl (
  • the alkyl group is a substituted C1-12 alkyl (such as substituted C1-6 alkyl, e.g., -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, or benzyl (Bn)).
  • substituted C1-6 alkyl e.g., -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, or benzyl (Bn)
  • haloalkyl is a substituted alkyl group, wherein one or more of the hydrogen atoms are independently replaced by a halogen, e.g., fluoro, bromo, chloro, or iodo.
  • Perhaloalkyl is a subset of haloalkyl and refers to an alkyl group wherein all of the hydrogen atoms are independently replaced by a halogen, e.g., fluoro, bromo, chloro, or iodo.
  • the haloalkyl moiety has 1 to 20 carbon atoms (“C1-20 haloalkyl”).
  • the haloalkyl moiety has 1 to 10 carbon atoms (“C1-10 haloalkyl”). In some embodiments, the haloalkyl moiety has 1 to 9 carbon atoms (“C1-9 haloalkyl”). In some embodiments, the haloalkyl moiety has 1 to 8 carbon atoms (“C1-8 haloalkyl”). In some embodiments, the haloalkyl moiety has 1 to 7 carbon atoms (“C1-7 haloalkyl”). In some embodiments, the haloalkyl moiety has 1 to 6 carbon atoms (“C1-6 haloalkyl”).
  • the haloalkyl moiety has 1 to 5 carbon atoms (“C1-5 haloalkyl”). In some embodiments, the haloalkyl moiety has 1 to 4 carbon atoms (“C 1-4 haloalkyl”). In some embodiments, the haloalkyl moiety has 1 to 3 carbon atoms (“C 1-3 haloalkyl”). In some embodiments, the haloalkyl moiety has 1 to 2 carbon atoms (“C 1-2 haloalkyl”). In some embodiments, all of the haloalkyl hydrogen atoms are independently replaced with fluoro to provide a “perfluoroalkyl” group.
  • haloalkyl hydrogen atoms are independently replaced with chloro to provide a “perchloroalkyl” group.
  • haloalkyl groups include -CHF 2 , -CH 2 F, -CF 3 , -CH 2 CF 3 , -CF 2 CF 3 , -CF 2 CF 2 CF 3 , -CCl 3 , -CFCl 2 , -CF 2 Cl, and the like.
  • heteroalkyl refers to an alkyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, sulfur, silicon, boron, and phosphorous within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain.
  • the heteroalkyl group is an alkyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, and sulfur within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain.
  • a heteroalkyl group refers to a saturated group having from 1 to 20 carbon atoms and 1 or more heteroatoms within the parent chain (“C1-20 heteroalkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 12 carbon atoms and 1 or more heteroatoms within the parent chain (“C1-12 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 11 carbon atoms and 1 or more heteroatoms within the parent chain (“C1-11 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 10 carbon atoms and 1 or more heteroatoms within the parent chain (“C1-10 heteroalkyl”).
  • a heteroalkyl group is a saturated group having 1 to 9 carbon atoms and 1 or more heteroatoms within the parent chain (“C1-9 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 8 carbon atoms and 1 or more heteroatoms within the parent chain (“C1-8 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 7 carbon atoms and 1 or more heteroatoms within the parent chain (“C1-7 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 6 carbon atoms and 1 or more heteroatoms within the parent chain (“C1-6 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 5 carbon atoms and 1 or 2 heteroatoms within the parent chain (“C1-5).
  • a heteroalkyl group is a saturated group having 1 to 4 carbon atoms and 1 or 2 heteroatoms within the parent chain (“C1-4 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 3 carbon atoms and 1 heteroatom within the parent chain (“C1-3 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 2 carbon atoms and 1 heteroatom within the parent chain (“C1-2 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 carbon atom and 1 heteroatom (“C1 heteroalkyl”).
  • a heteroalkyl group is a saturated group having 2 to 6 carbon atoms and 1 or 2 heteroatoms within the parent chain (“C2-6 heteroalkyl”). Unless otherwise specified, each instance of a heteroalkyl group is independently unsubstituted (an “unsubstituted heteroalkyl”) or substituted (a “substituted heteroalkyl”) with one or more substituents.
  • alkenyl refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 20 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds).
  • an alkenyl group has 2 to 20 carbon atoms (“C 2-20 alkenyl”). In some embodiments, an alkenyl group has 2 to 12 carbon atoms (“C 2-12 alkenyl”). In some embodiments, an alkenyl group has 2 to 11 carbon atoms (“C 2-11 alkenyl”). In some embodiments, an alkenyl group has 2 to 10 carbon atoms (“C 2-10 alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms (“C 2-9 alkenyl”). In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C 2-8 alkenyl”).
  • an alkenyl group has 2 to 7 carbon atoms (“C 2-7 alkenyl”). In some embodiments, an alkenyl group has 2 to 6 carbon atoms (“C 2-6 alkenyl”). In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C 2-5 alkenyl”). In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C 2-4 alkenyl”). In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C 2-3 alkenyl”). In some embodiments, an alkenyl group has 2 carbon atom (“C 2 alkenyl”).
  • the one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl).
  • Examples of C 2-4 alkenyl groups include ethenyl (C 2 ), 1-propenyl (C 3 ), 2-propenyl (C 3 ), 1-butenyl (C 4 ), 2-butenyl (C 4 ), butadienyl (C4), and the like.
  • Examples of C2-6 alkenyl groups include the aforementioned C2-4 alkenyl groups as well as pentenyl (C 5 ), pentadienyl (C 5 ), hexenyl (C 6 ), and the like.
  • alkenyl examples include heptenyl (C 7 ), octenyl (C 8 ), octatrienyl (C 8 ), and the like. Unless otherwise specified, each instance of an alkenyl group is independently unsubstituted (an “unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) with one or more substituents.
  • a heteroalkenyl group refers to a group having from 2 to 10 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“C2-10 heteroalkenyl”). In some embodiments, a heteroalkenyl group has 2 to 9 carbon atoms at least one double bond, and 1 or more heteroatoms within the parent chain (“C2-9 heteroalkenyl”). In some embodiments, a heteroalkenyl group has 2 to 8 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“C 2-8 heteroalkenyl”).
  • a heteroalkenyl group has 2 to 7 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“C 2-7 heteroalkenyl”). In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“C 2-6 heteroalkenyl”). In some embodiments, a heteroalkenyl group has 2 to 5 carbon atoms, at least one double bond, and 1 or 2 heteroatoms within the parent chain (“C 2-5 heteroalkenyl”).
  • a heteroalkenyl group has 2 to 4 carbon atoms, at least one double bond, and 1 or 2 heteroatoms within the parent chain (“C 2-4 heteroalkenyl”). In some embodiments, a heteroalkenyl group has 2 to 3 carbon atoms, at least one double bond, and 1 heteroatom within the parent chain (“C 2-3 heteroalkenyl”). In some embodiments, a heteroalkenyl group has 2 carbon atoms, at least one double bond, and 1 heteroatom within the parent chain (“C 2 heteroalkenyl”). In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double bond, and 1 or 2 heteroatoms within the parent chain (“C 2-6 heteroalkenyl”).
  • each instance of a heteroalkenyl group is independently unsubstituted (an “unsubstituted heteroalkenyl”) or substituted (a “substituted heteroalkenyl”) with one or more substituents.
  • alkynyl refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 20 carbon atoms and one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds) (“C2-20 alkynyl”). In some embodiments, an alkynyl group has 2 to 10 carbon atoms (“C2-10 alkynyl”).
  • an alkynyl group has 2 to 9 carbon atoms (“C2-9 alkynyl”). In some embodiments, an alkynyl group has 2 to 8 carbon atoms (“C2-8 alkynyl”). In some embodiments, an alkynyl group has 2 to 7 carbon atoms (“C2-7 alkynyl”). In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C2-6 alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C2-5 alkynyl”). In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C2-4 alkynyl”).
  • an alkynyl group has 2 to 3 carbon atoms (“C2-3 alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“C2 alkynyl”).
  • the one or more carbon-carbon triple bonds can be internal (such as in 2- butynyl) or terminal (such as in 1-butynyl).
  • Examples of C2-4 alkynyl groups include, without limitation, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like.
  • Examples of C2-6 alkenyl groups include the aforementioned C2-4 alkynyl groups as well as pentynyl (C5), hexynyl
  • alkynyl examples include heptynyl (C7), octynyl (C8), and the like. Unless otherwise specified, each instance of an alkynyl group is independently unsubstituted (an “unsubstituted alkynyl”) or substituted (a “substituted alkynyl”) with one or more substituents.
  • heteroalkynyl refers to an alkynyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, sulfur, silicon, boron, and phosphorous within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain.
  • the heteroalkynyl group is an alkynyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, and sulfur within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain.
  • a heteroalkynyl group refers to a group having from 2 to 20 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“C 2-20 heteroalkynyl”). In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 10 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“C 2-10 heteroalkynyl”). In some embodiments, a heteroalkynyl group has 2 to 9 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“C 2-9 heteroalkynyl”).
  • a heteroalkynyl group has 2 to 8 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“C 2-8 heteroalkynyl”). In some embodiments, a heteroalkynyl group has 2 to 7 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“C 2-7 heteroalkynyl”). In some embodiments, a heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“C 2-6 heteroalkynyl”).
  • a heteroalkynyl group has 2 to 5 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms within the parent chain (“C 2-5 heteroalkynyl”). In some embodiments, a heteroalkynyl group has 2 to 4 carbon atoms, at least one triple bond, and 1or 2 heteroatoms within the parent chain (“C 2-4 heteroalkynyl”). In some embodiments, a heteroalkynyl group has 2 to 3 carbon atoms, at least one triple bond, and 1 heteroatom within the parent chain (“C2-3 heteroalkynyl”).
  • a heteroalkynyl group has 2 carbon atoms, at least one triple bond, and 1 heteroatom within the parent chain (“C 2 heteroalkynyl”). In some embodiments, a heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms within the parent chain (“C1-6 heteroalkynyl”). Unless otherwise specified, each instance of a heteroalkynyl group is independently unsubstituted (an “unsubstituted heteroalkynyl”) or substituted (a “substituted heteroalkynyl”) with one or more substituents.
  • carbocyclyl refers to a radical of a non-aromatic cyclic hydrocarbon group having from 3 to 14 ring carbon atoms (“C3-14 carbocyclyl”) and zero heteroatoms in the non- aromatic ring system.
  • a carbocyclyl group has 3 to 14 ring carbon atoms (“C3-14 carbocyclyl”).
  • a carbocyclyl group has 3 to 13 ring carbon atoms (“C3-13 carbocyclyl”).
  • a carbocyclyl group has 3 to 12 ring carbon atoms (“C3-12 carbocyclyl”).
  • a carbocyclyl group has 3 to 11 ring carbon atoms (“C3-11 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 10 ring carbon atoms (“C3-10 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms (“C3-8
  • a carbocyclyl has 3 to 7 ring carbon atoms (“C3-7 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C3-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 4 to 6 ring carbon atoms (“C4-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 6 ring carbon atoms (“C5-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms (“C5-10 carbocyclyl”).
  • Exemplary C3-6 carbocyclyl groups include cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like.
  • Exemplary C3-8 carbocyclyl groups include the aforementioned C3-6 carbocyclyl groups as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptanyl (C7), bicyclo[2.2.2]octanyl (C8), and the like.
  • Exemplary C3-10 carbocyclyl groups include the aforementioned C 3-8 carbocyclyl groups as well as cyclononyl (C 9 ), cyclononenyl (C 9 ), cyclodecyl (C 10 ), cyclodecenyl (C 10 ), octahydro-1H-indenyl (C 9 ), decahydronaphthalenyl (C 10 ), spiro[4.5]decanyl (C 10 ), and the like.
  • Exemplary C 3-8 carbocyclyl groups include the aforementioned C 3-10 carbocyclyl groups as well as cycloundecyl (C 11 ), spiro[5.5]undecanyl (C 11 ), cyclododecyl (C 12 ), cyclododecenyl (C 12 ), cyclotridecane (C 13 ), cyclotetradecane (C 14 ), and the like.
  • the carbocyclyl group is either monocyclic (“monocyclic carbocyclyl”) or polycyclic (e.g., containing a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic carbocyclyl”) or tricyclic system (“tricyclic carbocyclyl”)) and can be saturated or can contain one or more carbon-carbon double or triple bonds.
  • Carbocyclyl also includes ring systems wherein the carbocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups wherein the point of attachment is on the carbocyclyl ring, and in such instances, the number of carbons continue to designate the number of carbons in the carbocyclic ring system.
  • each instance of a carbocyclyl group is independently unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a “substituted carbocyclyl”) with one or more substituents.
  • Cycloalkyl refers to a saturated carbocyclyl group.
  • a cycloalkyl group has from 3 to 14 ring carbon atoms (“C3-14 cycloalkyl”).
  • a cycloalkyl group has 3 to 10 ring carbon atoms (“C3-10 cycloalkyl”).
  • a cycloalkyl group has 3 to 8 ring carbon atoms (“C3-8 cycloalkyl”).
  • a cycloalkyl group has 3 to 7 ring carbon atoms (“C3-7 cycloalkyl”).
  • a cycloalkyl group has 3 to 6 ring carbon atoms (“C3-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 4 to 6 ring carbon atoms (“C4-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C5-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C5-10 cycloalkyl”). Examples of C5-6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5).
  • C3-6 cycloalkyl groups include the aforementioned C5-6 cycloalkyl groups as well as cyclopropyl (C3) and cyclobutyl (C4).
  • C3-8 cycloalkyl groups include the aforementioned C3-6 cycloalkyl groups as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each instance of a cycloalkyl group is
  • heterocyclyl refers to a radical of a 3- to 14-membered non- aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, silicon, boron, and phosphorous (“3-14 membered heterocyclyl”).
  • the heterocyclyl group is a radical of a 3- to 14-membered non- aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur.
  • the point of attachment can be either to a ring carbon atom or a ring heteroatom of the heterocyclyl group, as valency permits.
  • the point of attachment can be a carbon or nitrogen atom, as valency permits.
  • a heterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”) or polycyclic (e.g., a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”) or tricyclic system (“tricyclic heterocyclyl”)), and can be saturated or can contain one or more carbon-carbon double or triple bonds.
  • Heterocyclyl polycyclic ring systems can include one or more heteroatoms in one or both rings.
  • Heterocyclyl also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more carbocyclyl groups wherein the point of attachment is either on the carbocyclyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system.
  • each instance of heterocyclyl is independently unsubstituted (an “unsubstituted heterocyclyl”) or substituted (a “substituted heterocyclyl”) with one or more substituents.
  • the heterocyclyl is substituted or unsubstituted, 3- to 8-membered, monocyclic heterocyclyl, wherein 1, 2, or 3 atoms in the heterocyclic ring system are independently oxygen, nitrogen, or sulfur, as valency permits.
  • a heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-10 membered heterocyclyl”).
  • a heterocyclyl group is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heterocyclyl”).
  • a heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”).
  • the 5-6 membered heterocyclyl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the 5-6 membered heterocyclyl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the 5-6 membered heterocyclyl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur.
  • Exemplary 5-membered heterocyclyl groups containing 1 heteroatom include tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione.
  • Exemplary 5-membered heterocyclyl groups containing 2 heteroatoms include dioxolanyl, oxathiolanyl and dithiolanyl.
  • Exemplary 5-membered heterocyclyl groups containing 3 heteroatoms include triazolinyl, oxadiazolinyl, and thiadiazolinyl.
  • Exemplary 6- membered heterocyclyl groups containing 1 heteroatom include piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl.
  • Exemplary 6-membered heterocyclyl groups containing 2 heteroatoms include piperazinyl, morpholinyl, dithianyl, and dioxanyl.
  • Exemplary 6-membered heterocyclyl groups containing 3 heteroatoms include triazinyl.
  • Exemplary 7-membered heterocyclyl groups containing 1 heteroatom include azepanyl, oxepanyl and thiepanyl.
  • Exemplary 8-membered heterocyclyl groups containing 1 heteroatom include azocanyl, oxecanyl and thiocanyl.
  • Exemplary bicyclic heterocyclyl groups include indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, tetrahydrobenzo- thienyl, tetrahydrobenzofuranyl, tetrahydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridinyl, decahydro-1,8-naphthyridinyl, octahydropyrrolo[3,2-b]pyrrole, indoliny
  • aryl refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 ⁇ electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C 6-14 aryl”).
  • an aryl group has 6-10 ring carbon atoms (“C 6-10 aryl”).
  • an aryl group has 6 ring carbon atoms (“C 6 aryl”; e.g., phenyl).
  • an aryl group has 10 ring carbon atoms (“C 10 aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl).
  • an aryl group has 14 ring carbon atoms (“C 14 aryl”; e.g., anthracyl).
  • Aryl also includes ring systems wherein the aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continue to designate the number of carbon atoms in the aryl ring system.
  • each instance of an aryl group is independently unsubstituted (an “unsubstituted aryl”) or substituted (a “substituted aryl”) with one or more substituents.
  • the term “heteroaryl” refers to a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 ⁇ electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, silicon, boron, and phosphorous (“5-14 membered heteroaryl”).
  • the heteroaryl group is a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring system (e.g., having 6,
  • each heteroatom is independently selected from nitrogen, oxygen, and sulfur.
  • the point of attachment can be either to a ring carbon atom or a ring heteroatom of the heteroaryl group, as valency permits.
  • the point of attachment can be a carbon or nitrogen atom, as valency permits.
  • Heteroaryl polycyclic ring systems can include one or more heteroatoms in one or both rings.
  • Heteroaryl includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the point of attachment is on the heteroaryl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heteroaryl ring system. “Heteroaryl” also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused polycyclic (aryl/heteroaryl) ring system.
  • Polycyclic heteroaryl groups wherein one ring does not contain a heteroatom e.g., indolyl, quinolinyl, carbazolyl, and the like
  • the point of attachment can be on either ring, e.g., either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom (e.g., 5-indolyl).
  • the heteroaryl is substituted or unsubstituted, 5- or 6-membered, monocyclic heteroaryl, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently oxygen, nitrogen, or sulfur.
  • the heteroaryl is substituted or unsubstituted, 9- or 10-membered, bicyclic heteroaryl, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently oxygen, nitrogen, or sulfur.
  • a heteroaryl group is a 5-10 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”).
  • a heteroaryl group is a 5-8 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”).
  • a heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”).
  • the 5-6 membered heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the 5-6 membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, each instance of a heteroaryl group is independently unsubstituted (an “unsubstituted heteroaryl”) or substituted (a “substituted heteroaryl”) with one or more substituents. [86] Exemplary 5-membered heteroaryl groups containing 1 heteroatom include pyrrolyl, furanyl, and thiophenyl.
  • Exemplary 5-membered heteroaryl groups containing 2 heteroatoms include imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl.
  • Exemplary 5-membered heteroaryl groups containing 2 heteroatoms include imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl.
  • heteroatom containing 3 heteroatoms include triazolyl, oxadiazolyl, and thiadiazolyl.
  • Exemplary 5-membered heteroaryl groups containing 4 heteroatoms include tetrazolyl.
  • Exemplary 6-membered heteroaryl groups containing 1 heteroatom include pyridinyl.
  • Exemplary 6-membered heteroaryl groups containing 2 heteroatoms include pyridazinyl, pyrimidinyl, and pyrazinyl.
  • Exemplary 6-membered heteroaryl groups containing 3 or 4 heteroatoms include triazinyl and tetrazinyl, respectively.
  • Exemplary 7-membered heteroaryl groups containing 1 heteroatom include azepinyl, oxepinyl, and thiepinyl.
  • Exemplary 5,6- bicyclic heteroaryl groups include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl.
  • Exemplary 6,6-bicyclic heteroaryl groups include naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.
  • Exemplary tricyclic heteroaryl groups include phenanthridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl, and phenazinyl.
  • acyl groups include aldehydes (-CHO), carboxylic acids (-CO 2 H), ketones, acyl halides, esters, amides, imines, carbonates, carbamates, and ureas.
  • a group is optionally substituted unless expressly provided otherwise.
  • the term “optionally substituted” refers to being substituted or unsubstituted.
  • the term “substituted” when referring to a chemical group means that at least one hydrogen present on the group is replaced with a permissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction.
  • a “substituted” group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position.
  • the invention is not limited in any manner by the exemplary substituents described herein.
  • exemplary substituents include halogen, -CN, -NO2, -N3, -SO2H, -SO3H, P(R cc )3 + X-, -P(OR cc )3 + X-, -P(R cc )4, -P(OR cc )4, -OP(R cc )2, -OP(R cc )3 + X-, -OP(OR cc )2, -OP(OR cc )3 + X-, - OP(R cc )4, -OP(OR cc )4, -B(R aa )2, -B(OR cc )2, -BR aa (OR cc ), C1-20 alkyl, C1-20 haloalkyl, C2-20 alkenyl, C2-20 alkynyl, C1-20 heteroalkyl, C2-20 heteroalkeny
  • R aa is, independently, selected from C1-20 alkyl, C1-20 haloalkyl, C2-20 alkenyl, C2- 20 alkynyl, C1-20 heteroalkyl, C2-20 heteroalkenyl, C2-20 heteroalkynyl, C3-14 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two R aa groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring; each instance of R bb is, independently, independently,
  • the molecular weight of a substituent is lower than 250, lower than 200, lower than 150, lower than 100, or lower than 50 g/mol.
  • a substituent consists of carbon, hydrogen, fluorine, chlorine, bromine, iodine, oxygen, sulfur, nitrogen, and/or silicon atoms.
  • a substituent consists of carbon, hydrogen, fluorine, chlorine, bromine, iodine, oxygen, sulfur, and/or nitrogen atoms.
  • a substituent consists of carbon, hydrogen, fluorine, chlorine, bromine, and/or iodine atoms.
  • a substituent consists of carbon, hydrogen, fluorine, and/or chlorine atoms.
  • each nitrogen atom substituent is independently substituted (e.g., substituted with one or more halogen) or unsubstituted C1-6 alkyl or a nitrogen protecting group.
  • the substituent present on the nitrogen atom is a nitrogen protecting group. Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3 rd edition, John Wiley & Sons, 1999, incorporated herein by reference.
  • Non-limiting examples of nitrogen protecting groups include benzyl (Bn), tert-butyloxycarbonyl (BOC), carbobenzyloxy (Cbz), 9-flurenylmethyloxycarbonyl (Fmoc), trifluoroacetyl, triphenylmethyl, acetyl (Ac), benzoyl (Bz), p-methoxybenzyl (PMB), 3,4- dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP), 2,2,2-trichloroethyloxycarbonyl (Troc), triphenylmethyl (Tr), tosyl (Ts), brosyl (Bs), nosyl (Ns), mesyl (Ms), triflyl (Tf), or dansyl (Ds).
  • Bn benzyl
  • BOC tert-butyloxycarbonyl
  • Cbz carbobenzyloxy
  • Fmoc 9-flurenylmethyl
  • At least one nitrogen protecting group is Bn, Boc, Cbz, Fmoc, trifluoroacetyl, triphenylmethyl, acetyl, or Ts.
  • each oxygen atom substituent is independently substituted (e.g., substituted with one or more halogen) or unsubstituted C1-6 alkyl or an oxygen protecting group.
  • the substituent present on an oxygen atom is an oxygen protecting group.
  • Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3 rd edition, John Wiley & Sons, 1999, incorporated herein by reference.
  • an oxygen protecting group is a silyl group.
  • oxygen protecting groups include t-butyldiphenylsilyl (TBDPS), t- butyldimethylsilyl (TBDMS), triisoproylsilyl (TIPS), triphenylsilyl (TPS), triethylsilyl (TES), trimethylsilyl (TMS), triisopropylsiloxymethyl (TOM), acetyl (Ac), benzoyl (Bz), allyl carbonate, 2,2,2- trichloroethyl carbonate (Troc), 2-trimethylsilylethyl carbonate, methoxymethyl (MOM), 1-ethoxyethyl
  • At least one oxygen protecting group is silyl, TBDPS, TBDMS, TIPS, TES, TMS, MOM, THP, t-Bu, Bn, allyl, acetyl, pivaloyl, or benzoyl.
  • each sulfur atom substituent is independently substituted (e.g., substituted with one or more halogen) or unsubstituted C 1-6 alkyl or a sulfur protecting group.
  • the substituent present on a sulfur atom is a sulfur protecting group.
  • Sulfur protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3 rd edition, John Wiley & Sons, 1999, incorporated herein by reference.
  • a sulfur protecting group is acetamidomethyl, t-Bu, 3-nitro-2-pyridine sulfenyl, 2-pyridine-sulfenyl, or triphenylmethyl.
  • a “counterion” or “anionic counterion” is a negatively charged group associated with a positively charged group in order to maintain electronic neutrality.
  • An anionic counterion may be monovalent (e.g., including one formal negative charge).
  • An anionic counterion may also be multivalent (e.g., including more than one formal negative charge), such as divalent or trivalent.
  • Exemplary counterions include halide ions (e.g., F-, Cl-, Br-, I-), NO3-, ClO4-, OH-, H2PO4-, HCO3-, HSO4-, sulfonate ions (e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10- camphor sulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonic acid-2-sulfonate, and the like), carboxylate ions (e.g., acetate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, gluconate, and the like), BF4-, PF4-, PF6-, AsF6-, SbF6
  • Exemplary counterions which may be multivalent include CO3 2- , HPO4 2- , PO4 3- , B4O7 2- , SO4 2- , S2O3 2- , carboxylate anions (e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelate, sebacate, salicylate, phthalates, aspartate, glutamate, and the like), and carboranes.
  • carboxylate anions e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelate, sebacate, salicylate, phthalates, aspartate, glutamate, and the like
  • carboxylate anions e.g., tartrate, citrate, fumarate, maleate, malate, malonate
  • Amino acid sidechains include side chains selected from those of the following amino acids: glycine, alanine, valine, cysteine, leucine, iso leucine, serine, threonine, methionine, glutamic acid, aspartic acid, glutamine, asparagine, lysine, arginine, proline, histidine, phenylalanine, tyrosine, and tryptophan, and those amino acids and amino acid analogs which have been identified as constituents of peptidylglycan bacterial cell walls.
  • Amino acid residues having “basic sidechains” include Arg, Lys and His.
  • Amino acid residues having “acidic sidechains” include Glu and Asp.
  • Amino acid residues having “neutral polar sidechains” include Ser, Thr, Asn, Gln, Cys and Tyr.
  • Amino acid residues having “neutral non-polar sidechains” include Gly, Ala, Val, Ile, Leu, Met, Pro, Trp and Phe.
  • Amino acid residues having “non-polar aliphatic sidechains” include Gly, Ala, Val, Ile and Leu.
  • Amino acid residues having “hydrophobic sidechains” include Ala, Val, Ile, Leu, Met, Phe, Tyr and Trp.
  • Amino acid residues having “small hydrophobic sidechains” include Ala and Val.
  • Amino acid residues having “aromatic sidechains” include Tyr, Trp and Phe.
  • aromatic sidechains include Tyr, Trp and Phe.
  • These and other exemplary substituents are described in more detail in the Detailed Description, Drawings, Examples, and Claims. The embodiments provided herein are not limited in any manner by the above exemplary listing of substituents.
  • Compounds and Constructs [101] As described herein, provided herein are compounds and constructs comprising a camptothecin conjugated to a FAP-cleavable moiety.
  • CAM is a camptothecin
  • alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, carbocyclylene, heterocyclylene, arylene, heteroarylene, PEG, or PSar is independently optionally substituted;
  • L 1 is a bond, C3-10 carbocyclylene, C6-10 arylene, 3- to 10-membered heterocyclylene, or 5- to 10- membered heteroarylene, wherein the carbocyclylene, arylene, heterocyclylene, or heteroarylene is optionally substituted;
  • R 2 and each instance of R 2A are independently hydrogen or optionally substituted C1-C6 alkyl; or optionally wherein L A and R 2 are joined together, or L A and R 2A are joined together, with the intervening atoms to form a 5- to 10-membered heterocyclic ring, wherein the heterocyclic ring is optionally substituted;
  • R 3 is hydrogen or optionally substituted C1-6 alkyl; each instance of R 3A is hydrogen, optionally substituted C1-6 al
  • the compound of Formula (I′), is of Formula (I), or a pharmaceutically acceptable salt thereof.
  • CAM is a camptothecin
  • R A is a reactive handle
  • L A is a bond, C1-30 alkylene, C1-30 haloalkylene, C2-30 alkenylene, C2-30 alkynylene, C1-30 heteroalkylene, C2-30 heteroalkenylene, C2-30 heteroalkynylene, C3-10 carbocyclylene, 3- to 10-membered heterocyclylene, C6-10 arylene, 5- to 10-membered heteroarylene, polyethylene glycol (PEG), or any combination thereof, wherein each alkylene, haloalkylene alkenylene, alkynylene, heteroalkylene,
  • heteroalkenylene, heteroalkynylene, carbocyclylene, heterocyclylene, arylene, heteroarylene, and PEG is independently optionally substituted;
  • L 1 is a bond, C3-10 carbocyclylene, C6-10 arylene, 3- to 10-membered heterocyclylene, or 5- to 10- membered heteroarylene, wherein the carbocyclylene, arylene, heterocyclylene, or heteroarylene is optionally substituted;
  • R 2 and each instance of R 2A are independently hydrogen or optionally substituted C1-C6 alkyl; or optionally wherein L A and R 2 are joined together, or L A and R 2A are joined together, with the intervening atoms to form a 5- to 10-membered heterocyclic ring, wherein the heterocyclic ring is optionally substituted;
  • R 3 is hydrogen or optionally substituted C1-6 alkyl; each instance of R 3A is hydrogen, optionally substituted C1-6 alkyl, or an amino acid sidechain; each instance of R
  • the compound is of Formula (I-C): or a pharmaceutically acceptable salt thereof, wherein -N(H)-L 2 - is a bond or a self-immolative linker.
  • the compound is of Formula (I-C): or a pharmaceutically acceptable salt thereof, wherein -N(H)-L 2 - is a bond or a self-immolative linker.
  • the compound is of Formula (I-D): , or a pharmaceutically acceptable salt thereof.
  • the compound is of Formula (I-E): , or a pharmaceutically acceptable salt thereof.
  • the compound is of Formula (I-F): or a pharmaceutically acceptable salt thereof.
  • the compound is of Formula (I-G): , or a pharmaceutically acceptable salt thereof.
  • the compound is of Formula (I-H): , or a pharmaceutically acceptable salt thereof, wherein -N(H)-L 2 - is a bond or a self-immolative linker.
  • the compound is of Formula (I-I): , or a pharmaceutically acceptable salt thereof, wherein -N(H)-L 2 - is a bond or a self-immolative linker.
  • the construct of Formula (II′), is of Formula (II), or a pharmaceutically acceptable salt thereof.
  • constructs of Formula (II): or a pharmaceutically acceptable salt thereof, wherein: Z is a binding moiety; R B is a diradical of a reactive handle; CAM is a camptothecin; L A is a bond, C1-30 alkylene, C1-30 haloalkylene, C2-30 alkenylene, C2-30 alkynylene, C1-30 heteroalkylene, C2-30 heteroalkenylene, C2-30 heteroalkynylene, C3-10 carbocyclylene, 3- to 10-membered heterocyclylene, C6-10 arylene, 5- to 10-membered heteroarylene, polyethylene glycol (PEG), or any combination thereof, wherein each alkylene, haloalkylene alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroal
  • L A and R 2 are joined together, or L A and R 2A are joined together, with the intervening atoms to form a 5- to 10-membered heterocyclic ring, wherein the heterocyclic ring is optionally substituted;
  • R 3 is hydrogen or optionally substituted C1-6 alkyl; each instance of R 3A is hydrogen, optionally substituted C1-6 alkyl, or an amino acid sidechain;
  • each instance of R 4 is independently halogen, C1-6 alkyl, C1-6 haloalkyl, -OR O , or -N(R N )2, wherein the alkyl or haloalkyl is optionally substituted;
  • m is 0, 1, 2, 3, 4, 5, 6, or 7;
  • n is 0, 1, or 2;
  • L 2 is
  • the construct is of Formula (II-A): , or a pharmaceutically acceptable salt thereof.
  • the construct is of Formula (II-B): or a pharmaceutically acceptable salt thereof, wherein -N(H)-L 2 - is a bond or a self-immolative linker.
  • the construct is of Formula (II-C): , or a pharmaceutically acceptable salt thereof, wherein -N(H)-L 2 - is a bond or a self-immolative linker.
  • the construct is of Formula (II-D): , or a pharmaceutically acceptable salt thereof.
  • the construct is of Formula (II-E): , or a pharmaceutically acceptable salt thereof.
  • the construct is of Formula (II-F): , or a pharmaceutically acceptable salt thereof.
  • the construct is of Formula (II-G): , or a pharmaceutically acceptable salt thereof.
  • the construct is of Formula (II-H): , or a pharmaceutically acceptable salt thereof, wherein -N(H)-L 2 - is a bond or a self-immolative linker.
  • the construct is of Formula (II-I): or a pharmaceutically acceptable salt thereof, wherein -N(H)-L 2 - is a bond or a self-immolative linker.
  • Camptothecin (CAM) [126] As generally defined herein, CAM is a camptothecin. In certain embodiments, CAM is any camptothecin provided herein. In certain embodiments, CAM is exatecan, SN-38, Dxd, belotecan, or topotecan. In certain embodiments, CAM is exatecan, SN-38, Dxd, or belotecan. In certain embodiments,
  • R′ is hydrogen, C 1 - 30 alkyl, C 1-30 haloalkyl, C 1-30 heteroalkyl, C 2-30 alkenyl, C 2-30 heteroalkenyl, C 2-30 alkynyl, C 2-30 heteroalkynyl, C 3-10 carbocyclyl, 3- to 10-membered heterocyclyl, C 6-10 aryl, 5- to 10-membered heteroaryl, PEG, or PSar or any combination thereof, wherein each alkyl, haloalkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, PEG, or PSar is independently optionally substituted.
  • R′ is hydrogen, R 1A , C 1 - 20 alkyl, C 1-20 haloalkyl, C 1-20 heteroalkyl, C 2-20 alkenyl, C 2-20 alkynyl, PEG, or PSar, wherein the alkyl, haloalkyl, heteroalkyl, alkenyl, alkynyl, PEG, or PSar, is optionally substituted.
  • R′ is C 1 - 20 alkyl, C 1-20 haloalkyl, C 1-20 heteroalkyl, C 2-20 alkenyl, C 2-20 alkynyl, PEG, or PSar, wherein the alkyl, haloalkyl, heteroalkyl, alkenyl,
  • R′ is C1-20 alkyl, C1-20 haloalkyl, C1-20 heteroalkyl, C2-20 alkenyl, C2-20 alkynyl, PEG, or PSar, wherein the alkyl, haloalkyl, heteroalkyl, alkenyl, or alkynyl, is optionally substituted.
  • R′ is C1-10 alkyl, C2-10 alkenyl, or C2-10 alkynyl, wherein the alkyl, alkenyl, or alkynyl is optionally substituted.
  • R′ is C1-10 alkyl or C1-10 haloalkyl.
  • R′ is -Me, -Et, -Pr, or -Bu. In some embodiments, R′ is - Me or -Et. In some embodiments, R′ is -Me or -CF3. In some embodiments, R′ is C1-10 heteroalkyl, C2-10 heteroalkenyl, or C2-10 heteroalkynyl, wherein the heteroalkyl, heteroalkenyl, or heteroalkynyl is optionally substituted. [129] In some embodiments, R′ is hydrogen or halogen. In some embodiments, R′ is halogen. In some embodiments, R′ is -F, -Cl, or -Br. In some embodiments, R′ is hydrogen or -F.
  • R′ is C 3-10 carbocyclyl or 3- to 10-membered heterocyclyl, wherein each carbocyclyl or heterocyclyl is independently optionally substituted.
  • R′ is C3-7 carbocyclyl, wherein each carbocyclyl is optionally substituted.
  • R′ is C 5-6 carbocyclyl, wherein each carbocyclyl is optionally substituted.
  • R′ is 3- to 7- membered heterocyclyl, wherein each heterocyclyl is optionally substituted.
  • R′ is 5- to 6-membered heterocyclyl, wherein each heterocyclyl is optionally substituted.
  • R′ is C6-10 aryl or 5- to 10-membered heteroaryl, wherein each aryl or heteroaryl is optionally substituted.
  • R′ is optionally substituted phenyl.
  • R′ is optionally substituted napthyl.
  • R′ is 5- to 6-membered heteroaryl, wherein each heteroaryl is optionally substituted.
  • R′ is 9- to 10- membered heteroaryl, wherein each heteroaryl is optionally substituted.
  • R′ is PEG or PSar, wherein the PEG or PSar is independently optionally substituted. In some embodiments, R′ is PEG or PSar. In some embodiments, R′ is PEG, wherein the PEG is optionally substituted. In some embodiments, R′ is PEG. In some embodiments, R′ is . In some embodiments, R′ is PSar, wherein the PSar is optionally substituted. In some embodiments, R′ is PSar. [134] In some embodiments, R′ is -L A -R A or hydrogen. In some embodiments, R′ is -L A -R A . In some embodiments, R′ is hydrogen.
  • R′ is hydrogen; and L 2 is a self-immolative linker substituted with -L A -R A .
  • R′′ is hydrogen, C1-30 alkyl, C1-30 haloalkyl, C 1-30 heteroalkyl, C 2-30 alkenyl, C 2-30 heteroalkenyl, C 2-30 alkynyl, C 2-30 heteroalkynyl, C 3-10 carbocyclyl, 3- to 10-membered heterocyclyl, C 6-10 aryl, 5- to 10-membered heteroaryl, PEG, or PSar or any combination thereof, wherein each alkyl, haloalkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, PEG, or PSar is independently optionally substituted.
  • R′′ is hydrogen, R 1A , C1-20 alkyl, C1-20 haloalkyl, C1-20 heteroalkyl, C2-20 alkenyl, C2-20 alkynyl, PEG, or PSar, wherein the alkyl, haloalkyl, heteroalkyl, alkenyl, alkynyl, PEG, or PSar, is optionally substituted.
  • R′′ is C1-20 alkyl, C1-20 haloalkyl, C1-20 heteroalkyl, C2-20 alkenyl, C2-20 alkynyl, PEG, or PSar, wherein the alkyl, haloalkyl, heteroalkyl, alkenyl, or alkynyl, is optionally substituted.
  • R′′ is C1-20 alkyl, C1-20 haloalkyl, C1-20 heteroalkyl, C2-20 alkenyl, C2-20 alkynyl, PEG, or PSar, wherein the alkyl, haloalkyl, heteroalkyl, alkenyl, or alkynyl, is optionally substituted.
  • R′′ is C1-10 alkyl, C2-10 alkenyl, or C2-10 alkynyl, wherein the alkyl, alkenyl, or alkynyl is optionally substituted. In some embodiments, R′′ is C1-10 alkyl or C1-10 haloalkyl. In some embodiments, R′′ is -Me, -Et, -Pr, or -Bu. In some embodiments, R′′ is - Me or -Et. In some embodiments, R′′ is -Me or -CF3.
  • R′′ is C1-10 heteroalkyl, C2-10 heteroalkenyl, or C2-10 heteroalkynyl, wherein the heteroalkyl, heteroalkenyl, or heteroalkynyl is optionally substituted.
  • R′′ is hydrogen or halogen. In some embodiments, R′′ is halogen. In some embodiments, R′′ is -F, -Cl, or -Br. In some embodiments, R′′ is hydrogen or -F.
  • R′′ is -OR O or - CO2R O .
  • R′′ is C 3-10 carbocyclyl or 3- to 10-membered heterocyclyl, wherein each carbocyclyl or heterocyclyl is independently optionally substituted. In some embodiments, R′′ is C 3-7 carbocyclyl, wherein each carbocyclyl is optionally substituted. In some embodiments, R′′ is C 5-6 carbocyclyl, wherein each carbocyclyl is optionally substituted. In some embodiments, R′′ is 3- to 7- membered heterocyclyl, wherein each heterocyclyl is optionally substituted. In some embodiments, R′′ is 5- to 6-membered heterocyclyl, wherein each heterocyclyl is optionally substituted.
  • R′′ is C 6-10 aryl or 5- to 10-membered heteroaryl, wherein each aryl or heteroaryl is optionally substituted. In some embodiments, R′′ is optionally substituted phenyl. In some embodiments, R′′ is optionally substituted napthyl. In some embodiments, R′′ is 5- to 6-membered heteroaryl, wherein each heteroaryl is optionally substituted. In some embodiments, R′′ is 9- to 10- membered heteroaryl, wherein each heteroaryl is optionally substituted. [142] In some embodiments, R′′ is PEG or PSar, wherein the PEG or PSar is independently optionally substituted. In some embodiments, R′′ is PEG or PSar.
  • R′′ is PEG, wherein the PEG is optionally substituted. In some embodiments, R′′ is PEG. In some embodiments, R′′ is . In some embodiments, R′′ is PSar, wherein the PSar is optionally substituted. In some embodiments, R′′ is PSar. [143] In some embodiments, R′′ is -L A -R B -Z or hydrogen. In some embodiments, R′′ is -L A -R B -Z. In some embodiments, R′′ is hydrogen.
  • L 2 is a bond, or -N(H)-L 2 - is a bond or a self-immolative linker. In some embodiments, L 2 is a bond, or -N(H)-L 2 - is a bond or a self-immolative linker, wherein the self- immolative linker is optionally substituted with -L A -R A . In some embodiments, L 2 is a bond, or -N(H)-L 2 - is a bond or a self-immolative linker, wherein the self-immolative linker is optionally substituted with - L A -R B -Z.
  • L 2 is a bond, or -N(H)L 2 - is a bond. In certain embodiments, L 2 is a bond, or -N(H)-L 2 - is a self-immolative linker. In certain embodiments, L 2 is a bond, or -N(H)-L 2 - is a self-immolative linker, wherein the self-immolative linker is optionally substituted with -L A -R A . In certain embodiments, L 2 is a bond, or -N(H)-L 2 - is a self-immolative linker, wherein the self-immolative linker is optionally substituted with -L A -R B -Z.
  • -N(H)-L 2 - is a bond or a self- immolative linker. In certain embodiments, -N(H)-L 2 - is a bond or a self-immolative linker optionally substituted with -L A -R A . In certain embodiments, -N(H)-L 2 - is a bond or a self-immolative linker optionally substituted with -L A -R B -Z. In certain embodiments, L 2 is a bond. In certain embodiments, - N(H)-L 2 - is a bond. In certain embodiments, -N(H)-L 2 - is a self-immolative linker.
  • -N(H)-L 2 - is a self-immolative linker optionally substituted with -L A -R A . In certain embodiments, -N(H)-L 2 - is a self-immolative linker optionally substituted with -L A -R B -Z. In certain embodiments, the self-immolativel linker is not substituted with -L A -R A . In some embodiments, the self- immolative linker is substituted with -L A -R A . In certain embodiments, the self-immolative linker is not substituted with -L A -R B -Z.
  • the self-immolative linker is substituted with -L A -R B - Z. In certain embodiments, the self-immolative linker is cleaved to release CAM.
  • each instance of Ring A is independently 5- or 6-membered heterocyclyl, 5- or 6-membered heteroaryl, or phenyl. In certain embodiments, each instance of Ring A is independently 5- or 6-membered heterocyclyl or 5- or 6-membered heteroaryl. In certain embodiments, each instance of Ring A is independently 5- or 6-membered heteroaryl or phenyl. In certain embodiments, at least one instance of Ring A is phenyl. In certain embodiments, at least one instance of Ring A is 5- or 6-membered heteroaryl. In certain embodiments, at least one instance of Ring A is 5- or 6- membered N-heteroaryl.
  • At least one instance of Ring A is pyridyl, pyrrolyl, or thiazolyl. In certain embodiments, at least one instance of Ring A is 5- to 6-membered heterocyclyl. In certain embodiments, at least one instance of Ring A is 5- to 6-membered N-heterocyclyl. In certain embodiments, at least one instance of Ring A is pyrrolidinyl. [150] As generally defined herein, r is 0, 1, 2, or 3. In certain embodiments, r is 1 or 2. In certain embodiments, r is 1. In certain embodiments, r is 2.
  • each instance of Y is independently a bond, optionally substituted C 1-8 alkylene, or optionally substituted C 1-8 heteroalkylene. In certain embodiments, each instance of Y is optionally substituted C 1-8 alkylene or optionally substituted C 1-8 heteroalkylene. In certain embodiments, each instance of Y is optionally substituted C 1-4 alkylene or optionally substituted C 1-4 heteroalkylene.
  • at least one instance of Y is a bond.
  • -N(H)-L 2 - is of the formula: , p .
  • -N(H)-L 2 - is of the formula: erein * denotes the point of attachment to CAM.
  • mula: or wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula:
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM. In certain embodiments, -N(H)-L 2 - is of the formula: denotes the point of attachment to CAM. In certain embodiments, -N(H)-L 2 - is of the formula:
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM. In certain embodiments, -N(H)-L 2 - is of the formula: , wherein * denotes the point
  • -N(H)-L 2 - is of the formula: denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: , wherein * denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: denotes the point of attachment to CAM.
  • -N(H)-L 2 - is of the formula: denotes the point of attachment to CAM.
  • p is 0, 1, 2, 3, or 4. In certain embodiments, p is 0, 1, 2, or 3. In certain embodiments, p is 0, 1, or 2. In certain embodiments, p is 0. In certain embodiments, p is 1. In certain embodiments, p is 2. In certain embodiments, p is 3. In certain embodiments, p is 4.
  • At least one instance of R 5 is of the formula: , wherein q1 is an integer from 1-25, inclusive, and each instance of R N is independently H, optionally substituted C1-6 alkyl, optionally substituted C3-7 carbocyclyl, or optionally substituted C1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • R 5 is -L 5 -R 5A .
  • L 5 is a bond, C1-20 alkylene, C1-20 haloalkylene, C1-20 heteroalkylene, PEG, or PSar, wherein the alkylene, haloalkylene, or heteroalkylene is optionally substituted.
  • L 5 is a bond, C1-20 alkylene, C1-20 haloalkylene, C1-20 heteroalkylene, or PEG, wherein the alkylene, haloalkylene, or heteroalkylene is optionally substituted.
  • L 5 is a bond.
  • L 5 is C1-10 alkylene, C1-10 haloalkylene, C1-10 heteroalkylene, or PEG.
  • L5 is of the formula: or , wherein q1 is an integer from 1-25,
  • At least one instance of R 5A is PEG. In certain embodiments, at least one instance of R 5A is of the formula: , wherein q1 is an integer from 1-25, inclusive, and R O is H, optionally substituted C1- 6 alkyl, optionally substituted C1-6 haloalkyl, optionally substituted C3-7 carbocyclyl, or optionally substituted C1-6 acyl.
  • At least one instance of R 5A is of the formula: , wherein q1 is an integer from 1-25, inclusive, and each instance of R N is independently H, optionally substituted C 1-6 alkyl, optionally substituted C 3-7 carbocyclyl, or optionally substituted C 1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • at least one instance of R 5 is of the formula: or , wherein q1 is an integer from 1-25, inclusive.
  • at least one instance of R 5 is of the formula: or , wherein q1 is an integer from 1-15, inclusive.
  • at least one instance of R 5 is .
  • at least one instance . [164] In certain embodiments, at least one instance of R 5 is PSar. In certain embodiments, at least one
  • R 5 is of the formula , wherein q3 is an integer from 1-25, inclusive, and each instance of R N is H, optionally substituted C1-6 alkyl, optionally substituted C3-7 carbocyclyl, optionally substituted C1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • At least one instance of R 5 is of the formula , wherein q3 is an integer from 1-25, inclusive, and each instance of R N is H, optionally substituted C 1-6 alkyl, optionally substituted C 3-7 carbocyclyl, optionally substituted C 1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • at least one instance of R 5 is of the formula t e o ua .
  • R 4 , m [165] As generally defined herein, m is 0, 1, 2, 3, 4, 5, 6, or 7. In certain embodiments, m is 0, 1, 2, 3, 4, 5, or 6. In certain embodiments, m is 0, 1, 2, 3, 4, or 5.
  • m is 0, 1, 2, 3, or 4. In certain embodiments, m is 0, 1, 2, or 3. In certain embodiments, m is 0, 1, or 2. In certain embodiments, m is 0 or 1. In certain embodiments, m is 0. In certain embodiments, m is 1. In certain embodiments, m is 2. In certain embodiments, m is 3. In certain embodiments, m is 4. In certain embodiments, m is 5. In certain embodiments, m is 6. In certain embodiments, m is 7.
  • each instance of R 4 is independently halogen, C1-6 alkyl, C1-6 haloalkyl, -OR O , or -N(R N )2, wherein the alkyl or haloalkyl is optionally substituted.
  • at least one instance of R 4 is Br, Cl, F, Me, Et, -CF3, -OEt, -OMe, -OH, -NMe2, -NHMe, or -NH2.
  • at least one instance of R 4 is optionally substituted C1-4 alkyl.
  • At least one instance of R 4 is Me, Et, n-Pr, i-Pr, n-Bu, i-Bu, or t-Bu. In certain embodiments, at least one instance of R 4 is Me or Et. In certain embodiments, at least one instance of R 4 is optionally substituted C 1-4 haloalkyl. In certain embodiments, at least one instance of R 4 is -CF 3 . In certain embodiments, at least one instance of R 4 is halogen. In certain embodiments, at least one instance of R 4 is Br, Cl, or F. In certain embodiments, at least one instance of R 4 is F. In certain embodiments,
  • each instance of R 4 is the same. In certain embodiments, at least one instance of R 4 is different. In certain embodiments, each instance of R 4 is different.
  • L 1 is a bond, C 3-10 carbocyclylene, C 6-10 arylene, 3- to 10-membered heterocyclylene, or 5- to 10-membered heteroarylene, wherein the carbocyclylene, arylene, heterocyclylene, or heteroarylene is optionally substituted.
  • L 1 is a bond.
  • L 1 is C 3-10 carbocyclylene, C 6-10 arylene, 3- to 10-membered heterocyclylene, or 5- to 10-membered heteroarylene, wherein the carbocyclylene, arylene, heterocyclylene, or heteroarylene is optionally substituted.
  • L 1 is C 3-10 carbocyclylene or C 6-10 arylene, wherein the carbocyclylene or arylene are optionally substituted. In certain embodiments, L 1 is C 3-10 carbocyclylene or 3- to 10-membered heterocyclylene, wherein the carbocyclylene or heterocyclylene are optionally substituted. In certain embodiments, L 1 is C 6-10 arylene or 5- to 10-membered heteroarylene, wherein the arylene or heteroarylene are optionally substituted. In certain embodiments, L 1 is 5- to 10-membered heteroarylene or 5- to 6-membered heterocyclylene, wherein the heteroarylene or heterocyclylene are optionally substituted.
  • L 1 is phenylene, wherein the phenylene is optionally substituted. In certain embodiments, L 1 is naphthylene, wherein the phenylene is optionally substituted. In certain embodiments, L 1 is C3-C7 carbocyclylene, wherein the carbocyclylene is optionally substituted. In certain embodiments, L 1 is 5- to 10-membered heteroarylene, wherein the heteroarylene is optionally substituted. In certain embodiments, L 1 is 5- or 6-membered heteroarylene, wherein the heteroarylene is optionally substituted. In certain embodiments, L 1 is 3- to 7-membered heterocyclylene, wherein the heterocyclylene is optionally substituted. , each of which is independently optionally substituted, wherein * denotes the point of
  • L 1 is of the formula: attachment to L A . In certain embodiments, L 1 is of the formula: denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: denotes the point of attachment to L A . In certain embodiments, L 1 is of the , wherein * denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: , wherein * denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: , wherein * denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: , wherein * denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: , wherein * denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: , wherein * denotes the point of attachment to L
  • L 1 is of the formula: , wherein * denotes the point of attachment to LA. In certain embodiments, L1 is of the formula: , wherein * denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: , wherein * denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: , wherein * denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: , wherein * denotes the point of attachment to L A . In certain embodiments, L 1 is of the formula: , wherein * denotes the point of attachment to L A .
  • L A is a bond, C 1-30 alkylene, C 1-30 haloalkylene, C 2-30 alkenylene, C 2- 30 alkynylene, C 1-30 heteroalkylene, C 2-30 heteroalkenylene, C 2-30 heteroalkynylene, C 3-10 carbocyclylene, 3- to 10-membered heterocyclylene, C 6-10 arylene, 5- to 10-membered heteroarylene, polyethylene glycol (PEG), polysarcosine (PSar), or any combination thereof, wherein each alkylene, haloalkylene alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, carbocyclylene, heterocyclylene, arylene, heteroarylene, PEG, or PSar is independently optionally substituted; optionally wherein L A and R 2 are joined together, or L A and R 2A are joined together, with the intervening atoms to form a 5- to 10-membered heterocycl
  • L A is a bond, C 1-30 alkylene, C 1-30 haloalkylene, C 2-30 alkenylene, C 2-30 alkynylene, C 1-30 heteroalkylene, C 2-30 heteroalkenylene, C 2-30 heteroalkynylene, C 3-10 carbocyclylene, 3- to 10- membered heterocyclylene, C 6-10 arylene, 5- to 10-membered heteroarylene, polyethylene glycol (PEG), or any combination thereof, wherein each alkylene, haloalkylene alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, carbocyclylene, heterocyclylene, arylene, heteroarylene, and PEG is independently optionally substituted; optionally wherein L A and R 2 are joined together, or L A and R 2A are joined together, with the intervening atoms to form a 5- to 10-membered heterocyclic ring, wherein the heterocyclic ring is optionally substituted.
  • L A is a bond, C1-30 alkylene, C1-30 haloalkylene, C2-30 alkenylene, C2-30 alkynylene, C1-30 heteroalkylene, C2-30 heteroalkenylene, C2-30 heteroalkynylene, C3-10 carbocyclylene, 3- to 10-membered heterocyclylene, C6-10 arylene, 5- to 10-
  • heteroarylene polyethylene glycol (PEG), or any combination thereof, wherein each alkylene, haloalkylene alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, carbocyclylene, heterocyclylene, arylene, heteroarylene, and PEG is independently optionally substituted.
  • PEG polyethylene glycol
  • L A is a bond, C1-20 alkylene, C1-20 haloalkylene, C2-20 alkenylene, C2-20 alkynylene, C1-20 heteroalkylene, C2-20 heteroalkenylene, C2-20 heteroalkynylene, C3-10 carbocyclylene, 3- to 10-membered heterocyclylene, C6-10 arylene, 5- to 10-membered heteroarylene, polyethylene glycol (PEG), or any combination thereof, wherein each alkylene, haloalkylene alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, carbocyclylene, heterocyclylene, arylene, heteroarylene, and PEG is independently optionally substituted.
  • L A is C1-20 alkylene, C1-20 haloalkylene, C1-20 heteroalkylene, C2-20 alkenylene, C2-20 alkynylene, polyethylene glycol (PEG), or any combination thereof, wherein the alkylene, haloalkylene, heteroalkylene, alkenylene, or alkynylene is optionally substituted.
  • L A is a bond.
  • L A comprises C 1-10 alkylene, C 2-10 alkenylene, or C 2-10 alkynylene, wherein the alkylene, alkenylene, or alkynylene is optionally substituted.
  • L A comprises C 1-4 alkylene.
  • L A comprises -CH 2 -. In certain embodiments, L A comprises C 1-6 haloalkylene. [173] In certain embodiments, L A comprises C1-10 heteroalkylene, C2-10 heteroalkenylene, C2-10 heteroalkynylene. In certain embodiments, L A comprises C 1-10 heteroalkylene, C 2-10 heteroalkenylene, C 2- 10 heteroalkynylene, wherein the heteroalkylene, heteroalkenylene, or heteroalkynylene comprises at least one oxygen atom.
  • L A comprises C 1-10 heteroalkylene, C 2-10 heteroalkenylene, C 2- 10 heteroalkynylene, wherein the heteroalkylene, heteroalkenylene, or heteroalkynylene comprises at least one nitrogen atom.
  • L A comprises -O-.
  • L A comprises - CO2-.
  • L A comprises -N(R N )-, wherein R N is H, optionally substituted C1-6 alkyl, optionally substituted C3-7 carbocyclyl, or optionally substituted C1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • L A comprises C3-10 carbocyclylene, 3- to 10-membered heterocyclylene, C6-10 arylene, or 5- to 10-membered heteroarylene.
  • L A comprises C3-7 carbocyclylene.
  • L A comprises cyclohexylene.
  • L A comprises .
  • L A comprises 3- to 7-membered heterocyclylene. In certain embodiments, L A comprises 5- to 7-membered heterocyclylene. In certain embodiments, L A comprises phenylene. In certain embodiments, L A comprises . In certain embodiments, L A comprises naphthylene. In certain embodiments, L A comprises 5- to 6-membered heteroarylene.
  • L A comprises 10-membered heteroarylene. In certain embodiments, L A comprises 5- to 10-membered heteroarylene comprising at least one nitrogen atom. In certain embodiments, L A comprises pyrrolylene, pyrazolylene, imidazolylene, or triazolylene. In certain embodiments, L A comprises . In certain embodiments, L A comprises pyridinylene, pyrazinylene, or pyrimidinylene. In certain embodiments, L A comprises quinolinylene. [175] In certain embodiments, L A comprises PEG or PSar. In certain embodiments, L A comprises PEG. In certain embodiments, L A comprises the formula: , wherein q1 is an integer from 1-25. In certain embodiments, L A comprises PEG.
  • L A comprises the formula: wherein q1 is an integer from 1-15. In certain embodiments, L A comprises the formula: wherein q1 is an integer from 5-10. [176] In certain embodiments, L A comprises PSar. In certain embodiments, L A comprises the formula , wherein q3 is an integer from 1-25, inclusive, and each instance of R N is H, optionally substituted C 1-6 alkyl, optionally substituted C 3-7 carbocyclyl, optionally substituted C 1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl. In certain embodiments, L A comprises the formula , wherein q3 is an integer from 1-25, inclusive. In certain embodiments, L A comprises the formula , wherein q3 is an integer from 1-10, inclusive. In certain embodiments, L A comprises the formula . certain embodiments, L A comprises the formula .
  • L A comprises the formula from 1-25, inclusive, and each instance of R N is H, optionally substituted C1-6 alkyl, optionally substituted C3-7 carbocyclyl, optionally substituted C1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • L A comprises the formula , wherein q3 is an integer from 1-25, inclusive, and each instance of R N is H, optionally substituted C 1-6 alkyl, optionally substituted C 3-7 carbocyclyl, optionally substituted C 1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • L A comprises the formula e o ua .
  • L A comprises the formula , wherein q3 is an integer from 1-25, inclusive, and each instance of R N is H, optionally substituted C1-6 alkyl, optionally substituted C 3-7 carbocyclyl, optionally substituted C 1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • LA comprises the formula , wherein q3 is an integer from 1-25, inclusive, and each instance of R N is H, optionally substituted C1-6 alkyl, optionally substituted C3-7 carbocyclyl, optionally substituted C1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • L A comprises the
  • L A comprises the formula . [179] In certain embodiments, L A comprises: . certain embodiments, L A is of the formula: , wherein q1 is an integer from 5-10. [180] As generally defined herein, q1 is an integer from 1-25, inclusive. In certain embodiments, q1 is an integer from 1-20, inclusive. In certain embodiments, q1 is an integer from 1-15, inclusive. In certain embodiments, q1 is an integer from 1-15, inclusive. In certain embodiments, q1 is an integer from 1-10, inclusive. In certain embodiments, q1 is an integer from 1-5, inclusive. In certain embodiments, q1 is an integer from 2-25, inclusive.
  • q1 is an integer from 2-20, inclusive. In certain embodiments, q1 is an integer from 2-15, inclusive. In certain embodiments, q1 is an integer from 2-10, inclusive. In certain embodiments, q1 is an integer from 2-5, inclusive. In certain embodiments, q1 is an integer from 5-25, inclusive. In certain embodiments, q1 is an integer from 5-20, inclusive. In certain embodiments, q1 is an integer from 5-15, inclusive. In certain embodiments, q1 is an integer from 5-10, inclusive. In certain embodiments, q1 is 1. In certain embodiments, q1 is 2. In certain embodiments, q1 is 3. In certain embodiments, q1 is 4. In certain embodiments, q1 is 5.
  • R A is a reactive handle.
  • R B is a diradical of a reactive handle.
  • reactive handle refers to any chemical moiety capable of reacting with another chemical moiety to form one or more covalent bonds.
  • the reactive handle is any reactive handle capable of conjugating a compound provided herein (e.g., a compound of Formula (I′) or (I)), or a pharmaceutically acceptable salt thereof, to a binding moiety to form a construct as provided herein (e.g., a construct of Formula (II′) or (II)), or a pharmaceutically acceptable salt thereof.
  • the reactive handle undergoes bioconjugation, such as to a protein, e.g., an antibody.
  • the reactive handle is capable of reacting with a cysteine residue (cysteine bioconjugation).
  • the reactive handle is capable of reacting with a lysine residue (lysine bioconjugation).
  • the reactive handle is capable of reacting with a tyrosine or tryptophan residue (tyrosine or tryptophan bioconjugation).
  • the reactive handle is site specific.
  • the reactive handle is a cysteine-selective reactive handle.
  • the reactive handle is a lysine-selective reactive handle. In certain embodiments, the reactive handle is a tyrosine-selective or tryptophan-selective reactive handle.
  • Reactive handles are disclosed, for example, in Rostovtsev, V. V. et al. Angew. Chem.2002, 114, 2708- 2711, Bernardin, B. et al. Nat. Protocol.2019, 14, 86-99, Szijj, P. A. et al. Org. Biomol. Chem.2020, 18, 9018-9028, and Yang, Q. et al. Research 2024, 7, 0410, each of which is incorporated herein by reference in its entirety.
  • Non-limiting examples of reactive moieties include alkenes, alkynes, alcohols, amines, thiols, azides, esters, amides, halogens, and the like.
  • the reactive handle comprises a maleimide, NHS-ester, amine, isocyanate, isothiocyanate, benzoyl fluoride, iodoacetamide, thiol, propionitrile, diazonium salt, aldehyde, aniline, ketone, iodoacetamide, alkyne, or azide moiety.
  • the reactive handle comprises a maleimide.
  • the reactive handle comprises an azide.
  • the reactive handle comprises an alkyne.
  • the reactive handle undergoes click chemistry.
  • Click chemistry is a describes chemistry tailored to generate substances quickly and reliably by joining small units together. See, e.g., Kolb, Finn and Sharpless Angewandte Chemie International Edition 2001, 40, 2004–2021; Evans, Australian Journal of Chemistry, 2007, 60, 384–395.
  • Exemplary coupling reactions include, but are not limited to, formation of esters, thioesters, amides (e.g., such as peptide coupling) from activated acids or acyl halides; nucleophilic displacement reactions (e.g., such as nucleophilic displacement of a halide or ring opening of strained ring systems); azide-alkyne Huisgen cycloaddition; thiol-yne addition; imine formation; Michael additions (e.g., maleimide addition); and Diels-Alder reactions (e.g., tetrazine [4 + 2] cycloaddition).
  • nucleophilic displacement reactions e.g., such as nucleophilic displacement of a halide or ring opening of strained ring systems
  • azide-alkyne Huisgen cycloaddition thiol-yne addition
  • imine formation Michael additions (e.g., maleimide addition)
  • alkyne-azide 1,3-cycloadditions may be used (e.g., the Huisgen alkyne-azide cycloaddition).
  • the alkyne-azide cycloaddition is copper-catalyzed.
  • the alkyne-azide cycloaddition is strain-promoted.
  • alkyne-azide reactions can be found in, e.g., Kolb, Finn and Sharpless Angewandte Chemie International Edition 2001, 40, 2004-2021; Kolb and Sharpless, Drug Discov Today 2003, 24, 1128-1137; and Evans, Australian Journal of Chemistry 2007, 60, 384–395.
  • the reactive handle comprises a halogen, alkene, alkyne, azide, tetrazine, or a moiety of one of the following formulae: [185]
  • the table below shows examples of reactive handles and their associated selectivity, e.g., for different amino acid residues.
  • R A comprises . certain embodiments, R B comprises in * denotes the point of attachment to the binding moiety. iments, R A comprises . some embodiments, R A comprises . some embodiments, R A comprises . some embodiments, R A comprises wherein * denotes the point of attachment to the binding moiety. In some embodiments, R A comprises , wherein * denotes the point of attachment to the binding moiety. In some embodiments, R A comprises , wherein * denotes the point of attachment to the binding moiety.
  • R 2 is hydrogen or optionally substituted C1-C6 alkyl; or optionally wherein L A and R 2 are joined together, with the intervening atoms to form a 5- to 10-membered heterocyclic ring, wherein the heterocyclic ring is optionally substituted.
  • R 2 is hydrogen or optionally substituted C1-C6 alkyl.
  • R 2 is H.
  • R 2 is optionally substituted C1-4 alkyl.
  • R 2 is Me, Et, n-Pr, i-Pr, n-Bu, i-Bu, or t- Bu.
  • R 2 is -(CH 2 ) 2 NMe 2 .
  • L A and R 2 are joined together with the intervening atoms to form a 5- to 10-membered heterocyclic ring, wherein the heterocyclic ring is optionally substituted.
  • L A and R 2 are joined together with the intervening atoms to form an optionally 5- to 6- membered heterocyclyl ring.
  • L A and R 2 are joined together with the intervening atoms to form an optionally substituted 5- to 6-membered heterocyclic ring fused to an aryl ring.
  • L A and R 2 are joined together with the intervening atoms to form optionally substituted isoindolinyl.
  • R 3 is hydrogen or optionally substituted C1-6 alkyl. In certain embodiments, R 3 is hydrogen or unsubstituted C1-6 alkyl. In certain embodiments, R 3 is hydrogen or optionally substituted C1-4 alkyl. In certain embodiments, R 3 is Me, Et, n-Pr, i- Pr, n-Bu, i-Bu, or t-Bu. In certain embodiments, R 3 is H, Me, or Et. In certain embodiments, R 3 is Et or Me. In certain embodiments, R 3 is H or Me. In certain embodiments, R 3 is H. In certain embodiments, R 3 is Me.
  • n is 0, 1, or 2. In certain embodiments, n is 0 or 1. In certain embodiments, n is 1 or 2. In certain embodiments, n is 0. In certain embodiments, n is 1. In certain embodiments, n is 2. [192] As generally defined herein, each instance of R 2A is independently hydrogen or optionally substituted C1-C6 alkyl; or optionally wherein L A and R 2A are joined together, with the intervening atoms to form a 5- to 10-membered heterocyclic ring, wherein the heterocyclic ring is optionally substituted. In certain embodiments, each instance of R 2A is independently hydrogen or optionally substituted C1-C6 alkyl.
  • At least one instance of R 2A is H. In certain embodiments, at least one instance of R 2A is optionally substituted C1-4 alkyl. In certain embodiments, at least one instance of R 2A is Me, Et, n-Pr, i-Pr, n-Bu, i-Bu, or t-Bu. In certain embodiments, at least one instance of R 2A is - (CH2)2NMe2. [193] In certain embodiments, L A and R 2A are joined together with the intervening atoms to form a 5- to 10-membered heterocyclic ring, wherein the heterocyclic ring is optionally substituted.
  • L A and R 2A are joined together with the intervening atoms to form an optionally 5- to 6- membered heterocyclyl ring. In certain embodiments, L A and R 2A are joined together with the intervening atoms to form an optionally substituted 5- to 6-membered heterocyclic ring fused to an aryl ring. In certain embodiments, L A and R 2A are joined together with the intervening atoms to form optionally substituted isoindolinyl. In certain embodiments, L A and R 2A are joined together with the intervening atoms to form .
  • each instance of R 3A is hydrogen, optionally substituted C1-6 alkyl, or an amino acid sidechain. In certain embodiments, each instance of R 3A is hydrogen or an amino acid side chain. In certain embodiments, each instance of R 3A is optionally substituted C1-6 alkyl or an amino acid side chain. In certain embodiments, at least one instance of R 3A is an amino acid side chain. In certain embodiments, at least one instance of R 3A is a basic amino acid sidechain. In certain embodiments, at least one instance of R 3A is an acidic amino acid sidechain. In certain embodiments, at least one instance of R 3A is a neutral amino acid sidechain.
  • At least one instance of R 3A is a hydrophobic amino acid sidechain. In certain embodiments, each instance of R 3A is hydrogen or optionally substituted C1-6 alkyl. In certain embodiments, at least one instance of R 3A is hydrogen or optionally substituted C1-4 alkyl. In certain embodiments, at least one instance of R 3A is Me, Et, n-Pr, i- Pr, n-Bu, i-Bu, or t-Bu. In certain embodiments, at least one instance of R 3A is H, Me, or -CH 2 OR O . In certain embodiments, at least one instance of R 3A is H, Me, or -CH 2 OH.
  • At least one instance of R 3A is H, Me, or Et. In certain embodiments, at least one instance of R 3A is Et or Me. In certain embodiments, at least one instance of R 3A is H or Me. In certain embodiments, at least one instance of R 3A is -CH 2 OR O or Me. In certain embodiments, at least one instance of R 3A is -CH 2 OH or Me. In certain embodiments, at least one instance of R 3A is H. In certain embodiments, at least one instance of R 3A is Me. In certain embodiments, at least one instance of R 3A is -CH 2 OR O . In certain embodiments, at least one instance of R 3A is -CH 2 OH.
  • each instance of R O is independently H, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 haloalkyl, optionally substituted C 3-7 carbocyclyl, or optionally substituted C 1-6 acyl.
  • at least one instance of R O is H.
  • each instance of R O is H.
  • at least one instance of R O is optionally substituted C 1-6 alkyl.
  • at least one instance of R O is optionally substituted C3-7 carbocyclyl.
  • at least one instance of R O is optionally substituted C1-6 acyl.
  • each instance of R N is independently H, optionally substituted C1-6 alkyl, optionally substituted C3-7 carbocyclyl, or optionally substituted C1-6 acyl, or two R N bonded to the same nitrogen are joined together to form optionally substituted 3-7 membered heterocyclyl.
  • at least one instance of R N is H.
  • each instance of R N is H.
  • at least one instance of R N is optionally substituted C1-6 alkyl.
  • at least one instance of R N is Me.
  • each instance of R N is Me.
  • at least one instance of R N is optionally substituted C3-7 carbocyclyl.
  • at least one instance of R N is optionally substituted C1-6 acyl.
  • two R N bonded to the same nitrogen are taken together to form optionally substituted 3-7 membered heterocyclyl.
  • Z is a binding moiety.
  • a binding moiety is polypeptide, for example one or more proteins (e.g., full length and/or peptide).
  • a binding moiety specifically binds to a target molecule, such as an antigen.
  • a binding moiety is an antigen-binding moiety, for example, a cell-binding moiety.
  • a binding moiety is an antibody.
  • an antibody is a full-length antibody.
  • an antibody is a chimeric antibody.
  • an antibody is a human antibody.
  • an antibody is a humanized antibody.
  • an antibody is a Fab fragment, a F(ab')2 fragment, a Fv fragment or a scFv fragment.
  • an antibody is a single domain antibody (e.g., NANOBODY®) derived from a camelid antibody or a single domain antibody (e.g., NANOBODY®) derived from shark antibody.
  • an antibody is a diabody.
  • an antibody comprises a framework having a human germline sequence.
  • an antibody comprises a heavy chain constant domain selected from the group consisting of IgG, IgG1, IgG2, IgG2A, IgG2B, IgG2C, IgG3, IgG4, IgA1, IgA2, IgD, IgM, and IgE constant domains.
  • an antibody comprises a heavy (H) chain variable region (abbreviated herein as VH) and/or a light (L) chain variable region (abbreviated herein as VL).
  • an antibody comprises an immunoglobulin constant domain, e.g., an Fc domain.
  • An immunoglobulin constant domain includes a heavy or light chain constant domain.
  • the heavy chain of an antibody described herein is an alpha ( ⁇ ), delta ( ⁇ ), epsilon ( ⁇ ), gamma ( ⁇ ) or mu ( ⁇ ) heavy chain.
  • the heavy chain of an antibody described herein comprises a human alpha ( ⁇ ), delta ( ⁇ ), epsilon ( ⁇ ), gamma ( ⁇ ) or mu ( ⁇ ) heavy chain.
  • an antibody described herein comprises a human gamma 1 CH1, CH2, and/or CH3 domain.
  • the amino acid sequence of the VH domain comprises the amino acid sequence of a human gamma ( ⁇ ) heavy chain constant domain.
  • a VH domain comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or at least 99% identical to any variable chain constant domains provided herein.
  • an antibody is modified, e.g., modified via glycosylation, phosphorylation, sumoylation, and/or methylation.
  • an antibody is a glycosylated antibody, which is conjugated to at least one sugar or carbohydrate molecule.
  • At least one sugar or carbohydrate molecule is conjugated to an antibody via N-glycosylation, O- glycosylation, C-glycosylation, glypiation (GPI anchor attachment), and/or phosphoglycosylation.
  • at least one sugar or carbohydrate molecule is a monosaccharide, disaccharide, oligosaccharide, or glycan.
  • at least one sugar or carbohydrate molecule is a
  • an antibody comprises a linker polypeptide.
  • a linker polypeptide comprises at least two amino acid residues joined by at least one peptide bond and is used to link two molecules (e.g., two peptides or polypeptides) to each other. Examples of linker polypeptides have been reported (see e.g., Holliger, P., et al. (1993) Proc. Natl. Acad. Sci.
  • an antibody can be part of a larger immunoadhesion molecule, formed by covalent or noncovalent association of the antibody or antibody portion with one or more other proteins or peptides.
  • immunoadhesion molecules include use of the streptavidin core region to make a tetrameric scFv molecule (Kipriyanov, S. M., et al.
  • an antibody is a bispecific antibody.
  • a bispecific antibody includes a polypeptide or a complex (e.g., two covalently linked polypeptides or monospecific antibodies) that includes two different antigen binding sites, e.g., paratopes, that have different antigen binding specificities.
  • a bispecific antibody is a polypeptide that includes two different antigen binding sites in which each antigen binding site binds to a different epitope of the same antigen.
  • a bispecific antibody is a polypeptide that includes two different antigen binding sites in which each site binds to a different antigen.
  • a bispecific antibody comprises two different sets of immunoglobulin variable domains, each set binding to a different epitope or group of epitopes.
  • an antibody is a multispecific antibody.
  • a multispecific antibody includes a polypeptide or a complex (e.g., two or more covalently linked polypeptides) that includes at least two different immunoglobulin variable domains or at least two different sites, e.g., paratopes, that specifically bind to one or more antigens.
  • a multispecific antibody includes a polypeptide that comprise at least two different sites in which each site binds to a different epitope of the same antigen.
  • a multispecific antibody includes a polypeptide that comprises at least two different sites in which each site binds to a different antigen.
  • an antibody is a chimeric antibody, which includes antibodies that comprise a sequence from two or more species; for example, antibodies that comprise heavy and light chain variable region sequences and/or constant domain sequences from two or more species, such as antibodies having murine heavy and light chain variable regions linked to human constant domains.
  • an antibody is a human antibody.
  • a human antibody includes antibodies that comprise variable and constant domains derived from human germline immunoglobulin sequences. The human antibodies of the disclosure can include amino acid residues not encoded by human germline
  • immunoglobulin sequences e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo
  • CDRs and, in particular, CDR3 for example in the CDRs and, in particular, CDR3.
  • Human antibody is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. [207] In certain embodiments, an antibody is a humanized antibody.
  • a humanized antibody includes antibodies that comprise heavy and/or light chain variable region sequences from a non-human species (e.g., a mouse) but in which at least a portion of the VH and/or VL sequence has been altered to be more “human-like,” i.e., more like human germline variable sequences.
  • a non-human species e.g., a mouse
  • One type of humanized antibody is a CDR-grafted antibody, in which human CDR sequences are introduced into non-human VH and/or VL sequences to replace the corresponding nonhuman CDR sequences.
  • humanized anti-VH4-34 antibodies and antigen binding portions are provided.
  • an antibody is an antibody fragment.
  • antibody fragments include Fab, Fab’, scFv (single-chain variable fragments), Fc (fragment crystallizable regions), VHHs (e.g., camelid heavy-chain antibody), and other sdAbs (single-domain antibodies, e.g., NANOBODIES®).
  • a binding moiety is an AFFIMER® polypeptide.
  • the AFFIMER® polypeptides provided herein are based on a human Stefin A scaffold that has been modified to prevent amino terminal acetylation/oxidation and increase thermal stability.
  • a protein scaffold includes a framework or structure that provides a stable platform for displaying functional protein domains or peptides, for example. These scaffolds are typically engineered proteins that have a robust and stable tertiary structure, allowing them to tolerate insertions, deletions, or substitutions without losing their overall fold and stability.
  • a scaffold comprises a sequence having at least 90% (e.g., at least 95%, at least 96%, at least 97%, or at least 98%) identity to the amino acid sequence of SEQ ID NO: 1 (human Stefin A): MIPGGLSEAK PATPEIQEIV DKVKPQLEEK TNETYGKLEA VQYKTQVVAG TNYYIKVRAG DNKYMHLKVF KSLPGQNEDL VLTGYQVDKN KDDELTGF (SEQ ID NO: 1) [210]
  • various aspects herein relate to polypeptides comprising a scaffold that comprises a sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 1, wherein the scaffold sequence includes (a) an insertion of an amino acid between amino acid positions corresponding to M1 and I2 of the amino acid sequence of SEQ ID NO: 1 and (b) a mutation at an amino acid position corresponding to N32 of the amino acid sequence of SEQ ID NO: 1 and a heterolog
  • the insertion of an amino acid between amino acid positions corresponding to M1 and I2 of the amino acid sequence of SEQ ID NO: 1 assists in N-terminal methionine cleavage, reducing N- terminal heterogeneity (e.g., presence or absence of methionine and/or acetylation of methionine).
  • a scaffold provided herein includes a modification that prevents amino terminal acetylation/oxidation relative to the naturally occurring human Stefin A protein.
  • a scaffold includes a modification that increases thermal stability (e.g., by at least 20%, at least 30%, at least 40%, or at least 50%) of the scaffold (or of a polypeptide containing the scaffold), relative to the naturally occurring human Stefin A protein.
  • a scaffold (or scaffold sequence, i.e., an amino acid sequence within a scaffold) includes (a) an insertion of an amino acid between amino acid positions corresponding to M1 and I2 of the amino acid sequence of SEQ ID NO: 1 and (b) a mutation at an amino acid position corresponding to N32 of the amino acid sequence of SEQ ID NO: 1, as shown in the amino acid sequence of SEQ ID NO: 2: MGIPGGLSEA KPATPEIQEI VDKVKPQLEE KTGETYGKLE AVQYKTQV-X1-TN YYIKVRAGDN KYMHLKVFKS L-X2-EDLVLTGYQ VDKNKDDELT GF (SEQ ID NO: 2), wherein X1 is any heterologous peptide (e.g., having a length of about 4 to 16 amino acids) and X2 is any heterologous peptide (e.g., having a length of about 4 to 16 amino acids).
  • X1 is any heterologous
  • a polypeptide comprises a scaffold comprising (a) a sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 1, wherein the scaffold sequence includes (i) an insertion of an amino acid (e.g., glycine) between amino acid positions corresponding to M1 and I2 of the amino acid sequence of SEQ ID NO: 1 and (ii) a mutation at an amino acid position corresponding to N32 (e.g., N32G) of the amino acid sequence of SEQ ID NO: 1, and (b) a heterologous peptide (e.g., two heterologous peptides).
  • an amino acid e.g., glycine
  • a polypeptide comprises a scaffold comprising (a) a sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 1, wherein the scaffold sequence includes (i) an insertion of an amino acid (e.g., glycine) between amino acid positions corresponding to M1 and I2 of the amino acid sequence of SEQ ID NO: 1 and (ii) a mutation at an amino acid position corresponding to N32 (e.g., N32G) of the amino acid sequence of SEQ ID NO: 1, and (b) a heterologous peptide (e.g., two heterologous peptides).
  • an amino acid e.g., glycine
  • a polypeptide comprises a scaffold comprising (a) a sequence having at least 98% identity to the amino acid sequence of SEQ ID NO: 1, wherein the scaffold sequence includes (i) an insertion of an amino acid (e.g., glycine) between amino acid positions corresponding to M1 and I2 of the amino acid sequence of SEQ ID NO: 1 and (ii) a mutation at an amino acid position corresponding to N32 (e.g., N32G) of the amino acid sequence of SEQ ID NO: 1, and (b) a heterologous peptide (e.g., two heterologous peptides).
  • an amino acid e.g., glycine
  • the insertion of an amino acid between amino acid positions corresponding to M1 and I2 of the amino acid sequence of SEQ ID NO: 1 is a glycine (G). In some embodiments, the insertion of an amino acid between amino acid positions corresponding to M1 and I2 of the amino acid sequence of SEQ ID NO: 1 is an alanine (A). In some embodiments, the insertion of an amino acid between amino acid positions corresponding to M1 and I2 of the amino acid sequence of SEQ
  • the sequence of a scaffold has at least 95% identity to the amino acid sequence of SEQ ID NO: 2.
  • a sequence of a scaffold has 95% identity to about 98% identity to the amino acid sequence of SEQ ID NO: 2. In some embodiments, a sequence of a scaffold has about 98% identity to the amino acid sequence of SEQ ID NO: 2. In some embodiments, a scaffold comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, a scaffold consists of the amino acid sequence of SEQ ID NO: 2. [218] Percent identity in the context of DNA, RNA, or protein sequences refers to the percentage of positions (nucleotides in DNA/RNA or amino acids in proteins) that are identical in two aligned sequences when compared.
  • Global and local alignment are two methods used in bioinformatics to align sequences, such as DNA, RNA, or proteins, to identify regions of similarity that may indicate functional, structural, or evolutionary relationships among the sequences.
  • a global alignment aligns two sequences from beginning to end, attempting to align every residue in each sequence. This method typically uses the Needleman-Wunsch algorithm and is ideal for sequences of roughly equal size and when you expect that the sequences are similar over their entire length.
  • a global alignment can introduce gaps as necessary to align as much of the sequences as possible.
  • a local alignment finds the most similar subsequence(s) between two sequences. It does not require that the entirety of either sequence align.
  • Cysteine Modifications [219] Any one or more of the polypeptides (e.g., AFFIMER ® polypeptides) described herein can be modified to include one or more cysteine (C). Due to the unique reactivity of cysteine's thiol (-SH) group, these amino acids can be used to enable, for example, conjugation (e.g., via maleimide chemistry) of moieties of interest (e.g., drugs, fluorophores, PEG).
  • C cysteine
  • -SH thiol
  • one or more C is in an N-terminal region of the polypeptide, for example, within 10 amino acids (e.g., within 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acids) of the N terminus of the polypeptide.
  • a C-terminal region of the polypeptide for example, within 10 amino acids (e.g., within 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acids) of the C terminus of the polypeptide.
  • one or more C is in a Loop 3 region of the polypeptide between a first heterologous peptide (Loop 2) and a second heterologous peptide (Loop 4), for example, a first peptide and a second peptide that specifically binds to FAP or HSA.
  • Fusion proteins can contain combinations of any polypeptide (e.g., AFFIMER ® polypeptide), for example, combinations of FAP binders, combinations of HSA binders, or FAP-HSA binder combinations).
  • a fusion protein comprises two polypeptides (e.g., AFFIMER ® polypeptides).
  • a fusion protein comprises three polypeptides (e.g., AFFIMER ® polypeptides).
  • a fusion protein comprises four polypeptides (e.g., AFFIMER ® polypeptides).
  • a fusion protein comprises five polypeptides (e.g., AFFIMER ® polypeptides).
  • polypeptides can include one or more cysteine.
  • Each polypeptide e.g., AFFIMER ® polypeptide
  • a polypeptide binds specifically to FAP and comprises the amino acid sequence of any one of SEQ ID NOs: 213-316 or 335-351.
  • a fusion protein comprises a polypeptide that binds specifically to FAP, wherein the polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 317-334 or 353- 388.
  • a polypeptide binds specifically to HSA and comprises the amino acid sequence of SEQ ID NO: 389-442, 453 or 454.
  • a fusion protein comprises a polypeptide that binds specifically to HSA, wherein the polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 443-452 or 455- 461.
  • a fusion protein comprises (i) a polypeptide that binds specifically to FAP and a polypeptide that binds to HSA, wherein the fusion protein comprises the amino acid sequence of any one of SEQ ID NOs: 462-501 or 553-567.
  • the scaffold further comprises a cysteine.
  • the cysteine is introduced at the C-terminus of the scaffold.
  • the cysteine is introduced at the N-terminus of the scaffold.
  • the cysteine is introduced between two heterologous proteins peptides (e.g., between a variable Loop 1 peptide and a variable Loop 2 peptide, e.g., between positions T51 and L73 of the amino acid sequence of SEQ ID NO: 1, or between positions T49 and L71 of the amino acid sequence of SEQ ID NO: 4).
  • the cysteine is introduced at a position in the scaffold corresponding to position D61 of the amino acid sequence of SEQ ID NO: 1.
  • cysteines are introduced: a first cysteine at the C-terminus of the scaffold and a second cysteine at a position corresponding to the region between positions T51 and L73 of the amino acid sequence of SEQ ID NO: 1 (e.g., a position corresponding to position D61 of the amino acid sequence of SEQ ID NO: 1).
  • the cysteine in some embodiments, is used for conjugation (e.g., in the context of protein conjugates, described below).
  • the use of cysteine for conjugation in AFFIMER® molecules also sets them apart from antibodies. In antibodies, cysteines are often part of the framework and/or variable regions of the antibodies, required to create and maintain the structure of the antibody (e.g., through disulfide bonds). Therefore, the use of cysteine conjugation in antibodies may cause
  • AFFIMER® polypeptides are not limited in this manner and have significantly more flexibility with respect to conjugation.
  • Heterologous Peptides include one or more heterologous peptide (heterologous to the modified human Stefin A scaffold) that enables the polypeptides to bind to a target with high affinity and extraordinarivity.
  • a heterologous peptide may be referred to herein as a variable Loop 2 peptide (comprising a variable Loop 2 sequence) or a variable Loop 4 peptide (comprising a variable Loop 4 sequence).
  • a heterologous peptide is considered an “insertion” in a scaffold sequence, for example, of an AFFIMER® polypeptide, such that when the percent identity of the amino acid sequence of a scaffold is calculated relative to human Stefin A (wild-type human Stefin A), the heterologous peptide (or heterologous peptides) is excluded from the calculation.
  • a polypeptide comprising a scaffold comprising a sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 1 does not factor a heterologous peptide into the percent identity calculation.
  • AFFIMER® polypeptide having the following full-length sequence: MGIPGGLSEA KPATPEIQEI VDKVKPQLEE KTGETYGKLE AVQYKTQVXX XXXXXTNY YIKVRAGDNK YMHLKVFKSL XXXXXXXE DLVLTGYQVD KNKDDELTGF(SEQ ID NO: 3) [230]
  • This sequence includes a scaffold sequence (SEQ ID NO: 4) as well as two heterologous peptide sequences (each designated “XXXXXXXX”), each inserted into the scaffold sequence.
  • a heterologous peptide can comprise an amino acid sequence having a length of about 8 to about 16 amino acids.
  • a heterologous peptide comprises an amino acid sequence having a length of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids.
  • a heterologous peptide comprises an amino acid sequence having a length of 9 amino acids.
  • a heterologous peptide comprises an amino acid sequence having a length of 12 amino acids.
  • a heterologous peptide comprises an amino acid sequence having a length of 15 amino acids.
  • a heterologous peptide comprises an amino acid sequence having a length of 4 to 12, 5 to 12, 6 to 12, 4 to 10, 6 to 10, 8 to 10, 5 to 15, 6 to 15, 7 to 15, 5 to 13, 7 to 13, 9 to 13, 11 to 13, 8 to 18, 9 to 18, 10 to 18, 10 to 16, 12 to 16, or 14 to 16 amino acids.
  • an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 4 to 12, 5 to 12, 6 to 12, 4 to 10, 6 to 10, or 8 to 10 amino acids and an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 4 to 12, 5 to 12, 6 to 12, 4 to 10, 6 to 10, or 8 to 10 amino acids.
  • an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 9 amino acids and an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 9 amino acids.
  • an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 9 amino acids and the polypeptide does not comprise an AFFIMER® variable Loop 2 peptide.
  • an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 5 to 15, 6 to 15, 7 to 15, 5 to 13, 7 to 13, 9 to 13, or 11 to 13 amino acids and an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 5 to 15, 6 to 15, 7 to 15, 5 to 13, 7 to 13, 9 to 13, or 11 to 13 amino acids.
  • an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 12 amino acids and an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 12 amino acids.
  • an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 8 to 18, 9 to 18, 10 to 18, 10 to 16, 12 to 16, or 14 to 16 amino acids and an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 8 to 18, 9 to 18, 10 to 18, 10 to 16, 12 to 16, or 14 to 16 amino acids.
  • an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 15 amino acids and an AFFIMER® variable Loop 2 peptide comprises an amino acid sequence having a length of 15 amino acids.
  • a heterologous peptide e.g., an AFFIMER® variable Loop 2 peptide
  • positions V47 and T51 of the amino acid sequence of SEQ ID NO: 1 correspond respectively to positions V48 and T49 of the amino acid sequence of SEQ ID NO: 4, which is the scaffold sequence (minus any heterologous peptide) of, for example, an AFFIMER polypeptide comprising the amino acid sequence of SEQ ID NO: 2, wherein each of X1 and X2 is an independent heterologous peptide.
  • a heterologous peptide e.g., an AFFIMER® variable Loop 2 peptide
  • a heterologous peptide e.g., an AFFIMER® variable Loop 4 peptide
  • AFFIMER® variable Loop 4 peptide is located in a sequence of a scaffold between amino acid positions corresponding to positions L73 and E78 of the amino acid sequence of SEQ ID NO: 1. As shown in the above alignment, positions L73 and E78
  • a heterologous peptide e.g., an AFFIMER® variable Loop 4 peptide
  • a heterologous peptide is located in a sequence of a scaffold between amino acid positions corresponding to positions L71 and E72 of the amino acid sequence of SEQ ID NO: 4.
  • a polypeptide comprises a scaffold comprising the amino acid sequence of SEQ ID NO: 2, wherein X1 is a heterologous peptide having a length of 8 amino acids, and X2 is a heterologous peptide having a length of 8 amino acids.
  • a polypeptide comprises a scaffold comprising the amino acid sequence of SEQ ID NO: 2, wherein X1 is a heterologous peptide having a length of 9 amino acids, and X2 is a heterologous peptide having a length of 9 amino acids.
  • a polypeptide comprises a scaffold comprising the amino acid sequence of SEQ ID NO: 2, wherein X1 is a heterologous peptide having a length of 9 amino acids, and X2 is a heterologous peptide having a length of 0 amino acids (i.e., there is no second heterologous peptide).
  • a polypeptide comprises a scaffold comprising the amino acid sequence of SEQ ID NO: 2, wherein X1 is a heterologous peptide having a length of 10 amino acids, and X2 is a heterologous peptide having a length of 10 amino acids.
  • a polypeptide comprises a scaffold comprising the amino acid sequence of SEQ ID NO: 2, wherein X1 is a heterologous peptide having a length of 12 amino acids, and X2 is a heterologous peptide having a length of 12 amino acids. In some embodiments, a polypeptide comprises a scaffold comprising the amino acid sequence of SEQ ID NO: 2, wherein X1 is a heterologous peptide having a length of 15 amino acids, and X2 is a heterologous peptide having a length of 15 amino acids. [240] Exemplary variable Loop 2 sequences and variable Loop 4 sequences are provided in Table 1 below. Table 1. AFFIMER® Loop 2 and Loop 4 Sequences
  • a heterologous peptide comprises a sequence selected from the amino acid sequence of any one of SEQ ID NOs: 5-108.
  • a heterologous peptide comprises a sequence selected from the amino acid sequence of any one of SEQ ID NOs: 109-212.
  • a polypeptide e.g., an AFFIMER® polypeptide
  • a polypeptide (e.g., an AFFIMER® polypeptide) comprises a variable Loop 2 peptide comprising the amino acid sequence of SEQ ID NO: 6 and a variable Loop 4 peptide comprising the amino acid sequence of SEQ ID NO: 110.
  • exemplary polypeptide sequences described herein are provided in Table 2. Table 2. Polypeptide Sequences
  • a polypeptide comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to any one of SEQ ID NOs: 213-567 (Table 2). [246] In some embodiments, the polypeptide comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 213. In some embodiments, the polypeptide comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 213.
  • the polypeptide comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 213. In some embodiments, the polypeptide comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 213. In some embodiments, the polypeptide comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 213. In some embodiments, the polypeptide comprises SEQ ID NO: 213. In some embodiments, the polypeptide consists of SEQ ID NO: 213. [247] In some embodiments, the polypeptide comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 214. In some embodiments, the polypeptide comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 214.
  • the polypeptide comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 214. In some embodiments, the polypeptide comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 214. In some embodiments, the polypeptide comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 214. In some embodiments, the polypeptide comprises SEQ ID NO: 214. In some embodiments, the polypeptide consists of SEQ ID NO: 214. [248] In some embodiments, the polypeptide comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 389. In some embodiments, the polypeptide comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 389. In some embodiments, the polypeptide comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 389. In some embodiments,
  • the polypeptide comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 389. In some embodiments, the polypeptide comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 389. In some embodiments, the polypeptide comprises SEQ ID NO: 389. In some embodiments, the polypeptide consists of SEQ ID NO: 389. [249] In some embodiments, the polypeptide comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 390. In some embodiments, the polypeptide comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 390. In some embodiments, the polypeptide comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 390.
  • the polypeptide comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 390. In some embodiments, the polypeptide comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 390. In some embodiments, the polypeptide comprises SEQ ID NO: 390. In some embodiments, the polypeptide consists of SEQ ID NO: 390. [250] In some embodiments, the polypeptide comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 391. In some embodiments, the polypeptide comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 391. In some embodiments, the polypeptide comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 391.
  • the polypeptide comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 391. In some embodiments, the polypeptide comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 391. In some embodiments, the polypeptide comprises SEQ ID NO: 391. In some embodiments, the polypeptide consists of SEQ ID NO: 391. [251] In some embodiments, the polypeptide comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 441. In some embodiments, the polypeptide comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 441. In some embodiments, the polypeptide comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 441.
  • the polypeptide comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 441. In some embodiments, the polypeptide comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 441. In some embodiments, the polypeptide comprises SEQ ID NO: 441. In some embodiments, the polypeptide consists of SEQ ID NO: 441. [252] In some embodiments, the polypeptide comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 442. In some embodiments, the polypeptide comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 442. In some embodiments, the polypeptide comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 442.
  • the polypeptide comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 442. In some embodiments, the polypeptide comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 442. In some embodiments, the polypeptide comprises SEQ ID NO: 442. In some embodiments, the polypeptide consists of SEQ ID NO: 442. [253] In some embodiments, the polypeptide comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 452. In some embodiments, the polypeptide comprises an amino acid sequence
  • the polypeptide comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 452. In some embodiments, the polypeptide comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 452. In some embodiments, the polypeptide comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 452. In some embodiments, the polypeptide comprises an amino acid sequence having at least 98% identity to SEQ ID NO: 452. In some embodiments, the polypeptide comprises SEQ ID NO: 452. In some embodiments, the polypeptide consists of SEQ ID NO: 452. [254] Non-limiting examples of other AFFIMER ® polypeptides that can be used in accordance with the disclosure include those described in International Publication Nos.
  • binding moieties include aptamers, ligands, receptor-binding domains, receptors, and small molecules.
  • a binding moiety is an aptamer.
  • a binding moiety is ligand.
  • a binding moiety is a receptor-binding domain. In some embodiments, a binding moiety is a receptor. In some embodiments, a binding moiety is a small molecule (e.g., a low molecular weight organic compound, typically less than 1,000 Daltons).
  • a binding moiety e.g., antibody or AFFIMER ® polypeptide
  • a binding moiety of the disclosure can target (specifically bind to) any cancer target described herein.
  • a binding moiety can target disease antigens, such as cancer antigens (e.g., expressed on a solid tumor).
  • a cancer antigen is expressed by head and neck cancer cells (e.g., salivary gland cancer, squamous cell carcinoma of the head and neck (SCCHN), and/or adenoid cystic carcinoma), soft tissue sarcoma cells (e.g., undifferentiated pleomorphic sarcoma and/or dedifferentiated liposarcoma), breast cancer cells (e.g., triple-negative breast cancer), lung cancer cells (e.g., non-small cell lung cancer (NSCLC)), gastric cancer cells, colorectal cancer cells, and pancreatic ductal adenocarcinoma cells.
  • head and neck cancer cells e.g., salivary gland cancer, squamous cell carcinoma of the head and neck (SCCHN), and/or adenoid cystic carcinoma
  • soft tissue sarcoma cells e.g., undifferentiated pleomorphic sarcoma and/or dedifferentiated liposarcoma
  • breast cancer cells
  • a cancer antigen is expressed by a cancer selected from pancreatic cancer, esophageal cancer, sarcoma, colorectal cancer, breast cancer (e.g., HR+ breast cancer or TNBC), NSCLC, SCLC, gastric cancer, ovarian cancer, and cholangiocarcinoma.
  • a cancer selected from pancreatic cancer, esophageal cancer, sarcoma, colorectal cancer, breast cancer (e.g., HR+ breast cancer or TNBC), NSCLC, SCLC, gastric cancer, ovarian cancer, and cholangiocarcinoma.
  • Non-limiting examples of cancer antigens expressed by head and neck cancer cells include [interlukin-8 (IL-8), melanoma associated antigens (MAGE), cytokeratin, E48 antigen, cathepsin D, pS2, P-glycoprotein, proliferating cell nuclear antigen (PCNA), TGF-a, TGG-b, E-cadherin, membrane type 1 matrix metalloprotease (MT1-MMP), CK19, CK8,Beta 2-microglobulin,CD 44,CD 80,1- ACT,CA125,Cyfra21-1, Cyclin D1, Ki6758, CKD2,MIB, C-erb2, and TGF- ⁇ .
  • IL-8 interlukin-8
  • MAGE melanoma associated antigens
  • E48 antigen E48 antigen
  • cathepsin D cathepsin D
  • pS2 P-glycoprotein
  • PCNA proliferating cell nuclear antigen
  • TGF-a
  • a binding moiety is an antibody that binds to a head and neck cancer antigen selected from etuximab, nivolumab, pembrolizumab, ramucirumab, durvalumab, avelumab, trastuzumab, panitumumab, ipilimumab, necitumumab, zalutumumab, ofatumumab, blinatumomab, dostarlimab, elotuzumab, tisotumab vedotin, margetuximab, glembatumumab vedotin, enfortumab vedotin, cemiplimab, loncastuximab tesirine, trastuzumab deruxtecan, ipilimumab-nivolumab combination therapy,
  • a binding moiety is an AFFIMER ® polypeptide that binds to a head and neck cancer antigen.
  • cancer antigens expressed by soft tissue sarcoma cells include MYF4, MYF3, FLI1, erythroblast transformation-specific transcription factor (ERG), Brachyury, SOX10, SATB2, ⁇ -catenin, MDM2, CDK4, SMARCB1, SDHB, TFE3, ALK, STAT6, DOG1, TLE1, MUC4, GRIA2, CD34, desmin, epithelial membrane antigen (EMA), keratin cocktail AE1/AE3, S100 protein, and alpha smooth muscle actin ( ⁇ -SMA), CD31, KIT, Ano-1, ⁇ -fetoprotein, OCT3/4, SALL4m, CD30, placental alkaline phosphatase (PLAP), cytokeratin
  • a binding moiety is an antibody that binds to a soft tissue sarcoma antigen selected from olaratumab, pembrolizumab, nivolumab, atezolizumab, durvalumab, avelumab, trastuzumab, cetuximab, panitumumab, ramucirumab, ipilimumab, necitumumab, blinatumomab, dostarlimab, enfortumab vedotin, tisotumab vedotin, margetuximab, glembatumumab vedotin, elotuzumab, onartuzumab, trastuzumab deruxtecan, anetumab ravtansine, nivolumab-ipilimumab combination therapy, spartalizumab, rilotumumab,
  • a binding moiety is an AFFIMER ® polypeptide that binds to a soft tissue sarcoma antigen.
  • cancer antigens expressed by breast cancer cells include HER2, estrogen receptor (ER), progesterone receptor (PR), MUC1, EGFR, NY-ESO-1, Mammaglobin-A, BRCA1, BRCA2, p53, CD24, CD44, GD2, B7-H4, CEA, TROP2, CXCR4, ERBB3, mesothelin, and cyclin D1.
  • a binding moiety is an antibody that binds to a breast cancer antigen selected from trastuzumab, pertuzumab, ado-trastuzumab emtansine, trastuzumab deruxtecan, bevacizumab, sacituzumab govitecan, margetuximab, pembrolizumab, nivolumab, ipilimumab, atezolizumab, durvalumab, cetuximab, catumaxomab, bevacizumab-awwb, trastuzumab-dttb, trastuzumab-qyyp, rituximab, tisotumab vedotin, elotuzumab, ibritumomab tiuxetan, panitumumab, brentuximab vedotin, polatuzumab, trast
  • a binding moiety is an AFFIMER ® polypeptide that binds to a breast cancer antigen.
  • cancer antigens expressed by lung cancer cells include GFR, ALK, KRAS, PD-L1, ROS1, MET, HER2, BRAF, RET, TP53, CEA, MUC1, NY-ESO-1, WT1, Mesothelin, Survivin, CA-125, TROP2, CD56, and GD2.
  • a binding moiety is an antibody that binds to a lung cancer antigen selected from pembrolizumab, nivolumab, atezolizumab, durvalumab,
  • a binding moiety is an AFFIMER ® polypeptide that binds to a lung cancer antigen.
  • cancer antigens expressed by gastric cancer cells include HER2, EGFR, CEA, MUC1, PD-L1, VEGFR2, Claudin 18.2, MAGE-A3, NY-ESO-1, FGFR2, Mesothelin, Survivin, CD133, CD44, CA-72-4, p53, CEACAM5, Cyclin D1, AFP, and ALDH1.
  • a binding moiety is an antibody that binds to a gastric cancer antigen selected from trastuzumab, ramucirumab, pembrolizumab, nivolumab, ipilimumab, cetuximab, bevacizumab, trastuzumab deruxtecan, zolbetuximab, avelumab, durvalumab, atezolizumab, necitumumab, margetuximab, tremelimumab, panitumumab, rituximab, tisotumab vedotin, elotuzumab, blinatumomab, inotuzumab ozogamicin, gemtuzumab ozogamicin, glembatumumab vedotin, trastuzumab-qyyp, trastuzumab-dttb, bevacizum
  • a binding moiety is an AFFIMER ® polypeptide that binds to a gastric cancer antigen.
  • cancer antigens expressed by colorectal cancer cells include CEA, EGFR, HER2, MUC1, KRAS, BRAF, TP53, PD-L1, VEGF, CD133, CD44, CA-19-9, TROP2, NY-ESO- 1, MAGE-A3, Mesothelin, CEACAM5, ALDH1, Claudin 18.2, and Survivin.
  • a binding moiety is an antibody that binds to a colorectal cancer antigen selected from cetuximab, panitumumab, bevacizumab, ramucirumab, trastuzumab, nivolumab, pembrolizumab, atezolizumab, durvalumab, avelumab, necitumumab, zalutumumab, ipilimumab, dostarlimab, enfortumab vedotin, elotuzumab, margetuximab, glembatumumab vedotin, trastuzumab deruxtecan, onartuzumab, rilotumumab, figitumumab, blinatumomab, loncastuximab tesirine, spartalizumab, cemiplimab, ibritumomab, cemi
  • a binding moiety is an AFFIMER ® polypeptide that binds to a colorectal cancer antigen.
  • cancer antigens expressed by pancreatic ductal adenocarcinoma cells include CA19-9, CEA, MUC1, MUC4, KRAS, p53, EGFR, PD-L1, Mesothelin, CEACAM6, Claudin 18.2, VEGF, TROP2, CD44, CD133, ALDH1, WT1, NY-ESO-1, Survivin, and Glypican-1.
  • a binding moiety is an antibody that binds to a pancreatic ductal adenocarcinoma antigen selected from rastuzumab, pembrolizumab, nivolumab, atezolizumab, durvalumab, cetuximab, ramucirumab, necitumumab, ipilimumab, panitumumab, elotuzumab, dostarlimab, avelumab, margetuximab, glembatumumab vedotin, onartuzumab, figitumumab, blinatumomab, enfortumab
  • a binding moiety is an AFFIMER ® polypeptide that binds to a pancreatic ductal adenocarcinoma antigen.
  • a binding moiety such as an AFFIMER® polypeptide or antibody, in some embodiments, binds to an antigen selected from the following antigens: 5T4, ADAM17, ADAM9, ALK, angiopoietin2, Axl, AXL, B7H3, B7H4, BAFF, BCMA, BSG, c-kit, CA-IX, CA125, CA6, CAIX, CCR5, CCR7, CD123, CD134, CD137, CD138, CD152, CD184, CD19, CD20, CD200, CD205, CD22, CD221, CD228, CD23, CD24, CD25, CD27, CD276, CD279, CD28, CD30, CD319, CD33, CD37, CD38, CD40, CD44, CD45, CD46, CD47, CD51, CD52, CD56, CD7, CD70, CD73, CD74, CD79B, CD79b, CD80, CD99, CDH3, CDH6, CEACAM5, CEA
  • a binding moiety such as an AFFIMER® polypeptide or antibody, in some embodiments, binds to an antigen selected from the following antigens: CEACAM5, CLAUDIN18.2, CLAUDIN4, CMET, DLL3, EDB-FN, FAP, FR ⁇ , HER2, HER3, LLRC15, Nectin-4, TF (Tissue Factor), and TROP2.
  • a binding moiety specifically binds to 5T4.
  • a binding moiety specifically binds to ADAM17.
  • a binding moiety specifically binds to ADAM9.
  • a binding moiety specifically binds to ALK.
  • a binding moiety specifically binds to angiopoietin2. In some embodiments, a binding moiety specifically binds to Axl. In some embodiments, a binding moiety specifically binds to AXL. In some embodiments, a binding moiety specifically binds to B7H3. In some embodiments, a binding moiety specifically binds to B7H4. In some embodiments, a binding moiety specifically binds to BAFF. In some embodiments, a binding moiety specifically binds to BCMA. In some embodiments, a binding moiety specifically binds to BSG. In some embodiments, a binding moiety specifically binds to c-kit.
  • a binding moiety specifically binds to CA-IX. In some embodiments, a binding moiety specifically binds to CA125. In some embodiments, a binding moiety specifically binds to CA6. In some embodiments, a binding moiety specifically binds to CAIX. In some embodiments, a binding moiety specifically binds to CCR5. In some embodiments, a binding moiety specifically binds to CCR7. In some embodiments, a binding moiety specifically binds to CD123. In some embodiments, a binding
  • a binding moiety specifically binds to CD134. In some embodiments, a binding moiety specifically binds to CD137. In some embodiments, a binding moiety specifically binds to CD138. In some embodiments, a binding moiety specifically binds to CD152. In some embodiments, a binding moiety specifically binds to CD184. In some embodiments, a binding moiety specifically binds to CD19. In some embodiments, a binding moiety specifically binds to CD20. In some embodiments, a binding moiety specifically binds to CD200. In some embodiments, a binding moiety specifically binds to CD205. In some embodiments, a binding moiety specifically binds to CD22. In some embodiments, a binding moiety specifically binds to CD221.
  • a binding moiety specifically binds to CD228. In some embodiments, a binding moiety specifically binds to CD23. In some embodiments, a binding moiety specifically binds to CD24. In some embodiments, a binding moiety specifically binds to CD25. In some embodiments, a binding moiety specifically binds to CD27. In some embodiments, a binding moiety specifically binds to CD276. In some embodiments, a binding moiety specifically binds to CD279. In some embodiments, a binding moiety specifically binds to CD28. In some embodiments, a binding moiety specifically binds to CD30. In some embodiments, a binding moiety specifically binds to CD319.
  • a binding moiety specifically binds to CD33. In some embodiments, a binding moiety specifically binds to CD37. In some embodiments, a binding moiety specifically binds to CD38. In some embodiments, a binding moiety specifically binds to CD40. In some embodiments, a binding moiety specifically binds to CD44. In some embodiments, a binding moiety specifically binds to CD45. In some embodiments, a binding moiety specifically binds to CD46. In some embodiments, a binding moiety specifically binds to CD47. In some embodiments, a binding moiety specifically binds to CD51. In some embodiments, a binding moiety specifically binds to CD52.
  • a binding moiety specifically binds to CD56. In some embodiments, a binding moiety specifically binds to CD7. In some embodiments, a binding moiety specifically binds to CD70. In some embodiments, a binding moiety specifically binds to CD73. In some embodiments, a binding moiety specifically binds to CD74. In some embodiments, a binding moiety specifically binds to CD79b. In some embodiments, a binding moiety specifically binds to CD79B. In some embodiments, a binding moiety specifically binds to CD80. In some embodiments, a binding moiety specifically binds to CD99. In some embodiments, a binding moiety specifically binds to CDH3.
  • a binding moiety specifically binds to CDH6. In some embodiments, a binding moiety specifically binds to CEACAM5. In some embodiments, a binding moiety specifically binds to CEACAM6. In some embodiments, a binding moiety specifically binds to CLAUDIN18.2. In some embodiments, a binding moiety specifically binds to CLDN6. In some embodiments, a binding moiety specifically binds to CLDN9. In some embodiments, a binding moiety specifically binds to CLL- 1. In some embodiments, a binding moiety specifically binds to cMET. In some embodiments, a binding moiety specifically binds to CSF-R1. In some embodiments, a binding moiety specifically binds to CSF2.
  • a binding moiety specifically binds to CTGF. In some embodiments, a binding moiety specifically binds to CTLA4. In some embodiments, a binding moiety specifically binds to CXCR4. In some embodiments, a binding moiety specifically binds to DCLK1. In some embodiments, a binding moiety specifically binds to DDR1. In some embodiments, a binding moiety specifically binds to
  • a binding moiety specifically binds to DLL3. In some embodiments, a binding moiety specifically binds to DLL4. In some embodiments, a binding moiety specifically binds to DPEP3. In some embodiments, a binding moiety specifically binds to DR5. In some embodiments, a binding moiety specifically binds to DSG2. In some embodiments, a binding moiety specifically binds to EDB-Fn. In some embodiments, a binding moiety specifically binds to EFNA4. In some embodiments, a binding moiety specifically binds to EGFL7. In some embodiments, a binding moiety specifically binds to EGFR.
  • a binding moiety specifically binds to ENB-FN. In some embodiments, a binding moiety specifically binds to ENO1. In some embodiments, a binding moiety specifically binds to ENPP3. In some embodiments, a binding moiety specifically binds to EpCAM. In some embodiments, a binding moiety specifically binds to EphA2. In some embodiments, a binding moiety specifically binds to EphA3. In some embodiments, a binding moiety specifically binds to ETB. In some embodiments, a binding moiety specifically binds to FAP. In some embodiments, a binding moiety specifically binds to FCRL5. In some embodiments, a binding moiety specifically binds to FGFR2.
  • a binding moiety specifically binds to FGFR3. In some embodiments, a binding moiety specifically binds to Flt3. In some embodiments, a binding moiety specifically binds to FOLR. In some embodiments, a binding moiety specifically binds to FR1. In some embodiments, a binding moiety specifically binds to FR ⁇ . In some embodiments, a binding moiety specifically binds to FUT3. In some embodiments, a binding moiety specifically binds to GC-C. In some embodiments, a binding moiety specifically binds to GD3. In some embodiments, a binding moiety specifically binds to gelatinase B. In some embodiments, a binding moiety specifically binds to Globo H.
  • a binding moiety specifically binds to GLUT1. In some embodiments, a binding moiety specifically binds to glypican3. In some embodiments, a binding moiety specifically binds to GPNMB. In some embodiments, a binding moiety specifically binds to GPR20. In some embodiments, a binding moiety specifically binds to GPRC5D. In some embodiments, a binding moiety specifically binds to GUCY2C. In some embodiments, a binding moiety specifically binds to HER1. In some embodiments, a binding moiety specifically binds to HER2. In some embodiments, a binding moiety specifically binds to HER3.
  • a binding moiety specifically binds to HGFR. In some embodiments, a binding moiety specifically binds to HLA- DR. In some embodiments, a binding moiety specifically binds to ICOSL. In some embodiments, a binding moiety specifically binds to IGF-1R. In some embodiments, a binding moiety specifically binds to IGF1. In some embodiments, a binding moiety specifically binds to IGF2. In some embodiments, a binding moiety specifically binds to IL17A. In some embodiments, a binding moiety specifically binds to IL17F. In some embodiments, a binding moiety specifically binds to IL1RAP.
  • a binding moiety specifically binds to IL2. In some embodiments, a binding moiety specifically binds to IL6. In some embodiments, a binding moiety specifically binds to ITGB6. In some embodiments, a binding moiety specifically binds to KAAG1. In some embodiments, a binding moiety specifically binds to KIR2D. In some embodiments, a binding moiety specifically binds to LAG3. In some embodiments, a binding moiety specifically binds to LAMP-1. In some embodiments, a binding moiety specifically binds to Lewis-Y antigen. In some embodiments, a binding moiety specifically binds to LIV-1. In some embodiments,
  • a binding moiety specifically binds to LIV1. In some embodiments, a binding moiety specifically binds to LRRC15. In some embodiments, a binding moiety specifically binds to LT ⁇ . In some embodiments, a binding moiety specifically binds to Ly6E. In some embodiments, a binding moiety specifically binds to LYPD3. In some embodiments, a binding moiety specifically binds to MIF. In some embodiments, a binding moiety specifically binds to MSLN. In some embodiments, a binding moiety specifically binds to Muc1. In some embodiments, a binding moiety specifically binds to MUC1. In some embodiments, a binding moiety specifically binds to Muc16.
  • a binding moiety specifically binds to MUC5AC. In some embodiments, a binding moiety specifically binds to Nectin-4. In some embodiments, a binding moiety specifically binds to Notch-3. In some embodiments, a binding moiety specifically binds to Notch1. In some embodiments, a binding moiety specifically binds to OAcGD2. In some embodiments, a binding moiety specifically binds to p53. In some embodiments, a binding moiety specifically binds to PCDP1. In some embodiments, a binding moiety specifically binds to PDL1. In some embodiments, a binding moiety specifically binds to PDL2. In some embodiments, a binding moiety specifically binds to PRL receptor.
  • a binding moiety specifically binds to PSMA. In some embodiments, a binding moiety specifically binds to PTK7. In some embodiments, a binding moiety specifically binds to RNF43. In some embodiments, a binding moiety specifically binds to RON. In some embodiments, a binding moiety specifically binds to ROR1. In some embodiments, a binding moiety specifically binds to ROR2. In some embodiments, a binding moiety specifically binds to SDC1. In some embodiments, a binding moiety specifically binds to SEZ6. In some embodiments, a binding moiety specifically binds to SLAMF2. In some embodiments, a binding moiety specifically binds to SLAMF6.
  • a binding moiety specifically binds to SLAMF7. In some embodiments, a binding moiety specifically binds to SLITRK6. In some embodiments, a binding moiety specifically binds to SSEA-4. In some embodiments, a binding moiety specifically binds to STEAP1. In some embodiments, a binding moiety specifically binds to sTn. In some embodiments, a binding moiety specifically binds to TAA. In some embodiments, a binding moiety specifically binds to TAG72. In some embodiments, a binding moiety specifically binds to TDGF1. In some embodiments, a binding moiety specifically binds to TEM1.
  • a binding moiety specifically binds to Tenascin C. In some embodiments, a binding moiety specifically binds to TF (Tissue Factor). In some embodiments, a binding moiety specifically binds to TGFb. In some embodiments, a binding moiety specifically binds to TIGIT. In some embodiments, a binding moiety specifically binds to TIM1. In some embodiments, a binding moiety specifically binds to TNF- ⁇ . In some embodiments, a binding moiety specifically binds to TNFR. In some embodiments, a binding moiety specifically binds to TRAIL. In some embodiments, a binding moiety specifically binds to TRAIL-R2.
  • a binding moiety specifically binds to TROP2. In some embodiments, a binding moiety specifically binds to TWEAKR. In some embodiments, a binding moiety specifically binds to TYRP1. In some embodiments, a binding moiety specifically binds to VEGF2. In some embodiments, a binding moiety specifically binds to VEGFR2. In some embodiments, a binding moiety specifically binds to vimentin. In some embodiments, a binding moiety specifically binds to VISTA.
  • Some aspects of the disclosure related a method of treating cancer, such as a solid tumor, comprising administering to the subject a therapeutically effective amount of a construct (e.g., an antibody-drug conjugate or AFFIMER®-drug conjugate) provided herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof wherein the binding moiety (Z) binds to an antigen selected from CEACAM5, CLAUDIN18.2, CLAUDIN4, CMET, DLL3, EDB-FN, FAP, FR ⁇ , HER2, HER3, LLRC15, Nectin-4, TF (Tissue Factor), TROP2.
  • a construct e.g., an antibody-drug conjugate or AFFIMER®-drug conjugate
  • Z binds to an antigen selected from CEACAM5, CLAUDIN18.2, CLAUDIN4, CMET, DLL3, EDB-FN, FAP, FR ⁇ , HER2, HER3, LLRC15, Nectin-4, TF (Tissue
  • Some aspects relate to a method of treating a tumor in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a construct provided herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof wherein the binding moiety (Z) binds to CEACAM5.
  • a binding moiety comprises labetuzumab, M9140, cibisatamab, LM-24C5, NEO-201, precemtabart tocentecan, actinium 225 labeled anti-CEA antibody, BA-1202, BGC-477, BGC-477, EBC-129, IBI-3020, NILK-2301, NILK-2401, PF-08046050, ABC-101, ATOR-4066, CEA ISAC, LM-004, NI-3301, PM-4008, TBADC-02, or GB-7012.
  • a binding moiety comprises tusamitamab (see, e.g., US Patent No.11,332,542).
  • Tusamitamab [270] Heavy Chain: EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWVAYISSGGGITYAPSTVK GRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCAAHYFGSSGPFAYWGQGTLVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSK
  • a binding moiety comprises zolbetuximab (see, e.g., Davies M, Nucleic Acids Res.2015; 43(W1):W612-20).
  • a binding moiety comprises LM-302.
  • a binding moiety comprises IBI343. [273] Zolbetuximab [274] Heavy Chain:
  • a binding moiety comprises ASP1002.
  • a binding moiety comprises KM3900 (see, e.g., US Patent No.8,076,458).
  • KM3900 [278] Heavy Chain: MGYSYIFLLSGTAGGLSEVQLQQSGPELVKPGASVKISCKASGYTFTDYYMNWVKQSHGKSLEY IGAVVPNNGVPTYNQKFKGKATLTVDKSSSTAYMELRSLTSEDSAVYYCARPHYYYAGRSGAMD YWGQGTSVTVSS (SEQ ID NO: 508) [279] Light Chain: MDFQVQIFSFLLISASVIMSRGQIVLTQSPAIMSASLGERVTMTCTASSTVSSTYLHWYQQKPGSSP KLYIYSTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCHQYHRSPPTFGGGTKLEIK (SEQ ID NO: 509) [280]
  • a binding moiety comprises telisotuzumab. In some embodiments, a binding moiety comprises REGN5093. In some embodiments, a binding moiety comprises MYTX-011. In some embodiments, a binding moiety comprises rilotumumab (see, e.g., Kanehusa, M et al. Nucleic Acids Research.2016, 44(D1), D457–D462). In some embodiments, a binding moiety comprises farletuzumab (see, e.g., Davies M, Nucleic Acids Res.2015; 43(W1):W612- 20).
  • a binding moiety comprises rovalpituzumab (see, e.g., Kanehusa, M et al. Nucleic Acids Research.2016, 44(D1), D457–D462). In some embodiments, a binding moiety comprises tisotumab (see, e.g., Kanehusa, M et al. Nucleic Acids Research.2016, 44(D1), D457–D462).
  • a binding moiety comprises tisotumab.
  • Some aspects relate to a method of treating a tumor in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a construct provided herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof wherein the binding moiety (Z) binds to EDB-FN.
  • a binding moiety comprises radretumab, for example, formatted on hIgG1 (“L19”: see, e.g., Pini A, et al. J Biol Chem.1998 Aug 21;273(34):21769- 76).
  • a binding moiety comprises OMTX-705. In some embodiments, a binding moiety comprises BI-765179. In some embodiments, a binding moiety comprises GEN-1057. In some embodiments, a binding moiety comprises sibrotuzumab (see, e.g., US Patent No.20,090,304,718).
  • Sibrotuzumab [300] Heavy Chain: MGWSGVFIFILSGTAGVQSQVQLQQSGAELARPGASVNLSCKASGYTFTNNGINWLKQRTGQGL EWIGEIYPRSTNTLYNEKFKGKATLTADRSSNTAYMGLRSKTSGDSAVYFLVTVSAAKTTAPSVYP LAP (SEQ ID NO: 520) [301] Light Chain: MDFQVQIFSFLLISASVIISRGQIVLTQSPAIMSASPGEKVTMTCSASSGVNFMHWYQQKSGTSPK RWIFDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSFNPPTFGGGTKLEIKRAD AAPTVS (SEQ ID NO: 521) [302] Some aspects relate to a method of treating a tumor in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a construct provided herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof
  • a binding moiety comprises mirvetuximab. In some embodiments, a binding moiety comprises farletuzumab. [303] Some aspects relate to a method of treating a tumor in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a construct provided herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof wherein the binding moiety (Z) binds to HER2.
  • a binding moiety comprises an antibody of Kadcyla, Enhertu, or RC48.
  • a binding moiety comprises trastuzumab (see, e.g., Ménard S., et al.
  • a binding moiety comprises pertuzumab (see, e.g., Adams C, Cancer Immunol Immunother.2006 Jun;55(6):717-27).
  • a binding moiety comprises zenocutuzumab. In some embodiments, a binding moiety comprises patritumab (see, e.g., Davies M, Nucleic Acids Res.2015; 43(W1):W612-20). In some embodiments, a binding moiety comprises seribantumab. In some embodiments, a binding moiety comprises lumretuzumab.
  • a binding moiety comprises LNTH-2403. In some embodiments, a binding moiety comprises SOT-106. In some embodiments, a binding moiety comprises ZL-6201. In some embodiments, a binding moiety comprises LRRC15-CD3. In some embodiments, a binding moiety comprises mAb211. In some embodiments, a binding moiety comprises samrotamab (see, e.g., US Patent No.11,045,480). In some embodiments, a binding moiety comprises ABBV-085 (see, e.g., International Publication No. WO2017/095805).
  • a binding moiety comprises enfortumab (see, e.g., US Patent No.9,314, 538). In some embodiments, a binding moiety comprises ADRX ⁇ 0706. In some embodiments, a binding moiety comprises LY4052031.
  • Some aspects relate to a method of treating a tumor in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a construct provided herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof wherein the binding moiety (Z) binds to TF (Tissue Factor).
  • a binding moiety comprises tisotumab.
  • Some aspects relate to a method of treating a tumor in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a construct provided herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof wherein the binding moiety (Z) binds to TROP2.
  • a binding moiety comprises sacituzumab (see, e.g., Zangard M, et al. Expert Opin Investig Drugs.2019 Feb;28(2):107-112). In some embodiments, a binding moiety comprises datopotamab. In some embodiments, a binding moiety comprises BNT325/DB-1305.
  • Sacituzumab [328] Heavy Chain: QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYT DDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVT VPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSD
  • a binding moiety can be any one of the foregoing antibodies, AFFIMER ® polypeptides or any antibody or AFFIMER ® polypeptide that binds to any one or more of the foregoing cancer antigens.
  • the compound is selected from those in Table 4:
  • the construct is selected from those in Table 5A, and pharmaceutically acceptable salts thereof:
  • the binding moiety comprises FAP-1 or FAP-2 (SEQ ID NO: 213 or 214, respectively) comprising one or more cysteine modifications (e.g., cysteine substitutions) for conjugation to the maleimide reactive handle of LP1 or LP2.
  • cysteine modifications e.g., cysteine substitutions
  • the compound is selected from those in Table 5B:
  • compositions comprising a construct described herein (e.g., a construct of Formula (II′) or (II)) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers and/or excipients.
  • a pharmaceutical composition comprising a construct described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • a construct described herein is provided in an effective amount in the pharmaceutical composition. In certain embodiments, the effective amount is a therapeutically effective amount.
  • Pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology.
  • Such preparatory methods include bringing the construct described herein (i.e., the “active ingredient”) into association with a carrier or excipient, and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping, and/or packaging the product into a desired single- or multi-dose unit.
  • Pharmaceutical compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses.
  • a “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient.
  • the amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage.
  • Relative amounts of the active ingredient, the pharmaceutically acceptable carrier or excipient, and/or any additional ingredients in a pharmaceutical composition described herein will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered.
  • compositions used in the manufacture of provided pharmaceutical compositions include inert diluents, solvents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, oils, butters, and/or waxes.
  • Excipients such as coloring agents, coating agents, sweetening agents, flavoring agents, and fragrances may also be present in the composition.
  • constructs and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, intradermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), mucosal, nasal, buccal, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol.
  • enteral e.g., oral
  • parenteral intravenous, intramuscular, intra-arterial, intramedullary
  • intrathecal subcutaneous, intraventricular, transdermal, intradermal, rectal, intravaginal, intraperitoneal
  • topical as by powders, ointments, creams, and/or drops
  • mucosal nasal,
  • routes of administration include intravenous administration (e.g., systemic intravenous injection) and direct intra-tumoral administration.
  • intravenous administration e.g., systemic intravenous injection
  • direct intra-tumoral administration e.g., direct intra-tumoral administration.
  • the most appropriate route of administration will depend upon a variety of factors including the nature of the agent (e.g., its stability in the environment of the gastrointestinal tract), and/or the condition of the subject (e.g., whether the subject is able to tolerate oral administration).
  • kits described herein further includes instructions for using the kit.
  • a kit described herein may also include information as required by a regulatory agency such as the U.S. Food and Drug Administration (FDA).
  • the information included in the kits is prescribing information.
  • the kits provide instructions for treating a disease (e.g., cancer) in a subject in need thereof.
  • a kit described herein may include one or more additional pharmaceutical agents described herein as a separate composition.
  • constructs described herein e.g., constructs of Formula (II′) or (II)
  • pharmaceutically acceptable salts thereof for use in treating cancer in a subject in need thereof.
  • uses of constructs described herein e.g., constructs of Formula (II′) or (II)
  • pharmaceutically acceptable salts thereof for use in treating cancer in a subject in need thereof.
  • methods comprising administering to a subject a construct described herein (e.g., constructs of Formula (II′) or (II)), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the subject has a disease characterized by FAP upregulation.
  • the subject has cancer.
  • the construct, pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof is administered by intravenous injection.
  • the disorder characterized by FAP upregulation is cancer, fibrosis, or inflammation.
  • the disorder characterized by FAP upregulation is cancer.
  • the cancer is selected from head and neck cancer, soft tissue sarcoma, breast cancer, lung
  • the cancer is selected from head and neck cancer, soft tissue sarcoma, breast cancer, lung cancer, gastric cancer, colorectal cancer, pancreatic cancer, uterine cancer, ovarian cancer, and cervical cancer. In some embodiments, the cancer is selected from breast cancer, lung cancer, gastric cancer, pancreatic cancer, uterine cancer, ovarian cancer, and cervical cancer.
  • the cancer is triple-negative breast cancer (TNBC), gastric cancer, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), uterine cancer, ovarian cancer, pancreatic cancer, or cervical cancer.
  • the cancer is selected from head and neck cancer, soft tissue sarcoma, breast cancer, lung cancer, gastric cancer, colorectal cancer, and pancreatic ductal adenocarcinoma.
  • the cancer is head and neck cancer.
  • the head and neck cancer is salivary gland cancer.
  • the cancer is sarcoma.
  • the cancer is soft tissue sarcoma.
  • the soft tissue sarcoma is undifferentiated pleomorphic sarcoma or dedifferentiated liposarcoma.
  • the cancer is breast cancer. In some embodiments, the cancer is triple-negative breast cancer (TNBC). In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is pancreatic ductal adenocarcinoma. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is small cell lung cancer (SCLC). In some embodiments, the cancer is non-small cell lung cancer (NSCLC). In certain embodiments, the cancer is gastric cancer. In some embodiments, the cancer is uterine cancer.
  • the cancer is ovarian cancer. In some embodiments, the cancer is cervical cancer. In some embodiments, the cancer is melanoma. [353]
  • the term “cancer” refers to a class of diseases characterized by the development of abnormal cells that proliferate uncontrollably and have the ability to infiltrate and destroy normal body tissues. In certain embodiments, the cancer is a solid cancer. In certain embodiments, the cancer is a hematopoietic cancer (i.e., hematological cancer).
  • the cancer is a hematopoietic cancer (e.g., leukemia (e.g., acute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myelocytic leukemia (AML) (e.g., B- cell AML, T-cell AML), chronic myelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL)); lymphoma (e.g., Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL)), non-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma)), f
  • ALL acute lymphocy
  • enteropathy type T-cell lymphoma subcutaneous panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma
  • heavy chain disease e.g., alpha chain disease, gamma chain disease, mu chain disease
  • MPD myeloproliferative disorder
  • PV polycythemia vera
  • ET essential thrombocytosis
  • AMM agnogenic myeloid metaplasia
  • the cancer is leukemia.
  • the cancer is acute lymphoblastic leukemia (ALL).
  • the cancer is early T-cell precursor (ETP)-acute lymphoblastic leukemia (ALL).
  • the cancer is lung cancer (e.g., bronchus cancer (e.g., bronchogenic carcinoma, bronchial adenoma), alveolar carcinoma, mesothelioma, small cell lung cancer (SCLC), non- small cell lung cancer (NSCLC), lung adenocarcinoma, chondromatous hamartoma, papillary adenocarcinoma).
  • lung cancer e.g., bronchus cancer (e.g., bronchogenic carcinoma, bronchial adenoma), alveolar carcinoma, mesothelioma, small cell lung cancer (SCLC), non- small cell lung cancer (NSCLC), lung adenocarcinoma, chondromatous hamartoma, papillary adenocarcinoma).
  • the cancer is a genitourinary cancer (e.g., bladder cancer (e.g., urothelial carcinoma), urethral cancer, kidney cancer (e.g
  • certain “benign” tumors may later give rise to malignant neoplasms, which may result from additional genetic changes in a subpopulation of the tumor’s neoplastic cells, and these tumors are referred to as “pre-malignant neoplasms.”
  • An exemplary pre-malignant neoplasm is a teratoma.
  • a “malignant neoplasm” is generally poorly differentiated (anaplasia) and has characteristically rapid growth accompanied by progressive infiltration, invasion, and destruction of the surrounding tissue.
  • a malignant neoplasm generally has the capacity to metastasize to distant sites.
  • anti-FAP AFFIMER polypeptides include a cysteine (C) (underlined either in Loop 3 or at the C terminus) to which the LP1 or LP2 was conjugated: ) Production, analysis and in vitro testing of AFFIMER® conjugates with multiple cysteines for multi-drug conjugation [503] PEG-Malemide (5kDa) was conjugated to AFFIMER® constructs on available thiol groups.
  • AFFIMER® constructs contained Cys residues at the Cterminus, within Loop3, and/or within Loop7 (Loop7 is only present in two-domain fusion proteins). These Cys enabled Malemide conjugation at each of the sites individually, or in combination.
  • Figure 9 shows the analytical SEC-HPLC profiles of unmodified protein, and 1x, 2x, or 3x PEG conjugated constructs. Single domain (FAP-2) and two-domain (FAP-2-FAP-1-H) AFFIMER® exemplars are shown. These data demonstrate that FAP binding AFFIMERS® can be conjugated to Malemide at multiple sites within the protein scaffold.
  • AFFIMER® protein internalization was measured using two techniques (FIGs.12A-2B). As shown in FIG.12A, AFFIMER® proteins were evaluated for internalization based on down-regulation of AFFIMER® protein at the cell surface of FAP-expressing cells at 37°C vs.4°C by flow cytometry. Antibodies (right-hand three columns, FIG.12A) were used as positive internalizing controls. [507] To confirm non-internalization, as shown in FIG.12B, lead AFFIMER® proteins containing an engineered C-terminal cysteine residue were conjugated to the pH-sensitive pHrodoTM red dye and incubated with HEK-FAP cells.
  • FAP-2 single-domain (FIG.12D, left) and FAP-2-FAP-1 two-domain (FIG.12D, right) AFFIMER® proteins were incubated at 4°C, 37°C, or 45°C for 1 week.
  • Protein aggregation in the samples was assessed by size exclusion chromatography (SEC). SEC profiles for the temperature-stressed samples are shown in FIG.12D with the percent purity for the monomeric species indicated. Results show tested AFFIMER® proteins are stable under stress temperature conditions.
  • SLFN11 expression leads to replication fork destabilization and impaired DNA repair, making SLFN11-positive tumors more susceptible to insult from DNA-damaging agents.
  • the clinical utility of SLFN11 has been demonstrated in several studies [512] A potential synergy emerges when FAP and SLFN11 expression profiles are considered together as tumors co-expressing high levels of FAP and SLFN11 are most likely to respond to pre
  • high FAP expression enables efficient localisation and cleavage of pre
  • Target binding The target-binding ability of the antibody conjugates was assessed by flow cytometry to target positive cells.
  • U87MG cells EMB-F +ve, LRRC15+ve
  • LS174T cells CEACAM5+ve
  • Antibody conjugates retained ability to bind their respective targets to equivalent levels as non- conjugated naked antibodies (FIG.15).
  • Anti-RSV conjugates were used as isotype controls and did not show binding as expected (data not shown).
  • Cytotoxicity assays [521] Cytotoxicity assays were performed using HEK293T cells overexpressing FAP and compared to parent cells (FAP-negative).
  • LP2, LP3, and LP4 were cleaved to varying extents, resulting in cytotoxicity levels proportional to their FAP sensitivity, with LP3 exhibiting the lowest, LP2 intermediate, and LP4 the highest activity.
  • LP3 Upon exposure to exogenous FAP or in cells expressing
  • Antibodies containing LP2 or LP3 retained FAP-dependent cytotoxicity, demonstrating enhanced activity in FAP-positive cells relative to FAP-negative controls (FIGs.16 and 17). Antibodies containing LP4 also retained FAP-dependent activity, as evidenced by reduced cytotoxicity in the presence of a FAP inhibitor; however, due to the high FAP sensitivity of LP4 and the presence of low FAP in the assay conditions, elevated cytotoxicity was observed in both FAP-positive and FAP-negative cells (FIG.18).
  • LP5 conjugates showed minimal cytotoxicity, which was independent of FAP, as expected (Fig.19).
  • Bystander cytotoxicity via FAP-dependent activation in tumor-fibroblast co-culture models [524] Bystander effect was assessed using 2D or 3D co-culture models, using GFP-expressing tumor cells (e.g., colorectal LS174T-GFP or triple negative breast cancer MDA-MB-231-GFP cells) and matched fibroblast cells (which express FAP to levels typically observed in tumors).
  • GFP-expressing tumor cells e.g., colorectal LS174T-GFP or triple negative breast cancer MDA-MB-231-GFP cells
  • Anti-CEACAM5-LP2 exhibited FAP dependent cytotoxicity, where it shows minimal cytotoxicity as a single agent and is only fully active in the presence of recombinant FAP in mono-culture when compared to FAP inhibited and warhead alone conditions.
  • fibroblast-derived endogenous FAP partially activated anti-CEACAM5-LP2 conjugate, resulting in measurable tumor cell killing consistent with a bystander effect. This partial activation achieved cytotoxicity levels within approximately 5-fold of those observed with the free warhead. Full activation, matching the potency of the warhead alone, was observed in the presence of exogenous recombinant FAP.
  • 3D spheroid cytotoxicity assays were performed using MDA-MB-231-GFP tumour cells (FAP negative) as mono-culture or co-culture with human mammary fibroblasts (FAP positive) grown in ultra-low attachment plates. Spheroids were incubated with anti-CEACAM5-LP2 conjugates for 7 days in complete MammoCult medium (chemically defined with no exogenous FAP). Tumour specific cytotoxicity was measured as % inhibition relative to vehicle control by GFP fluorescence using the IncuCyte (FIG. 21).
  • Anti-CEACAM5-LP2 exhibited FAP dependent cytotoxicity, where it is inert as a single agent and only fully active in the presence of recombinant FAP in mono-culture when compared to FAP inhibited and warhead alone conditions. In the coculture, the presence of FAP-expressing fibroblasts was sufficient to fully activate the conjugate, resulting in tumor cell killing equivalent to that observed with the free cytotoxic payload.
  • anti-CEACAM5 antibody alone did not show significant tumour growth inhibition compared to the vehicle control group.
  • the anti-CEACAM5 conjugates showed various degree of efficacy: anti-CEACAM5-LP3 showed low tumour growth inhibition, anti-CEACAM5-LP4 showed higher efficacy and anti- CEACAM5-LP2 showed the best response with tumours starting to grow above ⁇ 250mm3 from day 39 only (FIG.22A). No signs of toxicity were observed with anti-CEACAM5-LP2. Very minor (less than 5%) and minor (less than 10%) body weight loss was seen with anti-CEACAM5-LP3 and anti- CEACAM5-LP4 respectively (FIG.22B).
  • CDX cell-line derived xenograft
  • the present disclosure includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process.
  • the present disclosure includes embodiments in which more than one, or all the group members are present in, employed in, or otherwise relevant to a given product or process.
  • the present disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can

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Abstract

L'invention concerne des composés, y compris des composés de formule (I') et (I), et des sels pharmaceutiquement acceptables de ceux-ci, et des constructions, y compris des constructions de formule (II') et (II), et des sels pharmaceutiquement acceptables de celles-ci, qui comprennent une fraction clivable par une protéine d'activation des fibroblastes (FAP) et sont capables d'administrer une camptothécine à des tissus exprimant FAP (par exemple, des cancers). L'invention concerne également des compositions pharmaceutiques et des kits les comprenant, ainsi que des procédés d'utilisation de celles-ci.
PCT/EP2025/067754 2024-06-24 2025-06-24 Conjugués inhibiteurs de topoisomérase activés par fap spécifiques à une cible et leurs utilisations Pending WO2026002995A1 (fr)

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